Validation of Analytical Method of Irbesartan Plasma in Vitro by High Performance Liquid Chromatography-Fluorescence

Validation of Analytical Method of Irbesartan Plasma in Vitro by High Performance Liquid Chromatography-Fluorescence

Harmita, Yahdiana Harahap and I. Kadek Arya M. Department of Pharmacy, University of Indonesia, Depok, Jakarta 16424, Indonesia

Received: November 29, 2011 / Accepted: February 01, 2012 / Published: July 30, 2012.

Abstract: Irbesartan is an antihypertensive drug whose concentration in blood is very small so it requires a sensitive method of analysis, selective and valid for analysis. In this study, it is carried out optimization of analytical conditions and validation for the analysis of irbesartan in plasma. Chromatography was performed on a C18 column (250 × 4.6 mm, 5 μm) under isocratic elution with acetonitrile-0.1% formic acid (46:54 v/v), pH 3.75. Detection was made at excitation 250 nm and emission 370 nm and analyses were run at a flow-rate of 1.0 mL/min at a temperature of 40 ºC. Losartan potassium was used as internal standard. Plasma extraction was done by deproteination with acetonitrile, mixed with vortex for 30 seconds, then centrifuged it at 10,000 rpm for 10 min. In plasma validation, the recovery was 96.22%, and the lower limit of quantification (LLOQ) in plasma was 2 ng/mL. The method also fulfill the criteria for accuracy and precision intra and inter day by normal values (%Diff) not exceed ± 15%. On the stability study, irbesartan in plasma temperature –20 °C has been stable for 28 days.

Key words: HPLC, fluorescence, irbesartan, validation, human plasma.

1. Introduction  on the kidneys especially in diabetic patients [2].

Irbesartan is a non-peptide compounds, with the Hypertension is a common health problem in chemical name 2-butyl-3-[[29-(1H-tetrazole-5-yl) developed countries and developing countries. The [1,19-biphenyl]-4yl]methyl]-1,3-diazaspiro[4,4]non-1- cause of hypertension is diverse due to genetic factors, en-4-one. Irbesartan is a hypotensive agent that does lifestyle, and stress. Uncontrolled hypertension can not require biotransformation to become active form lead to various diseases such as stroke, heart failure, [3]. Peroral drug absorption is rapid, bioavaibilitasnya diabetic nephropathy, myocardial infarction, kidney about 60-80% and 90% protein bound. At therapeutic failure and even death. According to the guidline of doses of irbesartan (75-300 mg), maximum the National Joint Committee, the management of concentration in plasma will be obtained about 1.5-2 patients with hypertension should be precise and fast, hours after dosing [4]. The maximum concentration in with drugs given singly or in combination therapy plasma after administration of a dose of 150 mg drugs [1]. Irbesartan is a class of hypertension drugs irbesartan is about 1.5 ± 0.29 g/mL [5]. called angiotensin II receptor blockers that work on Numerous HPLC methods for the quantification of the renin-angiotensin-aldosterone system. Besides irbesartan in biological samples have been reported. useful for lowering blood pressure, drugs known as These methods involved the use of protein angiotensin II receptor blockers have a protective effect precipitation, liquid-liquid extraction and solid-phase

extraction coupled with fluorescence detection or Corresponding author: Harmita, Ph.D., research field: pharmaceutical chemistry. E-mail: harmita21@yahoo.co.uk.

ultraviolet chromatography or high performance liquid

Validation of Analytical Method of Irbesartan Plasma in Vitro by

High Performance Liquid Chromatography-Fluorescence

chromatography tandem mass spectrometry method

certain concentration.

(HPLC⁄MS⁄MS) [6-8]. Human plasma calibration standards of irbesartan In bioequivalence studies, the proposed method

were prepared by spiking an appropriate amount of the should be simple and able to process hundreds of

working standard solutions into drug-free human samples in a limited time. This paper describes a

plasma. The concentration of irbesartan in calibration simple, rapid, precise, and accurate HPLC method for

curve was 2.05 ng/mL, 5.12 ng/mL, 10.24 ng/mL, determining irbesartan in human plasma in vitro.

51.20 ng/mL, 256 ng/mL, 512 ng/mL, 1,024 ng/mL, and 5,120 ng/mL. Quality control (QC) samples were

2. Materials and Methods

prepared at three concentrations that were low (6.14

2.1 Chemicals and Reagents ng/mL), medium (2,048 ng/mL), and high (4,096 ng/mL).

Irbesartan (99.7% on assay) were obtained from Hetero Labs Limited. Losartan Potassium (99.6% on

2.4 Sample Preparation

assay) were obtained Ipca Labs Limited. Acetonitrile

0.25 mL of plasma containing certain and methanol were HPLC-grade and were purchased

concentrations of irbesartan were added 25.0 μL of the from Merck. The other chemicals and reagents were

internal standard working solution (6 μg/mL). Three analytical grade. Human plasma was provided by

part of acetonitril (750.0 μL) was added to precipitate Indonesian blood bank (Palang Merah Indonesia).

protein in plasma, vortex-mixed for 30 sec and

2.2 Chromatographic Conditions centrifuged at 10,000 rpm for 10 min. A 20 μL aliquot of the supernatant was injected into the HPLC system.

The HPLC system (Shimadzu, Japan) used consisted of a model LC-10AD pump, a fixed manual

2.5 Validation of This Method

injection loop of 20 μL, and a model RF-10AXL The validation parameters obtained were specificity,

Fluorescence detector; data acquisition was performed linearity, sensitivity, accuracy, precision, recovery and with the SCL-10A processor. The analytical column

stability. The method was validated according to employed was a Kromasil C18 column (250 × 4.6 mm,

USFDA guidance for bioanalytical method validation i.d., 5 μm).

The mobile phase consisted of acetonitrile-0.1% Six randomly selected blank plasma samples [9] formic acid (46:54 v/v). The mobile phase was

were processed by a similar extraction procedure and adjusted to pH 3.75 ± 0.01 with 1 N NaOH or dilute

analyzed to determine the extent to which endogenous phosphate acid (85% v/v), filtered through a 0.45 μm

plasma components may contribute to interference at cellulose membrane filter (Whatman) and degassed

the retention time of irbesartan, and losartan before use (Elmasonic S60H). The detection potassium. wavelength was set at excitation 250 nm and emission

The calibration curves were constructed each day 370 nm. Chromatography separation was performed at

before the analysis of the samples by plotting the temperature 40 °C and flow rate was maintained at 1

peak-area ratio versus the drug concentrations. The mL/min.

lower limit of quantification (LLOQ) was defined as

2.3 Standard Solutions and Quality Control Samples the lowest concentration with a coefficient of variation

(CV) of less than 20% and accuracy of 80-120%. The Primary stock solutions of irbesartan (1 mg/mL)

accuracy and precision of the method were assessed and losartan potassium (1 mg/mL) were prepared in

intra-day and inter-day during 5 days by determining methanol. Then, diluted with methanol to obtain a

QC plasma samples at three concentrations which

Validation of Analytical Method of Irbesartan Plasma in Vitro by

High Performance Liquid Chromatography-Fluorescence

were low (6.14 ng/mL), medium (2,048 ng/mL), and losartan potassium were well resolved with respective high (4,096 ng/mL) concentrations, accompanying by

retention times of 6.2 min and 10.4 min.

a standard calibration curve on each analytical run.

3.2 Calibration Curve and Limit of Quantification The recovery of irbesartan was evaluated by

comparing measured concentration obtained from The calibration curves were linear over the peak areas of pre-treated quality control plasma

concentration range of 2.05-5,120 ng/mL with a samples (n = 5) with mean measured concentration of

correlation coefficient of 0.9999. The correlation those spiked-after extraction samples at the same

coefficient from replicate calibration curves on nominal concentrations. Stability quality control different days was more than 0.9995. The lower limit plasma samples were conducted at low and high

of quantification with a coefficient of variation of less concentrations and were subjected to short-term (6 h

than 20% was 2 ng/mL.

and 24 h) incubation at room temperature, three freeze/thaw cycles, and storage for 28 days (–20 ºC). 3.3 Precision and Accuracy

The stability of primary stock solutions were also The coefficient variation values of both inter- and being conducted for 25 days (5 ºC).

intraday analysis for 5 days at three concentrations

3. Results which each concentration is conducted at 5 replicates

were less than 6.37% whereas the %Diff were less

3.1 Specificity than 13.43%. The inter- and intra-day precision and

The current method showed excellent accuracy values of the assay method are presented in chromatographic specificity with no endogenous

Table 1.

plasma interference at the retention times of irbesartan

3.4 Recovery

and losartan potassium as internal standard. Chromatograms obtained from human blank plasma

The mean extraction recoveries of irbesartan at and human blank plasma spiked with irbesartan (0.5

three concentrations (low, mid, high) were μg/mL) and losartan potassium (20 μg/mL) are shown

86.23%-113.45%, 85.37%-99.31%, and 85.73%- in Figs. 1A and 1B, respectively. Irbesartan and

103.44%, respectively.

B Retention Time Detector A (Ex:250nm, Em:370nm) BLANK4555-012 Retention Time UKS05-006

0.5 A 0.5 Detector A (Ex:250nm, Em:370nm)

0.2 0.2 Area (µV/s) rbesar

(µV/s)

0.0 2.5 5.0 7.5 10.0 12.5 15.0 0.0 6. Minutes 0.0 0 2 4 6 Minutes 8 10 12

Retention time (min)

Retention time (min)

Fig. 1 Representative chromatogram of human blank plasma (A), human blank plasma spiked with irbesartan (1) and losartan potassium (2) (B).

Validation of Analytical Method of Irbesartan Plasma in Vitro by

High Performance Liquid Chromatography-Fluorescence

3.5 Stability not less than the volume of blood. It is also usually become the component of mobile phase with the result

Treated plasma samples were found to be stable at that the system will be able to accept. Results least 24 h when the samples were kept at room indicated that direct protein precipitation with temperature (%Diff < 15%). The concentrations of acetonitrile was simple and rapid and good separation irbesartan in plasma which underwent three of the drug and I.S. was achieved using the freeze-thaw cycles or storage at –20 ºC for 28 days precipitation method. The centrifugator was used were found to be stable with % differentiation less

optimally at 10,000 rpm for 10 min. The aim was to than 15%. The stability data of irbesartan stored under

obtain the pure supernatant which was ready to be various conditions and subjected to freeze-thaw cycles

injected.

are shown in Table 2. The primary stock solutions were also found to be stable for 25 days when were

4.2 Optimization of Mobile Phase kept at 5 ºC. The stability data of irbesartan stored

The chromatographic conditions were optimized by under various conditions and subjected to freeze-thaw

injecting analytes with mobile phase containing cycles are shown in Table 2.

varying percentages of organic phase and flow rates of

4. Discussion

mobile phase to achieve good resolution and symmetric peak shapes for irbesartan and losartan

4.1 Preparation of Plasma Samples potassium, as well as a short retention time. As Protein precipitation has the advantages of expected, the retention times increased with simplicity and universality, so it was used to prepare

decreasing acetonitrile percentage and system flow the plasma samples [8]. The reason of choosing

rates. The chosen mobile phase pH was 3.75 ± 0.01 by acetonitrile as the precipitation agent was caused by

giving the most symmetric peak shapes for irbesartan its ability to precipitate protein, especially when given

and losartan potassium.

Table 1 Accuracy and precision from the determination of irbesartan in human plasma (n = 25/concentration).

Concentration

Mean ± SD (ng/mL)

6.85 Min. = –13.77% Max. = 14.67% Mid

3.41 Min. = –14.63% Max. = 2.81% Min. = –14.28%

Table 2 Stability data of irbesartan in human plasma.

Concentration CV (%) %Diff Short term stability for 24 h in plasma at room temperature Low

Long term storage at – 20 °C for 28 days Low

Three freeze/thaw cycles Low

4.64 –14.46 - 9.02%

High

6.46 –5.67 - 9.37%

Validation of Analytical Method of Irbesartan Plasma in Vitro by

High Performance Liquid Chromatography-Fluorescence

A Detector A (Ex:250nm, Em:370nm) Detector A (Ex:250nm, Em:370nm)

Retention Time UKS05-006

Retention Time STD013763-001

B Name

(µV/s)

Irb

Vo

(µV/s)

ir besa

Retention time (min)

Retention time (min)

Name Retention Time std4060-014 Detector A (Ex:250nm, Em:370nm) (1)

(µV/s)

lium a 7 K

Retention time (min)

Fig. 2 Chromatogram of irbesartan (1) and losartan potassium (2) in varying percentages of organic phase. The chromatograms

representative system in acetonitrile -0.1% formic acid (46:54, v/v) (A); (37:63, v/v) (B); and (40:60, v/v) (C).

Optimal conditions were a mobile phase consisting Thus the assay is suitable for routine analysis when of acetonitrile: 0.1% formic acid (46:54, v/v) pH 3.75

determining assay on biological samples to perform ± 0.01 arranged by 1 N NaOH and 85% phosphate

bioequivalence studies. A simple, rapid, precise, and acid. Under optimum conditions, the chromatographic

accurate HPLC method for determining irbesartan in run time for each sample was completed within 14

human plasma has been presented. Although lower min.

sensitivity was obtained in comparison to previously

4.3 Advantages of the Method published LC methods with mass spectrometry

detection, the resulting LLOQ (2 ng/mL) was In comparison to previously published HPLC

sufficient for human pharmacokinetic studies. methods for separation and quantitation of irbesartan,

5. Conclusion

the major modifications incorporated into the current

method include: simple sample preparation procedures, An analytical method developed for irbesartan common and cheap HPLC equipment and mobile

quantification in plasma samples showed good phase additives, and a relative short analysis time as

specificity, sensitivity, linearity, precision, and well.

accuracy over the entire range of clinically significant

Validation of Analytical Method of Irbesartan Plasma in Vitro by

731

High Performance Liquid Chromatography-Fluorescence

and therapeutically achievable plasma concentrations, irbesartan and hydrochlorothiazide in human plasma by liquid chromatography, Departement of Analytical

thereby enabling its use in bioequivalence trials. Chemistry, Faculty of Pharmacy, University of Ankara,

References Turkey, 2002, available online at: http://www.

sciencedirect.com/science?_ob=ArticleURL&_udi=B6 [1] Gilman and Goodman, The Pharmacological Basis of

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1548849389&_rerunOrigin=google&_acct=C00005022 [3] Martindale, The Complete Drug Reference,

1&_version=1&_urlVersion=0&_userid=10&md5=39f Pharmaceutical Press, USA, 2007.

a6759c63b9431803608ed5a1149b4&searchtype=a#m4. [4]

C.F. Lacy, Drug Information Handbook, 13th ed., Lexi

com.

Comp., USA, 2000, pp. 826-827. [8] N. Ganesan, M. Gomathi, Method development and [5] H.R. Brunner, Hypertens, Lippincott Williams & Wilkins

validation of irbesartan using LCMS/MS: Application to Inc., USA, 1997.

pharmacokinetic studies, A.J. College of Pharmacy, [6] S.K. Bae, J.K. Min, J.S. Eon, Y.C. Doo, HPLC

Chennai, India, 2010, available online at: Determination of Irbesartan in Human Plasma: Its

http://jocpr.com/vol2-iss4-2010/ JCPR-2- 4-740-746.pdf.

Application to Pharmacokinetic Studies, Wiley [9] Food and Drug Administration, Guidance for industry: Interscience, USA, 2008, pp. 568-572, available online at:

Bioanalytical method validation, 2001, available http://onlinelibrary.wiley.com/doi/10.1002/bmc.1154/abstr

online at http://www.fda.gov/downloads/Drugs act 2.

GuidanceComplianceRegulatoryInformation/Guidance [7] N. Erk, R. Bischoff, Simultaneous determination of

/UCM070107.pdf.

Journal of Life Sciences 6 (2012) 732-739

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

1 2 2 Hala Abou Shady 1 , Mona Kilany Abd El Gawad , Qadria Genedi and Wafaa Farouk 1. Microbiology Dept., Faculty of Science, Ain Shams University, Cairo 11566, Egypt

2. Microbiology Dept., National Organization for Drug Control and Research (NODCAR), Cairo 12553, Egypt

Received: October 13, 2011 / Accepted: January 12, 2012 / Published: July 30, 2012.

Abstract: Soil oxalate degrading bacteria is an important source for removing of toxic oxalate compounds. Isolation, screening, characterization, identification and fingerprint of the soil oxalate degrading bacteria from Egyptian fertile soil was aimed in this work. Isolated bacteria were first characterized using API 20 NE system. Nine RAPD primers were used in genetic fingerprinting of the characterized bacteria using RAPD-PCR. The preliminary characterization and polyphasic taxonomy for mesophilic, aerobic oxalate-degrading bacteria isolated from Egyptian fertile soil including auxanography using API galleries confirmed that the isolate belonged to the genus Burkholderia cepacia, the percentage of identity was 99.8% for 5167777 profiles. Fingerprinting by RAPD-PCR showed a DNA fingerprint ranging from 0 to 9 bands, over a size range of 344 bp to 3,280 bp.

Key words: Burkholderia cepacia, oxalotrophic, RAPD-PCR, fingerprint, API 20 NE, isolation, identification.

1. Introduction  useful in bioremediation, a practice using biological compounds to remove hazardous compounds and

Walter Burkholder was first isolated and described pollutants. Chlorinated phenols and phenoxyacetates

Burkholderia cepacia as the causative agent of are commonly used in agriculture as pesticides, bacterial rot of onion bulbs [1]. One area of difficulty

herbicides, and preservatives. They compose a major in the field has been the changing nomenclature and

group of recalcitrant environmental pollutants. B. older studies referring to ‘Pseudomonas cepacia’ or

cepacia has the ability to use these compounds as a ‘B. cepacia’ [2]. B. cepacia complex species are

source of carbon and energy, therefore breaking them soil-dwelling bacteria commonly found on plant roots

down and removing them from the environment [5, [3]. Pandoraea sp. OXJ-11 was isolated from soil

6]. B. cepacia is recognized for its abilities to samples, which can grow in the medium with oxalate

promote maize growth [7], to enhance crop yields [8, as the sole carbon and energy source [4].

9], and to suppress many soil borne plant pathogens

B. cepacia complex species are of significant [7, 10, 11]. Commercial systems for bacterial

environmental interest, along with their identification have been offered for approximately 30

antinematodal and antifungal properties to control years. Of the original manual systems, only the API

soil pathogens. They can also degrade a large variety

20 NE (bioMe’rieux, Marcy l’Etoile, France) has of toxic compounds. This makes them extremely

remained available. The API 20NE test outperformed the other systems in identifying B. cepacia [12].

Corresponding author: Mona Kilany Abd El Gawad, Ph.D., Seventeen B. cepacia isolates from environment in lecturer, research field: microbiology. E-mail:

mona.kilany@yahoo.com. Hong Kong were correctly identified using API 20

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of

Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

NE system [13]. Genotypic identification methods typed by RAPD. The RAPD typing method was found are emerging as an alternative or complement to

to be reproducible, discriminatory, and more sensitive established phenotypic identification procedures [14].

than PCR ribotyping and it is versatile fingerprinting For bacteria, 16S rRNA gene sequence analysis is a

method for studying the epidemiology of B. cepacia widely accepted tool for molecular identification

[27]. Analysis by RAPD fingerprinting demonstrated [15-18]. Polyphasic taxonomic studies have that the strains selected genetically heterogenous,

discovered that B. cepacia is a complex of nine representing 24 different B. cepacia complex strain genetically distinct species which defined as types [28]. A large collection of B. cenocepacia

genomovars [19]. After that, B. cepacia are isolates were genotyped by means of randomly recognized to be representative of a family of at least

amplified polymorphic DNA typing (RAPD) and

10 closely related, but genotypically distinct, strains rep-PCR using the BOX-A1R primer (BOX-PCR) known as the Burkholderia cepacia complex (Bcc)

[20]. B. cepacia adapted to live in the soil or Oxalotrophs may be used to remove oxalates from sediments may encounter divergent strains in their

wastewater and soil and to protect a host plant from particular niche more often than pathogenic bacteria

infection by oxalate-producing fungal pathogens (e.g., forms, which may never occur as a mixed infection, so

Sclerotinia sclerotiorum and Phytophtora infenstans), single bacterial species may be able to exhibit

oxalotrophic bacteria in the gastrointestinal tract of differing types of population structures depending on

man and animals and may be used in the treatment of environmental conditions. Further studies evaluating

kidney stones [30].

the temporal variation of B. cepacia genotypes are Thus in this report, isolation and use of API 20NE needed to draw final conclusions on the prevalence and

system in order to phenotypic identification of B. frequency of genetic exchange in this species [21].

cepacia isolate were conducted. Besides, the isolate There are several molecular methods that help

was evaluated for RAPD strain type by RAPD-PCR laboratory characterization of these types. Presently,

analysis.

the formerly recognized genomovars of ‘B. cepacia’ include B. cepacia (genomovar I), B. multivorans

2. Materials and Methods

(formerly genomovar II), B. cenocepacia (formerly

2.1 Enrichment and Isolation

genomovar III), B. stabilis (formerly genomovar IV),

B. vietnamiensis (formerly B. cepacia genomovar V), Several isolates of mesophilic oxalotrophs have

B. dolosa (genomovar VI), B. ambifaria (genomovar been isolated from Egyptian fertile soil and litter close VII), B. anthina (genomovar VIII), B. pyrrocinia

to the oxalate excreting plants such as sorrel, spinach, (genomovar IX) and B. ubonensis (genomovar X) [22].

tomato, potato, mango, cabbage and carrot which were More recently, Au Coin et al. (2010) [23] investigated

collected from different governorates; Giza, Kalubeia that Burkholderia cepacia complex (Bcc) bacteria

and Menofeia. Selective enrichment cultures were include at least 17 separate species producing

used to obtain mixed bacterial populations that were lipopolysaccharide (LPS) structures that are quite

capable of oxalate utilization according to Aragno and different. Three PCR-based methods for typing B.

Schlegel (1992) [31]. Morphologically distinct cepacia have also been reported: (PCR ribotyping)

colonies that developed clear zones on the CaOx [24], randomly amplified polymorphic DNA medium were selected. Clear zones around colonies (AP-PCR or RAPD) [25], and enterobacterial could be due to the oxalate being degraded and/or repetitive intergenic consensus sequence PCR [26].

transformed and/or mineralized. The isolates were All B. cepacia complex isolates were genetically

purified by streaking and checked microscopically.

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

2.2 Maintenance of the Culture manufacturer’s instructions with including RNase A (200 µg/mL) in extraction buffer. Extracted genomic

The purified cultures were maintained on Schlegel’s

DNA was kept at –70 °C till use.

basal mineral medium agar slants and stored at 4 °C or as glycerol suspensions (20%, v/v) of cells at –20 °C [32].

2.8 B. cepacia DNA Amplification by RAPD-PCR

2.3 Strain Characterization B. cepacia was genetically typed by RAPD, using nine 10-base oligonucleotide primers as described by

Preliminary characterization of strains was done by Mahenthiralingam et al. (1996b) [37] with some

colony morphology, cell morphology, motility and modifications. The procedure was performed using

Gram stain using standard methods [33]. QIAGEN ® Fast Cycling PCR Kit (Qiagen). Each

2.4 Resistance to Antibiotics reaction was carried out in a final volume of 25 μL Resistance to antibiotics of the selected isolate was

with the final concentration containing 1 × reaction determined by the agar diffusion method using

buffer including the concentration of 50 ng genomic standard antibiotic disks (Oxoid). The inhibition zones

DNA, 4.0 mM Magnesium Chloride, 2 units Taq were examined after 24 and 48 h. Selected isolate was

DNA Polymerase, 0.4 mM dNTP-mixture and 10 pM considered susceptible when the inhibition zone was

of each 10-base primer separately. Universally used

12 mm or more in diameter [34]. primers were Primers I (Pharmacia), OPA-1 (GE Healthcare Limited, Amersham Place Little Chalfont,

2.5 Heavy Metal Ion Resistance and Buckinghamshire) and Gibco-BRL 3 The resistance to heavy metal ions (Zn 2+ , Ni 2+ , Pb 2+ ,

(Gibco-BRL). In addition, 6 different newly designed

Cr 2+

2 O 7 , Co , Cu , and Hg ) was evaluated by the primers were used as shown in Table 1. Cycling agar-diffusion method [35]. Clear zone surrounding a

procedure was performed using PTC 200 Peltier disk indicated that the bacteria were sensitive to that

Thermal Cycler (MJ Research-USA) with the metal.

following profile: Pre-denaturation of DNA at 95 °C for 5 min followed by 45 cycles of denaturation at

2.6 Identification Using the API 20 NE System

95 °C for 1 min, annealing at 36 °C for 1 min and The API 20 NE (bioMe´rieux, Marcy-I’Etoile, France)

extension at 72 °C for 2 min and final extension of 2 testing was performed according to the instructions of

minutes at 72 °C.

the manufacturer. Substrate assimilations and

2.9 Gel Analysis

biochemical tests were read after 24 and 48 h. Interpretation of the results was done using the

The PCR products were run in 1.3% gel. The gel identification software version 6.0 [14]. Oxidase,

was run at appropriate distance (short run) and then peroxidase and catalase activities were tested. Growth

photographed. The gel was left to run more distance

on MacConkey agar without crystal violet was observed. Table 1 Primers used in RAPD -PCR amplification of B.

cepacia genomic DNA.

2.7 Preparation of Genomic DNA from Bacterial Culture

Primer name

Sequence (5’ to 3’) PROBEDB_PUID

Saad-2 GTGCTACGTC Isolation of genomic DNA from B. cepacia was 10554779

Osama-5 GACCATCGTC 10554777 done according to McDowell et al. (2001) [36] with

Osama-6 CGCTC 10554778 brief modification using DNA easy Blood & Tissue

Essam-7 GATCGGACAC 10554774 Kit from QIAGEN. Kit buffers preparations and

Essam-8 GCTCGAACTT 10554775 Essam-9 CCACGCGCAA genomic DNA extraction was done according to the 10554776

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of

Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

(long run) to separate overlapped bands. Pictures were the descending order of metal toxicity observed was photographed using gel documentation system Hg > Co > Cu > Cr 2 O 7 > Ni > Zn > Pb. (ProXima AQ-4). Migration distance was manually

3.5 API 20 NE

calculated and bands sizes were calculated according The commercial API 20 NE system showed an

to Raghava (2001) [38] using online program excellent identification for Burkholderia cepacia

(http://imtech.res.in/raghava/dnasize/). where the percentage of identity was 99.8% for

3. Results

5167777 profiles. The results of the conventional

3.1 Isolation and Screening of Bacterial Isolates biochemical and the assimilation tests are presented in

Table 4.

Oxalate-enrichment cultures from the soil and in

3.6 Genotype Characterization Using RAPD-PCR the litter below oxalate-bearing plants of different

fertile regions yielded numerous oxalate-degrading The polymorphisms amplified by the nine RAPD isolates that could catabolize oxalate aerobically.

primers from DNA extracted from B. cepacia showed Screening revealed 16 isolates were the most potent

a DNA fingerprint (Figs. 1a & 1b) ranging from 0 to 9 to utilize oxalate as a sole carbon source. Bacteria

bands, over a size range of 344 bp to 3280 bp (Table 5). isolated from soil adjacent to the sorrel plant found

4. Discussion

in Giza governorate (So-G) showed the highest potential of oxalate utilization, thus it was chosen for

The bacteria, which are capable of using oxalate as further study.

a sole carbon and energy source, are described as

Table 2 Antibiotic sensitivity of the isolates So-G.

3.2 Isolate Characterization Inhibition zone after

Antibiotic

It is rod-shaped, free-living, motile, Gram-negative 24 h (mm) 48 h (mm)

4.0 bacilli, catalase & oxidase positive, ranging from 3.6

Ofloxacin (5 µg )

3.0 3.5 1.6-3.2 μm forming visible pinpoint non pigmented

Ceftazidime (30 µg)

2.8 3.0 colonies within 72 hours and the colonies appear to be

Cefoperazone (75 µg)

2.6 3.5 smooth and somewhat elevated.

Claforan cefotaxime (30 µg)

Ciprofloxacin (5 µg)

Cefepime (30 µg)

3.3 Antibiotics Sensitivity

Tobramycin(10 µg)

2.0 2.6 Sulphamethxazole trimethoprime (25 µg) 2.3

2.5 Isolate So-G was highly sensitive to Ofloxacin (5 µg),

2.0 2.0 Ceftazidime (30 µg), Claforan cefotaxime (30 µg),

Erythromycin (15 µg)

1.3 1.5 Cefoperazone (75 µg), Ciprofloxacin (5 µg), moderately

Amoxicillin- clavulanic acid (30 µg)

Ampicillin sulbactam (20 µg)

0 sensitive to Tobramycin (10 µg), Sulphamethxazole 0

Cefexime (5 µg)

trimethoprime (25 µg), Cefepime (30 µg), Table 3 Heavy metal ion tolerance of selected isolate.

Erythromycin (15 µg), Amoxicillin- clavulanic acid

Inhibition zone (mm) (30 µg) and resistant to Ampicillin sulbactam (20 µg)

Heavy metals

Hg 2+ 23 Co and Cefexime (5 µg) as shown in Table 2. 2+ 16 Cu 2+ 15

3.4 Heavy Metal Tolerance

Cr 2 O 7 15

The isolate So-G showed varied degrees of

Ni 2+ 13 Zn 2+ 12

tolerance to the seven metals tested (Table 3). So-G

Pb 2+ 11

isolate was found to be highly sensitive to mercury,

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

Table 4 API 20 NE identification table for B. cepacia.

Table 5 Products of RAPD -PCR.

Test Result Test Result Test Result

RAPD-PCR products (bp) NO3 + PNPG + GNTa +

Primer#

P1 3080 2274 1349 1013 714 344 - - - TRP -

P2 2023 -------- GLU + ARAa + ADIa +

GLUa +

CAPa +

P3 2502 2056 1312 998 --- - - ADH + MNEa + MLTa +

URE - MANa +

-------- ESC -

CITa +

P4 -

P5 2185 -------- GEL -

1779 1649 1505 1363 1020 810 693 488 P7 1829 1405 886 704 --- - - P8 2463.76 1446 1160 750 --- - - P9 2398 1925 1350 977 876 740 653 527 -

from a freshwater stream that grew with oxamate or oxalate in the presence of sulfate and yeast extract. This strain was described as a new subspecies of Desulfovibrio as D. vulgaris sub sp. oxamicus [43]. Among aerobic oxalotrophic bacteria, Methylobacterium extorquens strain AM1 [44, 45] have been completely described from the physiological and biochemical points of view. The aerobic oxalotrophic bacteria are taxonomically extremely heterogeneous. Most species can be classified with the Gram-negative genera, Pseudomonas, Alcaligenes, Ralstonia and Xanthobacter . Starkeya novella (formerly Thiobacillus novellus ) is a strictly aerobic, facultative chemolithoautotrophic, methylotrophic Gram- negative, and rod-shaped sulfur bacterium [46].

A limited number of aerobic oxalotrophic bacteria have been completely described. Most of them are

Fig. 1 Electrophoresis pattern of RAPD-PCR products.

facultative methylotrophs and/or facultative hydrogen- a: short run; b: long run yellow arrows indicate sizes of

oxidizing chemolithoautotrophs [30]. GelPilot Wide Range Ladder (Qiagen) while green arrows

Antibiotic sensitivity of the isolated bacteria was indicate separated overlapped bands. studied. Several antibiotics were used in this study.

being “oxalotrophic”. Oxalotrophic bacteria are Antibiotic sensitivity of Burkholderia cepacia was numerous and widespread in soil and that are

also studied by other workers [47, 48]. Sahin (2002) relationship exists between the presence of the

[34] reported that Bulkholdaria cepacia was resistant oxalogenic plants and the relative abundance of

to two or more of the studied antibiotics. In the current oxalotrophic guilds in the total bacterial communities

study, Burkholderia cepacia was shown to possess [39]. In the present study oxalotrophic bacteria were

resistance to some antibiotics such as ampicillin. isolated from different Egyptian fertile soils. Many

Sahin (2005) [32] also showed that Burkholderia workers [34, 40, 41] also isolated several strains of

cepacia possess multiple antibiotic resistance such as mesophilic oxalotrophs from soil and litter close to the

ampicillin. In our study Burkholderia cepacia was oxalate excreting plants.

shown to be sensitive to ceftazidime, this result is in Postgate (1963) [42] isolated Desulfovibrio strains

agreement with that result obtained by Gautam et al.

Isolation, Phenotypic Identification and Fingerprint by RAPD-PCR of

Oxalotrophoic Burkholderia cepacia from Egyptian Fertile Soil

(2009) [49]. to separate these compact bands. These compact Mercury was found to be the most toxic metal to all

RAPD-PCR products were shown as a result of the three types of (yellow pigmented, pink pigmented

amplification of genomic DNA using primer P3 and and non-pigmented) oxalotrophic bacteria. It was

P6. Amplification of genomic DNA using other previously shown that the descending order of metal

primers did not require further migrations of the toxicity observed was Hg > Cr > Cu = Zn = Pb > Co >

RAPD-PCR product bands.

Ni. Our results showed the same pattern indicated by

5. Conclusion

Sahin (2002) [34]. Comparison of tolerance to mercury and chromium indicated that several of the

Mesophilic oxalotrophic bacteria isolated from studied isolates by Langenbach (1988) [50] were

Egyptian fertile soil close to the oxalate excreting sensitive to these metal ions, but each isolate exhibited

plants were identified using API 20 NE system. The

a characteristic level of sensitivity to each metal. isolate was confirmed to be Burkholderia cepacia. For the API 20 NE, a result seems reliable when a

Genetic fingerprint of Burkholderia cepacia was done species assignment is reported as an excellent or very

using RAPD-PCR.

good identification [14]. In the current study, API 20

Acknowledgments

NE was used to identify isolated bacteria and showed excellent identification of the isolated bacteria. Other

The authors gratefully acknowledge the National studies showed that API 20 NE correctly identified

Organization for Drug Control and Research for 792 of 800 isolates of Burkholderia pseudomallei

providing financial support. The authors also thank isolates (sensitivity 99%) and of the B. cepacia

Prof. Dr. Essam I. Hassan from National Organization isolates, nine profiles were obtained and 17 (89%)

for Research & Control of Biologicals, for his kindly were correctly identified with more than 98% ID [13].

suggestions for the experiment, and verification of the Further, one out of 10 known species (formerly,

species identifications obtained by the PCR procedure genomovar) of the genus Burkholderia can be labeled

described and Dr. Khalid El Gayar, VACSERA, for without molecular processing. Many laboratories are

helping in API NE20 identification of the isolate. identifying the organisms by automated systems.

