Journal of Life Sciences Volume 10 Numbe (1)
J LS
Journal of Life Sciences
Volume 10, Number 1, January 2016 (Serial Number 92)
Contents
Zoology and Botany
1 Teat Length and Lactation Period as A Predisposition Factor of Subclinical Mastitis in Dairy Cattle in Bandung, Indonesia
Herwin Pisestyani, RP Agus Lelana and Yeti Nevia Septiani
7 The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
Mehmet Ufuk Kasim and Rezzan Kasim
16 The Cytochrome b Polymorphism of Meat Lines Rabbits
Vladimir Parkanyi, Lubomir Ondruska and Dusan Vasicek
Ecology
21 Using Magnetic Technique to Increase Efficiency of Organic Pollutants Biodegradation in Wastewater
Khalid Falih Hassan, Suadad Awad Kadhim, Noor Nihad Baqer and Elaaf Safa Al-deen Hassan
33 The Impact of Life Sciences Developments on the UK Urban Environment
Peter Jones, Martin Wynn, David Hillier and Daphne Comfort
Interdisciplinary Researches
39 Serpentine Design on Forest Roads by the Internal Circular Curve Method: A Case Study in Serbia
Bogdan Ž. Stefanovi ć
48 The Role of Homocysteine and Other Clinical Laboratory Markers in Assessing Cardiovascular Risk in Patients on Hemodialysis
Irena Ivanova Gencheva-Angelova, Adelaida Lazarova Ruseva and Pavlina Dimitrova Yordanova-Laleva
54 Epidemiology of ADRs in Internal Medicine, HCC
Alba Delia Campaña S., Anthon Álvarez Arredondo, J. Alfredo Contreras G., S. Saúl Irizar S. and A. Miroslava Flores R.
Journal of Life Sciences 10 (2016) 1-6
doi: 10.17265/1934-7391/2016.01.001
DAVID PUBLISHING
Teat Length and Lactation Period as A Predisposition Factor of Subclinical Mastitis in Dairy Cattle in Bandung, Indonesia
1 2 Herwin Pisestyani 2 , RP Agus Lelana and Yeti Nevia Septiani
1. Veterinary Public Health Division, Department of Animal Health and Veterinary Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor 16680, Indonesia
2. Clinic Division, Department of Clinic Reproduction and Pathology, Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor 16680, Indonesia
Abstract: In order to improve the practical approach to clinical diagnosis of subclinical mastitis, 72 dairy cattle from KPSBU Lembang Bandung Regency was used in this research. The purpose of this research was to study the correlation between udder length and lactation period with the profile of subclinical mastitis. Lactation period data was collected from interviews with farmers. Subclinical mastitis testing by IPB-1 Mastitis Test reagent and udder length measured using measuring tape (cm). Data was analyzed using Duncan test and logistic regression. The highest correlation of subclinical mastitis occurred to dairy cattle with average udder length of 7.5 cm and was in the third and forth lactation period. This result shows that udder length and lactation period of dairy cattle is a predisposing factor for subclinical mastitis. This finding is important to improve the milking management as well as a way to diagnose subclinical mastitis.
Key words: Lactation period, subclinical mastitis, udder length.
1. Introduction the farmers accessibility to obtain a subclinical mastitis diagnosis kit. These conditions are also faced by the
Subclinical mastitis is a disease in dairy cattle that livestock in West Java with business scale of 5.8 heads has not been handled well. Subclinical mastitis is per business unit and production capability of inflammation of the internal tissue of the udder [1]. approximately 11.6 liter/head/day [5]. Therefore a Losses caused by subclinical mastitis are decreased in more practical alternative solution is needed, such as milk production, decreased of milk quality, expensive studying the predisposing factors that can be used as an cost for treatment and medication, early culling of
indicator of subclinical mastitis.
cattle as well as purchasing of new dairy cattle [2]. An attempt to identify subclinical mastitis These threats are faced by 127,211 farmers in predisposing factors among others is to assess udder Indonesia whose cattle population reached 475 health. According to Blakely, J. [6], udder health of thousand heads with 19 million liter of milk production dairy cattle in relation to milk production is affected by [3]. It will be hard for Indonesia to reach the cows’ physical condition, milking habits, body self-sufficiency in milk if it does not pay attention to condition score and udder scoring. Udder scoring these threats are. Indonesia is currently only able to covers teats length, position of the front teat, front fulfill 20-30% of its domestic demand [4]. udder linkage, udder depth, rear udder height, middle One of the problems faced in handling subclinical ligament and position of rear teat [6]. Saragih, B. [7] mastitis lies on the ability to diagnose in the field and stated that other than the anatomical factor of the udder,
cleanliness of the floor, floor surface and hygienic Corresponding author: Herwin Pisestyani, DVM, research
field: veterinary public health. milking of udder also affect udder health. Udder health
2 Teat Length and Lactation Period as A Predisposition Factor of Subclinical Mastitis in Dairy Cattle in Bandung, Indonesia
can be achieved by maintaining personal hygiene, farmer and cow in normal lactation condition. milking hygiene, good shed management, good
Farmers’ information includes: name of farmer, milking management, vaccination program, and village, and number of cow in a shed. Cow administrating antibiotic periodically [8].
information includes: cow number, age and lactation BIF [9] assign a standard that average size teat
period, and amount of milk production per day. length for both front teats of Frisian Holstein (FH) cow
Somatic Cell Count (JSS) testing is done by indirect should be 6 cm in length and 2.9 cm in diameter, while
method. The indirect method used is subclinical the rear has an average size of 5 cm in length and 2.6
mastitis diagnosis using IPB-1 Mastitis Test reagent. cm in diameter. Prihadi, S. [10] stated that Indonesian
Measuring the length of teats is done using tape FH cow is different then it’s origin, it has the ability to
measure expressed in centimeters (cm). produce an average of 10 liter per day with 12-15 month
2.4 Determination of Sample Size and Withdrawal calving interval and lactation period approximately 10
month or an average milk production of 2,500-3,000 Determination of sample size begin with recording liter per lactation. The average age of dairy cows first
the number of dairy cattle in Sukajaya Village based lambing is 2-2.5 years and this is the first lactation
on udder health testing requirements which are; cows period [11]. Lactation period is the period the cow
is in its normal lactation condition. The amounts of produces milk between birth and dry period, therefore
sample size obtained are 72 dairy cows. duration of lactation is between 8-10 months [2].
2.5 Measuring of Teat Length
There have not been many studies of predisposing factors for subclinical mastitis in dairy cattle. Therefore
Teat length is measured using measuring tape (cm). this research is needed to know teat length and lactation
Measuring is performed on each quarter after cow is period as one of the predisposition factor for
bathed.
subclinical mastitis in the livestock in KPSBU
2.6 Subclinical Mastitis Testing
Lembang Bandung Regancy. Subclinical mastitis testing is done by indirect method
2. Material and Methods
using IPB-1 Mastitis Test reagent [12]. Paddle is filled
2.1 Time and Place of Research with 2 mL of milk from a cow that was previously The research is conducted in the month of
bathed, udder cleaned, and then milk is milked from July-December 2012, on the livestock who are
the teat. First and second spray is disposed, and then members of KPBSU Lembang in Sukajaya Village,
the next spray of milk is collected in the paddle. IPB-1 District Lembang, Bandung Regency, West Java.
Mastitis Test reagent is added in the sample with a 1:1 ratio. Sample and reagent is homogenized horizontally
2.2 Tools and Materials for 15-30 s. Test principles of IPB-1 Mastitis Test is Tools used in this research are paddle, measuring
that the reagent will bind with somatic cell nucleus tape (cm), cloth and stationery. Materials used are IPB-1
DNA which will form a think gelatin like mass. A reagent mastitis test and milk sample from each quarter.
thicker mass shows more somatic cell nucleus DNAs that are bind with IPB-1 Mastitis Test reagent.