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Journal of Life Sciences 6 (2012) 740-746

The Effect of a Dietary Supplement Spirulina

and Bifidobacterium adolescentis on the

Cholesterol-Lowering in Vitro and in Vivo

Amel Doumandji, Dahmane Alili and Abderrahmen Benzaiche Department of Agricultural Sciences, Faculty of Agro-Veterinary, Saâd DAHLAB University, Blida 270-9000, Algeria

Received: November 25, 2011 / Accepted: January 16, 2012 / Published: July 30, 2012.

Abstract: The survey’s results showed a significant percentage of women and especially men having an above normal cholesterol. In order to help reduce excessive rate of blood cholesterol, we used a local strain of Spirulina (Tamanrasset, in south of Algeria) associated with probiotic bacteria (Bf. adolescentis). Experiments in vitro showed a significant degradation of total cholesterol by the combination of Spirulina and Bf. adolescentis (74.5%) after 72 hours incubation at 37 °C. A cholesterol is added to the standard diet mice in order to increase the total cholesterol for three lots. However, the rates of total cholesterol in mice receiving lower fermented milk with Bf. adolescentis enriched by dry Spirulina. This shows that the decrease of cholesterol rate is closely related to the presence of Bf. adolescentis and Spirulina. In vitro and in vivo results show, however, that it is possible to obtain a natural product (Spirulina) and a pseudo-strain lactic (Bf. adolescentis) and to participate in the prevention of cardiovascular disease risk factor whose hand is cholesterol.

Key words: Spirulina, Bifidobacterium adolescentis, hypocholesterolemic activity, in vitro, in vivo.

1. Introduction  Safety of Health Products, Mar. 2005).

Spirulina , blue-green, has now become a medical Cholesterol is essential for the functioning of the food. The world of medical research has discovered cell. It is a substance that plays a fundamental role, that the nutrients of Spirulina (good quality proteins, either as parent compound of hormones, bile acids and balanced fatty acids, vitamins, antioxidants and vitamin D, either as a structural component of cell minerals) contributed to the fight against many health membranes or where it promotes their fluidity and problems like high cholesterol [4]. permeability [1]. But the excess of cholesterol is In Algeria, we are at the stage of the craft dangerous; it is implicated as a risk factor for (home-made) and experimental production. The cardiovascular disease [2]. These diseases are the unique Algerian which knows perfectly the process of leading cause of death worldwide. They are production of this seaweed is Mister HIRI. He responsible each year of the death of more than 17 succeded to make it move from its natural million people, or 30% of all deaths worldwide and 25 environment (El Guelta) to a pool. He has a pond of a million deaths are expected in 2020 [3]. So, the

surface slightly over 20 m 2 and produces 20 kg of reduction of the cardiovascular risk depends on the

Spirulina dry per year. It is in the region of reduction in the concentration of the cholesterol, Tamanrasset that it takes place [5]. especially LDL-cholesterol (French Agency for the The probiotics are living microorganisms that

Corresponding author: Amel Doumandji, Ph.D., professor, favorably impact on the host by improving its research field: agricultural sciences . E-mail: corino147@yahoo.fr.

intestinal balance. They are among the

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

microorganisms that interest researchers for their The test tubes of the three combinations were cholesterol-lowering actions [6].

incubated at 37 °C.

This study is conducted in this way, in order to Cholesterol rate is measured at different times [after prove the cholesterol lowering effect of

24 h (T1), 48 h (T2) and 72 h (T3)].

Spirulina- associated Bifidobacterium adolescentis.

2.2.1.2 Analysis of Total Cholesterol in Vitro

The present study is divided into four steps: For the analysis of total cholesterol we used  In vitro study of the effect of strain Bf.

enzymatic colorimetric method.

adolescentis on cholesterol rates; Free cholesterol and esterified cholesterol in the  In vitro study of the effect of Spirulina on

sample, give as coupled reactions described below, a cholesterol rates;

colored complex quantified by spectrophotometry.  In vitro study of the effect of Spirulina associated

Cholesterol esterase

with strain Bf. adolescentis on cholesterol rates;

Cholesterol ester → fatty acids + Cholesterol

 In vivo study of the effect of Spirulina strain

Cholesterol oxydase

associated with Bf. adolescentis on cholesterol levels.

Cholesterol +1/2 O 2 +H 2 O 2 → Cholestenone + H 2 O

Peroxydase

2. Materials and Methods

H 2 O 2 + 4-Aminoantipyrine + Phenol → quinoneimine + 4 H 2 O

2.1 Biological Material

1 mL of each combination removed and centrifuged at 3000 towers/min for 10 min. The supernatants were

 The strain of Bifidobacterium adolescentis was collected for the determination of total cholesterol isolated and identified from the stool of healthy (Fig. 1): Prepare the test to be analyzed as indicated in infants breastfed by their mothers. It is preserved in

Table 1.

skim milk at a temperature of –20 °C; After the preparation of the tests, the tubes are well  The dry Spirulina harvested and dried in the shaken and incubated for 5 min at 37 °C. You have to region of Tamanrasset (south of Algeria); read the absorbance of the standard and sample in  The mouse comes from the Pasteur Institute of comparison with the white (reagent A) at 500 nm. Algiers.

Expression of results

2.2 Methods

2.2.1 The in Vitro Study

2.2.1.1 Preparation of Combinations From the precultures of Bf. adolescentis, dried Spirulina and standard cholesterol, the authors realize the following combinations:

 First combination: 1 mL pre-culture of Bf. adolescentis + 1 mL cholesterol standard + 9 mL of MRS medium;

Fig. 1 The supernatants recovered after centrifugation.  Second combination: 0.9 mg of Spirulina (it is

added at a rate of 10%) + 1 mL cholesterol standard + Table 1 Method of analysis of total cholesterol in vitro.

9 mL of sterile distilled water; Blanc Standard Sample  Third combination: 1 mL of the preculture + 0.9

Standard cholesterol

10 µL

Sample - - 10 mg of Spirulina (it is added at 10 %) + 1 mL µL

1 mL cholesterol standard + 9 mL of MRS medium.

Reagent (A)

1 mL

1 mL

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

Cholesterol concentration of the sample is calculated as follows:

C ch =A Sample /A Standard *C Srandard

where C: concentration of cholesterol; A sample : absorbance of the sample; A standard : absorbance of the standard; C standard : standard concentration (2 g/L).

2.2.2 The in Vivo Study The significant results of the cholesterol-lowering effect of Spirulina associated with Bf. adolescentis observed in vitro have generated an in vivo study

Fig. 3 The milk powder.

performed on NMRI mice. For this we have provided two batches of four mice

5 min. Then the tubes were stored at –20 °C until the and a set of 4 control mice, 12 in total these mice are

day of their uses.

bred NMRI Swiss, aged 45 days and each weighing In order to confirm the effectiveness of

32 g on average. pasteurization, a tube containing reconstituted milk The mice are placed in metal cages of 30 cm square,

and pasteurized straw is left on for 24 h at room kept in a well veterinarian ventilated cabinet, at a

temperature in order to note the presence or absence temperature of 27 °C. With a 12 h light continuously

of coagulation of the environment. from 8 am to 20 pm the food and water are distributed

The reconstituted milk is homogenized and regularly and cleaning of cages is every three days

distributed in sterile tubes at a rate of 9 mL in each (Fig. 2).

tube.

2.2.3 Diet Preparation

2.2.3.2 Development of Fermented Milk Enriched

2.2.3.1 Reconstitution of Milk Powder with Bifidobacterium adolescentis and Spirulina The milk is produced in December 2009 in New

A colony is removed and inoculated into 9 mL of Zealand (Fig. 3).

reconstituted milk and Spirulina-enriched at 10% and The milk reconstitution is made according to the

an incubation at 37 °C for 18 hours. instructions on the packaging or 144.44 g of powdered

2.2.3.3 Constitution of Experimental Groups whole milk powder which are dissolved in 1 L of

We have 12 mice that were fed previously with a sterile distilled water.

standard diet (137 g of granules per day feed and water) The pasteurization of milk is produced at 90 °C. for

during an adjustment period of 7 days. Then the following 7 days, the mice diet was supplemented with

other constituents listed in Table 2. The cholesterol is added to the diet to raise blood cholesterol rates in mice.

The preparation of the diet is made every day. After 7 days, sampling of blood is realized for three

prizes (lots) to measure total cholesterols (mice have to

be on an empty stomach during 12 h). The blood is collected by puncture at the retro-orbital sinus and collected in dry tubes, it can take Fig. 2 The mice used in the study in vivo .

up to 0.5 mL of blood [7] (Fig. 4).

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

Table 2 The experimental groups.

RS + Ch + Al RS: Standard diet 137 g/day. Ch: Cholesterol standard 0.1 mL/day. Al: Nutriceutical (fermented milk to Bf. adolescentis enriched with Spirulina) 1 mL/day.

adolescentis , because we note an important decrease of 94% to QT4. Secondly, the Spirulina for which we observe a decrease of the rate of 69.5% cholesterol. The presence of Bf. adolescentis stimulates the hypocholesterolemic action of the Spirulina with a rate of decrease of the rate of 74.5% cholesterol (Table 3).

The objective of this study is to follow the effect hypocholesterolemic of the Spirulina alone, of Bf.

Fig. 4 Removal of blood.

adolescentis as well as their association.

The obtained results indicate that there is an

3. Results and Discussion

important degradation of the cholesterol for three

3.1 The Study of Cholesterol-lowering Effect of Spirulina Table 3 Results of in vitro cholesterol -lowering effect

associated Spirulina with Bf. adolescentis.

Table 3 and Fig. 5 are reminiscent of the results of QT0 QT1 QT2 QT4

the assay of total cholesterol in vitro at different times

2 0.61 0.44 0.24 of incubation.

Spirulina

2 0.12 0.10 0.04 According to our results, the hypocholesterolemic

Bf. adolescentis

2 0.51 0.32 0.19 effect is optimal under the effect of only Bf.

Spirulina + Bf. adolescentis

QT: Quantity of cholesterol at different times (g/L).

Fig. 5 Results of in vitro cholesterol -lowering effect associated Spirulina with Bf. Adolescentis.

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

combinations (overalls). But that of Bf. adolescentis of hypercholesterol level responsible for coronary is the most important (94%), while that of the

diseases. As an example, Scientifics brought to light a Spirulina is 69.5% since the association both gives a

decrease of the rate of blood cholesterol to subjects 74.5% rate. The assimilated quantities increase

subjected to a diet supplemented with Lactobacillus according to time.

plantarum 299 v [8].

It was reported that consumption of fermented milk

3.2. The Study of the Cholesterol Lowering Effect of with Lactobacillus decreased greatly the risk of Spirulina Associated with Bf. adolescentis in Vivo cardiovascular disease in Maasai people in East Africa

According to the results obtained in vitro it is noticed (Kenya) despite the atherogenic diet [9]. Decreases in that the Spirulina associated with Bf. adolescentis

blood cholesterol by the administration of yogurt or contributes to decrease the rate of 74% cholesterol. To

fermented milk with lactic acid bacteria were confirm our results in vivo study was realized.

observed in chickens [10], humans [11], rabbit [12] The dosage of the total cholesterol of mice showed

and rats [13]. The administration of Lactobacillus that the daily grip (taking) of 1 mL of milk fermented

reuteri CRL 1098 (104/day) to hypercholesterolemic with Bf. adolescentis enriched there Spirulina at the

mice for 7 days decreased the concentration of total rate of 10% lowers the rate of blood cholesterol (Table

blood cholesterol by 38%. This becomes equivalent to

4, Fig. 6). the concentration of control mice (67.4 mg/mL) [11]. The results of the activity hypocholesterolemic

This low dose of L. reuteri has introduced a 40% observed in vitro and in vivo in this present study

reduction in triglycerides and a 20% increase on the confirm those exposed in the scientific literature.

rate of HDLch/LDLch without translocation of Indeed, preliminary studies revealed that the bacteria of the microflora to naive spleen or to the consumption of yogourt or fermented milk containing

liver. The authors concluded that administration of probiotics pull a decrease of the rate of cholesterol in

probiotics contributed to the normalization of blood the blood, and, consequently, the reduction of the risks

cholesterol.

Table 4 Percentage of total cholesterol in each batch.

3rd lot Average of the rate of cholesterol g/L for three lots

Fig. 6 T he determination of total cholesterol in mice in each group .

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

However, this effect is not always observed [14] the latter case, the uptake depends on the pH, the [15]. For example, studies that made use of concentration of esterified cholesterol and the nature Lactobacillus acidophilus L1 in hypercholesterolemic

of the bile salt used [20].

men conclude that a small reduction (3%) in LDL

4. Conclusions

cholesterol and no significant change in total cholesterol, HDL cholesterol or level of blood

The cholesterol is an elementary molecule essential triglycerides [16]. In another study, of 17 for the well functioning of our body. Excess in the blood, normocholesterolemic young women, consumption of

it becomes responsible of cardiovascular diseases. yogurt containing probiotics (100 g/day) significantly

The Spirulina is a cyanobacteria traditionally used increased the HDLch not the LDLch [17]. It is

for centuries by some populations. This Seaweed is however necessary to note that origins used in these

characterized by its beneficial effects on human health, studies are different.

including cholesterol-lowering effect. The

Spirulina is endowed with a Bf. adolescentis is a strain that was isolated from cholesterol-lowering activity: polyunsaturated fatty

the stools of infants exclusively breastfed. The MRS acids (PUFA), omega-3 and Omega-6 of Spirulina

medium is a good culture environment for the growth would prevent the accumulation of cholesterol in the

of our strain of interest. This bacterium degrades body. This could partly explain the decreased levels of

cholesterol metabolites found in the faces. cholesterol and triglycerides observed in experiments.

We investigated the cholesterol-lowering effect of These experiments on humans, however, are made

Spirulina associated with Bf. adolescentis, in vitro and with low numbers [18, 19].

in vivo . The test results show an in vivo degradation of The preliminary study of the behavior of the cells of

cholesterol by Bf. adolescentis associated with bifidobacteria towards the cholesterol showed that

Spirulina.

these microorganisms can move the cholesterol The results of experiments in vivo indicate that milk contained in an environment of culture. This fermented with Bf. adolescentis enriched Spirulina has movement of cholesterol takes place in the presence of

a cholesterol-lowering effect (a decrease of 37.5%). bile. The study of the fate of cholesterol moved has

In perspectives, it would be interesting to consider: shown that it can be picked up by the growing cells

 Optimize the culture conditions of Spirulina; and precipitate with deconjugated bile salts at acidic

 Explore the synergy or antagonism between the pH (< 5.4). Among the various bile salts bile

lactic flora, pseudo-lactic acid and Spirulina; component. The trihydroxy conjugated appear to be

 To study the hypocholesterolemic action of Bf. responsible for the capture. The non-proliferating cells

adolescentis and Spirulina on other animals; of bifidobacteria are unable to capture the original

 To study in vivo cholesterol-lowering action of Bf. serum cholesterol. The activity of deconjugation of

adolescentis only;

bile salts is necessary for the uptake of cholesterol.  To study the in vivo hypocholesterolemic action Strains of bifidobacteria no conjugated are unable to

of Spirulina alone;

capture cholesterol even in the presence of bile salts.  To dose the LDL-cholesterol and HDL In addition, the products of deconjugation of bile salts

cholesterol as part of an in vitro and in vivo. (deconjugated bile salts and amino acids) have no

effect on the uptake of cholesterol. The esterified

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A. Dilmi-Bouras, Suivi de Streptococcus thermophilus et by growing cells but also by non-proliferating cells. In

de Lactobacillus bulgaricus sur le métabolisme du

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on

the Cholesterol-Lowering in Vitro and in Vivo

cholestérol, Thèse de Doctorat, INA, El Harrach, Alger, tolerance and cholesterol reduction by Enterococcus 2002, p. 99.

faecium , a candidate microorganism for the use as a [2] J. Fábregas, L. Ferrón, J. Abalde, B. Cabezas, A. Otero,

dietary adjunct in milk products, Milchwissenschaft 51 Changes in the gross chemical composition of mass

(1998) 383-385.

cultures of the marine microalgae Dunaliella tertiolecta

C. Thakur, A. Jha, Influence of milk, yoghurt and with different aeration rates, Biores. Technol. 53 (1995)

calcium on cholesterol-induced atherosclerosis in rabbits, 185-188.

Atherosclerosis 39 (1981) 211-215. [3] R. Saïle, H. Taki, Cholestérol, lipoprotéines et

[13] K.K. Grunewald, Serum cholesterol levels in rats fed athérosclérose: De la biochimie à la physiopathologie,

skim milk fermented by Lactobacillus acidophilus, Les Technologies de Laboratoires 2 (2007) 4-11.

Journal of Food Science 47 (1982) 2078-2079. [4] U.V. Mani, S. Desai, U.M. Iyer, Studies on the long-term

G. Kiessling, J. Schneider, G. Jahreis, Long-term effect of Spirulina supplementation on serum lipid profile

consumption of fermented dairy products over 6 months and glycated proteins in NIDDM patients, Journal of

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Nutrition 56 (2002) 843-849.

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Cardiovascular Diseases 16 (2006) 531-535. [6] C.J. Saloff-Coste, Lactobacillus acidophilus, World

[16] J.W. Anderson, S.E. Gilliland, Effect of fermented milk Newsletter Danone 13 (1997) 2-7.

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serum cholesterol in hypercholesterolemic humans, Biologie et techniques expérimentales, Thèse de

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H. Bukowska, J. Pieczul-Mroz, M. Jastrzebska, K. probiotic and conventional yoghurt on the plasma lipid Chelstowski, M. Naruszewicz, Decrease in fibrinogen

profile in young healthy women, Annals of Nutrition and and LDL-cholesterol levels upon supplementation of

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supplementation of Spirulina on hypercholesterolemic (1998) 437-438.

patients, Journal of Food Science and Technology 33 [9]

G.V. Mann, Studies of a surfactant and cholesteremia in

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Journal of Life Sciences 6 (2012) 747-753

Attitudes and Practices of Health Care Providers Regarding the Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

1 1 1 2 Frederic Nogbou Ello 1 , Thomas Yapo Aba , Ismael Ouattara , Didier Koumavi Ekouévi , Offoue Kra , Pierre

3 3 1 Aimé Assemian 1 , Amath Wade , Serge-Paul Eholie and Emmanuel Bissagnene 1. Department of Infectious and Tropical Diseases in Abidjan, University Hospital of Treichville, 01 BP V3 Abidjan 01, Cote d’Ivoire

2. PAC-CI Program, Abidjan, University Hospital of Treichville, USAC Extension, 18 BP 1954 Abidjan 18, Cote d’Ivoire 3. Laboratoire Sanofi Aventis, Sanofi-Aventis, Dakar, BP 3529, Sénégal

Received: December 06, 2011 / Accepted: March 06, 2012 / Published: July 30, 2012.

Abstract: Malaria is a major public health problem in the Cote d’Ivoire where it is the leading cause of morbidity and mortality. The objective of this study was to assess the knowledge, attitudes and practices of health care providers on the diagnosis and therapeutic procedures used to ensure the management of uncomplicated malaria in the health district of Abidjan. A cross-sectional study was conducted from November 2008 to December 2008 among 169 caregivers who treated 1,691 cases of uncomplicated malaria. Results: The knowledge of the caregivers of the national guidelines regarding malaria was sufficient (89%). A diagnosis of uncomplicated malaria was established only on clinical arguments (70%). The percentage of patients who were prescribed artemisinin-based combination therapy (ACT) was 86%. However, 25% of the antimalarials drugs that were prescribed to patients (oral quinine, artemisinin derivatives monotherapy, and herbal medicine) did not meet the recommendations on first choice treatment as suggested by the national program against malaria. These results indicate sufficient knowledge of the national guidelines for the treatment of malaria by health care providers as evidenced by acceptable prescriptions for ACTs despite an incomplete knowledge of the guidelines.

Key words: Malaria, supplier of service, ACT, knowledge, Africa.

1. Introduction a top priority. Significant efforts have been made, but the clinical management of malaria is still impeded by

Malaria is a major public health problem in tropical the chemo resistance of Plasmodium falciparum to the countries because of its frequency, severity and antimalarials drugs that are available in Africa [4, 5]. economic consequences. According to WHO, 247 This emergence of resistance and the loss of million cases of malaria were reported worldwide in effectiveness of antimalarials drugs have challenged 2006 with 881,000 deaths in Africa, and 91% and the search for new treatments, including 85% of these deaths occurred in children under 5 artemisinin-based combination therapies (ACTs) that years of age [1-3]. Therefore, malaria is included on are considered to be the best treatment for the list of priority conditions of the National Health uncomplicated malaria [6-8]. In Sub-Saharan Africa, Agendas in each country in Africa, which two ACTs are widely known and prescribed: demonstrates the political will to make malaria control artesunate-amodiaquine (ASAQ) and

artemether-lumefantrine (AM/LF) [9-12]. However, Corresponding author : Ello Nogbou Frédéric, Ph.D.,

research fields: infectious and tropical diseases. E-mail: their use in clinical practice raises the following ellonogboufrdric@yahoo.fr.

Attitudes and Practices of Health Care Providers Regarding the Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

difficulties: 1) tolerance or deemed tolerance; 2) the  Patients (children and adults) who underwent lack of human resources for their prescription; 3) the

counseling for signs of uncomplicated malaria. profile of patient eligibility to receive such care; and 4)

Informed consent was obtained from both the the sequence of their prescription in case of treatment

practitioners and the patients who were seen in the failure of one of the other CTA molecules. In Africa,

consultations. The study was approved by the the management of malaria is performed by doctors,

National Ethics Committee for the Life and Health nurses, midwives, community caregivers and other

Sciences of Cote d’Ivoire.

relay community members from remote health centers To calculate the size of our study population, we serve as caregivers. Under these conditions, the lack

used the following hypothesis instead. We estimated of knowledge and attitudes about treatment strategies

that 60% of the patients were seen within 24 hours can lead to noncompliant practices from the various

and adhered to the appropriate treatment according to providers.

the Abuja Declaration [15]. Using an alpha risk of This study aimed to evaluate the knowledge and

Type (5%) and an accuracy of ± 3%, the number of practices of caregivers on the prescription of subjects required for inclusion was estimated to be anti-malaria drugs and investigate the methods used to

1,025. We estimated that 10% of the patients refused diagnose uncomplicated malaria.

to participate in this study, and a total of 1,130 subjects with signs of uncomplicated malaria were

2. Materials and Methods

included in this study.

2.1 Study Design

2.3 Data Collection

This was a descriptive cross-sectional study that took Data collection was performed by 13 doctors who place from Nov. 14, 2008 to Dec. 26, 2008 in health

were previously trained for a week in the Department facilities in the city of Abidjan. Abidjan is the economic

of Infectious and Tropical Diseases (DITD) at the capital of the Cote d’Ivoire and an agglomeration

University Hospital of Treichville. Data was collected comprising 11 communes: Port-Bouet Vridi, Kumasi,

with two standardized questionnaires: one for Marcory, Treichville, Plateau, Cocody, Adjame, caregivers and one for patients. The following variables

Williamsville, Abobo, and Attiécoubé Yopougon. The were collected: demographics (age and gender); job estimated population is 4,000,000 inhabitants [13]. The

tenure; knowledge; attitudes and practices of the health weather conditions were greater than average rainfall in

care providers; and training for the prescription of ACT 2008 and temperature of 1,200 mm [800 to 1,800] and

and the therapeutics used to treat malaria.

27 °C, respectively. The city is entirely built on the lagoon Ebrié between the Atlantic Ocean to the South, 2.4 Statistical Analysis

the town of Bingerville to the East, and Banco forest to

A database was created with the software program the North and West. As a result, malaria is endemic

EpiData, and data were analyzed using the software there all year and represents 50% to 60% of the reasons

program Stata, Version 9.2 (StataCorp, College for consultations in health centers [14].

Station, Texas). The results were presented as the frequencies or means with standard deviations. A

2.2 Study Population comparison of the frequencies was performed with a The study population consisted of:

Chi-squared test, and a comparison of the mean or  Health care providers who were involved in the

median values was performed with Student’s t-test or management of malaria, including the diagnosis and

a nonparametric test. The difference was statistically treatment;

significant if the P < 0.05.

Attitudes and Practices of Health Care Providers Regarding the

Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

3. Results

than among nurses and midwives (92.6% vs. 82.4%, P = 0.013) (Table 1).

3.1 Demographic Characteristics of the Study In addition, knowledge of the guidelines was Participants statistically different depending on the length of

In total, 254 (98%) out of 260 caregivers professional experience (P = 0.045). Among the participated in the survey. There were 163 physicians

health care providers with knowledge of the (64.2%), 77 (30.3%) nurses and 14 (5.5%) midwives.

guidelines, 89.6% and 95.2% had work experience of We collected information on 1,691 cases of less than 5 years and 5 to 10 years, respectively. The uncomplicated malaria, and 1,587 (93.8%) were

training of the health care providers was associated analyzed. The average age of the 254 caregivers was

with knowledge of the national guidelines on malaria.

36 years (range 25 to 59). The average age was 35 Among the 226 (43.3%) caregivers who knew the years (range 25 to 59) for doctors and 38 years (range

instructions, 108 (98.2%) were trained, whereas 118

26 to 51) for the paramedical team that was composed

(82%) were not (P < 0.001).

of nurses and midwives. Age was not statistically significant between the occupational categories (P = 3.3 Malaria Diagnosis Practices for Uncomplicated

0.56). The sex ratio (male/female) was 2/1. Malaria Approximately 52% of the caregivers had practiced

The diagnosis of uncomplicated malaria was medicine for less than 5 years, and 48% had practiced

established by 176 (69.2%) of the providers based on medicine for over five years.

clinical criteria only, whereas 78 (31.2%) of the

3.2 Knowledge of National Guidelines by the Health providers used other clinical and biological criteria.

Care Providers Clinical and laboratory criteria were used by more

doctors than nurses and midwives (39.9% vs. 14.3%, P Of the 254 caregivers interviewed, 226 (88.9%)

< 0.001) and trained care staff compared to untrained positively responded about the national guidelines for

care staff (43.6% vs. 20.8%, P < 0.001). The location the management of malaria. The proportion of health

of the practice and the duration of professional care providers who claimed to know the national

experience were not associated with the use of clinical guidelines on malaria was higher among physicians

criteria (Table 2).

Table 1 Factors associated with the response of health professionals.

Knowledge of health care providers on national guidelines P

No (N = 28), n (%) Professional category

Total (N = 254), n (%)

Yes (N = 226), n (%)

Duration of practicing* 0.045 < 5 years

5-10 years

Work sector 0.367 Public

Training of health professional < 0.001 Yes

* Duration of professional experience.

Attitudes and Practices of Health Care Providers Regarding the Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

3.4 Prescription of Antimalarials

4. Discussion

The criteria that were used for choosing a treatment

This survey was conducted in the health district of

for malaria in clinical practice are in descending order:

Abidjan and reveals sufficient knowledge of health

the processing efficiency (39%), tolerance (21%), and

care delivery (89%) based on the national guidelines

duration of treatment (15%), number of decisions

for the treatment of uncomplicated malaria with a

(15%) and cost of treatment (10%). The first line of

significant difference between the medical and

therapy, artesunate-amodiaquine (ASAQ), was paramedical teams. This finding could be explained by prescribed the most (38%) and followed by the

the cascade training strategy that was adopted by the

combination of artemether-lumefantrine (AM/LF) National Program against Malaria (NMCP) in the (30%) and oral quinine (8%). Overall, 25% of the

Cote d’Ivoire, which first targeted the doctors who

prescribed treatments were not included in the

then trained the paramedical team. It must be

national guidelines: namely, artemisinin monotherapy

emphasized that this training was insufficient

(3%), other ACTs (19%) and herbal treatments (3%).

(approximately 43% in our study). A diagnosis of

ACTs were prescribed for 1,371 patients (86%), and the

uncomplicated malaria continues to be largely

ASAQ molecule was prescribed the most (Table 3).

determined using clinical criteria in 70% of cases

Table 2 Factors associated with malaria diagnosis practices.

Arguments for malaria diagnostic P Total (N = 254) n (%) Clinical (N = 176) n (%) Clinical et biological (N = 78) n (%)

Profession < 0.001 physicians

Duration of practicing*

5-10 ans

Location of pratices

Training of health professional

* Duration of professional experience; UH: University Hospital; UHCC: Urban Health Center Community Based; CHF: Community-Based Health Facility; GH: General Hospital.

Table 3 Antimalarials prescribed (N = 1587).

Artesunate-amodiaquine (ASAQ)

Artemether-lumefantrine (AM/LF)

Dihydroartemisin-piperaquine 197 12.4 Oral Quinin

Artésunate-mefloquine 58 3.7 Artesunate+Pyriméthamin-sulfamethoxypyrazin 49 3.1 Artemisin-based monotherapy 44 2.8 Herbal medecine

Others 37 2.3

Attitudes and Practices of Health Care Providers Regarding the Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

regardless of the professional experience of the provider and despite recommendations to treat only patients suffering from malaria (thick positive drop or positive TDR). The factors that influenced the knowledge and attitudes of the caregivers were professional category, job tenure and training of staff in the management of uncomplicated malaria. The percentage of patients who were prescribed ACT was 86%, and this rate is acceptable according to the realities on the ground. However, 25% of the antimalarials drugs prescribed (oral quinine, artemisinin derivatives monotherapy, and Die Kouadio ® ) did not meet the recommendations of the first choice treatments proposed by the NMCP. The results of our study were consistent with those studies reported in 2004 in the back country of Bouake, which indicates that the malaria treatment protocol recommended by the NMCP in the Ivory Coast and WHO is not always observed [16]. This finding supports the study conducted by the Medical Research and Social Science Aventis Center for French-speaking Africa in 1999, which indicated that 49% of caregivers (doctors, nurses and midwives) were familiar with the WHO protocol but did not always practice accordingly [17]. In cases of suspected malaria, parasitological confirmation was requested by relatively few medical and paramedical personnel. Whether all of the subjects that were malaria patients included in the study could not be confirmed. A cross-sectional survey that was conducted in pharmacies in the District of Guédiawaye, Senegal on the rational use of drugs against uncomplicated malaria revealed a significant deviation from the standard protocol for the curing of uncomplicated malaria [18]. In this study, 31% of the prescribers did not adhere to the national protocol. The implementation of the rapid diagnosis test (RDT) was generally low (14.1%), and chloroquine monotherapy accounted for 23.9% of the prescriptions [18]. In another study that was conducted in Madagascar, knowledge of the National Program

against Malaria (NMCP) by providers was low and estimated as 37.5%. Training in the management of uncomplicated malaria had not been conducted for approximately 18.8% of the providers and chloroquine accounted for 25% of the antimalarials prescriptions [18]. Little information is available on the factors that are associated with the knowledge and attitudes of health professionals; however, a study conducted in Dakar on female health care professionals indicated that there was a significant relationship between the level of knowledge and years of practice. The determinants of knowledge were age and education level. The practices of mothers were influenced by age, source of income and marital status [19]. This study was significant because it was multicenter (75 centers) and forward-looking with a typical sample size of 1,587 patients. In addition, there are few studies in West Africa on the determinants of the prescription of ACT. As a result, the NMCP now has updated data that could be used in training programs, and thus promote the rational use of ACT in the Ivory Coast. However, the study has limitations that are mainly due to information bias, particularly the non-collection and poor collection of certain variables by the investigators, which made the data unusable in 104 (6.2%) cases. Additionally, the study was conducted only in urban areas (Abidjan) for logistical reasons and cost saving objectives, whereas the management of uncomplicated malaria in rural areas is most often allotted to poorly trained paramedics whose prescriptions are based on personal experiences. The introduction of ACT has highlighted the need to improve the specificity of a malaria diagnosis. The relatively high cost of these drugs should lead to the treatment of only patients with positive parasitemia to reduce the cost of medical expenses. However, in addition to the economic considerations, a parasitological diagnosis has the following advantages:

1) better management of patients because of the accuracy of the diagnosis; 2) identification of non-malarial subjects for whom an alternative

Attitudes and Practices of Health Care Providers Regarding the Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

752 diagnosis should be sought; and 3) prevention of

unnecessary exposure to malaria and reduction of side effects, drug interactions and selection pressure [20]. In areas of high stable transmission, a malaria diagnosis in children older than 5 years of age and adults should be based on a parasitological confirmation (thick drop or TDR), which is particularly relevant in areas where the prevalence of HIV/AIDS is high and in areas with a high incidence of non-malarial febrile illness in patients infected with HIV [20]. The resistance to antimalarials drugs is growing and undermining the efforts of malaria control. When patients are treated with an artemisinin-based monotherapy, they may be tempted to stop treatment too quickly once the symptoms have disappeared, which results in partial treatment because of the persistence of parasites in the blood. Without the administration of a second drug in combination (as with ACT), the resistant parasites survive and can be transmitted to a mosquito and, thus, to another person. Monotherapy are the main cause of the developing resistance to artemisinin. The growth and expansion of resistance to other broad geographic areas, as it happened with chloroquine and sulfadoxine- pyrimethamine (SP) would result in serious public health consequences because no other antimalarials replacements will be available for another 5 to 10 years [20]. The prescription of oral quinine as a first-line therapy by several providers (8%) is applicable in cases of ACT-contraindication according to the WHO recommendations and national guidelines [21, 22], especially with pregnant women who cannot receive artemisinin derivatives in the first trimester of pregnancy. The prescription of combination therapies must be followed to prevent overprescription instead that could lead to the rapid development of resistance to this molecule and significantly increase existing resistance. The last point is noncompliance with national guidelines regarding the choice of therapeutic molecules that are recommended as first-line therapies. Artesunate-amodiaquine is

recommended by the NMCP; however, the fixed combination of artemether + lumefantrine represents 34% of the choices of health care providers in the first-line treatment of uncomplicated malaria. Amodiaquine is regarded by the population as well-tolerated antimalarials [23], and good tolerance of ASAQ has been demonstrated in many studies [24, 25]. In addition, the prescription of herbal medicines, such as “Die Kouadio ® ,” is a nonconforming use because little is known about this molecule regarding its effectiveness and potential toxicity. Training (initial and continuing) and objective information for prescribers and dispensers of the national guidelines of the NMCP should be continued and improved for distribution on a larger scale, and access to care by the poor should be facilitated by a grant of the cost of ACT and TDR. It is imperative that prescribers follow the WHO recommendations, including the latest directives on the management of malaria that propose the development of a rapid diagnostic test for all suspected cases of malaria to ensure adequate use of ACT only in patients who suffer from malaria [26]. This directive requires the development of rapid diagnostic tests, grant-aided appropriate training and the withdrawal of oral artemisinin-based monotherapy. Awareness campaigns should be improved to avoid self-medication and the use of herbal or traditional therapies to treat fever in endemic areas.