2.3 Research Method Results interpretation. The reaction is observed
This research is a field study of subclinical mastitis according to Sudarwanto M. [1], if reaction is negative, cases. Data is obtained through research and interview.
milk and reagent stays homogeneous, if positive one it Interview is conducted to obtain information regarding
will form a thin mucous, positive two will form a
Teat Length and Lactation Period as A Predisposition Factor of Subclinical
Mastitis in Dairy Cattle in Bandung, Indonesia
thicker mucous, positive three mucous is very thick
Table 1 Average of teat length in each period of lactation
like a gelatin mass. of dairy cattle in KPSBU Lembang, Bandung Regency.
Teat length (cm) Lactation
2.7 Data Analysis
average min
max 1 4.75 3.75 5.0
Data is analyzed descriptively, Gamma test and 2 5.6 5.0 6.2 logistic regression analysis. Descriptive analysis and
Gamma test is used to see relation or correlation 7.1 7.5 5 7.6 7.2 7.8
between variables in the study and to determine the 6 8 7.5 8.3
association variables that are ordinal [13]. Data is also analyzed by using Microsoft Office Excel 2007 and
Milking with two fingers is milking with the thumb and Duncan test.
forefinger that shifts from the base of the teat down while messaging, loosen and tighten to the top. The
3. Result and Discussion
weakness of milking this way is that the teat and udder
3.1 Influence of Teat Length Towards the Severity of are always wet, this can cause lesion on the udder and Subclinical Mastitis
can slowly change the anatomical shape of the teat to become longer [16]. This method of milking is applied
Udder or cows mammary gland consist of 4 separate on farms in KPSBU Lembang. Udder with weak
parts. The left and right part is separated by a sulcus ligaments will show an udder shape that falls and hangs.
called sulcus intermamaria that runs longitudinally. Weak Ligamentum suspensorium will cause the udder
Aspects that must be observed from the udder are size, to loose shape and loose its fold between quarters.
production capacity, udders’ height from floor surface Weakness of this ligament also causes the cows’ teat to
when cow is standing, and udder symmetry [14].
be prominent and long [15]. This study also shows the Ligamentum suspensorium is the divider from each
influence of teat length towards the degree of severity udder quarter. This ligament functions as a foundation
of subclinical mastitis from KPSBU Lembang dairy for the udder and a divider between quarters. The outer
farm that is presented in Table 2.
ligament is the Ligamentum lateralis [15]. This Data obtain from this study indicates that teat length
ligament functions as a buffer and gives the shape of significantly affects the level of severity of subclinical
the udder. Age can affect the strength of the ligament, mastitis (P < 0.05). Table 2 shows that teat with a 7.5
which separates each quarter of the udder. The higher cm length is more susceptible to subclinical mastitis
the lactation period shows the older the age of the cow. compared to teat with a 4.75 cm length, therefore
Average teat lengths of each lactation period of dairy teat length can be used as a predisposing factor of
cattle in KPSBU Lembang Bandung Regency are
shown in Table 1.
Table 2 Influence of teat length towards the degree of
Results of this study show an increase of teat length severity of subclinical mastitis from KPSBU Lembang
Bandung Regency.
in each lactation period. In the first lactation period teat
Degree of
Teat length (cm)
length is 4.75 cm, second lactation 5.6 cm, third
max lactation 6.5 cm, forth lactation 7.5 cm, fifth lactation
Severity of MSK*
average min
7.6 cm, and sixth lactation 8 cm. Teats that become + 5.75ª 5.5 6 longer is influenced by incorrect ways of milking and
++ 6.5 b 67 +++ 7.5 c 7 8.3
weakness of ligament. Description:*subclinical mastitis; different superscript letter in
Traditional milking by using two fingers (Strip the same column followed by the same letter are not Method) will affect the anatomical shape of the udder.
significantly different at the test level of P < 0.05.