5. Conclusion

This study reveals sufficient knowledge of the national guidelines for the treatment of malaria by health care providers. However, the diagnosis of malaria continues to be made solely on clinical criteria. The training of health care providers regarding the proposed national guidelines for malaria treatment and the value of the rational use of ACTs needs to be improved. Therefore, it is necessary that different health care professionals’ work together in the fight against malaria to curb these practices through

Attitudes and Practices of Health Care Providers Regarding the

Management of Uncomplicated Malaria in Abidjan, Cote d’Ivoire

awareness campaigns. Clark, D. Njama-Meya, Safety and tolerability of combination antimalarial therapies for uncomplicated

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Ministère de la santé et de l’Hygiène Publique, Directives [3] WHO, Antimalarials drugs combination therapy, Report

Nationales de prise en charge du paludisme, Arrêté N° of a WHO Technical Consultation, WHO/CDS/RBM/

024/CAB/MSHP du 12/01/07, 2007. 2001, 35, 2001.

[15] WHO, The African Summit on Roll Back Malaria Abuja, [4] A.J. Djaman, L.K. Basco, A. Mazabraud, Establishment of

WHO/CDS/RBM/2000.17, April 25, 2000. a monitoring system of drug resistance of Plasmodium

[16] C. Pulcherie Kiki-Barro, K.F. N’goran, W. Yavo, R. Kassi, falciparum in Yopougon (Abidjan): In vivo sensitivity to

E.I. Menan, V. Djohan, Délivrance des antipaludiques en chloroquine and evaluation of resistance to pyrimethamine

officine dans le traitement de l’accès palustre simple Étude after analysis of the mutation of the gene for dihydrofolate

de la qualité du traitement: Cas de la ville de Bouaké (Côte reductase, Cahiers Santé 12 (2002) 363-367.

d’Ivoire), Cahiers D'études et de Recherches [5] R.R. Kassi, W. Yavo, D.G.A. Molak, H.I.E. Menan, C.P.

Francophones/Santé, 14 (2) (2004) 75-79. Barro-Kiki, A. Bamba, Efficacité thérapeutique de la

[17] Centre de Recherches et d’Etudes Medico-Sociales chloroquine dans le traitement du paludisme à

Aventis Afrique Francophone, Premier Panel Ivoirien Plasmodium falciparum chez les enfants du dispensaire de

Pour une Utilisation Optimisée des Moyens Actuels de San Pedro en Côte d’Ivoire, Bull. Soc. Path. Exot. 97

Lutte Contre la Malaria, Proj. Optim. 1 (1999) 9-19. (2004) 129-130.

[18] www.remed.org/revue_remed_no39_p.pdf. [6] WHO, Facts on ACTs (Artemisinin-Based Combination

[19] I. Seck, I.S. Fall, A. Faye, O. Ba, A. Tal-Dia, Malaria Therapies), January 2006.

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the rural area of poponguine Senegal, Med. Trop. 68 (2008) White, International artemisinin study group, artesunate

629-633.

combinations for treatment of malaria: Meta-analysis, [20] OMS, Aide-mémoire No. 94, Révisé en Avril 2010. Lancet 363 (2004) 9-17.

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of uncomplicated Plasmodium falciparum malaria in P.A. Seck, Randomized comparative study of the efficacy

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736-738.

a single intake versus two daily intakes in the treatment of [22] A.A. Adegnika, L.P. Breitling, S.T. Agnandji, S.K. Chai, uncomplicated falciparum malaria, Malaria J. 7 (2008) 16.

D. Schütte, S. Oyakhirome, Effectiveness of quinine [9] D.H. Hamer, M. Ndhlovu, D. Zurovac, F. Matthew, Y.A.

monotherapy for the treatment of Plasmodium falciparum Kojo, P. Chanda, Improved diagnostic testing and malaria

infection in pregnant women in Lambaréné, Gabon, Am. J. treatment practices in Zambia, JAMA 297 (2007)

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[23] K.A. Neftel, W. Woodtly, M. Schmid, P.G. Frick, G. Fehr, [10] J.L. N’diaye, M. Randrianarivelojosia, I. Sagara, P.

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Journal of Life Sciences 6 (2012) 754-757

On the Reproduction Number and a Presentation of Results for Infectious Diseases Models

Valeriy Dmitriy Perminov “BioTeckFarm”, 8, Tvardovskogo str., Moscow 123458, Russia

Received: January 15, 2012 / Accepted: April 06, 2012 / Published: July 30, 2012.

Abstract: The classical Kermack-McKendrick homogeneous SIR (susceptible, infected and removed) model is well known. Its general solution is a function of the unique parameter (the reproduction number) that is equal to a mean number of secondary cases produced by a typical infected individual in a completely susceptible population. If the reproduction number is more than one (the threshold value) its value describes an epidemic scope: larger values correspond to more severe epidemics. In the more complex compartment SIR models the population is divided into several non-overlapping groups. It allows us to partly remove assumptions of

the classical model. It is well known that for this kind of models, just as for the classical model there is the threshold parameter R 0 . Usually it is called by the same name—the reproduction number—though the physical meaning of this parameter has changed. The main purpose of the paper is to show that this new parameter is a not unique measure of an epidemic severity for any compartment SIR model. In particular it means that for such models comparison of the severity of two epidemics by simple comparing values of their reproduction numbers is incorrect. For compartment models these statements were proved with the help of the corresponding ODEs analysis. Very popular now individual-based models (IBMs) are more complex in comparison with the compartment ones since they use overlapping groups (school children are members of families also, for example). In such a case Diekmann’s calculation method for the reproduction number used in many papers is inapplicable as well as a presentation the simulation results obtained as functions of this parameter.

Key words: Mathematical SIR and IBM models, epidemic, reproduction number.

1. Introduction  infected, removed) and , R

0 and id are a non-dimensional time, the reproduction number and

In few last years numerous papers have been initial data correspondingly. Moreover in published where the classical threshold criterion was homogeneous population the reproduction number has applied to individual-based models for infectious the threshold value: an epidemic ensures if R 0 > 1 and diseases and results obtained were presented as it does not evolve if R 0 < 1. Averaging over the whole

functions of the reproduction number R 0 only (for

population in the definition of the reproduction

example, [1-5]). The reproduction number R 0 is

number in a heterogeneous population prompts a usually defined as the mean number of secondary supposition that the classical threshold criterion might cases produced by typical infected individual in a prove invalid. Indeed for a population containing two completely susceptible population. It is well known unequal and non-overlapping groups it is possible to (for example, [6]) that for the classical assign the distribution of the secondary cases over Kermack-McKendrick SIR model any solution can be groups in such a way that the whole reproduction

presented in the vector functional form x = F( ; R 0 , id),

number will be less than 1 but an epidemic takes place where x = (non-dimensional numbers of susceptible, in the smaller group in which the number of the

Corresponding author: Valeriy Dmitriy Perminov, Ph.D., secondary cases for a typical infected individual is research field: infectious diseases modeling. E-mail: vdperm@yandex.ru.

more than 1. This fact was first noticed by Diekmann

755 et al. [7] as early as in 1990.

On the Reproduction Number and a Presentation of Results for Infectious Diseases Models

interacting non-overlapping groups in detail. Assume Main goal of this paper is to analyze a role and

that the infection rates in the group i as a consequence meaning the reproduction number in different SIR

of contact with members of the group j are given by a models.

matrix p ij , and the removed rates in the groups are given by positive constants a 1 and a 2 . Then by

2. Results and Discussion

analogy with the classical Kermack-McKendrick Let us study a simple SIR model with two

model we can write:

dS 1  

p 11 I 1 S 1  p 21 I 2 S 1 ,

S 1 ( 0 )  S 10  0

dt

dI 1 

p 11 I 1 S 1  p 21 I 2 S 1  a 1 I 1 ,

I 1 ( 0 )  I 10  0

dt

dR 1 

dt dS 2

S 2 ( 0 )  S 20  0 , S 0  S 10  S 20  0 dt

  p 12 I 1 S 2  p 22 I 2 S 2 ,

dI 2 

p 12 I 1 S 2  p 22 I 2 S 2  a 2 I 2 , I 2 ( 0 )  I 20  0

dt

dR 2 

dt

In physics and mechanics a non-dimensional form of model equations is generally used. It helps (before solving the equations!) to find non-dimensional parameters (or in other words “similarity parameters”) which the solution will have to depend on. Let us introduce new non-dimensional variables:

s k  S k / S 0 , i k  I k / S 0 , r k  R k / S 0 ,   a 1 t , p 0  max i , j p ij , i , j , k  1 , 2 (2)

Then our system can be rewritten in the following form:

S  10 

ds 1 *  p 11 p 21 

R  i 1 s 1 

i 2 s 1  ,

 s 10  0

S  20 

ds 2 *  p 12 p 22 

R 

i 2 s 2  ,

 s 20  0

I  10

di 1 *  p 11 p 21 

R  i 1 s

i 2 s 1   i 1 ,

 i 10

di 2 *  p 12 p 22  a 2 I 20

 R  i 1 s 2 

i 2 s 2   ,

 i 20

dr 1 

dr 2 a 2

On the Reproduction Number and a Presentation of Results for Infectious Diseases Models

Any solution of this ODE system can be threshold condition exists also for such models. presented in the vector functional form x = F( ;

However neither classical nor Diekmann’s definition p *

of the threshold condition can be used. Therefore an group are not those of another group (it is the case

ij /p 0 ,a 2 /a 1 ,R ,s 10 ,s 20 ,i 10 ,i 20 ). If members of one

existence of a threshold condition as well as any in our example) the threshold condition exists [7, 8])

complex similarity parameter including some

and can be written down in the form R 0 = f(p ij /p 0 ,

combination of model parameters (e.g. the

reproduction number in particular) has to be proved dominant eigenvalue of so call “next step matrix”.

2 /a 1 ,R ,s 10 ,s 20 ,i 10 ,i 20 ) > 1 where R 0 is the

numerically. Unfortunately such a proving is absent in Sometime this parameter R 0 is called the pioneer papers [1-5]. Authors assume that the reproduction number though it does not correspond

reproduction number R 0 (in classical or Dieckmann’s to its abovementioned epidemiological definition

definition) is only a unique similarity parameter in [7]: we can qualitatively interpret this threshold

general heterogeneous case and it submits to the condition as a requirement of an epidemic outbreak

classical threshold condition. The correctness of such

an assumption is doubtable. It is possible to propose a solution of our system can be rewritten in the

absence in all groups for R 0 < 1. Therefore any

disproving example for the general case. If the

following form x = F 1 ( ; p ij /p 0 ,a 2 /a 1 ,R 0 ,s 10 ,s 20 ,i 10 ,

influenza virus has an unqualified childish orientation

i 20 ). This functional form of the solution allows (i.e. the number of secondary cases among children is concluding that even for the quite simple model R 0 more than one while any other contacts are

is the threshold parameter only and its value can not characterized by a mean number of the secondary describe the solution unequivocally because it is

cases less than one) the average number of secondary necessary to know values of other parameters. In

cases can be less than one but an epidemic takes place

particular value of R 0 > 1 alone cannot be

among children.

considered as a measure of an epidemic scale. So one can make a conclusion that a use of the It is evident that these conclusions for a model of

classical reproduction number as the similarity this type are general and remain valid if the number of

parameter that measures the severity of an epidemic groups is more than 2.

and submits to the threshold condition R 0 > 1 is Nowadays individual-based models (IBMs; see, for

incorrect for general heterogeneous models. Using example, [9]) are more and more often used in

Diekmann’s definition is also incorrect because in a simulations of infectious diseases spreading. As a rule

general case it is impossible to divide the city there is no any formal mathematical presentation for

population into set of non-overlapping groups. such models and a modeler has to suppose a priori

3. Conclusions

that the simulation results are functions of all model parameters and initial data. In IBMs the probability

It is worth noting that 1) Most of papers cited above for a person to get infected strongly depends on

have been devoted to the problem of an epidemic contact network that may not be divided in spread across a country. In such a case a solution has non-overlapping groups (for example schoolchildren

to strongly depend on a population traffic between are members of families). It means that IBMs are

different sites that as a rule is not uniform; 2) In more complex models in comparison with the above

concordance with above-stated conclusions an idea of considered compartment SIR models and therefore

comparison the results obtained for different IBMs for their solutions have to be functions of a more number

the same values of the reproduction numbers [5] is of similarity parameters. It is likely that some

incorrect from mathematical point of view; 3) Authors

757 of [10, 11] using another approaches came to

On the Reproduction Number and a Presentation of Results for Infectious Diseases Models

[3] I.M. Longini Jr., A. Nizam, S. Xu, K. Ungchusak, W. conclusions similar to ours and 4) Number of papers Hanshaoworakul, D.A. Cummings, et al., Containing pandemic influenza at the source, Science 309 (5737)

using R 0 as an only similarity parameter in the

(2005) 1083-1087.

heterogeneous population continues to grow. [4] T.C. Germann, K. Kadau, I.M. Longini Jr., C.A. Macken, Mitigation strategies for pandemic influenza in the United

Acknowledgements

States, PNAS 103 (15) (2006) 5935-5940. [5] M.E. Halloran, N.M. Ferguson, S. Eubank, I.M. Longini This work was assisted through participation in

Jr., D.A.T. Cummings, B. Lewis, et al., Modeling targeted “Optimal Control and Optimization for Individual-

layered containment of an influenza pandemic in the based and Agent-based Models” Investigative

United States, Proc. Natl. Acad. Sci. USA 105(12) (2008) Workshop at the National Institute for Mathematical 4639-4644. [6] J.D. Murray, Mathematical Biology, Vol. 1, Springer,

and Biological Synthesis, sponsored by the National

New York, 2002.

Science Foundation, the U.S. Department of [7] O. Diekmann, J.A.P. Heesterbeek, J.A.J. Metz, On the Homeland Security, and the U.S. Department of

definition and computation of the basic reproduction ratio R0 in models for infectious diseases in

Agriculture through NSF Award #EF-0832858, with heterogeneous populations, J. Math. Biol. 28 (1990)

additional support from The University of Tennessee,

365-382.

Knoxville. [8] P. van den Driessche, P. Watmough, Reproduction number and some sub-threshold endemic equilibria for

References

compartmental models of disease transmission, Matematical Bioscience 180 (2002) 29-48.

[1] N.M. Ferguson, D.A. Cummings, S. Cauchemez, C. Fraser, [9] V. Grimm, S.F. Railsback, Individual-Based Modeling S. Riley, A. Meeyai, et al., Strategies for containing an

and Ecology, Princeton University Press, Princeton and emerging influenza pandemic in Southeast Asia, Nature

Oxford, 2005.

437 (7056) (2005) 209-214. [10] R. Breban, R. Vardavas, S. Blower, Theory versus Data: [2] N.M. Ferguson, D.A.T. Cummings, C. Fraser, J.C.

How to calculate R 0 ?, PloS One 3 (2007) 1-4. Cajka, P.C. Cooley, D.S. Burke, Strategies for

[11] J. Li, D. Blakeley, R.J. Smith, The Failure of R 0 , Hindawi mitigating an influenza pandemic, Nature, 442 (7101)

Publishing Corporation, Computational and Mathematical (2006) 448-452.

Methods in Medicine 2011, Article ID 527610, pp. 1-17.

Journal of Life Sciences 6 (2012) 758-765

High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

1 Leonor Deis 1, 2 , Maria Inés de Rosas and Juan Bruno Cavagnaro

1. Instituto de Biología Agrícola Mendoza, Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Alte, Brown 500, PC: 5507 Chacras de Coria, Argentina 2. Facultad de Ciencias Agrarias, Universidad Nacional de Cuyo, Alte, Brown 500, PC: 5507 Chacras de Coria, Argentina

Received: July 19, 2011 / Accepted: August 18, 2011 / Published: July 30, 2012.

Abstract: The predicted increase of temperature by effect of climatic change (1.5 °C to 4.5 °C), will affect some berry components. Anthocyanins and flavonols concentration are responsible of wine color. The level of anthocyanin is affected by light intensity, temperature, sugars, growth regulators and vineyard management. Abscisic acid (ABA) increases the synthesis of anthocyanin in grape. The object of study was to evaluate the effect of different temperature in berries in vitro, of two cultivars combined with ABA treatments. The treatments were control (C, water) and ABA treatment (1,000 ppm) and temperature: 25, 33 and 40 ºC. In Cabernet Sauvignon, 25 and 33 ºC did not affect anthocyanin total concentration but 40 °C produced a 30% decrease in anthocyanin. ABA treatment increased anthocyanin vs. C at 25 and 33 ºC, mainly due to glucosylated forms. But ABA + 40 ºC showed a 44-60% decrease in all anthocyanins forms compounds. Response of Malbec to the highest temperature (40 ºC), at the end of ripening, was different; higher temperature produced only slight decrease of total anthocyanins concentration (decrease of 7%). Combination of temperature + ABA at 20 °C and 33 ºC increased anthocyanin. But ABA + 40 ºC decreased glucosylated and cumarylated forms of anthocyanin.

Key word: Temperature, ABA, anthocyanins, grape, Malbec, Cabernet Sauvignon.

1. Introduction  ripening period and they have not had similar results [4, 6, 10, 23]. When Castellarini et al. [24] compared

Anthocyanins are predominant pigments of grape weather condition of two year and anthocyanin skins and they are the responsible for the color of the concentration, they showed what the years with more red wine. For winemakers, the anthocyanins and days with temperatures over 30 °C have the major polyphenols are very important, because those are anthocyanin content. But, other authors demonstrated transfer in wine. Their contents in wine determine the what higher temperatures during all days affected the organoleptic properties and the health benefit.

color of grape [25, 26].

Anthocyanins accumulation was affected by weather Anthocyanin accumulation is enhanced by a plant conditions, especially light intensity and temperature hormone ABA [14, 27, 28], cold temperature, and [1-6], growth regulators [7-14], water status [15, 16] high light intensity [23]. Different authors consider and vineyard management [17-21]. that for some varieties the optimal temperatures for Temperature is an external factor that affects the anthocyanins accumulation in grape are in 15-25 °C coloration of grapes [22]. Authors studied the effect at during the day and 10-20 °C at night. When differences temperature on skin color during the temperature between day/night are greater than 10 °C

generally depressed fruit coloration [1]. Experiments Corresponding author: Leonor Deis, Ph.D., research fields: plant physiology, viticulture and oenology. E-mail: with temperature-control were showed that exposing

ldeis@fca.uncu.edu.ar.

759 vines or clusters to high temperature (30 °C) inhibited

High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

5’ position. Flavonoid precursors are initially recruited anthocyanins accumulation [10, 23].

from the phenylpropanoid pathway by a small family Anthocyanins are synthesized via the flavonoid

of chalcone pathway. Parallel pathways downstream pathway in grapevine cultivars that have the wild-type

of F3’H and F3’5’H [30, 31] produce either cyaniding VvmybA1 transcription factor for the expression of

or delphinidin. The 3’ position of cyanidin and UFGT [29]. The encoded enzyme UFGT catalyse

delphinidin and sequentially the 5’ position of glycosylated unstable anthocyanidin aglycones into

delphinidin can be methoxylated by OMT that generate pigmented anthocyanins (Fig. 1). Two primary peonidin, petunidin and malvidin, respectively. anthocyanins (cyanidin and delphinidin) are Anthocyanins are delivered into the vacuole where they synthesized in the cytosol of berry epidermal cells.

are visible as coloured coalescences (anthocyanic Cyanidin has a B-ring di-hydroxylated at the 3’ and 4’

vacuolar inclusions). It still remains unknown whether positions, whereas delphinidin has a tri-hydroxylated

anthocyanins enter the vacuole as single molecules B-ring because of an additional hydroxyl group at the

and thereafter they aggregate or if cytoplasmic vesicles

Fig. 1 Key steps of the flavonoid pathway leading to anthocyanin biosynthesis. (Extracted to 24).

760 High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

containing coalesced anthocyanins interact with the

light-darkness 16/8 hours.

tonoplast [32]. Whatever the mechanism, some The + ABA treatments was obtained added 10 mL members of the GST protein family are believed to -1 of ABA solution 1 g·L for Petri dish (+ A) and the –

participate in vacuolar trafficking and sequestration of ABA treatments was obtained added 10 mL of water anthocyanins [33, 34].

deionized and sterilized for Petri dish (– A). These Total amount of anthocyanins and the relative

treatments were combined with different temperatures: abundance of single anthocyanins are extremely

25, 33 and 40 °C, being: 25 °C plus ABA (25 + A), variable among red- to blue-skinned cultivars. Both

25 °C without ABA (25 – A), 33 °C plus ABA (33 + traits are under genetic control and are A), 33 °C without ABA (33 – A), 40 °C plus ABA (40 developmentally regulated. Physiological studies + A), 40 °C without ABA (40 – A). show that incident radiation on bunches [35, 36], plant

2.1 Grape Extraction for UV-Vis Analysis and HPLC water status [37] and exogenous hormones [38, 39]

modify anthocyanin content, expanding the range of For determining anthocyanin concentration, the metabolite variation beyond that due to the genetic

skin was removed from the grape and placed in a synthases (CHS1, CHS2, CHS3) and enter the

methanol-HCl (99:1) solution for 48 h at –20 °C for flavonoid background of a given cultivar. In particular,

extraction. The liquid fraction was filtered through 0.4 little is known about whether or not, and if so, to what

mL cellulose acetate filter. Anthocyanin pigments extent the expression of anthocyanin biosynthetic

were quantified according to Maza et al. [40]. The genes is affected by seasonal water availability

determination of total polyphenols (TP) was made throughout the progress of ripening.

from the same extract but absorbance was measured at The objective of this study was to determine the

280 nm.

combined effect of ABA and high temperatures on UV-Vis analysis: A Varian Cary WinUV anthocyanin profile in two varieties of grape in vitro

spectrophotometer with 10 and 1 mm—optical path cultivation.

cells was used to perform the absorptiometric measurements. For anthocyanin content, the solution

was diluted 1:50 (v/v) with acidified distilled water Berries of Vitis vinifera cv. Cabernet Sauvignon

2. Materials and Methods

(1% v/v HCl) and absorbance was measured at 520 and Malbec were sampled randomly from four plants

nm against a blank of reagents.

at maturation and post-harvest. The berries were For polyphenol content, the solution was diluted washed with deionized water, sterilized in 15%

1:100 (v/v) with distilled water and absorbance was sodium hypochlorite for 30 min and rinsed with

measured at 280 nm against a blank of reagents. sterilized water. Following procedures were carried

HPLC: The measurements were made with HPLC out in a clean bench and all instruments used were

LKB 2152, Wavelength monitor 2141 and Integrator sterilized. Each berry was cut longitudinally into

HP 3395.

halves with a razor blade. The ten half-cut berries For HPLC analysis, these samples were filtered were placed on a filter paper in a sterilize Petri dish

using a hydrophilic PTFE membrane filter (0.45 mm) and 5 mL of test solution was added before filter this

before injection. Reversed phase HPLC separations by a membrane (0.45 µm). The Petri dish was covered

were conducted using the Lichrosorb RP-18 5 µm 250 with Parafilm “M” (Laboratory Film, Pechiney,

mm × 4.6 mm Cat.N°54949, Suppelco Inc. columns. Chicago, EEUU). Then the ABA added and the Petri

The flow rate was 0.8 mL/min, and the sample volume dish was maintained under the photoperiod was 20 µL. Anthocyanins were detected using the

761 SPD-10A UV-Vis detector (Shimadzu, Japan) with

High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

for methylation of di-hydroxiled anthocyanins. absorbance at 518 nm. For linear gradient elution, the

When the grape had 24°Brix, total anthocyanins method using 10% formic acid, 3% acetonitrile and

decreased vs the 22°Brix. Naturally, the anthocyanins 87% water bidistille as solvent A and 10% formic acid,

content maximum is produced before the 50% acetonitrile and 40% bidistilled water as solvent

optimal °Brix for the harvest. Not only total

B. the gradient was follows: 0 min, 6% B; 15 min, anthocyanins were minor, but also changed the profile. 30% B; 30 min, 50% B; 35 min, 60% B; 41 min, 6%

At 40 °C decreased the tri/di hydroxylated ratio for B; 45 min, 6% B.

decrease of tri-hydroxyled and increased of The individual anthocyanins were identified by

di-hidroxylated compounds, and this variation is comparing their retention time with those of standards

detrimental for stability of wine.

and published data. High temperature decreased methylation of delfinidin (malvidin) but did not affect the petunidin.

2.2 Statistical Analysis The methylation is favorable for winemaking because One way analyses of variance (ANOVA) and the

the compounds methyled are most stability. Fisher’s multiple comparison of means in order to

The ratio 3’,4’,5’-hydroxylated/3’,4’-hydroxylated discriminate between the averages by the minimum

anthocyanins was higher at 33 °C. This result can be difference, with a significance level of P < 0.05 were

understood for the activity or transcription of F3’H applied.

and F3’5’H regulated with temperature. Temperature on wine color has two effects: one is

directly for diminution of anthocyanin total content Fig. 2 shown temperature effects on total and another indirect because nonmethyled anthocyanins content (TA) in grape. In two moments,

3. Result and Discution

compound has less stability in wine. The result high temperatures (40 °C) decreased TA. Analysis to

showed the temperature effect on the enzymatic profile evidenced decrease tri-hydroxylated and activity of OMT. di-hydroxylated anthocyanins content when

At 22°Brix, ABA applied increased TA at 25 and temperature increases (Table 1). This ratio was higher

33 °C, but at 40 °C produced decreased of this at 33 °C because of the content of di-hydroxylated

compound. The main effect was increase to malvidin compounds was 33% minor and the tri-hydroxylated

3-O-glucoside in 58% at 25 °C and 35% at 33 °C. was only 3%. Finally, the color of these berries was

Also, ABA increased the tri-hydroxyled and in minor darker because it increases the ratio blue/red. For the

intensity the di-hydroxyled compounds. The tri/di- winemaking this is favorable due to tri-hydroxylated

hydroxyled ratio was similar for 25 and 40 °C, but for anthocyanins are most stable during the elaboration

33 °C it was higher than them. This effect is positive and conservation of wine.

for the colors of wine.

Methylation of tri-hydroxylic anthocyanins was Additionally, the ABA increased the derivates favored by high temperature which eventually compound (malvidin acetil, peonidin cumaril, outweighs the decrease of anthocyanins. However,

malvidin cumaril) giving stability of colors. In high temperature did not affect O-methyltransferase of

absolute values the largest increase experiment was di-hydroxylated anthocyanins. There are specific

for malvidin, in relative values, the ABA stimulates enzymes for each group (di or tri hydroxylated

the synthesis of petunidin.

compounds) [32], and they could be regulated by At 24°Brix, the anthocyanins content was lower than different factors. 33 °C was the optimum temperature

22°Brix, but malvidin 3-O-glucoside contents increased

762 High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

A Total anthocyanins in Malbec (22°Brix)

gr 37 33+A

of

d 40+A

Total anthocyanins

B Total anthocyanins in Malbec (24°Brix)

c / 25+A

of g 27

mg 22 33+A 40+A

7 Total anthocyanins

C Total anthocyanins in Cabernet Sauvignon

42 (24°Brix) a

25+A /g 22

7 Total anthocyanins

Fig. 2 Total anthocyanins from grapes subjected to different treatments: 25, 33 and 40 °C, being: 25 °C plus ABA (25 + A), 25 °C without ABA (25 - A), 33 °C plus ABA (33 + A), 33 °C without ABA (33 - A), 40 °C plus ABA (40 + A), 40 °C without ABA (40 - A).

A: Total anthocyanins in Malbec of 22°Brix; B: Total anthocyanins in Malbec of 24°Brix; C: Total anthocyanins in Cabernet Sauvignon of 24°Brix. Means affected by the same letter are not significantly different at P = 0.05.

High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

Table 1 Anthocyanins profile of grape at temperatures different combined with treatment of ABA.

Treatments Cya3-O-gl Peo3-O-gl Del3-O-gl Pet3-O-gl Mal3-O-gl acethyl acethyl Peo

Peo

Mal

cumaril cumari Tri/bi-hydroxylated OMT1 OMT2 OMT3

Malbec 22°Brix

Malbec 24°Brix

Cabernet Sauvignon 23°Brix

Cya3-O-gl: cyanindin 3-O-glucoside; Peo3-O-gl: peonidin 3-O-glucoside; Del3-O-gl: delphinidin 3-O-glucoside; Pet3-O-gl: petunidin3-O-glucoside; Mal3-O-gl: malvidin 3-O-glucoside; Peo acethyl glu: peonidin 3-acetylglucoside; Mal acethylgl: malvidin 3-acetylglucoside; Peo cumaril: peonidin 3-p-coumaroylglucoside; Mal cumaril: malvidin 3-p-coumaroylglucoside. OMT1: O-methyltransferase petunidin derivated of delphinidin; OMT2: O-methyltransferase malvidin derivated of delphinidin; OMT3: O-methyltransferase peonidin derivated of cyanindin.

in grape at 25° y 33°C when we supply ABA. compounds contents, favoring the colors blue in wine. The tri/di-hydroxyled ratio was superior only at

For the winemaking the decreased in TA produced

25 °C. At 33 °C was similar a without ABA because for high temperature (40 °C) is a negative economical increased two derived di-hydroxyled compound effect and it is of major intensity in Cabernet (petunidin 3-O-glucoside). Those effects evidenced

Sauvignon than Malbec.

the stimulation of F3’5’H and O-methyl transferase ABA supply incremented the TA and malvidin (derived of cyaniding 3-O-glucoside).

3-O-glucoside was the principal compound affected.

Cabernet Sauvignon was more sensible at Also, ABA favored the methylation of compounds temperature change. Grape at 33 °C showed a higher

which is reflected with greater color stability in content of TA. Tri/di-hydroxyled ratio was incremented

wine.

at 33 °C but decreased 22% at 40 °C (more important

4. Conclusion

than Malbec). When the temperature was 25 °C, tri/di-hycroxylad ratio did not show changes, and this

Global climate is changing and is predicted was due to increase of di and tri-hydroxyled increases in temperatures. Is necessary to know the anthocyanins. At 33 °C, the increase was due

response of different cultivars versus temperature principally at delphinidin and malvidin 3-O-glucoside.

increase for to redefine the regions for each cultivar This temperature modified the activity or expression of

and strategies used to compensate the adverse effects F3’5’H at increment the delphinidin and derivate

on the color of wines.

764 High Temperature and Abscisic Acid Modified the Profile of Anthocyanins in Grape (Vitis vinifera L.)

With regard to the wine industry, color is crucial to

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Journal of Life Sciences 6 (2012) 766-770

Substrate Type Affects Growth, Yield and Mineral Composition of Cucumber and Zucchini Squash

1 2 2 3 1 Mariateresa Cardarelli 1 , Youssef Rouphael , Salem Darwich , Elvira Rea , Antonio Fiorillo and Giuseppe Colla 1. Department of Agriculture, Forestry, Nature and Energy, University of Tuscia, via S. C. De Lellis snc, Viterbo 01100, Italy

2. Faculty of Agricultural and Veterinary Sciences, Lebanese University, Dekwaneh-El Maten, Beirut, Lebanon 3. CRA-Research Centre for Soil/Plant System, via della Navicella 2-4, Roma 00184, Italy

Received: November 24, 2011 / Accepted: January 31, 2012 / Published: July 30, 2012.

Abstract: The expansion of hydroponics in many countries of the world in the last few decades may be ascribed to the ability of soilless growing systems to avoid various problems arising from the use of the soil. Cucumber (Cucumis sativus L.) and zucchini (Cucurbita pepo L.) plants were grown in closed-soilless culture under unheated-greenhouse conditions at the experimental farm of University of Tuscia, Central Italy to evaluate the effects of four substrates (rockwool, pumice, perlite, and cocofiber) on growth, yield and plant mineral composition. For both cultures, plants grown in cocofiber, perlite and pumice yielded more than those grown in rockwool. The better temperature regime in cocofiber, perlite and pumice was due to the greater thermal inertia compared to rockwool slabs. The concentration of N in zucchini and cucumber leaves was significantly higher in cocofiber, perlite and pumice in comparison to the rockwool treatment. The concentration of K was significantly affected by the substrate only for the zucchini squash with the highest value recorded on the organic substrate (cocofiber), whereas the Ca concentration was significantly influenced by the growing media only for cucumber with the highest value observed on pumice. Finally, the lowest Mg concentration in leaf tissue was observed on plants grown with the rockwool substrate for both zucchini squash and cucumber. We can conclude that cocofiber, perlite and pumice are suitable for zucchini and cucumber production in closed soilless system, whereas the use of rockwool is more suitable for crops grown under heated greenhouse conditions.

Key words: Substrate type, yield, mineral composition, cucumber, zucchini squash.

1. Introduction represent a major proportion of cultivable land throughout the world [4]. A further advantage of

The revolutionary expansion of hydroponics in hydroponics is the precise control of nutrition. This is many countries of the world in the last three decades particularly true in crops grown either on inert may be ascribed to the ability of soilless growing substrates or in pure nutrient solution [5]. However, systems to be independent of the soil and hence of all even in soilless crops grown in chemically active problems related to it. Hydroponics has proved to be growing media, the nutrition of the plants can be an excellent alternative to soil sterilization, especially better controlled than in crops cultivated in the soil, in view of the fact that the use of chemical soil due to the limited volume of substrate per plant and its sterilants, such as methyl bromide [1-3]. Moreover, standard, homogeneous constitution, which is well the cultivation of greenhouse crops and the

known to the grower [2, 3].

achievement of high yields and good quality are The porous materials used as substrates in soilless possible with hydroponics even in saline or sodic soils, culture are distinguished as organic or inorganic or non-arable soils with poor structure, which growing media. The organic materials used in soilless

culture originate from plant residuals and are therefore Corresponding author: Giuseppe Colla, Ph.D., researcher,

research field: horticulture. E-mail: giucolla@unitus.it. subjected to biological degradation. The decomposed

767 organic materials are more or less chemically active,

Substrate Type Affects Growth, Yield and Mineral Composition of Cucumber and Zucchini Squash

“Afrodite” hybrid (Syngenta, Switzerland) and due to the presence of ion exchange sites, which may

cucumber (Cucumis sativus L.) “Edona” hybrid adsorb or release nutrients. In contrast, most inorganic

(Royal Sluis) was germinated in vermiculite on Mar. materials are chemically inactive (inert). Therefore,

10 and Aug. 15, 2001, respectively. Plants remained many authors use the terms “organic” and “inorganic”

in the seed pots until the two true leaf stages. The growing media as synonyms to “chemically active”

seedlings were transplanted on Mar. 23 and Sept. 1 for and “inert” substrates, respectively [1]. Zucchini

zucchini and cucumber, respectively, into cocofiber, squash and cucumber are important crops that have

pumice, perlite, and rockwool at a plant density of gained popularity for both open field and protected -2 2.1 plants·m .

cultivation in the Mediterranean region [6-8]. In both experiments, plants were fertilized with the However in the last 20 years, growing zucchini squash -1 following nutrient solution (mg·L ): N-NO

3 (160), S and cucumber in soilless culture have become

(26), P (13), Cl (57), K (150), Ca (122), Mg (59), Na increasingly popular among commercial growers.

(50), Fe (3), Mn (0.8), Cu (0.07), Zn (0.1), B (0.3), The aim of this study was to determine, yield,

Mo (0.05). The electrical conductivity (EC) values growth, and nutrient accumulation of two -1 were kept within the range of 1.8 to 2.0 dS·m , while

Cucurbitaceae species (zucchini and cucumber) in the pH of the solution was maintained between 5.8 relation to the type of substrate (organic vs. inorganic)

and 6.3 by adding an acid mixture. Nutrient solution in unheated greenhouse conditions.

was pumped from independent tanks through a drip irrigation system with one emitter per plant and an

2. Materials and Methods

emitter flow rate of 2 L·h -1 . The excess of the nutritive

Two experiments were conducted in two solution was recycled for the entire growing cycle. consecutive growing seasons (spring and fall) in a

Irrigation scheduling was performed using electronic

low-tension tensiometers (LT-Irrometer). During the experimental farm of Tuscia University, central Italy.

polyethylene 400 m 2 greenhouse situated at the

crop cycle, fruits of zucchini squash and cucumber Inside the greenhouse, the high temperature and

were harvested when they reached marketable size; relative humidity were controlled through ventilation.

fruits that were deformed or badly misshapen were

A randomized complete block design with four considered unmarketable. Fruits were dried in a replicates (ten plants per experimental unit) was used

forced-air oven at 80 °C for 72 hours, then ground in a to compare four substrates: perlite (Perlite Italiana),

Wiley mill to pass a 20-mesh screen and analyzed for pumice (Europumice), cocofiber (Cocco Ter), and

the following elements: N, P, K, Ca, and Mg. rockwool (Grodan).

At final harvest four plants of zucchini squash and In both experiments, soilless plants were placed in

cucumber per plot (experimental unit) were separated single plastic channel benches (section 26  12 cm;

into stems and leaves for biomass determination and length 5 m with 1.5% slope) which contained in the

subsequently ground for major and trace elements bottom a plastic drainage layer (1.5 cm height)

determination (N, P, K, Ca, Mg, Na, Cu, Fe, Mn, and, covered with geo-textile. Perlite, pumice and Zn). The nitrogen concentration in plant tissues cocofiber were added to fill the channels while

(leaves and fruits) was determined after mineralization rockwool was used as slab (100 cm length  15 cm

with sulfuric acid by “Regular Kjeldahl method” [9], width  7.5 cm depth). In all plot, the surface of

P, K, Ca, Mg, Na, Cu, Fe, Mn, and Zn were substrate was covered with white plastic film.

determined by dry ashing method at 400 °C for 24 Seeds of zucchini squash (Cucurbita pepo L.)

hours, dissolving the ash in 1:25 HCl, and assaying

Substrate Type Affects Growth, Yield and Mineral Composition of Cucumber and Zucchini Squash

the obtained solution using an inductively coupled substrate temperatures in the rockwool substrate was plasma emission spectrophotometer [10].

always lower than those recorded on cocofiber, perlite All data were statistically analyzed by ANOVA

and pumice (data not shown). In fact, the lowest air using the SPSS software package (SPSS 10 for

temperature recorded on Apr. 14 (–3.3 °C) and Nov. Windows, 2001). Duncan’s Multiple Range test was

10 (3 °C) in the zucchini and cucumber experiments, performed at P = 0.05 on each of the significant

respectively caused damage to the leaves especially variables measured.

for plants grown on rockwool due to the higher plant vigor and the low volumetric heat capacity of

3. Results and Discussion

substrate. The better temperature regime in cocofiber, The total and marketable yield of zucchini squash

perlite and pumice was due to the higher substrate and cucumber were significantly affected by the 3 volume per plant (9,375 vs. 5,625 cm /plant) which

substrates. For both zucchini and cucumber the leads to a greater thermal inertia compared to highest marketable yield was recorded on cocofiber,

rockwool slab. No significant difference among perlite and pumice, followed by rockwool, with no

treatments was observed for the total above ground significant differences observed between the first three

dry biomass of zucchini squash, whereas a significant treatments (Table 1). Moreover the lowest marketable

difference was recorded in cucumber, with the lowest zucchini yield observed on rockwool was mainly

values observed in the rockwool substrate. attributed to a reduction in the fruit number and not to

The macro- and microelements concentration in

a change in the fruit mean weight, whereas in zucchini and cucumber leaves as a function of the cucumber, the yield reduction on rockwool was

substrate are displayed in Tables 2 and 3. The related to a reduction in both fruit mean weight and

concentration of N in zucchini and cucumber leaves fruit number (Table 1). Our results are in line with

was significantly higher in cocofiber, perlite and previous researches carried out on cucumbers [11] and

pumice in comparison to the rockwool treatment on tomatoes [12].

(Table 2). Moreover, no significant differences were The above findings may be related to a low

observed for the P concentration in both crops. The temperature regime in rockwool slabs during the early

concentration of K was significantly affected by the part of the growing cycle for zucchini and the late part

substrate only for the zucchini squash with the highest of the cropping cycle for cucumber where the minimal

value recorded on the organic substrate (cocofiber),

Table 1 Effects of substrates on total and marketable yield (kg·plant -1 ), fruit mean weight (g·fruit -1 ), fruit number (No.·plant -1 ), and above-ground dry biomass (g·plant -1 ) of zucchini and cucumber plants.

Yield Marketable fruit Substrate

Total above ground Total Marketable Mean mass Number dry biomass

Zucchini

375.3 a Perlite

Cocofiber 2.34 a 1 2.30 a

113.9 a

20.1 a

364.3 a Pumice

341.8 a Rockwool

305.5 a Cucumber Cocofiber

271.1 a Perlite

287.0 a Pumice

224.7 a Rockwool

185.3 b 1 Means within columns separated using Duncan’s multiple range test, P = 0.05.

3.31 b

2.40 b

163.2 b

14.7 b

Substrate Type Affects Growth, Yield and Mineral Composition of Cucumber and Zucchini Squash

Table 2 Effects of substrates on major elements concentration of zucchini and cucumber leaves.

Major elements (g·kg -1 of dry weight)

Substrates N P K Ca Mg

Zucchini

14.8 a Perlite

Cocofiber 32.6 a 1 2.1 a

37.8 a

24.4 a

13.6 ab Pumice

12.9 b Rockwool

14.6 c Cucumber Cocofiber

8.7 a Perlite

6.9 bc Pumice

5.7 c 1 Means within columns separated using Duncan’s multiple range test, P = 0.05.

Rockwool 28.2 b 4.4 a 34.8 a

30.2 c

Table 3 Effects of substrates on sodium and trace element concentration of zucchini and cucumber leaves.

Trace elements (mg·kg -1 of dry weight)

Substrates Na Cu Fe Mn Zn

Zucchini

58.2 c Perlite

Cocofiber 257.2 b 8.7 a 57.8 a

116.7 b

87.0 b Pumice

111.6 a Cucumber Cocofiber

Rockwool 743.7 a 7.2 a 52.1 a

359.7 a

53.0 b Perlite

70.2 a Pumice

67.9 a 1 Means within columns separated using Duncan’s multiple range test, P = 0.05.

Rockwool 196.2 a 10.0 a 61.7 a

41.1 b

Table 4 Effects of substrates on major elements concentration of zucchini and cucumber fruits.

Major elements (g·kg -1 of dry weight)

Substrates N P K Ca Mg

Zucchini

5.5 a Perlite

Cocofiber 47.5 a 1 5.2 a

46.0 a

4.1 a

5.7 a Pumice

5.4 a Rockwool

5.7 a Cucumber

4.1 a Perlite

Cocofiber 26.3 a 4.9 a 87.9 a

3.8 b

3.8 ab Pumice

3.9 ab Rockwool

3.6 b 1 Means within columns separated using Duncan’s multiple range test, P = 0.05.

whereas the Ca concentration was significantly observed on plants grown with the rockwool substrate influenced by the growing media only for cucumber

for both zucchini squash and cucumber (Table 2). with the highest value observed on pumice. Finally,

The highest Na for both zucchini squash and the lowest Mg concentration in leaf tissue was

cucumber was recorded on rockwool, whereas no

Substrate Type Affects Growth, Yield and Mineral Composition of Cucumber and Zucchini Squash

significant differences were observed for the Cu and

Agronomy 23 (2005) 183-194.

Fe concentration (Table 3). Moreover, the highest [4] Y. Rouphael, M. Cardarelli, E. Rea, A. Battistelli, G. Colla, Comparison of the subirrigation and drip-irrigation system

value of Mn, and Zn were recorded on perlite and for greenhouse zucchini squash production using saline rockwool, respectively.

and non-saline nutrient solutions, Agricultural Water For the zucchini experiment, no significant

Management 82 (2006) 99-117. difference among treatments was observed for the [5] Y. Rouphael, G. Colla, The influence of drip-irrigation or subirrigation on zucchini squash grown in closed-loop

concentration of K, Ca, and Mg in fruit, whereas the substrate culture with high and low nutrient solution lowest value of N, and P was observed on rockwool

concentrations, HortScience 44 (2009) 306-311. substrate (Table 4). Finally, in the cucumber [6] Y. Rouphael, G. Colla, Modelling the transpiration of a

experiment no significant effect of substrate was greenhouse zucchini crop grown under a Mediterranean climate using the Penman-Monteith and its simplified

recorded for the N, and P concentration, while the version, Australian Journal of Agricultural Research 55 highest K and Mg concentration was observed in

(2004) 931-937.

fruits harvested from plants grown in the cocofiber [7] G. Colla, Y. Rouphael, M. Cardarelli, M. Tullio, C.M. substrate (Table 4), which is interesting from a Rivera, E. Rea, Alleviation of salt stress by arbuscular mycorrhizal in zucchini plants grown at low and high

nutritional point of view because fruits and vegetables phosphorus concentration, Biology and Fertility of Soils usually contribute to 35%, and 24% respectively of the

44 (2008) 501-509.

total K, and Mg to the dietary intake of humans [13]. [8] Y. Rouphael, M. Cardarelli, E. Di Mattia, M. Tullio, E. Rea, G. Colla, Enhancement of alkalinity tolerance in

4. Conclusions

two cucumber genotypes inoculated with an arbuscular mycorrhizal biofertilizer containing Glomus

To summarize, authors can conclude, that cocofiber, intraradices , Biology and Fertility of Soils 46 (2010) perlite and pumice are suitable for zucchini and

499-509.

cucumber production in closed soilless system, [9] J.M. Bremner, Total nitrogen, in: C.A. Black, D.D. Evans, whereas the use of rockwool is more suitable for crops I.L. White, L.E. Ensminger, F.E. Clark (Eds.), Methods of

Soil Analysis, Agronomy Monograph 9, 1965, pp. grown under heated greenhouse conditions.

1149-1178. [10] Y.P. Karla, Handbook of Reference Methods for Plant

References

Analysis, CRC Press Inc., Boca Raton, Fla., 1998, pp. [1] D. Savvas, Hydroponics: A modern technology supporting

165-170.

the application of integrated crop management in [11] T. Bas, Possibilities of using different organic and greenhouse, Journal of Food Agriculture & Environment 1

inorganic materials for greenhouse cucumber production, (2003) 80-86.

Thesis, Ege University, Izmir, 1991. [2] Y. Rouphael, G. Colla, Growth, yield, fruit quality and

[12] R. Choukrallan, N. Oujlouq, Etude comparative d’une nutrient uptake of hydroponically cultivated zucchini

culture de tomate conduite sur perlite et en plein sol sous squash as affected by irrigation systems and growing

serre, dans la region de souss Massa, These de Troisième seasons, Scientia Horticulturae 105 (2005) 177-195.

Cycle Agronomie, IAV Hassan II, Agadir, 1994. [3] Y. Rouphael, G. Colla, Radiation and water use

[13] O.A. Levander, Fruit and vegetable contributions to efficiencies of greenhouse zucchini squash in relation to

dietary mineral intake in human health and disease, Hort different climate parameters, European Journal of

Science 25 (1990) 1486-1488.

Journal of Life Sciences 6 (2012) 771-775

Effect of Compost Based Substrate and Mycorrhizal Inoculum in Potted Geranium Plants

Monica Tullio, Federico Calviello and Elvira Rea CRA, Agricultural Research Council, Research Centre for the Soil-Plant System, Via Della Navicella, 2-4, Rome 00184, Italy

Received: November 25, 2011 / Accepted: January 31, 2012 / Published: July 30, 2012.

Abstract: Two are the critical factors in the commercial production of ornamental plants: the substrate and the fertilization method, since the “sustainable floriculture” discourages the use of synthetic chemical fertilizers and peat-based substrates. The aim of this paper was to test a peat/compost based substrate and a guano + mycorrhizal inoculum fertilizer mixture, in order to obtain a sustainable quality yield of geranium plants. Geranium cuttings (Pelargonium zonale cv. real polaris and Pelargonium grandiflorum cv. lotus) were grown for two months in a glasshouse. Peat, in the growing substrate, was replaced with high quality compost (20%

and 40% as reduction percentages). The fertilizer mixture was made up of guano (3 g/L) and mycorrhizal inoculum (7.5 L/m 3 ). Plants of Pelargonium cv. Real polaris, grown on a substrate made up of peat and 20% of high quality compost, presented the better trade features, so to satisfy the “sustainable floriculture” need.

Key words: Compost-peat, horticultural products, pelargonium, guano, mycorrhizal inoculums.

1. Introduction  authorized in organic agriculture. Arbuscular mycorrhizal (AM) fungi are soil microrganisms living

The choices of growth substrate and fertilization in very intimate contact with roots of most plant method are two critical factors in the commercial species and they cannot be found in a compost-peat production of flowering ornamental plants. substrates. These fungi can help the plant to uptake Environmental and economical concerns have nutrients, such as P, N, Zn, Cu [3] and sometimes K generated interest in seeking alternatives for peat moss [4]. The symbiosis with these fungi may induce earlier because of the detrimental effects of peat harvesting flowering and increased flower number [5]. This trait on wetland ecosystems [1]. Composts have proved to of AM fungi is of particular interest to horticultural

be very promising such as a peat substitute [2]. production. Researchers have tested the interactive

However compost as a nutrient source for plants may effects of nutrient supply and mycorrhizal

require amendments to other organic nutrient sources colonization on flowering. On a peat substrate with

to meet the plant demand. A mixture made up of organic NPK fertilizer, mycorrhizal pelargonium

guano and a commercial mycorrhizal inoculum can be plants flowered earlier [6]. The effect of compost able to supply plants with the required nutrients and addition on mycorrhizal plants has been investigated minimize the environmental impact with respect to in a few studies only. So the aim of this work was to chemical fertilization. Guano is an organic NP the test a peat-compost substrate in association with fertilizer; its peculiarities are the fast nitrification and an organic fertilizer as media for potted pelargonium the absence of P insolubilization processes. It is production and to study the effect on plant quality

parameters.

Corresponding author: Elvira Rea, head of research, research field: plant growing system implementation. E-mail: elvira.rea@entecra.it.

Effect of Compost Based Substrate and Mycorrhizal Inoculum in Potted Geranium Plants

2. Materials and Methods

of compost and the fertilizer mixture substrate (CM20P and CM40P: P. × hortorum cv. Real polaris

High quality compost was used, obtained by the cuttings; CM20L and CM40L: P. grandiflorum cv

decomposing process of organic waste such as food Lotus cutting). All pots were watered to maintain

and vegetables residues from Rome local markets. It favourable water conditions in the substrates. The

was manufactured by AMA S.p.A. (Italy). In the plants were grown for two months in an automatic

present study, this compost was mixed with blonde greenhouse of “Vivai Albani & Ruggeri”. A rooting

peat (Technostrat srl, Sopram Association), to obtain a hormone (RIZOPON AA 0.5% 3-indolyboterzuur 0.5

compost based substrate with 20% or 40% compost by kg) was administrated to the Pelargonium

volume. The fertilizer mixture was made up of guano grandiflorum cuttings, at the cutting surface. The

(Guanito, 3 g/L; Italpollina S.p.A., Italia) and a fertilization plan was N:P:K= 1:0.6:1.3 for guanoless

commercially available mycorrhizal arbuscular crude theses and N:P:K= 1:0:1.3 for the other ones. At the

inoculum (Aegis granulo; contains Glomus intraradices

end of the experiment, shoots were separated from , 100 infective units per g inoculum;

3 administrated dose was 7.5 L/m roots, and shoot fresh weight (FW), shoot height (H), ; Italpollina S.p.A., leaves number and the branching values were

Italia). This mixture was mixed uniformly into the recorded. The mycorrhizal colonization of roots was

potting substrate before cuttings planting. On these determined following the method of Grace and

substrates, before the transplanting, the following Stribley (1991) [8]. Percentage of root colonization

parameters were detected: pH (CEN, EN 13037 was recorded by a stereomicroscope (Nikon

method), electrical conductivity (CEN, EN 13037 Instruments S.p.A., SMZ-U, Japan) at 100 × using the

method) and P and K concentration by incineration grid line intersection method [9]. Pots were arranged

and determination at the Inductively Coupled Plasma in a completely randomized design. Data were

Atomic Emission Spectrometric (ICP). The Total N analyzed by a two-factorial analysis of variance, with

concentration was determined by Kjeldhal method, cultivar and treatments as experimental factors. Mean

1883 [7]. Cuttings of pelargonium (Pelargonium × separation was carried out with the test LSD (P <

hortorum cv. Real polaris and Pelargonium 0.05). Data were analyzed using Statgraphics Plus 5.1

grandiflorum cv Lotus, Vivai Albani & Ruggeri, Italy) software (Statpoint techonologies, Inc., Warrenton,

were placed, singly, in 200-mL pots filled with the

Virginia, USA).

carried out substrates. The control theses had a blond peat substrate (TP: P. × hortorum cv. Real polaris

3. Results and Discussion

cuttings; TL: P. grandiflorum cv. Lotus cuttings); the theses C20 and C40 had a blond peat and 20% or 40%

The values of the chemical, physical parameters and v/v of compost substrate respectively (C20P and C40P:

the N, P and K concentration of the substrates were in P. × hortorum cv Real polaris cuttings; C20L and

according to a their use for pelargonium growing C40L: P. grandiflorum cv. Lotus cutting); the theses

(Table 1).

CM20 and CM40 had a blonde peat, 20% or 40% v/v Colonization of P. × hortorum cv. Real polaris

Table 1 pH and electrical conductibivity (E.C.) values and N, P, K contents of the substrates.

Potting substrates pH

K (% dw) Control 5.8

E. C. (mS/cm)

N (% ss)

P (% ss)

0.32 0.37 0.024 0.061 C20 6.9 0.54 0.76 0.097 0.27 C40 7.3 0.63 0.98 0.13 0.41 CM20

7.0 0.86 0.91 0.17 0.33 CM40

Effect of Compost Based Substrate and Mycorrhizal Inoculum in Potted Geranium Plants

roots by AM was not significant different between grandiflorum root colonization is available because the two compost addition percentages. Average root

these plants had a very little root system (1 or 2 little colonization percentage was 52.33% (a) for CM20

roots, few centimetres long) which did not allow any thesis and 47.33% (a) for CM40 thesis. determination. All biometric parameters recorded such Non-inoculated plants remained free of mycorrhizal

as shoot FW, shoot height and leaves number of colonization, although the substrates had not been

Pelargonium × hortorum plants, grown on 20% sterilized before use. No information about P.

compost based substrate with the fertilizer mixture,

resh (g

5 f fg g g

tf

Shoo CML40

pelargonium species

Fig. 1 Shoot fresh weight values (g) of the two pelargonium species.

Different letters denote significant differences between means of the treatments determined by Fisher’s least significant difference (LSD) procedure (P ≤ 0.05). P value: P (treatment × cultivar) = 0.

15 bc b

n 10 d

number

e pla

fg f

av Le

0 TP CP20 CP40 CMP20 CMP40 TL CL20 CL40 CML20 CML40

Leaves number/plant

Pelargonium species Pelargonium species

Fig. 2 Leaves number/plant of the two pelargonium species.

Different letters denote significant differences between means of the treatments determined by Fisher’s least significant difference (LSD) procedure (P ≤ 0.05). P value: P (treatment × cultivar) = 0.

Plant h

TP CP20 CP40 CMP20 CMP40 TL

CL20 CL40 CML20 CML40

Pelargonium species

Fig. 3 Plant high values (cm) of the two pelargonium species.

Different letters denote significant differences between means of the treatments determined by Fisher’s least significant difference (LSD) procedure (P ≤ 0.05). P value: P (treatment × cultivar) = 0.

Effect of Compost Based Substrate and Mycorrhizal Inoculum in Potted Geranium Plants

CP40 CMP20 CMP40

TL CL20

CL40 CML20 CML40

ran

ch

in

Pelargonium species

Fig. 4 Branching values of the two pelargonium species.

Different letters denote significant differences between means of the treatments determined by Fisher’s least significant difference (LSD) procedure (P ≤ 0.05). P value: P (treatment × cultivar) = 0.

had the highest values (Figs. 1-3). Pelargonium × hortorum plants, grown on 20% compost based substrate with the fertilizer mixture, had the highest branching values, too (Fig. 4).

Considered parameters for Pelargonium grandiflorum plants showed very little differences among them. In particular, the control and CL20 plants had the higher values of shoot FW and leaves number with respect to the other theses. The control plants had the highest values of shoot height. All theses had the same values of branching (Figs. 1-4). Taking into account that the aim of this research was to check a compost-based substrate with 20% or 40% compost by volume in association with a fertilizer mixture made up of guano and a commercial mycorrhizal crude inoculum, in potted pelargonium plants, the two compost-based substrates, used for the trial, resulted suitable for growing of potted pelargonium plants. Two were the pelargonium species considered: P. × hortorum cv. Real polaris and P. grandiflorum cv. Lotus. Roots of inoculated Pelargonium × hortorum , after two months from the transplant, were well colonized with mycorrhizal fungi and the root colonization percentage was about 50%. This was true at both compost addition rate (20% and 40%). The quality of P. × hortorum cv. Real polaris plants, grown on 20% compost based substrate in association with the fertilizer mixture, was the highest. A significant result of this study for

practical floriculture point of view was the increased branching of these plants because this parameter is the only economically important one for the plants, in this growth stage. In P. grandiflorum cv. Lotus, the increasing of the rate of compost in the substrate from 20% to 40% with or without the fertilizer mixture had

a very little effect on the growth. Biometric parameters were slightly higher in the control and in the plants grown on 20% compost-based substrate. Lopez showed that compost based substrates utilized as potting media for domestic use, produced an underdevelopment of Pelargonium zonale cv. Lucky Break F2 plants with respect to the control [10]. This behaviour was related to the inferior physical properties of the compost-based potting media, nitrogen immobilization due to the high C/N ratio of pine bark, and probably lack of available phosphorus originated by high calcium and high pH of the compost-based media. In this trial, the little size of the root system, due to the trouble doing rooting of this specie, has to be taken into consideration such as a possible factor causing the slight plant underdevelopment.

4. Conclusion

To summarize we can conclude that the AM colonization was well established in pelargonium plants on a horticultural substrate. The compost amount didn’t influence the AM capability to colonize

Pelargonium species

Branching

Effect of Compost Based Substrate and Mycorrhizal Inoculum in Potted Geranium Plants

775

the root system and there weren’t any differences Developmental Plant Biology 1 (2001) 1-15.

E. George, Nutrient uptake: Contribution of arbuscular between the two compost addition percentages. The mycorrhizal fungi to plant mineral nutrition, in: Y. growing technique of pelargonium plants using 20%

[4]

Kalpulnik, D.D. Jr. Doudds (Eds.), Arbuscular compost-peat substrate and an organic fertilizer made

Mycorrhizas: Physiology and Function, Kluwer, 2000, pp. up of guano and arbuscular mycorrhizae inoculum,

307-343.

A. Gaur, A. Adholeya, Diverse response of five can be used for obtaining high quality horticultural ornamental plant species to mixed indigenous and single

[5]

products. isolate arbuscular mycorrhizal inocula in marginal soil So this work has proven that compost-based

amended with organic matter, Journal of plant Nutrition 28 substrates resulted an alternative media for potted

(2005) 707-723. [6] J. Novak, Effects of arbuscular mycorrhizal fungi and

pelargonium plants production. A significant result of organic fertilization on growth, flowering, nutrient uptake,

this study for a practical point of view was the photosynthesis and transpiration of geranium increased branching of the plants because this

(Pelargonium hortorum L.H., Bailey “Tango Orange”), parameter is the only economically important one, in

Symbiosis 37 (2004) 259-266. [7] J. Kjeldhal, A new method for the determination of

this growth stage. nitrogen in organic matter, Zeitschreft Fur Analytische

References Chemie 22 (1883) 366.

C. Grace, D.P. Stribley, A safer procedure for routine [1] M.R. Evans, S. Konduru, R.H. Stamps, Source variation in

[8]

staining of vesicular-arbuscular mycorrhizal fungi, physical and chemical properties of coconut coir dust,

Mycological Research 95 (10) (1991) 1160-1162. HortScience 31 (1996) 965-967.

[9] M. Giovannetti, B. Mosse, An evaluation of techiques for [2] O. Verdonck, Compost from organic waste materials as

measuring vesicular arbuscular mycorrhizal infection in substitutes for the usual horticultural substrates, Biological

roots, New Phytology 71 (1980) 287-295. Wastes 26 (1988) 325-350.

[10] R. López, C. Durán, J.M. Murillo, F. Cabrera, Geranium’s [3]

E. Rea, M. Tullio, Arbuscular mycorrhizae: A natural response to compost based substrates, Acta Horticulturae answer to fertilisation need, Recent Research in

469 (1998)255-262.

Journal of Life Sciences 6 (2012) 776-782

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

1 2 Aymen Nefla 1 , Ridha Ouni and Saïd Nouira 1. Department of Biology, University of Sciences of Tunis, El Manar II, Tunis 2092, Tunisia

2. Association for the Protection of Environmental and Natural Heritage, Béjaoua 2 SidiThabet CP: 2020, Tunisia

Received: December 26, 2011 / Accepted: March 06, 2012 / Published: July 30, 2012.

Abstract: The study of the reproduction of the Glossy Ibis (Plegadis falcinellus) in Tunisia was undertaken from 2008 to 2010. Until the discovery of nesting in 2008, this species had only been considered as wintering in Tunisia. The Tunisian breeding population resides in the Lebna Dam in the north east of the country where it forms a mixed nesting colony with other species of the family Ardeidae ((Bubulcus ibis (Bi): 388 pairs, Ardeola ralloides (Ar): 17 pairs, Egretta garzetta (Eg): 27 pairs), (Bi: 300 pairs, Ar: 25 pairs, Eg: 40 pairs) and (Bi: 400 pairs, Ar: 30 pairs, Eg: 10 pairs) recorded in 2008, 2009 and 2010 respectively). All nests were constructed on Acacia horrida. Laying began in early May. The average clutch size over the three years of the study was 3.44 ± 0.73 eggs (N = 29 nests). Hatching success was 83 % (2.86 ± 1.18 eggs hatched/nest) and 2.65 ± 1.17 hatchlings/nest survived until the age of 10 to 12 days. Egg mortality was 17% during the incubation phase and chick mortality was 7.2%. No interannual variation was detected in these parameters.

Key words: Breeding parameters, Plegadis falcinellus, Lebna dam, Tunisia.

1. Introduction  Mediterranean European countries (continental Italy [9]; Sardinia [10]; the South of France [11]; Spain [12]).

The Glossy Ibis (Plegadis falcinellus) has a Conversely in Tunisia, the Glossy Ibis has always cosmopolitan distribution [1]. Its current reproduction been observed in double passing (pre and postnuptial) zone is vast and scattered. The most important and in winter in wetlands around Tunisia with the reproduction sites are situated in the south east of postnuptial passage being the most prominent [13]. Africa, Madagascar, the south of Europe, the Middle According to Shogolev [14], the majority of these East, South East Asia, Australia, as well as the coasts individuals are without a doubt from the Balkans, of North America and the the Caribbean [2]. In North while some Ukrainian populations that winter in the Africa, Glossy Ibis had nested commonly in Morocco Niger Basin also pass through Tunisia while crossing and Algeria in the 19th century [3, 4]. Subsequently, the Sahara. This bird had only ever been known as a recording of the reproduction of the species had halted wintering species [3, 13, 15, 16] until nesting was for almost a century only to begin again with new observed for the first time in 2008 at Lebna in Cap recordings of reproduction at the mouth of the Massa

Bon [17].

Wadi in 1994 [5], in the palm grove of Marrakech, Because of their vulnerability, their conservation Morocco in the 80’s [4, 6], and at Lake Tonga, Lake value and their quick response to environmental Fetzara, Dakhla, and Chatt in Algeria [7, 8]. This return disturbances, the waders, such as Glossy Ibis, are of Glossy Ibis has also concerned several regarded as bio-indicators water-birds [18, 19]. Then

Corresponding author: Aymen Nefla, Ph.D., assistant it was essential to study the reproduction of this professor, research fields: biodiversity and populations biology. E-mail: aymennefla2007@yahoo.fr.

important species on Lebna dam. In order to assess the

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

sustainability of settlement of Glossy Ibis in Lebna cereal crops. During the three study years, the Glossy dam and to determine its reproductive performance we

Ibis always nested within the same mixed heronry that propose to carry out a complete study of the breeding

contained the Cattle Egret (Bubulcus ibis; 2008: 388, parameters of this species. Thus we propose to do:

2009: 300 and 2010: 400 pairs) (the dominant species),

Estimate the Glossy Ibis populations size; the Little Egret (Egretta garzetta; 2008: 27, 2009: 40

Provide information on preliminary population and 2010: 10 pairs), and the Squacco Heron (Ardeola parameters (such as the nesting period, the average

ralloides ; 2008: 17, 2009: 25 and 2010: 30 pairs). The clutch size, hatching success, fledging success and

colony occupies an area of 100 m 2 on a small island breeding success.

(N: 36.74521, E: 10.91661) situated 100 m from the bank of the lake where (Acacia horrida) and (Tamarix

2. Materials and Methods

africana ) grow (Fig 2).

The study was undertaken at Lebna Dam in Cap The south-western part of the dam (at 5 km of Bon. The dam is located in the north east of Tunisia

nesting area) is one of the most important feeding sites and covers a surface of 724 ha with a reservoir of 30

of the species. All feeding areas are located on the million m 3 of water (Fig 1).

periphery of the colony. The most frequented one are The site is formed of two major parts, a lagoon and

Korba lagoon (at 19 km of nesting area), Lahjar dam Lebna Wadi. The latter feeds the lagoon with fresh

(at 15 km of nesting area) and Mlaabi dam (at 11 km water, which drains into the Mediterranean. The bank

of nesting area).

of the lagoon, the bank of the Wadi, and other shallow Visits to the study site were spaced in 2008 between parts of both provide favorable habitat for the growth

the end of May (the date of the discovery of the of (Phragmites australis) and (Typha latifolia), which

colony) and the end of July, and between mid-May extend for a surface of 40 ha. The flooded zone (20 ha)

and the end of July in 2009 and 2010 at intervals of is covered by herbaceous aquatic plants and the

one visit per week, during which nests were observed surrounding plains are cultivated by vegetable and

and all changes noted.

Mlaabi dam Lahajar dam

Tunisia

Korba lagoon

Cap Bon

Lebna dam

Islet position

Fig. 1 Map showing the position of the nesting area at the Lebna dam (Google Earth 2010).

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

Fig. 2 Vegetal composition of heronry.

The estimation of the total number of reproductive size (number of eggs layed/number of clutches) and pairs was done by a direct count of constructed nests.

breeding success (number of survivors/number of All constructed nests of Glossy Ibis were individually

eggs incubated) [20] are expressed by averages and marked by small numbred wooden labels placed

proportions so that these data can be compared with below the nest. We studied 8, 18 and 3 nests during

other studies.

the 3 years, respectively. Nests were weekly checked All Comparisons of averages of reproductive from the beginning of the egg laying period with the

parameters were done using the student t-test and P < aid of a mirror, fixed at the top of a pole. We recorded

0.05 was used as the level of significance. the number of eggs, hatchlings and number of chicks

3. Results and Discussion

surviving per nest until the age of 10 to 12 days (when, upon our approach, they were capable of climbing

The number of breeding pairs of the Glossy Ibis has from the nest).

shown a large fluctuation over the past three years We considered youngs who attain 10 to 12 days as

going from 18 couples in 2009 to only 3 in 2010 and survivors. The clutch was considered complete when

all nests were built on Acacia horrida. The observed there was no change after two consecutive visits. The

decline is probably due in part to the lowered water total mortality rate (number of eggs and young

levels in the dam which has allowed for the lost/number of eggs layed), the mortality rate of eggs

development of a passage allowing predators and (number of eggs lost/number of eggs layed), the

children to gain access to the island (Fig 3), and in mortality rate of young (number of lost young/number

another part to the increase in the numbers of ardeid of eggs hatched), the fledging success (number of

nesters on the acacia, especially the Cattle Egrets. survivors/number of nest), hatching rate ((number of

They are the most competitive species and reduce the eggs hatched/number of eggs laid) × 100), hatching

nesting space for the Glossy Ibis, which has always success (number of eggs hatched/nest) average clutch

shown a preference to nest on acacia, despite the

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

Fig. 3 Appearance of the passage leading to the nesting island. a: passage leading predators and children to the nesting island; b: water level of previous years.

availability of nearby tamarix (Tamarix africana). due to the earlier onset of spring and production of This hypothesis can be supported by Korschgen and

food that occurs at these latitudes [24]. With the Dahlgren [21], which showed that disturbance can

exception of 2009, a nest of 4 eggs with 3 young cause, in addition to increasing desertion of nests and

hatching was the most frequent case during the whole reducing hatching or fledging success, a decline in the

study period (Table 1).

number of breeding pairs. We recorded an average clutch size of 3.44 ± 0.73 Regardless of study year, nesting in Lebna dam has

(SD) eggs, this value was considerably important always begun in early May, early relative to what was

compared with other study (Table 2). The average noted by Heim de Balzac and Mayaud [3] in North

clutch size of many birds increases with increased Africa (June-July) and a bit later than was noted by

distance from the equator [25]. Although some Herons Boucheker [8] in Algeria (mid-April). This variation

and Egrets obey this model [26-29], no relation found is probably due to climate variables such as rainfall

between Glossy Ibis clutch size and distance from and temperature which play an important role in

equator (Table 2).

determination of onset of breeding [22]. The timing of The maximum clutch was 4 eggs whereas the breeding of some birds like Ammodramus maritimus

minimum of 1 or 2 eggs only represented 7% of the total mirabilis in the Florida Everglades was related to

nests. The majority of nests produced young and the rainfall preceding the breeding season, with females

maximum proportion was recorded in 2009 (Table 3). initiating nesting up to 1 month earlier in years with

Table 1 Number of pairs producing 1, 2, 3 or 4 eggs and/or

greater rainfall [23]. In the South-West of Spain, the

youngs.

earlier reproduction of Night Heron Nycticorax Eggs Youngs

Year

nycticorax occurred when winter temperatures were Total 123 4 1234 higher and winter rainfall lower [24]. Earlier

2008 8 0 0 2 6 0151 reproduction in southern areas has been shown for

2009 18 0 1 9 8 1475 other bird species, and has been interpreted as being

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

Table 2 Comparison between recorded reproductive parameters in lebna and other colonies.

Average clutch size

Hatching rate %

Fledging success

(egg/clutch)

(eggs hatched/eggs laid)

(survivor/nest)

(Lebna) Tunisia (present study)

North America [31-34]

2.4-2.9

0.4-1.0

The hatching success and the fledging success of species’ reproductive parameters (hatching success, young over the 3 study years attained respectively

fledging success and breeding success), which are

2.86 ± 1.18 (SD) eggs/nest and 2.65 ± 1.17 (SD)

essential for Assessment of nesting birds.

young/nest. The breeding success of the Glossy Ibis The measurement of reproductive parameters of the noted at Lebna was 72%, its maximum being in 2009

Glossy Ibis in Tunisia was not possible until 2008, (Table 3).

date of the first reproduction of the species in this The total mortality rate reached 23% over the two

country. It is in Lebna dam where she trained with phases of incubation and rearing (until 10 to 12 days

other herons a mixed nesting colony.

old young). The birds lost 17% of their eggs during the During the entire period of our study, the species first phase and 7.2% of their young during the second

has consistently nested on the same site within the phase. Otherwise stated, mortality touched 0.58 ± 0.94

same colony. Although always present, there was a (SD) eggs/nest and 0.31 ± 0.66 (SD) young/nest. In

wide fluctuation in the number of breeding couples. 2008, the low mortality rate of Glossy Ibis compared

This instability is probably due to changes in rainfall. with recorded for other ardeid nesters (Bubulcus ibis:

In fact, the elevation of the dam’s water level will 52.22% and Ardeola ralloides: 41.87%) [17] is

completely isolate the island. Thus, predators and probably due to site selection by this species, which at

small children will not be able to reach the colony at Lebna has always chosen to nest on Acacia horrida,

least during the first week of incubation and rearing, with long and dense spines, that provide more

which will avoid disturbances.

protection for eggs and nestlings, in contrast to some The very high rainfall of the year 2009 (unlike 2008 other species of ardeid birds which nest in the more

and 2010) allowed recording the highest breeding exposed branches of Tamarix africana.

parameters. The values recorded during this study Average clutch size, hatching success, fledging

Table 3 Breeding parameters of Glossy ibis Plegadis

success, and mortality rate did not show inter-annual

falcinellus.

variation (Table 4). 2008 2009 2010

Active nests

4. Conclusions

Average clutch size (egg laid/clutch)

66.7 Because of its vulnerability and conservation value, Hatching success (egg hatched/nest) 2.87 3 2 Glossy Ibis is ecologically considered as one of the

Percentage of hatching (%)

66.7 most remarkable species. In fact, considering its quick

Percentage of nests with nestlings (%) 87.5 94.4

66.7 response to environmental disturbances, it is regarded

Percentage of successful nest (containing at least 1 survivor) (%)

91.3 92.6 100 as one of water-birds bio-indicators. This response

Percentage of survivors (%)

30 18 33.3 may be manifested by a decrease in number of

Mortality (%)

2.62 2.77 2 breeding pairs which will automatically affect the

Fledging Success (survivor/nest)

Breeding success (%)

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

Table 4 Inter -annual variation of breeding parameters. Average clutch size (egg laid/clutch)

Hatching success (egg hatched/nest)

Fledging success (survivor/nest)

2009 2010 2009 2010 2009 2010 t

df 24 9 24 9 24 9 P 0.14 0.25 0.79 0.36 0.75 0.5 t

0.77 1.37 1.04 2009 df ―

19 P 0.44 0.18 0.3 t : a t-test is any statistical hypothesis test in which the test statistic follows a Student’s t distribution if the null hypothesis is

supported. It can be used to statistically test hypothesis of equality of two means; df: the number of observations (total number of egg or survivor concerned by comparison) minus the number of necessary relations among these observations; P : the P-value is the probability of obtaining a test statistic at least as extreme as the one that was actually observed, assuming that the null hypothesis is true. One often “rejects the null hypothesis” when the P-value is less than the significance level α (Greek alpha), which is often 0.05 or 0.01. When the null hypothesis is rejected, the result is said to be statistically significant.

were higher than those mentioned by other authors in of the World, Lynx Edicions, Barcelona, 1992.

H. Heim de Balzac, N. Mayaud, Les Oiseaux du Australia and North America. In fact, Lebna’s Nord-Ouest de l’Afrique, Paul Chevalier, Paris, France,

populations showed some reproductive affinities to

North African populations. [4] M. Thévenot, R. Vernon, P. Bergier, The birds of Glossy Ibis has been always regarded as wintering Morocco, British Ornithologists’ Union, Checklist Series

n°20, 2003.

in Tunisia. The reproductive performance recorded in

E. Rousseau, Nouveau cas de reproduction de l’Ibis this study indicates some environmental changes that

falcinelle (Plegadis falcinellus) au Maroc, Alauda 62 promoted its current installation. Further work must

(1994) 313-314.

D. Barreau, P. Bergier, L’avifaune de la région de

be done to highlight the effect of biotic and abiotic Marrakech (Haouz et Haut-Atlas de Marrakech, Maroc) 2:

factors on the eco-biology of this species on Lebna Non passeraux, Alauda 69 (2001) 167-202. dam.

G. Belhadj, B. Chalabi, Y. Chabi, Y. Kayser, M. Such work seems to be essential in order to take the

Gauthier-Clerc, Le retour de l’Ibis falcinelle (Plegadis falcinellus ) nicheur en Algérie, Aves 44 (2007) 29-36.

necessary measures of conservation and protection of

A. Boucheker, R. Nedjah, F. Samraoui, R. Menaï, B. this new nesting population. Otherwise, the permanent

Samraoui, Aspects of the breeding ecology and settlement of this species in Tunisia will be

conservation of the Glossy Ibis in Algeria, Waterbirds 32 threatened. (2) (2009) 345-351. [9] P. Brichetti, Nidificazione di nitticora nycticorax

Acknowledgments

nycticorax e mignattaio Plegadis falcinellus in Puglia, Avocetta 10 (1986) 59-60.

Our special thanks to Ms. Shauna Joy Hawkins [10] M. Grussu, Nidificazione e svernamento del mignattaio Regayeg and Ms. Wided Tlili Mrabet for their

Plegadis falcinellus e nidificazione della sgarza ciuffetto Ardeola ralloides in Sardegna, Rivista Italiana di

cooperation and help.

Ornitologia, Milano 57 (1987) 62-68.

References H. Heinzel, D. Martinoles, Nouvelle nidification de l’Ibis

falcinelle Plegadis falcinellus en France, Alauda 56 [1] S. Cramp, K.E.L. Simmons, The Birds of the Western

(1988) 429-430.

Palearctic, Vol. 1, Oxford University Press, Oxford, [12] F.J. Purroy, Atlas de las Aves de España (1975-1995), United Kingdom, 1977.

Sociedad Española de Ornitologia, Lynx Edicions, [2] J. Del Hoyo, A. Elliot, J. Sargatal, Handbook of the Birds

Barcelona, 1997.

The Breeding Status of the Glossy Ibis Plegadis falcinellus in the Lebna Dam in Cap Bon, Tunisia

[13] P. Isenmann, T. Gaultier, A. El Hili, H. Azafzaf, H. [23] R.L. Boulton, B. Baiser, M.J. Davis, T. Virzi, J.L. Dlensi, M. Smart, Les Oiseaux de la Tunisie—Birds of

Lockwood, Variation in laying date and clutch size: The Tunisia, Société d’Etudes Ornithologiques de France,

everglades environment and the endangered cape sable France, 2005.

Seaside Sparrow (Ammodramus Maritimus Mirabilis), [14]

I.V. Shogolev, Migration and wintering grounds of The Auk 128 (2) (2011) 374-384. Glossy Ibises (Plegadis falcinellus) ringed at the colonies

D. Parejo, J.M. Sanchez, J.M. Avilès, Breeding biology of the Dnestr delta, Ukraine, Black Sea, Colonial

of the Night Heron Nycticorax nycticorax in the Waterbirds 19 (1996) 152-158.

south-west of Spain, Ardeola 48 (1) (2001) 19-25. [15] R.D. Etchecopar, F. Hue, Les Oiseaux du Nord de

D. Lack, The Natural Regulation of Animal Numbers, l’Afrique, Boubée N. & Co, Paris, 1964.

Oxford University Press, London, 1954. [16] N. Mayaud. Les oiseaux du nord-ouest de l’Afrique,

D.A. Jenni, A study of the ecology of four species of notes complémentaires, Alauda 50 (1982) 45-67, 114-145

herons during the breeding season at Lake Alice, Alachua and 286-309.

County, Florida, Ecological Monographs 39 (1969) [17] R. Ouni, A. Nefla, A. El Hili, Nidification de l’ibis

243-270.

falcinelle Plegadis falcinellus au Cap-Bon, Tunisie, [27] H.M. Pratt, D.W. Winckler, Clutch size, timing of laying, Alauda 77 (2) (2009) 115-120.

and reproductive success in a colony of Great Blue [18] J.A. Kushlan, Colonial waterbirds as bioindicators of

Herons and Great Egrets, Auk 102 (1985) 49-63. environmental change, Colonial Waterbirds 16 (1993b)

[28] W.J. Arendt, A.I. Arendt, Aspects of the breeding biology 223-251.

of the Cattle Egret (Bubulcus ibis) in Montserrat, West [19] J.C. Ogden, A comparison of wading bird nesting colony

Indies, and its impact on nest vegetation, Colonial dynamics (1931-1946 and 1974-1989) as an indication of

Waterbirds 11 (1988) 72-84.

ecosystem conditions in the southern Everglades, in: S.M. [29] G.S. Ranglack, R.A. Angus K.R. Marion, Physical and Davis, J.C. Ogden (Eds.), In Everglades: The Ecosystem

temporal factors influencing breeding success of Cattle and Its Restoration, St. Lucie Press, Delray Beach,

Egret Bubulcus ibis in a west Alabama colony, Colonial Florida, 1994, pp. 1-38.

Waterbirds 14 (1991) 140-149.

[20] H. Hafner, Contribution a l’étude de quatre espéces de [30] K.W. Lowe, Egg size, clutch size and breeding success of herons (Egretta garzetta L., Ardeola ralloides Scop.,

the Glossy Ibis Plegadis falcinellus, Emu 83 (1983) Ardeola ibis L., Nycticorax nycticorax L.) pendant leur

31-34.

nidification en Camargue, Ph.D. Thesis, Université Paul [31] J. Bull, Birds of New York State, Doubleday, Garden Sabatier de Toulouse, 1977.

City, NewYork, 1974.

[21] C.E. Korschgen, R.B. Dahlgren, Human Disturbances of

B. Williams, Growth rate and nesting aspects for the Waterfowl: Causes, Effects, and Management, U.S. Fish

Glossy Ibis in Virginia, The Raven 46 (1975) 35-50. and Wildlife Service, Washington, 1992.

[33] J. Burger, Competition between Cattle Egret and native [22] K. Immelmann, Avian biology, in: D.S. Farner, J.R. King,

North American herons, egrets, and ibises, Condor 80 K.C. Parkes (Eds.), Ecological Aspects of Periodic

(1978) 15-23.

Reproduction, Academic Press, New York, 1971, pp. [34] L.M. Miller, J. Burger, Factors affecting nesting success 341-389.

of the Glossy Ibis, Auk 95 (1978) 353-361.

Journal of Life Sciences 6 (2012) 783-785

Survey on Parasites in Sparrows of Amol (Mazandaran

Province, Iran)

1 2 2 2 Sina Faghihzadeh Gorji 2 , Bahareh Shemshadi , Hamid Habibi , Sattar Jalali , Masoud Davary and

Mohammadreza Sepehri 2

1. Member of Young Researchers Company, Islamic Azad University, Garmsar Branch, Iran 2. Faculty of Veterinary Medicine, Islamic Azad University, Garmsar Branch 3581631167, Iran

Received: October 23, 2011 / Accepted: February 01, 2012 / Published: July 30, 2012.

Abstract: In order to survey on sparrows parasites (Passer domesticus), 100 samples were collected from the citrus gardens of Amol’s City. The surface of their body checked for presence of ectoparasites. The blood samples took from their wings for blood parasites. At the laboratory, the body of sparrows opened and the gastrointestinal checked for endoparasites. The intestine opened and scratched carefully to find worms. Worms kept in alcohol-glycerin (10%). Eggs & oocysts of the parasites checked, by fecal floatation test. The result shows that the infection with Railletina is 1%, Ascaridia galli (2%), Heterakis gallinarum (4%), eggs of A. galli (5%), oocyst of Eimeri (28%), Dermanyssus gallinae (1%). 11% of the sparrows infected with Plasmodium in their bloods.

Key words: Parasites infection, sparrow, Iran.

1. Introduction  6] but there is not any information in Iran about the presence of parasites in sparrows. Moreover, we do

The house sparrow Passer domesticus L. is a not know exactly which parasites can be transmitting sociable bird belonging to Passeridae [1]. This bird is by this bird to poultries and humans, and what we closely associated with human habitation, nests under should do to limit the detriments. Therefore, in this roof tiles, in holes, cavities and sometimes builds research we want to find out which parasites can nests in the open; loud noisy flocks gather in hedges

infect sparrows in Iran.

and bushes. Some people in the word use Sparrows as food. Furthermore, free birds, like Sparrows are the

2. Materials and Methods

guest of many places, so they can bring some viral, In the present study, the authors chased 100 samples bacterial, or parasitical diseases that can affect both from different regions of city (citrus gardens and humans and animals [2]. Sparrows can come in some farms of chickens) in winter of 2008. The surface of stores and stain the materials in these stores. Some of their bodies checked for presence of ectoparasites. our problems in poultry industries back to the Ectoparasites kept in alcohol-glycerin (10%). The pollution that these free birds bring to us [3, 4]. blood samples took from the vein of their wings, and According to these dangers, we should understand the the expansions of them colored with “Giemsa”. problems might be caused by sparrows. For finding the helminthes of gastrointestinal, the Numerous studies have addressed the occurrence of intestine opened, after scratching the mucous of the parasites in the house sparrows all over the world [3, 5, intestine the presence of parasites investigated. All

Corresponding author: Sina Faghihzadeh Gorji, Ph.D. contents poured into the plate and checked under the candidate, researcher, research fields: veterinary medicine,

lamp to find worms. Feces checked for eggs and avian diseases. E-mail: dvm_sinafg@yahoo.com.

Survey on Parasites in Sparrows of Amol (Mazandaran Province, Iran)

oocysts of parasites by fecal floatation test. transmit by them, we can be ready to prevent or treat In addition, the feces expansions colored with

people and animals.

“Trichrome” for avian protozoa and Giardia. Infection with Plasmodium and its dangers studied

The smears of the esophagus colored with in much research around the word, and it shows the

“Modified ziel-nelson” to find Cryptosporidium. important role of Sparrows to distribute of

Genus of all parasites performed under optic Plasmodium in avian [6-8]. Parasites of genus microscope. Plasmodium cause anemia, so scientists call it avian malaria. Mosquitoes transmit all species of

3. Results

Plasmodium. In our study, we obtain 11% pollution The result shows that infection with Railletina is

(Fig. 1), because of Mazandaran climate and humidity, 1%, Ascaridia galli is 2% and Heterakis gallinarum is

there are many mosquitoes that they can transmit this 4%. In addition, the examination of stool with the

parasite to poultries. As we know, avian Plasmodium fecal flotation method shows that the contamination

is not important for humans.

with eggs of A. galli and oocyst of Eimeria Infection with genus Eimeria in sparrows is very sequentially is 5% and 28%.

dangerous to spread coccidiosis in poultries [9], so we In our study, infection with Plasmodium was 11%.

should control our farms from this danger by good The review of ectoparasite shows 1% contamination

quarantine and prevention. This parasite has a short with scab Dermanyssus gallinae.

and direct life cycle so it is more dangerous than other parasites that need intermediate hosts. There is a

4. Conclusions

relationship between coccidiosis and other poultry Because of some people, use sparrows as food and

diseases like necrotic enteritis, Salmonella typhimurium the ability of sparrows to income in the aviary

and blackhead. Coccidiosis remains one of the most (traditional or industrial breeding) so they can scathe

expensive and common diseases of poultry production people directly or indirectly. Therefore, by in spite of advances in chemotherapy, management, understanding the diseases and parasites that can

nutrition, and genetics. In our study, pollution to this

Heminth of intestine % 15

Blood parasites

% 10 % 4 % % 5 5 Eggs or oocyst of Parasites

% 1 % 2 % 1 in feces % 0

Ectoparasites

Fig. 1 Rate of pollution in Sparrows.

Survey on Parasites in Sparrows of Amol (Mazandaran Province, Iran)

parasite was more than other parasites (28%). As we [2] H. Whitney, Salmonella in Songbirds, Government of know, infection to coccidiosis depends to the number Newfoundland and Labrador Dept of Natural Resources

Pub. AP033, July 27, 2004.

of invading organism, so we can control this parasite [3] R. Brad, LeaMaster, Parasites Important to Poultry in by vaccination, drug prevention, and good sanitation.

Hawaii and Their Control, Published by the Colledge of Mites can infect Sparrows to some disease like

Tropical Agriculture and Human Resources (CTAHR), April 2007, available online at:

erysipelas [10] and fowl pox [11]. Therefore, attention

http://ctahr.hawaii.edu/freepubs.

to the role of this bird for spreading these diseases is [4] W.L. Hoyle, Transmission of Poultry Parsites by Birds too important. Red mite cause anemia for Sparrows,

with Special References to the English or House Sparrow too. and Chickens, Kansa Academy of Science, Vol. 41, 1903, (Mar. 31 – Apr. 2, 1938), pp. 379-384.

Other parasites that we find need intermediate hosts, [5] C. Calegaro-Marques, S.B. Amato, Helminths of so they are important for sparrows and other free birds.

introduced house sparrows (Passer domesticus) in Brazil: Nevertheless, they are not so important for industrial

Does population age affect parasite richnes?, Iheringia, breeding. Ser. Zool. 100 (1) (2010) 73-78. [6] G. Valkiunas, T.A. Iezhova, C.V. Bolshakov, V. Kosarev,

Eliminate sparrows to enter farms is too important Blood parasites of the house sparrow Passer domesticus because we can control the harms of this bird with this

from northwestern Russia, with remarks on trends of method. global geographical distribution in this bird, Journal of

We should remind our farmers and breeders that the Naturral History 40 (29-31) (2006) 1709-1718. [7] E.D. Box, Blood and tissue protozoa of English sparrow best ways to control all diseases are good quarantine

(Passer domesticus) in Galveston, Texaa, Journal of and biosecurity. Therefore, we should remind,

Protozoology 13 (1996) 204-208.

“Prevention is better than treatment”. [8] C.G. Ludford, H.G. Puchase, H.W. Cox, Ducks infectious anemia virus associated with Plasmodium lophrae,

Acknowledgments

Experimental Parasitology 31 (1972) 29-38. [9] T. Smith, E.W. Smillie, A note on coccidia in sparrows This study supported by Garmsar University. The

and their assumed relation to Blackhead in turkys, Exp. authors thank Miss Sh.F. Gorji and everyone who

Med. 25 (3) (1917) 415-420.

helped them in the present study. [10] J. Chirico, H. Eriksson, O. Fossum, D. Jansson, The poultry red mite, Dermanyssus gallinae, a potentional

References vector of Erysipelothrix rhusiopathiae causing

erysipelas in hens, Medical and Vetrinary Entomology [1] S. Cramp, C.M. Perrins, Handbook of the birds of Europe,

17 (2003) 232-235.

the Middle East and North Africa: The birds of the [11] F.B. Shirinov, A.I. Ibragimova, Z.G. Misirov, Spread of Western Palaearctic, Vol. 8, Oxford University Press,

fowl pox virus by the mite Dermanyssus gallinae, Oxford, 1994, p. 899.

Vetrinarya (Moscow), Abst. Vet. Bull. 4 (1972) 48-49.

Journal of Life Sciences 6 (2012) 786-793

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech Republic in the Years 1999-2010 and Race Spectrum of This Population

Lubomir Vechet Crop Research Institute, Division of Plant Genetics, Breeding and Product Quality, Drnovska 507, Prague-Ruzyne, Czech Republic

Received: September 19, 2011 / Accepted: November 07, 2011 / Published: July 30, 2012.

Abstract: Powdery mildew (Blumeria graminis f.sp. tritici) is one of the most important leaf diseases of wheat on the worldwide. Evolutionary potential of a pathogen population is reflected in its population genetic structure. Severity of powdery mildew in twelve years of small-plot experiments was in individual years different. Incidence of powdery mildew was followed in 1999-2010 and powdery mildew development was studied in 2003 and 2004 in the Crop Research Institute (CRI). These experiments were made on the cultivar Kanzler, susceptible cultivar to powdery mildew. Disease severity was made as a percentage of typical symptoms of the disease. Structure of powdery mildew population was probed in 2009 and 2010 in CRI and in the locality Humpolec. The highest was in 2004 and the lowest in 2003. Development of the disease in two extreme years depended on progress of weather factors mainly to average daily temperature and photosynthetic active radiation. In the year 2009 was higher disease severity in CRI than in the CRI station in Humpolec but in 2010 vice versa higher disease severity was in Humpolec. Observation showed that in small distances of two localities can be differences in composition of powdery mildew population. Higher virulence in CRI was to the gene of specific resistance Pm1 whereas in Humpolec to the gene Pm17. In the year 2010 in CRI was the highest virulence to the gene Pm1 whereas in Humpolec the highest virulence was to the genes Pm3f and Pm4a. Differences of powdery mildew were in 12 years of observation were very distinct and showed difference in structure of powdery mildew population in near localities.

Key words: Powdery mildew, incidence, development, virulence, race spectrum.

1. Introduction  obligate biotrophic powdery mildew fungus Blumeria graminis f.sp. tritici have been genetically

Powdery mildew caused by the fungus Blumeria characterized [2]. Among them, the Pm3 gene was one graminis (DC) E.O. Speer f.sp. tritici Em. Marchal of the first described loci [3], and since then a total of (syn. Erysiphe graminis DC f.sp. tritici Marchal), is

10 different resistance specificities (Pm3a to Pm3j) one of the most important foliar diseases of wheat have been assigned to the Pm3 locus on the short arm worldwide [1]. It is an obligate parasite, growing only of wheat chromosome 1A [3]. We hypothesize that the on living tissue. Powdery mildew begins rapid growth evolutionary potential of a pathogen population is on the lower leaves and sheaths when plants begin to reflected in its population genetic structure [4]. joint. Pathogen populations with a high evolutionary In wheat, 30 Pm genes conferring resistance to the potential are more likely to overcome genetic

Corresponding author: Lubomir Vechet, Ph.D., research resistance than pathogen populations with a low fields: induced resistance of wheat to powdery mildew, septoria

evolutionary potential.

tritici blotch on wheat, powdery mildew. E-mail: vechet@vurv.cz.

Incidence of powdery mildew was followed for

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech

Republic in the Years 1999-2010 and Race Spectrum of This Population

twelve years (1999-2010) and powdery mildew 747 Pm6; Disponent Pm8; Amigo Pm17; Maris development was studied in the years 2003 and 2004.

Huntsman Pm2, 6; Appolo Pm2, 4b, 8; Maris Dove Structure of the powdery mildew population was

Pm2 + Mld; W 7984 Pm18). From individual wheat probed in two different localities in 2009 and 2010.

cultivars with specific genes of resistance were prepared leaf segments and placed on water

2. Materials and Methods

benzomidazole agar brimming into plastic boxes.

2.1 Plant Materials, Cultivars Boxes were put under sedimentation tower. Leaf segments were inoculated by relevant isolate of

Incidence and development of powdery mildew was powdery mildew. After inoculation plastic boxes followed in Crop Research Institute (CRI Prague) were placed into the climabox. Disease severity was small-plot experiments in the period from the second evaluated after 12 days as a percentage segment area half of May to the end of July on the cultivar Kanzler,

covered by powdery mildew.

susceptible standard to this disease. Kanzler is generally susceptible to powdery mildew, and

2.4 Statistical Evaluation

powdery mildew isolates are regularly propagated on Relationship among disease severity and individual line Kanzler [5]. weather factors (average daily temperature, relative

2.2 Evaluation of Powdery Mildew Severity humidity, precipitation, speed of wind, global radiation, photosynthetic active radiation,

Incidence of powdery mildew was evaluated as dehumidification of a leaf in minutes) obtained from disease severity and development of the disease by meteorological station in the years 2003 and 2004 measuring disease severity in several time intervals. were expressed as correlative coefficient—r by means Disease severity was expressed as cumulative of statistical programme Unistat 5.1. The dates were percentage of leaf area disease—CPLAD on the plant gathered in the tree terms of evaluation for the year where percentage of a leaf covered by powdery 2003 and in five terms for the year 2004. mildew for all lives leaves are counted up.

3. Results

2.3 Evaluation of Powdery Mildew Virulence

3.1 Disease Severity in 1999-2010

Development of powdery mildew on individual leaves of wheat plants was ascertained in the year

Disease severity of powdery mildew considerably 2004. Variability in virulence to the specific genes of

fluctuated in individual years of experiments (Fig. 1). wheat to powdery mildew was ascertained in two

The highest disease severity was in the year 2004 years (2009 and 2010) in the CRI (Crop Research

much more than in other years. On the second place Institute in Prague) and in the experimental station of

was powdery mildew severity in the year 2001 which the CRI Humpolec distant from CRI about one

however was much lower than in 2004. The lowest hundred kilometres. Obtained isolates were disease severity for twelve years of experiments was cultivated on the cultivar Kanzler under standard

in the year 2003. Very low disease severity was in the conditions (21 °C, 16 hour of light) in the climabox

years 2007, 2000, 2005, and 2008 also. Disease where wheat plants were tested on virulence to

severity in the rest years was among the years 2001 individual genes of resistance (Axminster Pm1; Ulka

and 2003. Nevertheless, all values of disease severities Pm2 ; Normandi Pm1, 2, 9; Asosan Pm3a; Transfed

were substantially lower than the value in the year Pm7 ; Chul Pm3b; Sonora Pm3c; Michigan Amber

2004. Disease severity in the years 2005 and 2008 was Pm 3f; Khapli Pm4a; Ronos Pm4b; Rektor Pm5; NK

very similar.

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech Republic in the Years 1999-2010 and Race Spectrum of This Population

3.2 Influence of Weather Factors radiation (PAR). Positive, significant, influence had Influence of individual weather factors in the years

dehumidification (DLM) of a leaf in minutes on the with maximum disease severity (2004) and minimum

disease severity. Disease severity in the year 2004 was disease severity (2003) on development of powdery

the highest. Development of the disease in this year mildew is expressed in Table 1. In the year 2003

was very speedy and it was influenced by weather development of the disease was the lowest for 12

factors. Because in this year was development of the years. In the period from 6th June till 13th June was

disease very fast it was followed influence of weather none growth of the disease. Development of the

factors about fifteen days formerly than in the year disease was strongly negatively, significantly

2003. Negative influence had precipitation (P). influenced by high average daily temperatures (ADT),

Positive and significant influence on development of global radiation (GR) and photosynthetic active

the disease had photosynthetic active radiation (PAR).

Fig. 1 Powdery mildew severity on the cultivar Kanzler Individual years of twelve years of small-plot experiments expressed as maximal values. (Crop Research Institute)

Table 1 Comparison of individual weather factors influence (ADT: average daily temperature °C; RH: relative humidity; P: precipitation; SW: speed of wind; GR: global radiation; PAR: photosynthetic active radiation; DLM: dehumidification of a leaf in minutes on disease severity (DS). Weather factors are counted for seven days before the date of evaluation; r: correlation coefficient.)

Date DS ADT RH P SW GR PAR DLM 2003

-0.6841 0.2585 0.0000 0.9006 -0.9762 -0.9551 0.7899 2004 21 May

0.5840 -0.2915 -0.6054 -0.0338

-0.0961 0.6975 -0.0805

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech

Republic in the Years 1999-2010 and Race Spectrum of This Population

But in the year 2004 with the highest disease severity end of June disease severity rapidly increased again. was positive medium correlation with average daily

temperature and negative with precipitation. 3.4 Severity and Race Spectrum of Powdery Mildew

Population in the Year 2009

3.3 Development of Powdery Mildew in the Year 2004 Seventeen wheat cultivars with specific genes of Development (Fig. 3) of the disease in late of May

resistance to powdery mildew indicated variability in 2004 to beginning of June wa s gradual but thereafter race spectrum inside population of this pathogen in

development of the disease picked up speed. Later the Crop Research Institute in 2009 (Fig. 4). Much from the beginning of June to the half of this month

more high disease severity in comparison with the the disease severity rapidly increased, subsequently

cultivar Kanzler, susceptible standard to powdery increasing of powdery mildew decelerated and to the

mildew, was on the cultivars Michigan Amber with

Fig. 3 Development of powdery mildew expressed as severity in five evaluations in the year 2004.

Fig. 4 Structure of race spectra powdery mildew population in the year 2009. (Crop Research Institute)

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech Republic in the Years 1999-2010 and Race Spectrum of This Population

specific gene of resistance Pm3f and Khapli with Crop Research Institute than in Humpolec particularly Pm4a . Lower values, than Kanzler, had the cultivars

to the gene Pm3f whereas in Humpolec to the gene Transfed (Pm7) and Ulka (Pm2) and the rest of

Pm1 . The lowest virulence in the Crop Research cultivars had more low disease severity. The lowest

Institute was to the gene Pm17 whereas in Humpolec disease severity was on the cultivar Amigo with the

the lowest virulence was to the genes Pm18, Pm3b specific gene of resistance Pm17. Very low these

and Pm5.

values had the cultivars Rector (Pm5), Maris Dove (Pm2 + Mld), Chul (Pm3b), Asosan (Pm3a), and 3.5 Severity and Race Spectrum of Powdery Mildew

Population in the Year 2010

Normandi (Pm 1,2,9) also. In the population of powdery mildew in Humpolec

Disease severity in the year 2010 in the Crop (Fig. 5) had the cultivar Axminster (Pm1) higher

Research Institute (Fig. 6) on the cultivar Axminster disease severity than the cultivar Kanzler, susceptible

(Pm1) was much higher than on Kanzler. A little standard. Nearly the same value of disease severity as

higher disease severity then Kanzler had the cultivar the cultivar Kanzler had the cultivar Khapli (Pm4a). A

Michigan Amber (Pm3f). Disease severity on others little less than Kanzler had disease severity the

cultivars with specific genes of resistance was lower cultivars Sonora (Pm3c) and Ulka (Pm2). Without any

in different values. The lowest disease severity had the symptoms of powdery mildew was the cultivar Amigo

cultivars Rektor (Pm5), Romos (Pm4b), Maris (Pm17), its value of disease severity was zero. Very

Huntsman (Pm2, 6), and Apollo (Pm2, 4b, 8). low disease severity had the cultivars Ralle (Pm3d),

In Humpolec (Fig. 7) the cultivars Michigan Amber Rektor (Pm5), and Maris Dove (Pm2 + Mld). Disease

(Pm3f) and Khapli (Pm4a) had higher disease severity severity of others cultivars fluctuated betwen these the

than the cultivar Kanzler. Lower disease severity than lowest and the highest values.

the cultivar Kanzler had cultivars Transfed (Pm7), In the year 2009 was higher disease severity in the

Ulka (Pm2) and Disponent (Pm8).

Fig. 5 Structure of race spectra powdery mildew population in the year 2009. (Crop Research Institute, The Experimental Station Humpolec)

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech

Republic in the Years 1999-2010 and Race Spectrum of This Population

Fig. 6 Structure of race spectra powdery mildew population in the year 2010. (Crop Research Institute)

Fig. 7 Structure of race spectra powdery mildew population in the year 2010. (Crop Research Institute, The Experimental Station Humpolec)

The others cultivars varried between high and the it was to the genes Pm17, Pm3b and Pm5. However lowest values of disease severity. The lowest disease

the lowest virulence was higher in Humpolec than in severity had cultivars Rektor (Pm5), Ronos (Pm4b),

the Crop Research Institute.

Maris Huntsman (Pm2, 6) and Apollo (Pm2, 4b, 8).

4. Discussions

In the year 2010 was the highest virulence in the Crop Research Institute against gene Pm1 and Pm18 4.1 Influence of Weather Factors on Disease Severity

whereas in Humpolec to the gene Pm3f and Pm4a. Race-specific resistance has been extensively used The lowest virulence in the Crop Research Institute

in wheat breeding programs, even though it is was to the genes Pm5, Pm4b and Pm2,6. In Humpolec

ephemeral. To determine which virulence is

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech Republic in the Years 1999-2010 and Race Spectrum of This Population

presented so that breeding strategies can be planned Pm4a , also. On the second place was virulence to to use most effective genes [6] regional surveys are

Pm4 in both areas. Much more virulence to Pm3, needed. Development of powdery mildew severity

Pm3f , and Pm8 was in Prague region than in for twelve year of monitoring very different from

Humpolec. Our observation showed that on small very high in the year 2004 right to minimum disease

distances can be differences in composition of severity in the year 2003. Strong dependence disease

powdery mildew population. Huszar et al. [8] found severity was on photosynthetic active radiation.

out in 1996-1998, according virulence analysis of Positive dependence was in the year 2004 with the

wheat powdery mildew the genes Pm4a, Pm1 + Pm2 highest disease severity and negative dependence

+ Pm9 were resistant to all isolates, Pm2 + Pm6 were was in the year 2003 with the lowest severity.

effective to all isolates only from western Slovakia. Similarly dependence was between disease severity

After a few years in Bohemia was in powdery and average daily temperature. In the year with

mildew population high diversity to the genes P1 and minimum of powdery mildew (2003) was disease

Pm4a versus finding in Slovakia. Effectiveness of severity influenced by speed of wind to conidia

Pm3d and Mld + mla was similar throughout transfer from lager distances and by dehumidification

Slovakia. Genes Pm2, Pm3a, Pm3b, Pm3c, Pm3f, of a leaf in minutes because higher dehumidification

Pm5 , Pm7 and Pm8 were completely ineffective. of a leaf can to a certain extend increase conidia

According to Costamilan [9] in Brazil gene Pm6 was germination and infection structures development.

highly effective (below 10% of susceptibility), Except negative high correlation between disease

whereas genes Pm3a and Pm8 were totally severity and photosynthetic active radiation in this

ineffective (susceptible to all isolates). Felsenstein year was the same dependence on photosynthetic

and Jaser [10] show that frequency of virulence to active radiation. In experiments dependence of

Pm1 is middle, to Pm3c offers average protection. disease severity on weather progress was determinate

Increase virulence toward Pm17, a widely used gene and there were demonstrated that from weather

from Amigo wheat. However in conditions of the factors mainly photosynthetic active radiation and

Czech Republic was virulence of powdery mildew average daily temperature have influence on population to the gene Pm17 very low. development of powder mildew severity.

5. Conclusions

4.2 Composition of Powdery Mildew Population Our monitoring powdery mildew in the Czech

Race-specific resistance and broad-spectrum Republic showed difference from structure of resistance (BSR) are two major types of disease

population in Slovakia and divergence in near resistance in plants. BSR refers to resistance against

localities. Differences in powdery mildew severity in two or more types of pathogen species or the

twelve years of monitoring in the same place were majority of races of the same pathogen species [7]. In

very distinctly.

the Czech Republic in both years (2009 and 2010)

Acknowledgments

was higher disease severity in Humpolec than in the Crop Research Institute. It is evident that in the year

The author would be like to thank E. Pribylova and 2009 in the region Prague predominated in P. Jirovec for technical help. The author gratefully population of powdery mildew virulence to specific

acknowledges the Research Plan of the VZ MZe genes of resistance Pm3f, Pm4a, whereas in the

(Project No. 0002700604) for providing financial locality Hunpolec the highest virulence to Pm1 and

support.

Incidence and Development of Powdery Mildew (Blumeria graminis f.sp. tritici) in the Czech

793

Republic in the Years 1999-2010 and Race Spectrum of This Population

References

locus, Euphytica 68 (1993) 223-229. [6] B.M. Cunfer, Powdery mildew, bread wheat improvement

[1] F.G.A. Bennett, Resistance to powdery mildew in wheat: and production, in: B.C. Curtis, S. Rajajaram, H. Gomez A review of its use in agriculture and breeding

(Eds.), FAO Plant Production and Protection Series, programmes, Plant Pathology 33 (1984) 279-300.

NO:30, Macpherson, Rome, 2002.

[2] S.L.K. Hsam, F.J. Zeller, Breeding for powdery mildew [7] Y. Kou, S. Wang, Broad-spectrum and durability: resistance in common wheat (Triticum aestivum L.), in:

Understanding of quantitative disease resistance, Current R.R. Bélanger, W.R. Bushnell, A.J. Dik, T.L. Carver (Eds.),

Opinion in Plant Biology 13 (2) (2010) 181-185. The Powdery Mildews: A Comprehensive Treatise, APS

[8] J. Huszar, P. Bartos, A. Hanzalova, Importance of wheat Press, St. Paul, Minnesota, 2002, pp. 219-238.

disease resistance for sustainable agriculture, Acta [3] L.W. Briggle, E.R. Sears, Linkage of resistance to

Fytotechnica at Zootechnica 4 (2001) 292-294. Erysiphe graminis f.sp. tritici (Pm3) and hairy glume (Hg)

[9] L.M. Costamilan, Variability of the wheat powdery on chromosome 1A of wheat, Crop Scientific 6 (1966)

mildew pathogen Blumeria graminis f.sp. tritici in the 559-562.

2003 crop season, Fitopatologia Brasileira 30 (4) (2005) [4] B.A. McDonald, C. Linde, Pathogen population genetics,

420-422.

evolutionary potential, and durable resistance, Annual [10] F.G. Felsenstein, B. Jaser, Effectiveness of qualitative Review of Phytopathology 40 (2002) 349-379.

powdery mildew resistance in wheat and barley and [5] F.J. Zeller, J. Lutz, U. Stephan, Chromosome location of

sensitivity of fungal cereal pathogens towards different genes for resistance to powdery mildew in common wheat

compounds, 2000, available online at: (Triticum aestivum L.), 1. Mlk and other alleles at the Pm3

http://www.epilogic.de/eng/ index.htm.

Journal of Life Sciences 6 (2012) 794-800

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens and Acacia auriculiformis in Godohou Village (Southern Benin)

1 2 Emile N. Houngbo 2 , Anne Floquet and Brice Sinsin 1. National Higher School of Agriculture (ENSTA-Ketou), University of Abomey-Calavi, 05 BP 774 Cotonou, Benin

2. Faculty of Agriculture, University of Abomey-Calavi, 05 BP 774 Cotonou, Benin

Received: January 27, 2012 / Accepted: April 06, 2012 / Published: July 30, 2012.

Abstract: The decomposition of the environmental degradation cost in Benin Republic revealed that the agricultural activities are responsible of the greatest amount of this cost (76.1%). This situation is strengthened by the fallow periods shortening (and even suppression) in the southern Benin in general, because of demographic pressure. The promotion of some improved fallow technologies (IFT) was then launched some decades ago. This study focused on two IFT of high agro-ecological value, Mucuna pruriens and Acacia auriculiformis, in order to determine the influence of the farmers’ wellbeing state on their farming systems in general, and on the adoption level of these two technologies in Godohou village (southern Benin). The data analysis revealed that: There was no significant difference between the farmers’ amount knowing the agronomic and ecological value of Mucuna pruriens and Acacia auriculiformis from a level of prosperity to another; Three wealth levels were distinguished: The poor class (Ayatonon class), the less poor class (Metchivo-Houedeka class) and the non poor class (Hotonon class) which represented respectively 35.4%, 55.9% and 8.7%;The adoption rate of Mucuna and Acacia fallow was globally low in Godohou village (25.7%), but this adoption rate was lesser in the poorest class than in all other classes. This trend was confirmed if we integrate a global agro-ecological analysis of the farming systems practiced by the farmers.

Key words: Godohou, poverty, environment, mucuna, acacia, adoption, level of prosperity.

1. Introduction  of the greatest amount of this cost (76.1%). This situation of environmental degradation by agriculture

In spite of high land pressure, the farmers of is strengthened by the fallow periods shortening, and southern Benin in general, and Godohou village in even suppression. That’s why the promotion of some particular, continue practicing mining agriculture. improved fallow technologies (IFT) was launched This kind of agriculture induces environmental some decades ago. But, unfortunately, the adoption degradation and threats the farming systems rate of that IFT still low. Given the view of Leach & sustainability. This reality does not respect the thesis Mearns [3] who established a theoretical causal of Boserup [1] who foresees agricultural relationship between poverty and environmental intensification in such a situation of land pressure. degradation, one could wonder if the rural poor According to Tovo [2], the decomposition of the cost farmers degrade the cultivated areas environment by of environmental degradation in Benin Republic the IFT rejection than the others. The case was applied revealed that the agricultural activities are responsible

to the special situation in Benin.

The main aim was to determine the influence of the Corresponding author: Emile N. Houngbo, Ph.D.,

agricultural economist, senior lecturer, researcher, research farmers’ wellbeing state on their farming systems in

field: socio-economics. E-mail: enomh2@yahoo.fr.

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens

and Acacia auriculiformis in Godohou Village (Southern Benin)

general, and on the adoption level of Mucuna and

2.3 Data Collection Method

Acacia in particular, in Godohou village (southern According to Aho et al. [6], the focus group method

Benin). It was supposed that the more the farmers are is an interesting qualitative method of social research poor, the lesser they adopt the IFT and then practice that can be very useful for analysis of needs to guide mining agriculture. development policies. Focus groups are composed of

2. Materials and Methods

eight to twelve persons chosen with regard to certain predetermined criteria based on the analysis needed.

2.1 Study Area The participants in focus group are invited to discuss The study was implemented in Godohou village, a

the topic of the study. The interviewer is a facilitator village located in the Adja plateau, the main

who guides the discussions on the topic using a occupied part of the Couffo Department (southern

semi-structured interview.

Benin). Adja plateau is recognized as the area of the In this study, the focus group method was used for highest land pressure in southern Benin, and then the

participatory analysis with two sub-groups of ten zone where the use of the IFT is necessary to main

persons; one sub-group of women and one sub-group soils’ productivity. The environmental degradation

of men. After the qualitative poverty analysis by the due to agriculture is very remarkable in the Adja

households’ classification through the Participatory plateau. Adja Plateau is also one of the major

Wealth Ranking Method of Grandin [7], a sample of agricultural production areas of Benin, characterized

70 households was selected and interviewed as far as by the concentration of more than 90% of the

their farming systems were concerned. These population in agriculture. The population density is

interviews were done through questionnaire from 350 inhabitants per km 2 [4] and 95% of the available

January 24 to February 2, 2005.

land is used each year for agriculture [5]. Godohou Mucuna pruriens and Acacia auriculiformis have village is located precisely in the District of

been targeted particularly in this research as improved Klouekanme, Department of Couffo. The local

fallow technologies (IFT) because their importance are language is Adja. Agriculture, and accessorily animal

well recognized both in agronomy and in terms of their husbandry, food processing and marketing of ability to reverse the current trend of the cultivated agricultural products are the most important areas degradation in southern Benin. These economic activities in Godohou village.

technologies are supposed to be very useful (technically) for the environment conservation under a

2.2 Approach of Poverty Measure hard land pressure zone, such as the Adja plateau where In this study, poverty was addressed from the

agriculture is still practiced by a very large proportion “non-welfarist” approach, with special attention to

of the population.

social perceptions which we believe are indicative not

2.4 Data Analysis

only of the monetary dimension, but also of the non-monetary dimension that is further fundamental

Data and information were analyzed using simple in poverty analysis. Another advantage of this

statistical parameters such as mean, proportion, approach is the appreciation of poverty in relation to

standard deviation and the calculation of confidence local realities, with “emic” analysis. Thus, the study

intervals. Descriptive analysis was then heavily used. approach was essentially qualitative. It highlights the

The comparison test of proportions was not found relevant dimensions of poverty as far as the farmers of

relevant in view of the structure of the three prosperity Godohou village are concerned.

levels identified.

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens

and Acacia auriculiformis in Godohou Village (Southern Benin)

3. Results

plastered with cement.

The Metchivo-Huedeka class represented 55.9% of

3.1 Levels of Prosperity of the Village and Their the farmers in Godohou village. 42.5% of the members Characterization of this class practice only agriculture and animal

Following the household heads wealth ranking breeding, and 57.5% of the farmers associate to these realized, it appeared that Godohou village people

two activities food processing and the marketing of appreciate the wealthy person (and thus the poor person)

agricultural products. The farmers of through the charges supported (number of children and

Metchivo-Huedeka class do not have money to build a women), the physical assets (motorcycles, buildings,

house with brick and plaster it with cement, but they machines, ...), the autonomy in satisfying the basic

can cover it of corrugated iron.

needs and the capacity to fulfill social obligations The Ayatonon class represented 35.4% of the (receiving guests, capacity to help others, ...). Based on

farmers in Godohou village. 75% of them practice these criteria, all the 229 households of the village were

strictly agriculture and animal breeding, and 25% of participatory classified into three welfare classes:

them associate food processing and the marketing of (i) Class 1: The “Hotonon” class;

agricultural products. The farmers of Ayatonon class (ii) Class 2: The “Metchivo-Huedeka” class;

have no money to build a good house. In the best case, (iii) Class 3: The “Ayatonon” or “Abonon” class.

they can build a house, never plastered with cement, Table 1 shows the rate of the different classes in the

but covered of the straw of cough grass. village. The Hotonon class represented 8.7% of the

On comparing the three classes of welfare, it farmers in Godohou village. All of the members of this

appears that the more the farmers practice other class practice several activities. Apart from agriculture

activities after agriculture and animal breeding, the and animal breeding, they practice food processing and

more they are not exposed to poverty. The quality of the marketing of agricultural products that bring them

the house owned is also an interesting indicator of a much money. The farmers of Hotonon class have

household welfare level in Godohou village. Figs. 1-3 money to build a beautiful house; a house covered of

show an example of house of households belonging to corrugated iron, and built with brick or not and/or

each of the three classes.

Table 1 Welfare classes in Godohou village.

Weight in the village Welfare classes

Characteristics

Number Percentage (%)

- The class of the non poor farmers - Farmers supporting a charge of 15 children and more

Hotonon class

- Farmers having more than 2 ha of land, a beautiful

house, a motorcycle and even a car, and sending their children to school - The medium welfare class - Farmers supporting less than 15 children

Metchivo-Huedeka class - Farmers having 1-2 ha of land, a house of mean 128 55.9 quality, a motorcycle, and sending their children to school - The class of the poor farmers - Farmers married or not

Ayatonon class

- Farmers having less than 1 ha of land, at most a

shattered house, a bicycle or not, and cannot send their children to school

Total 229 100

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens

and Acacia auriculiformis in Godohou Village (Southern Benin)

3.2 IFT Adoption, Farming Systems and Their Environmental Impacts

3.2.1 Adoption of Mucuna and Acacia All the respondents and all of the farmers are aware of Mucuna and Acacia, their beneficial effects in terms of agriculture and economic development. There is no difference between the proportions of farmers knowing the two IFT (Figs. 4 and 5). But,

very few farmers have adopted at least one of them.

Fig. 4 A fallow of Mucuna pruriens in Godohou village.

Fig. 1 House of a Hotonon. Fig. 5 Acacia based agroforestry in Godohou village.

More than 60% of the respondents have integrated neither Mucuna nor Acacia in their farming systems. However, there is a difference between the adoption rates of these technologies if we move from a welfare class to another. Table 2 shows the rates of adoption per welfare class:

This table indicates that the farmers belonging to the Hotonon class use more Mucuna and/or Acacia in their farming systems than all other classes. Statistical

analysis was not relevant for the adoption rates

Fig. 2 House of a Metchivo-Huedeka.

statistical comparison, because the products of the sample size by the adoption rates per welfare class are

not higher than 5 (np < 5), except for the Metchivo- Huedeka class. We therefore made the decisions only

Table 2 Adoption rates of Mucuna and/or Acacia in Godohou village.

Welfare class

Sample

Adoption rate

structure

Hotonon class

Metchivo-Huedeka class 40

Ayatonon class

All 70 25.7

Fig. 3 House of an Ayatonon.

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens

and Acacia auriculiformis in Godohou Village (Southern Benin)

with the data of Table 2. We concluded that the transfer of organic matter to the farm (animal waste, adoption rate of the IFTs (Mucuna and Acacia

compost, crop residue, ...), the use of Mucuna pruriens especially) increased from the lowest level of and Acacia auriculiformis in the farming systems. prosperity to the highest level of prosperity; from the

The practice of active transfer of organic matter to Ayatonon class to the Hotonon class.

the farm is not important. There is no developed animal

3.2.2 The Farmers’ Attitude of Soil Management breeding which could support that practice. Moreover, All of the respondents practiced food crops

the transfer of organic matter is operated only on a few polycropping. The practice of intercropping is farms closed to the habitations. The lack of organic widespread in all of the levels of prosperity. The

matter in the soil and the widespread practice of the use polycropping is practiced with maize, cassava, of fertilizers in the village could justify the tomatoes, peppers, yams, beans, pigeon peas, sweet

acidification of the soils exposed to erosion as stated by potatoes, peanuts. The crop succession, besides the

GRADID [8].

permanent presence of natural palm trees in all of farms, Even though the overall farming systems are is generally characterized mainly by a degrading in the village, with a global weak level of maize-cassava-groundnut in the first rainy season and a

agroforestry practice, there is a relationship between bean-peanut association in the second rainy season.

the welfare classes and agroforestry adoption. The The practice of fallow is approximately nonexistent.

table 3 shows the farming systems appreciation per Only 14.3% of the farmers practiced the fallow

welfare class as far as their capability of the technique, and they were found only in the classes of

environmental degradation is concerned. Hotonon and Metchivo-Huedeka. Permanent cropping

It is clear that apart from the differential was practiced by 85.7% of the respondents fully

integration of the two improved fallow technologies concentrated in the Ayatonon class. It follows that the

by the different levels of prosperity and the trend of fallow practice decreased from the Hotonon

differential practice of natural fallow, the other farm class (16.7%) to the Metchivo-Huedeka class (12.5%).

management behaviors were approximately the same The members of the Ayatonon class did not practice

for all the farmers. It appears that the farmers’ this agricultural technique.

farming systems are less and less degrading from the

3.2.3 Impact of Agricultural Practices on the Ayatonon class to the Hotonon class. Although the Environment

farming systems practiced in Godohou village are To assess the impact of the agricultural practices on

generally degrading, the highest rate of degrading the cultivated environment in Godohou village, we

farming systems was located in the Ayatonon class, analyzed more closely the practice of fallow, the

the poorest class.

Table 3 Synthesis of the farming systems analysis.

Deduction of the effects Welfare classes

Adoption of Mucuna

Use of organic

1 Fallow practice 2 and/or Acacia Crop rotation matter on the environment 3 Hotonon class

Good 4 Less degrading system Metchivo-Huedeka class

Less weak

Weak

Weak

Good Degrading system Ayatonon class

Weak

Very weak Weak

Very degrading system Whole

Very weak

Non-existent Weak

Good

Weak

Very weak

Weak

Good

Degrading system

1 Adoption: Less weak > Weak > Very weak; 2 Practice: Weak > Very weak > Non-existent; 3 Effects: Very degrading > Degrading > Less degrading; 4 There is the presence of cereal, root/tuber and leguminous such as bean and pigeon pea.

Poverty and Agroforestry Adoption: The Cases of Mucuna pruriens

and Acacia auriculiformis in Godohou Village (Southern Benin)

4. Discussion

technological innovations. Moreover, Floquet and Mongbo [15] remarked in southern Benin that despite

The findings of this study are in line with that of Reij their awareness of the soil degradation, the farmers and Waters-Bayer [9] who, analyzing the were adopting very weakly the improved fallow characteristics of innovative farmers identified through

technologies.

the programs ISWC (Indigenous soil and Water conservation) and PFI (Promoting Farmer Innovation

5. Conclusions

in Rainfed Agriculture) in East Africa, revealed that Mining agriculture is developed in Godohou most of them were richer than average. But, they also village. Though farmers know the positive noticed that Verhoeven and van der Kroon [10] stated agro-ecological effects of Mucuna pruriens and that there is no direct relationship between innovation Acacia auriculiformis , their adoption rate is globally capacity and farmers economic status. weak, in spite of the hard land pressure in the zone. Normally, agroforestry practice is necessary in The farmers of the poorest class adopt less that Godohou village on taking into account the technologies than all other farmers. Poverty appears classification of Ruthenberg [11] who defined the

to be a determinant obstacle to agroforestry adoption symbol R representing the proportion of cultivated

in the village. A fight against poverty is then area in relation to the area owned by the farmer.

necessary to improve agroforestry adoption and Estimating R = 77, say R > 66, in Godohou village, we

sustainable agriculture practice in Godohou village, concluded the permanent cultivation systems in the

and in southern Benin.

village. Unfortunately, the Cajanus cajan growing which is widespread in the village could not improve

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190.

Journal of Life Sciences 6 (2012) 801-816

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

1 2 3 Adrien Njepang Djomo 4 , Gode Gravenhorst , Anthony Kimaro and Marney Isaac 1. 43 Trillium Crt, Kingston, ON, K7K 6W9, Canada

2. Faculty of Forest Sciences and Forest Ecology, Centre for Tropical and Subtropical, Agriculture and Forestry (CeTSAF), Georg-August-Universität Goettingen, Buesgenweg 1, D-37077 Göttingen, Germany 3. World Agroforestry Centre (ICRAF), ICRAF Tanzania Programme, P. O. Box 6226, Dar-es-Salaam, Tanzania 4. Department of Physical and Environmental Sciences, University of Toronto, Toronto, Canada

Received: December 15, 2011 / Accepted: March 12, 2012 / Published: July 30, 2012.

Abstract: Forest degradation and biomass damage resulting from logging is currently difficult to evaluate with satellite images, but contributes substantially to carbon emissions in the tropics. To address this situation, we modelled how changes in the minimum felling diameter affect stem density, basal area and the related carbon biomass at the end of the felling cycle (30 years) in a semi-deciduous natural forest in Cameroon. With new MFDs estimates, at 7% logging damage rate, we found that the stem density of initially harvestable trees reduces from 12.3 (50.4 MgC·ha -1 ) to 6.7 (32.5 MgC·ha -1 ) trees per ha and the number of initial residual trees increases from 80 (18.9 MgC·ha -1 ) to 85.7 (36.8 MgC·ha -1 ) trees per ha. This corresponds to an avoided damage estimated at

17.9 MgC·ha -1 . We also found that increasing mortality and damage intensity also increases the damage on carbon biomass estimated to be 8.9 MgC·ha -1 at 10% or to be 17.4 MgC·ha -1 at 15% logging damage. Overall, our study shows that proper determination of MFD of logged species taking into consideration their capacity of reconstitution and the Reduced Impact Logging can avoid the loss of up to 35 MgC·ha -1 .

Key words: Carbon estimations, felling cycle, logging damage, minimum felling diameter (MFD), moist tropical forest, REDD, species reconstitution.

1. Introduction  trees in temperate and boreal forests [1, 2]. Therefore, an equivalent rate of deforestation or forest

About 30% of the land surface is covered by forests degradation will result generally in larger amounts of which store up to 80% of global aboveground carbon removed from the tropical forests than in the terrestrial carbon (C) and up to 40% of global

other forest types [3].

belowground terrestrial organic C [1]. Tropical forests Generally forest degradation refers to the loss of cover less than half of the entire world forest area and forest biomass without noticeable changes in the store as much carbon in their below and aboveground forest cover [4]. Selective logging in tropical forests biomass as in all the other forests outside the tropics. can result in forest degradation [5-7]. This logging This is due to the fact that in tropical forests trees hold, related forest degradation directly alters species on average, about 50% more carbon per hectare than composition, forest biomass, and micro-climate

[8-11]. Another consequence of this human activity Corresponding author: Adrien Njepang Djomo, Ph.D.,

research assistant, research fields: forest management and is carbon dioxide (CO 2 ) emission due to burning of environment. E-mail: adriendjomo@yahoo.com.

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

slash after logging or decomposition of unburned analyze the impact of harvest on stand dynamics and slash and/or organic matter. Such emissions are

the reconstitution or recovery of the forest at the end critical since part of the biomass which ends up

of the felling cycle (30 years); (ii) analyze the yield of stored in long-lasting structures (e.g. houses, wood

commercial species and determine the minimum materials) represents only a fraction of the biomass

felling diameter (MFD); (iii) calculate and compare initially held in a forest [12]. However, reducing CO 2 growth projection of the harvestable trees and the

emissions from deforestation and forest degradation residual stand at different logging damage intensities; in the tropics has been shown as one of the cost

and (iv) discuss a methodology for estimating carbon

emissions (sink or source) from forest degradation in atmosphere [13, 14]. Consequently, the concept of

effective options to reduce CO 2 emissions into the

the tropics.

Reducing Emissions from Deforestation and forest

2. Materials and Methods

Degradation (REDD) has been developed over the last few years to address this global problem. Under

2.1 Study Site

REDD, a wide range of policies and measures (PAM) and mitigation mechanisms, allows countries to

The data for this study were collected in Cameroon, claim financial compensation through international

in the eastern province, within the district of Boumba carbon markets for avoided deforestation associated

and Ngoko, sub-district of Yokadouma. Yokadouma with implementing forest conservation programs [15].

is a city situated 650 km east of the capital (Yaoundé), Today, through satellite images and field surveys, it

in the heart of the tropical rain forest (Fig. 1). The is possible to estimate deforestation. One of the

research area is located between latitude 3°17 ′33″ N challenges in the REDD mechanism is how to

and 3°25 ′9″ N and longitude 14°44′22″ E and determine credible policy and determine acceptable

14°53 ′33″ E. The geological formations in this area techniques for linking measurements of reduced

are of Precambrian origin and comprise sandstones damage from forest degradation to emission of

rich in quartz, schists and limestones. Ferralitic acid greenhouse gases [16]. Efforts for measuring forest

soils of red and red-brown colour with high clay degradation to date have focus on the detection of

content and a surface horizon characterized by a low selective logging and fire damage [7]. Since these

concentration of organic matter are dominant in this methods are still in the research and development

region. In some areas, the ferralitic soils are phase, it is questionable whether a small forest

interwoven with hydromorphic soils of brown-black clearance representing a harvest of 3 trees of 100 cm

colour, which are rich in organic matters. The dbh during which 150 neighbouring small trees are

topography is in general gently undulating with damaged, can be detected by satellite imagery.

valleys and ridges among flat basins, ranging in

This study develops techniques for linking elevation from 400 m to 700 m above sea level [17]. measurements of reduced damage from forest The climate is equatorial Guinean with four degradation to CO 2 emissions, which is one of the

distinguished seasons constituted of two rainy seasons main challenge for successful REDD implementation

and two dry seasons. From mid-March to mid-June in the tropics [16]. The objective of the present study

there is normally a shorter and less intensive rainy is to analyze the stand development of a typical forest

season; the period from mid-June to mid-August area in Cameroon and to discuss a methodology for

usually corresponds to a short dry season; in the estimating the potential carbon emissions reduction

period from mid-August to mid-November there is a from forest degradation under sustainable forest

long and heavy rainy season; during the period from management. The specific objectives are to: (i)

mid-November to mid-March there is usually a long

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

Fig. 1 Forest research area.

dry season. The mean air temperature fluctuates

2.3 Classification System

between 23 °C and 25 °C with very low variation For this study, four species groups were used for

during the year. The annual rainfall varies between the classification of the measured trees. Group 1

1600 mm and 2000 mm [18]. The river system is very stands for mature merchantable trees with diameter dense and belongs to the Congo Basin.

above the minimum felling diameter (MFD). Group 2

2.2 Sampling Design contains immature merchantable trees with diameter under the MFD. Group 3 stands for all non

The research forest is a semi-deciduous natural commercial trees with a prescribed MFD by forest forest of 22,000 ha comprising different ecological

administration. Group 4 contains all non commercial features (one logged, two logged, unlogged, low land

trees without a prescribed MFD. The future stand and moderate hills). For this research, 30 sample plots

projection analysis was made on commercial species of 20 m × 50 m were laid out uniformly in the

constituted by Group 1 and Group 2. The yield and different ecological features within the study area. In

determination of sustainable MFD were made for nine each plot, a subplot, of 20 m × 5 m was used for

commercial species contained in Group 1. Trees were seedling assessment. All trees with diameter at breast

grouped in diameter classes. The width of a diameter height (dbh) exceeding 20 cm were recorded in the

class used was 10 cm. For example, a diameter class plots. In the subplots, small trees with dbh ranging

35 contains all trees with dbh between 30 cm and 39.9 cm. between 10 cm and 19 cm were recorded. For each

2.4 Future Stand Projection

tree, the species name, the dbh (cm), the height (m), the stratum and the geographical coordinates x and y

The future stand projection was evaluated for 24 were assessed.

commercial species belonging to Group 1 and 2. The

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

number of survival N after t years in a forest stand hectare and also for the basal area per hectare of all initially constituted of No individuals was calculated

commercial trees in all diameter classes. To account using the formula [19]:

for the incoming natural regeneration of new trees into N

t = [N o (1– Δ)] × (1–a) the smallest diameter class, a constant value of 60 (1) where a is the mortality rate for each diameter class

trees ha -1 obtained from the analysis at initial year in (% year -1 ), N o the number of individuals at time 0, N t the smallest diameter class 15 cm at all time steps was

the number of individuals after t years, t the number

assumed.

of years and Δ the damage rate for each diameter

2.5 Determination of the Minimum Felling Diameter class (% of logging damage year ). Based on the

(MFD)

study [19] of de Madron et al. (1998a) in Dimako forest in Eastern Cameroon we considered a value of

The specifications of rules for harvesting and 1% of all trees for the mortality rate and 7% for

associated silviculture in a mixed tropical forest are damage rate due to forest exploitation in all diameter

complex and many variations are possible. The most classes. For estimation of the stand development, the

basic elements are generally felling cycle and a concept of a Diameter Class Projection Model

species-determined minimum felling breast height (DCPM) was used [20]. The ingrowths into class k

diameter [20]. The felling cycle is the period and outgrowths from the k th class are a result of the

between two successive harvests in the same forest combination of mean increment, mortality and

stand [23]. The optimum felling cycle is one that logging damage. Therefore formula (1) can be

ensures complete forest recovery and sufficient stem rewritten as follows:

recruitment. The MFD is the lower limit which will

retain enough trees to give a second harvest after the N k , t  1  N k , t 

 [ N k  1 , t ( 1   )]  (

felling cycle. It is also the legal diameter limit for

harvest. For the determination of the MFD, we used ( 1  a ) 

 [ N k , t ( 1   )]  ( 1  a )

w the values for different species suggested by the where N th

k-1,t is the number of trees in the k-1 class at forest administration in Cameroon (Table 1) as period t, i the mean annual growth (cm year -1 ), t the

starting point of the calculation of the percentage of projection period (year), w the width of the diameter

reconstitution. The percentage of reconstitution is class (cm), a the mortality rate for each diameter class

defined here as the rate of the original harvestable (% year -1 ) and Δ damage rate for each diameter class

tree population which will likely recover through (% of logging damage year -1 ). Many studies on tree

stem growth after harvest until the end of the felling growth of commercial species have been carried out in

cycle. For this calculation, we used the following Cameroon using the tree ring analysis method [21, 22],

formula:

successive measurements of commercial trees of the %Re = (i×t) × [N o (1– Δ)] × (1–a)/(w×N p ) (3)

stand in Cameroon and other countries in Central where %Re is the percentage of reconstitution of Africa [19, 20, 23-26]. For this research, the mean

individuals initially exploitable, N o the number of annual diameter growths used were obtained from

considered individuals below the MFD, N P the growth analysis conducted by these studies. The stand

number of trees initially exploitable, i the mean annual

projection was calculated for all commercial species growth (cm year ), t the projection period (year), w after each period of 5 years up to 30 years. We used

the width of the diameter class (cm), a the mortality

30 years as a limit because it corresponds to the -1 rate for each diameter class (% year ) and Δ the duration of felling cycle in Cameroon. The likely

damage rate for each diameter class (% of logging future stand was evaluated for the number of trees per

damage per year). The calculation was done for each

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

commercial species. The optimum MFD was obtained

2.6 Sensitivity Analysis

by repeating many times the calculation of the Mortality rate can be defined as the number of trees percentage of reconstitution, while changing the value

dying during a growth period due to internal or of the MFD. The optimum level was achieved when

external factors. The internal factors can be the lack of the new MFD was able to ensure a recovery of at least

nutrients, shading, pest and diseases, or age of trees. 80% of the original tree density and basal area at the

The external factors can be catastrophes such as end of the felling cycle (Fig. 2).

tropical storms or logging damages. Logging damage

Table 1 Reconstitution of each harvestable commercial species. The percentage of reconstitution (%R) of each species is calculated using the Administrative Minimum Felling Diameter (Adm. MFD) as starting point. Whenever the %R is < 100%, a heigher MFD is considered and a new percentage of reconstitution (%R) recalculated. The bold values represent the optimum MFD.

Scientific name Ø increment (cm/year) Adm. MFD %R New MFD %R with new MFD New MFD %R with new MFD Alstonia boonei

0.9 50 87 70 74 80 93 Ceiba pentandra

0.9 50 0 60 0 60 - Erythropleum ivorense

0.4 50 0 90 41 90 - Guarea cedrata

0.5 80 147 80 - 80 - Mansonia altissima

0.5 60 70 70 111 70 - Nesogordonia papaverifera

0.4 50 110 50 - 50 - Pycnanthus angolensis

0.7 60 108 60 - 60 - Terminalia superba

0.7 60 50 70 64 80 83 Triplochiton scleroxylon

Diameter class taken into consideration for reconstitution analysis: A, then A and B, then A and C

Number of stem A initially exploitable

Fig. 2 Analysis of reconstitution of initially exploitable trees by those below the minimum felling diameter.

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

is the damage or mortality of residual trees due to species, dbh ≥ MFD) we found 12 trees per hectare harvest of trees. In many studies and also in our case,

(3% of all measured trees) with a basal area of mortality refers to damage due to internal factors. Tree

7m 2 ·ha -1 (23% of total basal area) and a carbon pool mortality in undisturbed tropical rain forest falls on -1 of 75 MgC·ha (30% of total carbon stock). The low

average between 1% and 3% of stem number per year stem number in combination with a large basal area [27-29] in all diameter classes. These values are almost

indicates that trees in this category are large in similar to the typical values of 1-2% of stem number

diameter size. In Group 2 (all immature per year for the average tree mortality rates in primary

merchantable species, dbh < MFD) we found 80 trees

2 rain forests suggested by other authors [27, 30-34]. -1 per hectare with a basal area of only 4 m ·ha and a Many studies of tropical forests in Africa and other

carbon pool of 21 MgC·ha -1 (8% of total carbon tropical forests over the world have reported logging

stock) (Fig. 3). These values show that this group damage intensities exceeding 7% of stand trees per ha

generally contains small trees with dbh smaller than [35-37]. We considered three scenarios for stand

the MFD. Groups 3 and 4 which are constituted of projection of commercial species: low logging damage

species of non commercial value, unknown or less (7%), moderate logging damage (10%) and high

known commercial values represent in total 80% of logging damage (15%) of stand trees per ha associated

total stem number, 66% of total basal area and 62% with mortality rate of 1%, 3% and 5% of stand trees per

of total carbon stock of the research area.

ha respectively. For each of these scenarios the stand We have shown that the number of trees per projection was refined comparing the situation for the

hectare in the studied forest was unequally MFD as suggested by the forest administration to the

distributed for various species groups. Eighty percent situation of the newly estimated MFD.

of trees in a hectare were in group 3 and 4, representing non-commercial trees (Fig. 3b). Other

2.7 Carbon Estimations studies also reported unequal species group

For total aboveground biomass estimation of trees, distribution in tropical rain/moist forests [26]. we used a regression equation developed for moist

However, basal area distribution of 57% for Groups tropical forests [38], with 443 trees ranging from

1 and 2 and 33 for Groups 3 and 4 (Fig. 3b) was 1-148 cm with biomass data collected from the

smaller than the values given by Bibani and Jonkers authors [38] and other sources [39-42]. This equation

[26], possibly due to the difference in defining is as follows:

species groups. These authors classified timber M = exp(-2.0815+2.5624ln(D)) (4)

species of high future potential (species which are where M represents the total dry aboveground biomass

likely to be commercial after 20-30 years) in Group 2 of a tree in kg and D the diameter in cm. The

and tree species producing non-timber forest corresponding carbon content in biomass was products (NTFP), which are used by the local

estimated assuming 46.5% of carbon in the biomass population, in Group 3. High concentration of tree [43]. In this paper, the biomass stands for the carbon

species in Groups 3 and 4 which are of little or no content in total dry aboveground biomass and is

commercial use, suggests that more research is expressed in MgC·ha -1 .

needed to increase the knowledge in wood properties

3. Results and Discussion

which can change the status of a species from a non-commercial value to a commercial value. The

3.1 Composition of the Actual Forest valorization of secondary species can also change the In species Group 1 (all mature merchantable

status of certain species.

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

Fig. 3 Stem number N (black), basal area G (grey) and carbon pool C (white) in the four species groups. Group1 mature merchantable species with diameter above the minimum felling diameter (MFD). Group2 immature merchantable species with diameter smaller than the MFD. Group3 all non commercial tree species with a MFD prescribed by forest administration. Group4

all non commercial trees without a prescribed MFD. A: Number of stems per ha (N/ha), basal area per ha (m 2 /ha) and carbon pool (MgC/ha). B: Stem number N, basal area G and carbon pool C in % of all tree species in stand at year 0.

3.2 Stand Projection for Future Forest Structure diameter [44]. In Fig. 2, the logarithm scale has been used for the representation of Y scale (trees·ha -1 ) to

The distribution of the stem number in year 0 varies enable differentiation for bigger diameters with stem from 60 trees·ha -1 in diameter-class 15 to 0.3 trees·ha -1 number less than 0.5 tree·ha -1 . Because it was not

in diameter-class 135. A projection of the stand possible to predict the incoming regeneration of new development over a period of 30 years shows that the

trees into the smallest diameter class, a constant value shape of the distribution will change over time as a

of 60 trees·ha -1 in the smallest class diameter 15 cm at result of growth and ingrowths into the next all time steps was assumed. The projection predicts a diameter-class (Fig. 4). For each time step of 5 years,

high number increase in smaller diameter classes to a the stand development models maintain the J-shape

very small increase in bigger diameter classes. The frequently encountered in natural forests in the tropics:

average growth rate of stem density per 5 year period the number of trees decreases with increasing tree

moves from 6 trees·ha -1 (34 trees·ha -1 ) in the class 25

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

100 100 Year Year 0 0

DBH (cm) DBH (cm)

Fig. 4 Stand projections of number of trees per hectare (N/ha) of 24 commercial species in the research area over periods of 5 year up to 30 years (individual lines). All curves in general follow the J-shape: many trees with smaller diameters to very few with bigger diameters. Tree species have minimum felling diameter between 50 cm and 80 cm.

to 3 trees·ha -1 (17 trees·ha ) in the class 35, 1.1 distribution shows that some species are not big

trees·ha -1 (6.8 trees·ha ) in the class 45, 0.4 tree·ha enough to ensure the sustainability of tree stock in (2.2 trees·ha -1 ) in the class 55 and to between 0.1

-1 -1

the stand (Fig. 5). This will lead to genetic drift in

tree·ha -1 and 0.2 tree·ha in the other diameter classes. the future due to the lack of parent trees for those The values in brackets are the tree stem density after

species. The use of newly estimated MFDs (Table 1)

30 years. The predicted stem density distribution did decreases the stem density of harvestable trees and not change when the newly estimated MFD were used.

increases the density of the residual trees (dbh > 10 The average growth rate of basal area per 5 year

cm) in the stand (Fig. 6). The stem density for

period moves from 0.3 m -1 ·ha (1.7 m ·ha ) in the harvestable trees at initial age becomes 7 trees·ha

2 -1

2 -1

class 25 to 0.3 m 2 ·ha -1 (1.6 m 2 ·ha -1 ) in the class 35, 0.2

(5 m 2 ·ha -1 ) (47 MgC·ha -1 ). The projection suggests a

2 m -1 ·ha (1.1 m ·ha ) in the class 45, 0.1 m ·ha (0.5 constant increase of harvestable trees by on average

2 -1 2 -1

2 m -1 ·ha ) in the class 55 and to between 0 and 0.2 0.9 tree·ha (0.8 m ·ha ) each period of 5 years, m 2 ·ha -1 in the other diameter classes. The model

2 -1

moving from 7 trees·ha -1 (5 m 2 ·ha -1 ) (47 MgC·ha -1 )

predicts an increase of harvestable trees on average by -1 to 12 trees·ha (9 m ·ha ) (95 MgC·ha ) after 30

2 -1

0.1 tree·ha -1 during each period of 5 years years. With these new MFDs, the density of the corresponding to an increase of basal area of 0.7

residual stand (dbh > 10 cm) at initial age becomes

m -1 ·ha . The density of all harvestable trees increases 86 trees·ha (6 m ·ha ) (43 MgC·ha ). The

from 12 trees·ha -1 (7 m 2 ·ha -1 ) (70 MgC·ha -1 ) at initial

projection suggests a constant increase of residual

time to 18 trees·ha -1 (12 m ·ha ) (116 MgC·ha ) after stand (dbh > 10 cm) by on average 10 trees·ha (0.8

30 years. The model predicts an increase of residual m 2 ·ha -1 ) each period of 5 years, moving from 86

trees (dbh > 10 cm) on average by 10 trees·ha -1 at each trees·ha (6 m ·ha ) (43 MgC·ha ) to 145 trees·ha

period of 5 years corresponding to an increase of basal -1 (11 m ·ha ) (67 MgC·ha ) after 30 years. area of 0.8 m 2 ·ha -1 . The density of residual trees

2 -1

3.3 Yield and Determination of Minimum Felling (dbh > 10 cm) increases from 80 trees·ha (4 m ·ha )

2 -1

(20 MgC·ha -1

) at initial age to 139 trees·ha -1 (8

Diameter

m -1 ·ha ) (46 MgC·ha ) after 30 years. The diameter The mature merchantable species in Group 1 contain

2 -1

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

Fig. 5 Diameter distribution of 9 commercial species in the research sample forest (3 ha). The minimum felling diameter (MD) is the limit above which all trees could be cut to sustain a continuously productive forest. Species a) b) d) i) have enough smaller trees which can help to recover outgoing harvestable trees. For species c) e) f) g) h), further investigation on reconstitution needs to be carried out.

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

Fig. 6 Comparison of projections of the stem density (N·ha ) for 24 commercial species calculating with the administrative MFDs and the newly estimated MFDs. A) harvestable species with moderate damage (1% mortality rate and 7% logging damage intensity).

B) residual stand (dbh > 10 cm) after harvest. C) harvestable species with 10% logging damage intensity (3% mortality rate) and 15% (5% mortality rate). D) residual stands (dbh > 10 cm) after harvest.

12.3 harvestable trees per hectare and 21.3 residual smaller diameter classes suggests that in case of trees per hectare. For Ceiba pentandra (L.) Gaertn and

harvest, this species will not recover. For Erythropleum ivorense

A. Chev. no reconstitution was observed. Triplochiton scleroxylon K. Schum. has 3

A. Chev. no residual trees are

Erythropleum ivorense

observed by using the MFD of 50 cm which is harvestable trees per hectare and 1.3 residual trees

suggested by the traditional forest management per hectare. For other species more residual trees

system. With the MFD of 90 cm, this species will than harvestable trees were observed. The analysis of

have a percentage of reconstitution of 41% after 30 tree density reconstitution of Alstonia boonei De Wild

years. The distribution of stem numbers of this species shows that with the MFD of 70 cm, the percentage of

shows that it was not possible to consider a higher reconstitution (e.g. formula (3) ) after 30 years will be

MFD (Fig. 5). Therefore, 90 cm represents the 74%. With the MFD of 80 cm, the percentage of

optimum MFD of this species in our case study. reconstitution will be 93% which represents the

However, two different suggestions can be formulated optimum minimum diameter for this species. For

for Erythropleum ivorense A. Chev. using this new Ceiba pentandra (L.) Gaertn only 0.3 stems·ha -1 are

MFD: the first is that, it should be totally forbidden to found in the diameter class 55. The absence of trees in

harvest this species during the next 15 years and start

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

harvest only after this period. The second is that only estimated on i) the base of diameter distribution, ii) equal or less than half of the harvestable trees of this

the probable density of residual trees remaining after species should be selected. For Guarea cedrata (A.

human interventions, and iii) the availability of parent Chev.) Pellegr. a percentage of reconstitution of 147%

trees. The estimation should be species and also forest was estimated by using the MFD of 80 cm proposed

specific. It cannot be generalized for a region or a by the forest administration. Therefore, 80 cm

country since the conditions are not the same between represents the optimum MFD for this species. For

different forests within a region or country. Within the Mansonia altissima

same forest, the value of MFD needs to be updated reconstitution is only 70% with the MFD of 60 cm

A. Chev. the percentage of

after human interventions, probably after 5 or 10 years. specified in the traditional forest management system.

It is likely that the change of the length of the felling This value increases to 111% when the MFD of 70 cm

cycle will also affect the MFD. For species with low is used. Hence dbh of 70 cm represents the optimum

percentage of reconstitution like Triplochiton MFD of Mansonia altissima A. Chev.. Nesogordonia

scleroxylon K. Schum. or Terminalia superba Eng. & papaverifera (A. Chev.) R. and Pycnanthus Diels., increasing slightly or maintaining the lower angolensis (Welw.) Warb. obtain percentage of

administrative MFD and adopting a longer felling reconstitution of 110% and 108%, respectively (Table

cycle (50 or 60 years or even more) as suggested by

1) by using the MFD of 50 and 60 cm, respectively the study [45], can also be another alternative. which were suggested by the forest administration.

Another study recommendeds different length of For Terminalia superba Eng. & Diels. a percentage of

felling cycles for species with different percentage of reconstitution of 50% is resulted by using the MFD of

reconstitution [20]. This can be difficult to manage

60 cm. This value increases to 64% by calculating especially in situations where fixed annual allowable with the MFD of 70 cm and up to 83% by using the

cutting areas are preferred. A longer felling cycle can MFD of 80 cm. This last value represents the best

also result in smaller annual allowable cutting areas. MFD for the reconstitution of this species. For

For Triplochiton scleroxylon K. Schum., a MFD of Triplochiton scleroxylon K. Schum., the percentage of

110 cm has also been used in Ghana, which has a high reconstitution is 17% by using a MFD of 80 cm. The

reduction of the original forest cover [23] with a percentage increases up to 52% by using a MFD of

felling cycle similar to this study. Another authors 110 cm and up to 69% by using a MFD of 120 cm.

compared the percentage of reconstitution after The distribution of number of stems per hectare of this

logging and after logging followed by thinning [46]. species shows presence of bigger trees than smaller

They showed that logging followed by thinning will trees. Therefore, the optimum MFD of this species

double the percentage of reconstitution after 20 or 30 should be 110 cm. With this MFD, only half of the

years for tree species with diameter between 60 cm harvestable trees will be remaining at the end of the

and 80 cm. Above 80 cm, no significant changes were felling cycle. Therefore, it is appropriate to harvest

observed. This study does not include the effect of half of this stock meaning that one harvestable tree on

thinning on the diameter growth rate of the residual two to allow the reconstitution of this species.

trees. Since one cannot predict the future effect of The estimated MFD (Table 1) was similar to the

thinning, the first approximation is to adjust the MFD MFD of 50-110 cm reported for different tree species

so that the percentage of the modelled reconstitution is in West Africa [23]. This study [23] gave not enough

close to 100% as in this study. It is also important to information concerning the basis for determining

analyze the diameter distribution before taking MFD of the tree species. The MFD should be

decision. For example, in the case of Guarea cedrata

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

(A. Chev.) Pellegr. (Fig. 5), diameter distribution trees below 10 cm) which was numerically estimated coupled with the percentage of reconstitution, shows

in this study may probably play an important role on that the MFD can be shifted backwards from 80 cm to

the forest growth structure. Normally, natural

60 cm without degradation. Measurements of stem regeneration may be abundant or not depending densities 10 or 15 years after logging are necessary to

mostly of incoming light into the stand and other compare the modelled projection with the real reaction

factors such as precipitation and soil conditions. of the stand and adjust the MFD.

3.5 REDD and Carbon Estimations from Forest

3.4 Sensitivity Analysis

Degradation

Higher logging damage tested in this study affected Modelling stand dynamics of managed tropical the resulting stand structure distribution and the

forests can provide essential information for growth prediction (Fig. 6 b,d). Increasing the logging

estimating greenhouse gas emissions from forest damage intensity but keeping the diameter growth rate

degradation. For example, with the new MFDs constant resulted in a reduction of the stand density,

estimated in this study and with a low logging and the stand growth distribution. In comparison to

damage of 7% and moderate mortality rate of 1%, an equivalent carbon value of 23 MgC·ha the moderate logging damage (7%), the model -1 was not

predicts at 10% logging damage a reduction of 8 damaged (remained on the stand). This value may

trees·ha -1 (1.4 m ·ha ) (11 MgC·ha ) after 30 years. increase if there is a consideration of the effect of

-1 2 -1

2 This corresponds to a loss of 0.7 tree·ha -1 (0.6 m ·ha ) each avoided harvestable tree on the residual trees

(6 MgC·ha -1 ) on harvestable trees and 8 trees·ha (0.8 and the expected stand growth due to ameliorated

m -1 ·ha ) (5 MgC·ha ) on residual stand. At logging light and soil resources. In fact, selective logging damage of 15%, the model predicts a reduction of 17

2 -1

increases photosynthetic radiation and nutrient trees·ha -1 (3 m 2 ·ha -1 ) (22 MgC·ha -1 ) after 30 years,

supply, and therefore stimulates growth of tree compare to logging of 7%. This corresponds to a loss

species, especially light-demanding tree species [48,

of 1.5 trees·ha -1 (1.2 m 2 ·ha -1 ) (13 MgC·ha -1 ) on

49]. Normally, when trees are taken out from a forest

system, it reduces the nutrients potentially released MgC·ha -1 ) on residual stand. The mean annual growth

harvestable trees and 15 trees·ha -1 (1.5 m 2 ·ha -1 ) (9

from the stems, branches, roots, barks and leaves.

found was 2.2 trees·ha -1 (0.3 m 2 ·ha -1 ) (2.4 MgC·ha -1 )

But the thinning effect probably increases the

at 7% logging damage, 1.9 trees·ha -1 (0.3 m 2 ·ha -1 ) (2

mineralisation rate of below ground organic material

2 MgC·ha -1 ) at 10% and 1.6 trees·ha (0.2 m ·ha ) (1.7 and reduces the competition for nutrients by the roots. MgC·ha -1 ) at 15%.

Further analysis is necessary on whether the pools of

The increase in stem density by 2.2 trees·ha -1 ·year nutrients within the forest system can be kept

constant or even increase to maintain a sustainable commercial trees seems to be consistent with other

corresponds to a basal area of 0.3 m 2 ·ha -1 ·year -1 for all

forest. Solar radiation reaching individual trees of the studies [24, 47]. After 14 years of investigation in a

remaining tree population is increasing after natural forest in Ivory Cost, an increase of 3.2

disturbance and thus photosynthesis and tree growth

trees·ha -1 ·year was estimated in an undisturbed forest rate of the individual trees could change. Presumably and 5 trees·ha -1 ·year -1 in a thinned forest [24]. A lower

most of the remaining trees will grow faster so that

value of 0.6 tree·ha -1 ·year was observed in another the diameter increment after the selective logging undisturbed forest and 1.8 trees·ha -1 ·year -1 in another

will be larger than before logging. This study also thinned forest [47]. The natural regeneration (all new

shows that increasing mortality and damage intensity

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

also increases the damage on carbon biomass damage on the residual stand and the avoided damage estimated to be 11 MgC·ha -1 at 10% or to be 22

compared to the previous activities. The resulted MgC·ha -1 at 15% logging damage.

financial incentives could be invested in the reduced For monitoring carbon pools in tropical forests, it is

impact logging (RIL) and in the silvicultural important that estimations of national equivalent

techniques such as suggested in Table 2 to increase baseline carbon (sink or source) and the actual level

tree growth and to reduce trees mortality and damage

be derived from local estimations provided by on residual trees. Additional studies on biodiversity individual projects. The sum of local estimations will

and on wood properties of less used or unused trees lead to a national estimation. Thus the quantification

which represent in this case 80% of the total number of national baseline estimations will avoid (Fig. 3) could be promoted by means of these sub-national leakages; the local baseline estimations

financial incentives. A mean coefficient of will enable more accurate estimations. In this study,

commercialization (ratio between the volume of wood three logging damage intensities of 7%, 10% and 15%

effectively used and the volume harvested for a given with newly estimated MFDs (Table 1) taking into

species) of 51% was reported for 24 commercial consideration the reconstitution of the stand and the

species with values ranging between 25% and 70% length of the felling cycle, resulted in different growth

[19]. This coefficient could be improved with scenarios of the stand both for harvestable trees and

financial incentive and a real implementation of a residual trees (Fig. 6). It is possible for a particular

newly orientated forest policy.

forest management project involved in REDD to

4. Conclusions

assume a logging intensity with low impact, to determine new MFDs and length of felling cycle with

This study provides a future stand development of ecological considerations and to estimate on this base

commercial species of the stand up to the end of the

a growth scenario of the forest stand. It may then be first felling cycle (30 years). The projection for feasible to estimate the harvest intensity, the damage

harvestable trees suggests for this time span an on the residual stand, an avoidable damage compared

increase of the stem density from 12 to 19 trees per to the previous level and its corresponding carbon

hectare. The basal area for harvestable trees also emission reduction. The estimation could be projected

increased from 7 m 2 ·ha -1 to 11m 2 ·ha -1 at the end of the in a commitment period (if already defined) and

30 year felling cycle. This estimation can only be achieved compared to the local baseline issued. By this method,

if the remaining trees are not seriously damaged

a carbon trade can be realized: if the emissions are during harvest and no unpredictable event severely lower (higher) than the baseline, an individual project

deteriorates the forest stand. Sustainable management can sell (buy) carbon. It may be even the case that

of tropical forests for timber production requires an carbon is not emitted but absorbed because of the high

effective system for predicting future forest growth photosynthesis rate due to higher absorbed and yields [50]. Inventories have to be updated for

optimizing stand management practices and evaluate and deposition of fertilizer compounds. The resulting

photosynthetic radiation, higher CO 2 concentration

economics of future stands [51]. Future stand sequestered carbon is larger and more carbon bonds

developments may be derived from current structures could be sold. Policy and Measures (PAM) needs to

through stand projection, using estimated seedling

be combined with Cap and Trade (CAT) to make the survival rates over the entire logging cycle, diameter trade efficient and avoid leakage. Post harvest

increment rates, a mortality ratio and logging damage inventories have to be carried out to evaluate the

intensity.

Modelling Stand Dynamics after Selective Logging: Implications for REDD and Carbon Pools Estimations from Forest Degradation

Table 2 Proposed silvilcultural system scenario for monitoring forest management under REDD (Reducing Emissions from Deforestation and Forest Degradation) during a felling cycle of 30 years and for example a commitment period (period during which the carbon is estimated at the beginning and evaluated at the end, with a potential carbon trade) of 10 years.

Year Operations L–3

Stratification of the forest according to the ecological characteristics and past logging operations; planning of activities. Inventory of the entire forest at low intensity in transect lines. Use for example plots S of 20 m × 250 m

subdivided in two subplots S 1 of 20 m × 5 m and S 2 of 5 m × 5 m. In the plots S, all trees with dbh > 10 L–2

cm are counted. In the subplot S 1 , all trees with dbh between 5-10 cm are counted; in the subplot S 2 evaluate the natural regeneration with dbh < 5 cm. Diameter distribution and percentage of reconstitution analysis and estimation of MFD. Growth projection analysis and estimation of an avoidable damage reduction with the related carbon estimation. Inventory in at least 2 to 3 permanent plots of 100 m × 100 m located on different ecological zones. The

L–2 permanent plots should be subdivided in subplots S 1 and S 2 . Measure trees as mentioned above. -Inventory of 1/30 of the total forest. In this section of the forest, count all the trees ≥ MFD. Sampling

L–1 intensity: 100 %. -Marking of at least one future crop tree per ha and trees to be felled. Ensure that all commercial tree species to be harvested are represented in the future crop trees.

L -Felling of marked trees on the above mentioned 1/30 of the forest. -Post felling inventory; Thinning of the future crop trees. Logging damage analysis.

L+5 2nd inventory on the permanent plots. L + 10

3rd inventory on the permanent plots. End of the first commitment period. 2nd inventory of the entire forest at low intensity. Evaluation of avoided logging damage. New carbon pools estimations.

L + 15 4th inventory on the permanent plots. 5th inventory on the permanent plots. End of the second commitment period. 3rd inventory of the entire

L + 20 forest at low intensity. Evaluation of avoided logging damage. New carbon pools estimations. L + 25

6th inventory on the permanent plots L + 30

End of the felling cycle. Inventory on the permanent plots. End of the third commitment period. Inventory of the entire forest at low intensity. Evaluation. Start of a new cycle.

L: Year of logging; L – 1: one year before logging; L + 5: five years after logging. Logging takes place each year in a 1/30 section of the forest. Always repeat inventory before logging and post inventory after logging in each logged section of the forest.

With the MFDs estimated in this study, the stem facilitate a voluntary option to a low logging damage density of harvestable trees decreased from 12 to 7

intensity and to a proper estimation of MFD, which trees per ha and the number of residual trees increased

provides sustainable timber production during the next from 80 to 86 trees per ha. These changes correspond

felling cycle, reduces damage and allows regeneration to an estimated carbon stock of 23 MgC·ha -1 . An

of the original and probably the historical forest. estimation of the MFD of commercial species should

Funds from REDD should be re-invested in the take into consideration the diameter distribution of the

implementation of a silvicultural technique which will species in the stand, the reconstitution of the

allow reduced impact on the residual stand, provide harvestable trees at the end of the felling cycle and the

more information on tree growth and also increase length of the felling cycle. The tree growth rate used

research on technical, biophysical and biochemical to simulate the stand development after logging

properties of actually non used trees which in the should take into account the different growth present study represent 80% of forest tree species. conditions after logging.

Acknowledgments

Financial incentives expected from the REDD mechanism will probably enable forest

This study would not have been possible without administrations and other stakeholders to implement a

the financial support of the Deutsche Gesellschaft für forest policy which will reduce forest degradation.

Technische Zusammenarbeit (GTZ). My thanks are The expectation through this new orientation is to

also extended to Katholische Hochschulgemeinde

Modelling Stand Dynamics after Selective Logging: Implications for

REDD and Carbon Pools Estimations from Forest Degradation

(KHG) and the International Student Services for their [11] M. Schulze, J. Zweede, Canopy dynamics in unlogged and logged forest stands in the eastern Amazon, Forest

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greenhouse gas emissions, in: P. Moutinho, S. early stage of this study are highly acknowledged. We

Schwartzman (Eds.), Tropical Deforestation and Climate Change, Amazon Institute for Environmental Research,

also thank Prof. Dr. Putz Francis E., University of Ford Foundation, Belem, Brazil, 2005. Florida (USA) and Prof. Dr. Alexander Knohl,

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Journal of Life Sciences 6 (2012) 817-825

Aqua Forestry and Duck Integration in Tamil Nadu, India

Faroque Rahamathulla Sheriff Department of Poultry Science, Madras Veterinary College, Chennai 600007, India

Received: November 10, 2011 / Accepted: January 16, 2012 / Published: July 30, 2012.

Abstract:

A study was taken up on integration of ducks in private and community fish ponds in Tami Nadu. A total of 50 ha of land under private holding and community water bodies each were selected to implement this model. Ducks were integrated based on the water spread area. Fodder, fruit, nuts, fuel, timber trees and vegetables were grown on pond bunds. Control units of fish ponds of farmers holding without integration of ducks also were taken up. A control unit also was established in the university farm land with and without integration of ducks. In private water bodies coconut trees and banana fetched additional revenue. During the first year the average fish production from the private land model was 1,552 kg and 1,731 kg for the first and second year respectively per Hectare. In community ponds the yields were 1,610 and 1,337 kg for the first and second year respectively. In field control units there was no increase in fish yield over the years. In private water bodies the beneficiaries got egg yield of 148 and 90 eggs and community water bodies 144 and 127 eggs per bird were obtained during first and second year respectively.

Key words: Aqua forestry, duck integration, plant integration, duck egg production.

1. Introduction varieties of plant may help the farmers and women to get better economic returns. This will not only

Under Indian conditions the demand for green generate employment through out the year but also fodder is high and it is time to convert the waste lands provide quick returns, enrich soil, control soil erosion and marshy lands for fodder cultivation to tide over and bring about more economic sustainability. the situation. Many models such as Agrisiliviculture, Addition of organic manure increases soil porosity Agrisilvipasture, Hortipasture, Hortisilvipature, and

and retention of moisture.

Silvipasture are practiced. Many of these systems have long incubation period for generation of income for

2. Present Scenario of Duck Farming

the practicing farmers. Under agro forestry system Many landless poor farmers rear their ducks on aqua forestry is one of the accepted models. open post harvested paddy fields, in lakes, ponds and In this system trees are planted on the bunds of fish reservoirs; for meeting their feed requirements to get ponds or lakes or reservoirs. Fish harvest takes at least egg production. The duck growers mostly prefer to six months to one year time and trees will fetch stay in water logged areas for a longer duration to get revenue after five to ten years time only. The land better foraging facilities. When the water gets depleted owner suffers from want of money for his day to day they start migrating in search of post harvest paddy requirements even after possessing water bodies or fields or water bodies. They migrate miles at a stretch marshy/swampy land or hiring out of community in search of post harvested paddy fields thus leaving ponds or lakes or reservoirs. Integration of ducks in the family in lurch. They may become bankrupt also fish ponds and proper planning to plant some of the in due course and become bonded slaves for life to the short duration crops mixed with medium duration egg marketing people who advance money for rearing

Corresponding author: Faroque Rahamathulla Sheriff, their birds and earn revenue. A sustainable model of professor and head, research field: poultry science. E-mail: sheriff.fr@gmail.com.

this type is essential at present.

Aqua Forestry and Duck Integration in Tamil Nadu, India

3. Materials and Methods

hectare (Ha) of water body ranged from 24 to 240 based on the water spread and availability of natural

A study was taken up on Integration of ducks in feed resources. When the ponds were dug for growing

Aqua forestry in private and community lands in of fish a depth of 1.5 metres was taken at the centre

Tamil Nadu Veterinary and Animal Sciences University, Chennai, India with the objective of

and about 1 metre at the sides. The dug up soil was made use of for the formation of bunds. The width of

 To establish duck rearing in aqua forestry to suit the pond bund varied from 0.5 to 1.0 meters

the private and community Water bodies in depending up on the size of the pond. Approximately Tamilnadu;

 To study the problems and advantages of duck one fifth of the land was utilized for the formation of cum fish farming for generating better income to the

pond bunds. In one acre of pond area 20 cents was farmers.

utilized for the formation of the bund. This land mass In private land a total of 33 beneficiaries practicing

was lying fallow and was utilized for the development fresh water fish farming were selected from nine

of aqua forestry model in the present study. The pond Districts of Tamilnadu (Chennai, Kanchepuram, owners who were having small and large ruminants

Virdhachalam, Thanjavur, Ramanathapuram, were interested to rear grass and fodder trees and Tiruvallur, Vellore, Madurai and Villupuram) for

others were interested in fruit trees and vegetables. implementation of model. The details of beneficiaries

Most of the fish growers planted coconut trees (Cocos and the area of fish farm and duck integration are

nucifera ) on the pond bunds. The total plantations that shown in Table 1.

were taken up in 50 Ha of pond bunds in private lands The ponds where fish was grown for a period of 6

were as follows:

Fodder: Albezia lebeck (300), Neem (Azardica months and above only were taken for the study. In

Tamil Nadu the fish varieties prescribed to be grown indica ) (1000), Erythrina indica (4000), Glyricidia in composite fish culture were Rohu (Labeorhita)

sepium (3000), Jamun (Syzygium cumini 150), Mirgal (Cirrhina mirgala) Catla (Catla catla)

Thespesia populena (100) and Pithcolumbium dulce Common Carp (Cyprinus carpio) Silver Carp (150); (Hypophthalmichthys moliotrix) and Grass Carp

Fruit trees: Coconut (600), Mango (1000), Guava (cteno pharyngodanidella). A total of 50 Ha water

(600), Pomegranate (600), Acid lime (500), Anona bodies owned by private land owners were identified

(500), Jack fruit tree (100) and Papaya (1000); to establish this model where the fishing operation

Timber and fodder trees: Teak (150), Bamboo, alone was in progress. Number of ducks integrated per

Casurina, Tamarind (150) and Prosopis (600);

Table 1 District wise distribution of beneficiaries for implementing duck rearing in aqua forestry system in private water bodies.

S.No. District

No. ducks integrated/Ha of pond (Range) 1 Chennai

No. beneficiaries

Area of farm (Ha)

2 2.75 156 (60-96)

2 Kanchepuram 8 12.95 792 (24-240) 3 Virdhachalam

3 4.5 302 (96-120) 4 Thanjavur

10 15.15 912 (24-120) 5 Ramanathapuram

7 Vellore 6 11.75 660 (72-120) 8 Madurai

Total 33 50 2978 (24-240)

Aqua Forestry and Duck Integration in Tamil Nadu, India

Vegetables seeds: Tomato, Water melon, Pumpkin, community lakes/ponds seventeen of the beneficiaries Brinjal, Drumstick, Yam, Okra and Spinach;

were from Chennai and Kanchepuram Districts. This Grass slips or seeds: Para grass, Guinea grass,

is due to availability of Community ponds/lakes under Napier × Bajra hybrid, Desmanthus (5 kg), Stylo

the control of Fish farmer’s development agency. The haemata (10 kg) and Leuceana leucocephala (25 kg).

Fish Farmers Development Agency supplied Similar model was implemented in community

fingerlings and also helped in rearing and marketing water bodies such as lakes and reservoirs and ponds in

of fish. The beneficiaries the area of water body in the eight Districts of Tamil Nadu (Kanchepuram, Chennai,

lake during the peak summer was taken as the area of Tiruvallur, Ramanathapuram, Vellore, Thanjavur, implementation. Based on the area ducks were Thoothukudi and Pudukkottai). Most of the lakes and

distributed as shown in Tables 1 and 2 in private and reservoirs were leased out by the Panchayats and the

community water bodies respectively. The number of similar type of fishes culture mentioned for the private

birds was finalized with the spread and depth of water ponds was stocked and reared. After a period of 6 to 8

and its duration of for fish culture. 100 ducks per months or till the water was available in the water

hectare of water body would be needed to excrete bodies it was reared and then harvested. The details of

duck droppings (6000 kg/ha/yr), to meet the ‘safe’ beneficiaries’ number and the area of pond or lake and

level dose of the duck wastes for one year [1]. duck number integrated are furnished in Table 2.

Experiments conducted at fish ponds indicated 100

A total of 25 beneficiaries were selected who have percent recovery of test fish from duck waste treated taken community ponds or lakes on lease in eight

water and the estimated net and gross fish production Districts to implement this model. The pond bunds of

were high. We collected performance of 10 the community water bodies were utilized to grow

aquaculture farms with at least one farm in each Neem, Palmyra, Prosopis, Eucalyptus, Coconut, district where the duck integration was not taken up. Mango and Guava. The pond bunds are used as path

To serve as control a similarly model was established way for the villagers to reach their fields and the fruit

in our University farm in 0.5 Ha land with integration saplings thus planted were destroyed. Due to this

of ducks and without integration of ducks. reason many beneficiaries preferred to grow timber,

4. Results and Discussion

fuel and fodder trees on the pond/lake bunds. Cencrus ciliaris seeds were sown on the pond bunds during

The private land beneficiaries from Vellore, monsoon rains. This model was primarily developed

Ramanathapuram and Tiruvallur were able to maintain to utilize the partial waste land in pond bunds and

the ducks for a period of 6 to 7 months only while other dykes for cultivation of trees to reduce soil erosion. In

districts farmers were able to maintain the birds in their

Table 2 District wise distribution of beneficiaries in community water bodies.

S.No. District

No. of ducks/Ha integrated 1 Chennai

Beneficiaries (No.)

Area of implementation (Ha)

10 28.75 24-120 2 Kanchepuram

7 6.75 24-96 3 Tiruvallur

3 3.25 6-24 4 Ramanathapuram 1

Total 25 50.0 6-120

Aqua Forestry and Duck Integration in Tamil Nadu, India

ponds from 8 to 15 months period. This is due to shortage of water as they are highly drought prone districts. Hence the ducks had to be taken for foraging in the post harvested paddy fields and nearby marshy lands and also migrated to nearby districts or states in search of foraging facilities. Some of them had to adopt hand feeding of ducks with paddy chaff and broken grains and had to invest lot of expenditure. Coconut trees started fetching revenue from fourth year onwards.

4.1 Revenue from Trees Each tree fetched more than 80 to 90 nuts per year.

In every hectare of pond bunds about 40 trees were grown and fetched around 2,500 nuts per year and added to their revenue to a tune of Rs.7,500/- Many farmers have grown Banana trees (Musa paradisiaca) on the pond bunds. In each hectare of water body around 60 banana trees were grown. They were able to make money through the leaves, fruits and flower every day. The torn leaves also were used for feeding the grass carp fish and nothing was wasted from banana crop. From each plant a revenue of Rs.100/- and around Rs.6,000/- was earned from 1 Ha of pond bunds. Growing of mango (Mangifera indica), pomegranate, guava (Psidium guajava), Acid lime (Citrus aurantifolia) were also taken up by many fish growers. Pomegranate tree had low survivability in the study area. Some fish growers at Kanchepuram District had preference for growing vegetables on the pond bunds like water melon (Citrus lanatus), tomatoes (Solanium lycopersicum), cucumber, pumpkins (Qucurbita maxima ), brinjal, drumstick (Moringa oleifera) and spinaches. This gave additional revenue besides use of the vegetables for their home consumption.

Some farmers in community ponds and reservoirs planted teak on the pond bunds for long term benefits. These saplings had more than 90 percent survivability up to three years of age and have grown to three metres height. Similarly those maintaining small ruminants had grown Neem and Albezia lebbeck and

these tree saplings had good survivability up to three years and had grown up to three metres height with good canopy cover. Integration of ducks in aqua forestry model has its own advantages as there was daily income from eggs for the farmers. In addition to yield from fruit bearing trees, no soil erosion from pond bunds was observed. Addition of organic manure to the pond helped in the development of phytoplankton (Diatomacce,

Chlorphycea and Myxophycea ) and zooplankton (Rotifers, Cladocerans, Copepods and their larvae) which served as fish feed. Due to paddling of webbed feet of ducks there was increase in oxygen level in water body which helped in better growth of fish. Integration of ducks eliminated snails, crabs, trash fish earth worms and aquatic weeds from water bodies. Growing of Para grass and Guinea grass had served as feed for grass carp fish. Seedlings of Subabul were twined at early age and grown on the pond bunds and this method had served as live fence for protection fish against poaching. Growing of fruit bearing trees and vegetables had attracted women to take part in cultivation and also for plucking the fruits and spinach from their own land. Many farmers realised the importance of growing fodder grass by using the pond bunds for feeding grass carp fishes and also for their large and small ruminants and has started growing them on pond bunds. The Para grass (Brachaiaria mutica ) variety was able to survive and gave good yield in the stagnant water also. When the pond was partially dry the grass seeds were sown and thereby good growth was noticed in a period of one month. This grass was consumed by grass carp fingerlings as soon as it was stocked in the ponds. This grass grows through out the year on self seeding and had grown on bunds and was continuously providing yield for the fish and also for other Livestock. This grass cultivation has controlled the soil erosion from pond bunds too. Those who had established this grass on their pond bunds had not incurred expenditure on strengthening of the bunds for more than four years.

821 On the outer side of the bund the Napier × Bajra

Aqua Forestry and Duck Integration in Tamil Nadu, India

4.2 Problems and Revenue in Implementation in grass and Guinea grass were cultivated. From each

Community Ponds

acre of pond area about 3-5 tons of grass was The community lake/pond owners planted fodder cultivated per year thus providing green biomass to

fuel and timber varieties only on the bunds as the the fishes and livestock continuously. Due to

income was useful to the community but not to the planting of vegetable and fruit trees in private water

fish growers as the lake/Reservoir can be utilized for bodies and rearing of ducks the fish farmers engaged

the leased period of one or two years only. They also labourers to look after them. They found women

could not engage any labourers to look after and were more responsible in collection of eggs and

protect the saplings. Some additional advantages plucking of fruits and vegetables than men labourers.

noticed were the soil pH came down from 8.0 to 7.6 One of the beneficiaries had grown jasmine plants on

after implementation of this model which is more the pond bunds and engaged the women to pluck

conducive for fish growth in community as well as in flowers daily for marketing. Another one had utilized

the private water bodies. Problems of crabs, snails, his barren water pond for six months to grow chilly

trash fish and toads were effectively controlled. crop as it had enough natural manure by integration

During the first year the average fish production of duck in the farm. This model has provided

from the model developed by private farmers was employment opportunity, additional income through

1,552 kg and in the second year it went up to 1,731 kg. eggs, duck meat, fruits, vegetables, nuts, flowers, and

The fish harvest was higher in 2nd and 3rd year of

a major harvest of fish once in a year. A mixed production which may be due to higher residual enterprise had improved the productivity and manorial quantity, higher natural feed from planktons

profitability besides transforming the ecology with and higher availability of oxygen to fish in the ponds synergistic interactions among different farm as seen in Table 3. enterprises. This type of farm had provided a

In community ponds the average fish yield was sustainable integrated resource management 1,277 kg during the first year and during the second approach for livelihood security for small and

year was 1,624 kg only even though the feeding marginal farmers.

expenditure was almost same (Table 4). Here also the

Table 3 Production and income generation from duck integrated aqua forestry farm in private lands.

Fingerlings stocked Culture period

Feed expenses for

Receipt (Rs) Districts

Years (No)/Ha

(Months)

ducks and fish (Rs) Fish produced (kg)

I II I II I II I II I II Chennai 4000 4000 15 12 8000 80 1900 3000 47500 75000 Kancheepuram 4000 4000

8 9 10333 11218 1250 1557 31250 38925 Virdhachalam 6000 6000

8 10 21650 21930 2600 2742 65000 68550 Thanjavur 4000 4000 13 12 22780 27060 2170 2830 54250 70750 Ramanathapuram 1600/ac 1600/ac 5

15000 19250 Tiruvallur 4000 4000 8 9 19333 18218 1450 1657 36250 41425 Vellore - 4000 - 7 12000 1500 - 37500 Madurai - 4000 - 11 - 16720 - 1900 - 47500 Villupuram - 4000 - 7 - 12000 - 1450 - 36250 Total 4333 4222 8.1 9.2 15682 16193 1552 1731 38800 43275

Aqua Forestry and Duck Integration in Tamil Nadu, India

fish production increased in the second year as seen in oxygen in water in private water bodies in all the the private water bodies even though there was

Districts studied. The increased receipt is shown in poaching problem in the community lakes and

Table 3. When the culture period was increased the reservoirs.

yield of fish also increased. It may be concluded that based on the marketing preference of big sized fish the

4.3 Production in Control Ponds remuneration from fish is more. This can be seen from

The private ponds where there was no integration of those units where the ducks were not integrated and ducks was taken up there were no increase in fish

the profit margin decreased in the same Districts yields over the years although the expenditure was

reared for almost the same period (Table 5). almost same (Table 4). The second year average yield

The fish production did not show any increase of fish was higher over the first year and the receipt

when there was no integration of the ducks in private also increased. The fish culture period also increased

water bodies. When the ducks were integrated in fish by more than a month during second year and

ponds the fish growth was good. This indicated that consequently the feeding expenditure too increased.

the integration of ducks had given added economic The fish production increased in integrated duck cum

advantage to the farmers. This was again proved in an fish farming due to addition of duck manure and

experimental condition at Livestock Research Station

Table 4 Fish production in community water bodies (per ha).

Fish production Receipts S.No.

Fish stocked

Culture period

Duck and fish feed

(Rs) District

Year

(No)

in months

expenses (Rs)

(Kg)

I II I II I II I II I II 1 Chennai

1600 42875 40000 4 Ramanathapuram 4000 - 6 - 2000 - 1200 - 30000 - 5 Vellore

4000 - 7 - 2000 - 1000 - 30000 - 6 Thanjavur 4000 - 7 - 2400 - 1000 - 30000 - 7 Thoothukudi 4000 - 6 - 2460 - 1800 - 45000 - 8 Pudukkottai 3000 - 6 - 1700 - 700 - 17500 -

Table 5 Production and income generation from private aquaculture farm without integration of ducks in private lands (per ha).

Receipt Years

Fingerlings

Culture period

Fish feed expenses

Fish produced

(Rs) District

stocked (No)

I II III I II III I II III Chennai

I II III I II III

31500 36000 - Kancheepuram 4000 4000 -

15000 16500 - Virdhachalam 6000 6000 - 8 9 - 20000 20000 -

40000 40000 - Thanjavur

42000 40000 - Ramanathapura

m - 4000 - - 6 - - 10000 - - 300 - - 6000 - Tiruvallur 4500 4500 - 8 9 - 20000 20000 - 1200 1500 - 26400 33000 -

Vellore - 5000 5000 -

1000 1200 - 37000 36000 Madurai

- 800 - - 24000 - Villupuram

- 4500 - - 10 - -

400 - - 10000 Total

Aqua Forestry and Duck Integration in Tamil Nadu, India

with 1.25 ac of pond area with integration of ducks The deviation in different stocking density of ducks in where the revenue was higher by two times over the

the present study may be due to the period of the study control without integration of ducks (Table 6).

and the natural feed that gets accumulated in the water Input-output relationship, integrated fish farming

bodies. The difference in the duck egg yield may also with ducks is considered to be the best model of

be due to the breed of ducks reared and the availability integration [2]. In the case of integrated fish farming

of feed to them [4]. Among the varieties of ducks with chicken, there is no “symbiotic relationship”

studied such as Nageswari, Sylhet Meat, Indian which exists in the case of geese, but the quantity of

Runner and Sera Chameli the Khaki Campbell crossed goose eggs produced is comparatively small and the

with local Pati variety had higher production market demand is low. Hence, fish-cum-duck farming

performance in condition prevailed at Assam State not only had the best economic benefits but also

climatic conditions [5]. By duck integration a demonstrated a close integrated relationship. When

production of 3,500-4,000 kg of fish, 18,000-18,500 compared with the fish-cum-cow integrated farming,

eggs and 500-600 kg duck meat from 1 Ha of pond the economic efficiency is higher in the former. Duck

area in 1 year without any supplementary feed and cum fish culture using Khaki Campbell ducks in the

fertilizers was obtained. In the present study the yield integrated system has produced 270 eggs per duck/yr

was lower than that reported which may be due to the and 6,000 kg fish/Ha/yr and without addition of any

breed, feeding, age of the bird, fish varieties stocked other fertilizer in the pond [3]. Number of ducks

and feeding of fish followed.

required to fertilize a hector of water spread area is a Thanjavur beneficiaries owning private water matter of consideration. One duck voided about

bodies could get highest egg yield of 148 eggs per 125-150 g excreta in a day. Therefore by stocking

bird per year and the lowest was in Tiruvallur District 250-300 ducklings/Ha water spread area the required

with 90 eggs during the first year of laying. This quantity of duck excreta, i.e. 10,000-15,000 difference was due to availability of water for 10 kg/year/Ha water spread area, can be received. So it

months and availability of natural vegetation for has been found that about 200-300 ducks/Ha water

grazing by the birds in Thanjavur District and only 6 spread area is sufficient to produce manure to fertilize

months at Vellore District. In the second year of

a pond of 1 Ha water spread area under fish culture. laying also the farmers at Thanjavur had 118 eggs per

Table 6 Production performance of composite fish culture with integration of ducks at Livestock Research Station (control farm).

Control Treatment S.No.

Year Parameter studied

I II III IV I II III IV 1 Water spread area (ac.)

1.25 1.25 1.25 1.25 1.25 1.25 1.25 1.25 2 Water level (ft)

2.0 2.5 2.5 2.5 2.0 2.5 2.5 2.5 3 Fish culture period (days)

145 4 Total feed issued to fish (Kgs)

- 5 Fingerlings stocked (Nos.)

5,000 5,000 5,000 5,000 5,000 5,000 5,000 6 Average initial weight of Fingerlings (g)

0.76 0.66 0.82 0.96 0.76 0.82 7 Harvest weight (g)

80 194.2 258.75 253.75 8 Total No. of fish harvested

2,000 2,220 1,610 2,235 2,270 2,635 2,120 9 Total harvest (Kg)

347.7 592.65 521.00 10 Survival percentage

44.51 44.56 47.0 41.5 Total receipt (Rs)

Aqua Forestry and Duck Integration in Tamil Nadu, India

bird followed by Villupuram and Kanchepuram. The could maintain the birds for about three years in these district wise duck egg yields in private water bodies is

two Districts. The lowest fish yield was at Pudukkottai given in Table 7. The duck egg yield in the

District which is due to short period of culture community water bodies integrated with ducks is

maintenance. The yield from fish was also increased presented in Table 8.

as the years passed by in Chennai and Kanchepuram. In Thanjavur district the maximum egg production

In other districts the fish culture and integration was of 144 eggs per bird was recorded followed by

not done for the entire period due to shortage of Kancheepuram district with 127 eggs per bird. During

water. The second year average yield of fish was the monsoon rains heavy inflow of trash fish was

higher over the first year and the receipt also found in community ponds and these trash fish served

increased in community lakes also. The fish culture as a good source of feed for ducks. After the harvest

period was almost same during second year the feeding of fish the trash fish in the slushy water were

expenditure was increased due to more enthusiasm consumed by scavenging by the ducks and the egg

and planning to achieve higher gains. production was good. The beneficiaries did not show

The pond bunds of the community water bodies interest to feed the ducks with concentrates except for

were utilized to grow neem, Palmyra, prosopis, paddy chaff unlike the beneficiaries of private ponds.

eucalyptus, coconut, mango and guava. The pond The community pond owners were allowing the birds

bunds are used as path way for the villagers to reach for foraging in the nearby post harvested paddy fields

their fields and the fruit saplings got destroyed. Due to when the water bodies dried up. The beneficiaries

this reason many beneficiaries preferred to grow timber,

Table 7 Duck egg production under integrated farm in private water bodies.

No. of eggs produced per bird S.No.

District

No. of beneficiaries

Area (Ha)

Year 1995-96 1 Chennai

Year 1994-95

6 11.75 91.0 - 7 Madurai

97.0 Total/Average

Table 8 Duck egg production in community water bodies.

Duck egg production (No.) S.No

District

No. Beneficiaries Area of implementation (Ha)

1995-96 1996-97 1997-98 1 Chennai

3 3.25 70 - - 4 Ramanathapuram 1 0.4 70 55 -

5 Vellore 1 1.25 125 105 - 6 Thanjavur

1 1.1 144 120 - 7 Thoothukudi 1

7.5 90 70 40 8 Pudukkottai 1

1.0 70 40 - Total

Aqua Forestry and Duck Integration in Tamil Nadu, India

fuel and fodder trees on the pond/lake bunds. erosion. In community water bodies’ cenchrus ciliaris Cencrus ciliaris seeds were sown on the pond bunds

grass and para grass got established well. Under during monsoon rains and within a months time there

composite fish culture the lowest preference was for was good growth of green cover. This grass was

common carp and silver carp fishes. Other fishes had consumed by grass carp fish and had put on good

good acceptance. common carp and silver carp could weight. In addition to manuring by the birds the

fetch only Rs.20/-per kg while other fishes such as villagers bring their cattle for bathing and also allow

mirgal, rohu, catla and grass carp could fetch Rs.30 to their black cattle to wallow in the ponds and lakes

35/-per kg.

which added manure to the water body continuously.

Acknowledgments

This had resulted in the fish getting adequate amount of natural flora and fauna for meeting their feed

The author wishes to place his sincere gratitude to requirements. The production performance of the Ministry of Rural Development, Department of composite fish culture with integration of ducks at

Land Resources, New Delhi, Government of India for our control farm is given in Table 6. The fish harvest

financial assistance to carry out this project. His was higher in the integrated system and thereby high

thanks are due to the support extended by the farmers receipt was earned. The survival rate of fingerlings

in recording the data and furnishing the same during also showed remarkable improvement showing better

the visit. He also wishes to express his thanks to ecology created by the integration of ducks in

authorities of Tamilnadu Veterinary and Animal aquaculture.

Sciences University for providing all assistance in conducting this research work without any

5. Conclusions

interruption or hindrance.

In private and community water bodies 60 ducks could be stocked per ha for a period of 6 to 7 months

Reference

and the remaining period the birds have to be hand fed [1] N.K. Yadav, V. Bhatnagar, Integration duck, in: Fish or allowed for foraging in the post harvested paddy

Farming System, FAO Animal Production, Animal fields. Ducks have to be fed with concentrate feed at Health Paper 27, 1992. [2] B.T. Hu, H.Z. Yang, Integrated management of fish, in:

50-100 g per day containing grains 50 percent Duck Farming and Its Economic Efficiency and gingelly oil cake or sun flower oil cake 25 percent

Revenue, FAO Corporate Repository NACA/Wp/84/14, wheat bran 20 percent mineral mixture 3 percent and

shell grit 2 percent for continuous laying. In addition [3] College of Fisheries Sciences, Govind Ballabh Pant University of Agriculture and Technology, India, 2009, they can be fed with paddy chaff ad libitum. Birds can

available online at: http://www.gbpuat.ac.in/acads/cfsc/

be maintained up to two years of age and later on had

index.htm.

to be replaced with young stock for maintaining good [4] Assam State Census, Duck, Fish Farming, College of Fishery, Raha, Nagaon, 2009, available online at: http:/

egg yield. Under private water bodies coconuts and /assamagribusiness.nic.in/2ndoct/fishduck.pdf.

banana plantation gave highest profit in short span of [5] B. Kalita, A Training Manual on Polyculture and time. Mango, guava and pomegranate growing was

Integrated Fish Farming, Sponsored by: Assam not advantageous.

Agricultural Competiveness Project, Government of Assam Organised by College of Fisheries, Assam

Guinea grass and Para grass got established on the Agricultural University, Raha 782103, Nagaon Assam,

pond bunds in private water bodies and avoided soil

India, Dec. 26-30, 2006.

Journal of Life Sciences 6 (2012) 826-831

The Importance of Fiber Production for Conservation of Native Sheep and Goat Breeds and Silkworm Lines in Turkey

1 2 1 2 Gursel Dellal 2 , Zeynep Erdogan , Erkan Pehlivan , Feryal Soylemezoglu and Aysem Yanar 1. Department of Animal Science, Faculty of Agriculture, Ankara University, Ankara 06110, Turkey

2. Department of Handicraft, School of Home Economics, Ankara University, Ankara 06110, Turkey

Received: February 15, 2012 / Accepted: April 25, 2012 / Published: July 30, 2012.

Abstract: In Turkey, studies devoted to conservation of animal genetic resources are carried out by general directorate of agricultural research (TAGEM) under the ministry of food, agriculture and livestock. There are 9 sheep and 3 goat breeds and 3 silkworm lines which produce animal fiber have been put under conservation by TAGEM since 1996. A considerable amount of these genotypes are preserved as in situ. The success of this conservation method is directly associated with economically making the best of these genotypes which are preserved. Hence, making the most of fiber production alongside meat and milk production would make a great contribution to their conservation. For this purpose, determining production biology of mohair and textile features of Ankara goats at present, determining all features of traditional items (especially handicrafts) made of fibers obtained from these breeds and lines which have been put under protection and creating new usage areas have been issues worked upon. Along with this, in recent years, just as the world, especially EU countries, interests and studies for issues such as organic animal fiber production, rural development and geographical indications have been increasing in Turkey, as well. In the near future, this situation is supposed to have a great effect on increasing of animal fiber production in a positive way, too. Hence, to benefit more effectively from fibers obtained from these breeds which have been put under protection will make a great contribution to be protected by breeders. For this purpose, there will be studies in the near future devoted to determining production biology of fiber and determining features of textile in breeds and lines and increasing the consumption of products to be obtained from these fibers as well as Ankara goats. Therefore, in this article, it was emphasized on the more efficient of ways to be benefited from fiber production in these breeds and lines under protected.

Key words: Sheep, goat and silkworm genetic resources, conservation, fiber production.

1. Introduction  contributions to folkloric culture are higher than chicken and cattle breeds increases the significance of

According to 2009 year datum, the number of sheep our genetic resources. Besides, in Turkey, local and goat are respectively 21,749,508 and 5,128,285, genetic resources of sheep, goat and silkworm have and the number of silkworm boxes are 5,683 [1]. In been decreasing for many years. Decrease in sheep Turkey, the number of sheep and goat is composed of and goat occurred very fast especially in some breeds local breeds. The number of local silkworm lines is and these breeds encountered with the danger of low. On the other hand, these genetic resources make extinct. Therefore, silkworm lines have been put under

a contribution to folkloric culture including protection besides some sheep and goat breeds by production of handicraft depends to animal fibers and general directorate of agricultural research (TAGEM) to national economy at a high level. The fact that the under the ministry of food, agriculture and livestock

Corresponding author: Gursel Dellal, Ph.D., professor,

since 1996-1997 [2].

research field: animal breeding. E-mail: There are two methods as ex situ and in situ gdellal@agri.ankara.edu.tr.

The Importance of Fiber Production for Conservation of Native

Sheep and Goat Breeds and Silkworm Lines in Turkey

devoted to conservation of animal genetic resources

2. Local Sheep, Goat and Silkworm Genetic

in the world. Executions devoted to utilization of

Resources in Turkey

available efficiency of livestock breeds protected as

2.1 Sheep

in situ because of the danger of extinct (particularly the production of special products) make a

In Turkey, a considerable part of sheep population

contribution at a high level to the protection of

is composed of local breeds. According to 2009 year

genetic resources. This situation makes fleece, whose

statistics, sheep number in Turkey is 21,749,508 and

basic efficiency is fiber, more important for Angora

nearly 95% of this is composed of local breeds [1]. In

goat, Cashmere goat, silkworm lines and Angora

Table 1, local sheep genotypes of turkey and their risk

rabbit. In USA, studies devoted to benefit from sheep,

situation are demonstrated. As can be seen in the table,

goat, Angora rabbit and South American Camelids to

2 of these genotypes are extinct, 3 are under severe

produce thin fiber have been intensified recently. In

threat of extinction, 9 are under threat and 6 are not

these studies, it has been given weight to elaborating

under threat of extinction [3].

fibers produced in the company to traditional and

2.2 Goat

new handicraft products. Thus, contribution of production of fiber to company’s economy increases.

According to 2009 year statistics, goat existence in

Therefore, in Turkey, benefiting from the fibers

Turkey is 5,128,285: 4,981,299 of the population is

acquired from silkworm lines and angora rabbit

Hair goat and 146,986 is Angora goat [1]. Nonetheless,

besides local sheep and goat breeds whose number

local goat genotypes, called Kilis, Honamli and

decrease more effectively will make a major Norduz, are counted as Hair goats because their contribution to protection of genotype and thereby to

number is low. In Turkey, local goat genotype number

company’s economy.

has been decreasing in parallel with many factors and

Table 1 Turkey local sheep breeds and types [3].

Breed and type

Not under threat of extinction

Under threat

Under severe threat

Extinct

Akkaraman X Morkaraman X

Güney Karaman

Ula ş (Kangal) Akkaraman

X Da ğlıç X İvesi X Herik X Tuj X Kıvırcık X Karayaka X Sakız X

İmroz X Hem şin X

Ödemi ş X Karakaçan X Karaka ş X Çine Çaparı X

Halkalı X Norduz X Karya X

The Importance of Fiber Production for Conservation of Native

Sheep and Goat Breeds and Silkworm Lines in Turkey

some of these genotypes are under threat of extinction Nevertheless, Angora rabbit wool and cashmere (Table 2).

production are done at a low level at certain times [4]. As seen from Table 3, according to the data

2.3 Silkworm acquired in 2009 Turkey produces about 40 thousand

Turkey local silkworm lines are Bursa Beyazı Alaca, tones of dirty fleece from sheep a year. When the 18 Bursa Beyazı and Hatay Sarısı. These lines are

year interval of 1991 and 2009 is analyzed it can be preserved by TAGEM [2].

seen that there is a steady decrease in fleece production and this decrease reached approximately

3. Conservation Studies

33.4% in 2009. There was also a significant decrease In Turkey, studies for conservation of animal

in mohair production between the years 1991-2009. genetic resources are carried out by general directorate

This fiber’s production having decreased by 87.4%, of agricultural research (TAGEM) under the ministry

only yielded 174 tones.

of food, agriculture and livestock. Conservation The main source of Cashmere cannot be exploited studies are done in primarily 3 ways as ex situ in vivo

to its fullest extent, despite the fact that the main (in research institutes), ex situ in vitro (in genbanks)

sources of Cashmere in Turkey are the local hair goats. and in situ (in dorves). Protected sheep breeds are

Due to this, it was not possible to find more data about Sakız, Çine Çaparı, Gökçeada, Kıvırcık, Herik,

the Cashmere fiber production [4]. Karagül, Hem şin, Norduz and Dağlıç, on the other

The most important source of goat top coarse hair is hand goat breeds are Angora, Kilis and Honamlı.

once again the hair goat. A constant decline in top Local silkworm lines are Bursa Beyazı Alaca, Bursa

coarse hair can be observed between the years Beyazı and Hatay Sarısı [2].

1991-2009, and the precise value of production decrease of this fiber is 49.4%. The yield in 2009 was

4. Fleece, Goat Fibers and Silk Production in

only 2,002 tones (Table 3).

Turkey

Despite silk production having an important place Fleece, mohair, goat top layer hair and production

in Turkish history, there was a major decrease in damp of silk fiber are done commercially in Turkey.

cocoon production between the years 1991-2009

Table 2 Turkey local goat breeds and types [3].

Breed and type

Under severe threat Angora goat

Not under threat of extinction

Under threat

Hair goat

Kilis goat

Malta goat

Norduz goat

Honamlı goat

*: The view of scientists working goat production in Turkey.

Table 3 Animal fiber production in Turkey [1].

Change (%) Animal fibers 1991 1995 2000 2005 2009 1991-2009

Production (ton)

Fleece 60,492 50,777 43,141 46,175 40,270 - 33.4 Mohair 1,379 797 421 302 174 - 87.4

Hair 3,955 3,397 2,697 2,654 2,002 - 49.4 Cocoon

4,353 271 60 157 136 - 96.9

The Importance of Fiber Production for Conservation of Native

Sheep and Goat Breeds and Silkworm Lines in Turkey

(96.9%). The production amount of damp cocoons in 2009 was 136 tones (Table 3).

5. Studies Related to Increasing Fleece and Goat Fiber Production in the World and European Union

In recent years, there has been an important increase in interest towards natural fibers due to developments in organic agriculture. Along with this, there are several various studies taking place on an international scale to decrease the negative effects caused by the global economic crisis on the existing rivalry of synthetic fibers between business’ economical livelihoods that depend on production of natural fibers, which includes animal fibers. Therefore, in order to draw attention to the importance of natural fibers, the FAO, industrial and several other manufacturers declared 2009 the international natural fiber year [5]. There are still important studies being carried out in order to develop thin animal fiber production to aid small family businesses economically and evaluating non-agriculturally suitable countries in the European Union (EU) [4-8].

Projects and models that support changes in the EU agricultural policies for production hold an important place. Due to this, there have been changes in EU animal fiber production policies that could be considered important over the past few years and along with the existing fiber production continuation, studies related to thin fiber production development have gained emphasis. Although animal fiber has been produced in EU, at present, one of the most important factors caused this policy change is a great majority of fibers produced what are composed of medium and bad quality to be regarded as thin fiber and the other one is; EU has a significant level of land resources which is proper to develop different kinds of animal fiber productions. This situation would make a contribution to benefiting from non arable land resources particularly mountain regions,

so EU goes towards production of thin animal fibers as thin fleece, cashmere, mohair and Angora rabbit and Alpaca hair, in terms of animal fiber production policies [4-8].

6. The Importance of Fiber Production in the Protection of Local Turkish Sheep, Goat and silkworm Lines

Since the animals are being taken advantage of economically, and the animal’s productivity will be used to benefit profit margins, the protection of Local farm animal’s genetic resources in situ will affect the protection activity in a positive way. Despite the fact that the main benefits are gained from meat and milk productivity from the sheep races (Sakız, Çine Çaparı, Gökçeada, Kıvırcık, Herik, Karagül, Hem şin, Norduz,

Da ğlıç) taken under protection from TAGEM in 2005, fleece production is not evaluated economically. Whilst the meat and milk production is emphasized on the once again protected goat races Kilis and Honamlı, along with their low level of meat and milk production, the Ankara goats are mainly reared for their mohair production. However, the silkworm lines that have been taken under protection (Bursa Beyazı Alaca, Bursa Beyazı and Hatay Sarısı) are used for their silk production. Over the past few years, there has been a decline in sheep and goat numbers, with causes stemming from reasons such as economical, sociological and structural, along with many others. However this situation should be expected to negatively affect the protected sheep and goats too. Despite the fact that the meat and milk productivity being increased in order to ensure that agricultural businesses that have opened up space for the protected sheep and goats to lay claim on them, excluding the support that TAGEM provided, other factors related to direct production being improved and made permanent will provide important contributions. One of the factors that should be emphasized upon is the fibers obtained from these races being used in a more efficient and profitable way. Just as the rest of the

The Importance of Fiber Production for Conservation of Native

Sheep and Goat Breeds and Silkworm Lines in Turkey

world, within the past 40-50 years, despite the induced textile industry [10]. Therefore, efforts should be stress brought about by the natural animal fibers in

made to use the cashmere obtained from Kilis and Turkey being used in the textile industry, the interest

Honamlı goats to be used in the textile industry and in these fibers has begun to increase over the past 5

for handicrafts. Again, the obtained top coarse hair years. Thus, as stated above, plans involving the

should be evaluated for use in handicraft production. development of animal fiber production and rural

The importance of the obtained cashmere being used development in the EU have gained importance. Due

in handicraft production is increasing in there are of to this, Turkey abiding the following methods to

origin in the Mediterranean region due to the fact that protect the fiber producing animals protected by

local tourism of this area is very developed; TAGEM being evaluated more effectively will

 Just like the Ankara goat, the silkworm is also a contribute to the continuation of the protection

fiber animal. Due to this reason, the lines of the efforts:

protected silkworms should be protected more  Excluding Kıvırcık, the sheep races Sakız, Çine

efficiently in order to make silk production more Çaparı, Gökçeada, Herik, Karagül, Norduz, Da ğlıç

profitable. Due to this, the silk obtained from these fleeces being suitable for carpet and rug production,

lines should initially be used for carpet production and should be evaluated as they have been for the past few

then for special handicraft products and should be centuries in their local area they were raised in.

marketed in an efficient way;

Because of this, the fleeces of these sheep races  Especially in the business in which it is produced, should be prioritized for carpet and rug production,

Just as the value of the handicrafts is increased when whilst simultaneously designing new models related

silk from the protected races of animal fibers is to handicraft production evaluation and marketing;

incorporated in the handicrafts, it should contribute to  Beneficial research for using the Karagül lambs

women’s employment. Efforts to use fleece, mohair in the production of astrakhan should be started

and silk in handicrafts can be seen in accordance to its immediately;

identification [9, 11]. While taking advantage of these  Since the Ankara goats main productivity stems

efforts, attempts to develop the production of from its mohair, these sheep desire a higher level of

traditional and new handicrafts should be emphasized. importance in comparison to the other sheep races.

Along with the working upon certain handicraft On the other hand, due to many factors, the inability

models (e.g. color, pattern, product type, and to take advantage of the mohair production and

marketing methods) production and marketing Turkey exporting a significant amount of mohair

methods becoming possible, the products must include which the textile industry requires. Therefore, information indicating which race or line the fibers alongside emphasizing genetic and environmental

were obtained from.

efforts, efforts should be made for the current

7. Conclusion

acquired mohair to be evaluated for use in handicraft production [4, 9];

There are nine sheep and three goat breeds and  The top fiber (hair) and thin lower fiber

three silkworm lines which contribute to animal fibre (cashmere/down fiber) are produced from the production in Turkey have been put under protected Kilis and Honamlı goats. Despite the fact

conservation by general directorate of agricultural that the cashmere production of both of these goats is

research (TAGEM) under the ministry of food, considerably low, the attributes of the cashmere of the

agriculture and livestock on the grounds that they Kilis goats has been deemed suitable for use in the

encountered with the danger of extinct. There are

The Importance of Fiber Production for Conservation of Native

Sheep and Goat Breeds and Silkworm Lines in Turkey

mainly three different methods of conservation studies

References

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there is a possibility that in our day, many negative [2] M.I. Soysal, Native Animal Genetic Resources of Türkiye, ISBN: 978-9944-5405-5-1, Tekirda ğ, Türkiye,

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flocks in the UK, Agricultural System 39 (1993) 273-287. been developments in a positive way about developing [8] H.W. Hopkins, Speciality fibers and markets, In: A.J.F. Russell (Ed.), Alternative Animals for Fibre

natural animal fiber production in the world and EU Productionited, Commission of the European

countries in recent years and it should be supposed to Communities, Brussels, 1993, p. 5-10. continue increasingly in the near future. For this

[9] Z. Yıldırım, A research on some physical properties and reason, new researches in terms of social and

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Ankara University, 1995

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