4 Teat Length and Lactation Period as A Predisposition Factor of Subclinical
Mastitis in Dairy Cattle in Bandung, Indonesia
subclinical mastitis. Teat with a 7.5 cm length has a subclinical mastitis in dairy cattle in KPSBU level of subclinical mastitis severity of positive 3 (+++),
Lembang is presented in Figure 1. teat with a 6.5 cm has a level of subclinical mastitis
This study shows that lactation period significantly severity of positive 2 (++), and teat with a 5.75 cm has
affects the degree of severity of subclinical mastitis (P
a level of subclinical mastitis severity of positive 1 (+). < 0.05), meaning that the higher the lactation period Teat condition that does not have subclinical mastitis is
will increase the degree of severity of subclinical shown by teat that has a length of 4.75 cm. According
mastitis. Base on Figure 1 it appears that the incidence to the result of the study, farmers can use teat length as
of subclinical mastitis with different degree of severity
a diagnosis indicator for subclinical mastitis that is has increased on each lactation period. For cows in its dairy cattle with teat length of 6.5-7.5 cm may have
first lactation, age between 2.5-3 years and its first time subclinical mastitis.
giving birth, shows that the presentation of subclinical BIF [9] stated that subclinical mastitis cases occur in
mastitis incidence with a degree of severity ranging teats with a length category of 7-9 cm. Subronto [2] stated
between 0.7%-1.7%. Incidence of subclinical mastitis udders that hangs tremendously or udders that are long
with a positive three degree of severity continues to and have large teat opening is a predispose factor to
increase until its peak that is at the forth lactation udder inflammation in terms of the cattle. Long teats
period as high as 8.3%. This is likely due to cows that will make it easy for microbes to enter the internal
previously experienced subclinical mastitis with a tissue of the udder from the outside of the teat through
degree of severity of 1 and 2 whom later became worse the teat hole (ascendens) and lesions on the udder due
and became a positive 3. In cows that have reached the to friction with the floor [16]. Teat size that are to short
fifth and sixth lactation period with age range of 8-9 is also a predispose factor for subclinical mastitis. This
years, it is seen that the degree of severity of subclinical is because farmers have difficulty in milking therefore
mastitis of positive three experience a decrease. Cows the alveoli cannot secrete milk completely.
that initially experienced subclinical mastitis but because they were not handled well became a clinical
3.2 Influence of Lactation Period towards Severity mastitis case may have caused this. The decrease of Level of Subclinical Mastitis percentage of positive 3 degree of severity of subclinical
Mastitis is an inflammation of the udder, whether it mastitis can also be due to healing process because of is acute, sub acute or chronic, with the increase of
somatic cells in the milk and physical as well as the composition of milk, with or without pathological changes in the gland [2]. 95-98% of mastitis incidence is subclinical mastitis, while 2-3% incidence of clinical mastitis is detected [1]. Losess caused by subclinical mastitis are decreased in milk production, decreased of milk quality, expensive cost for treatment and medication, early culling of cattle as well as purchasing of new dairy cattle [2].
Length of the teat, ligament linkage, feed, weather, lactation period and genetic quality of the cow can
Fig. 1 Influence of lactation period towards the degree of
influence subclinical mastitis [17]. The influence of
severity of subclinical mastitis in KPSBU Lembang
lactation period towards the degree of severity of
Bandung Regency.
Teat Length and Lactation Period as A Predisposition Factor of Subclinical
Mastitis in Dairy Cattle in Bandung, Indonesia
good handling. This study is consistent to [18] study up 70% per head [1]. In the practices of dairy that stated that subclinical mastitis incidence will
farming, it is known that the correlation between increase until cow reaches the age of around eight years
lactation periods, teat length with subclinical and will continue to increase from the first lactation
mastitis cannot be separated from the affects of early until its peak that is in the third and forth lactation
selection [19]. Correlation between teat length and period which will then decrease according to the next
lactation period with subclinical mastitis incidence of lactation period.
dairy farms in KPSBU Lembang is presented in Increase of subclinical mastitis incidence may occur
Figure 2.
because the older the cow becomes the udders main Figure 2 shows a linear correlation between teat supporting tissue that is Ligamentum suspensatorium
length and lactation period towards degree of incidence lateral et medial weakens and nutrition that is given to
of subclinical mastitis in KPSBU Lembang. This the cow lacks guarantee which can lead to clinical
means there is a significant correlation between teat mastitis incidence [15]. The increase of lactation period
length and lactation period towards degree of incidence will cause the time needed for teat cistern to close
of subclinical mastitis. Higher lactation period and longer which will facilitate the entrance of longer teats will increase the percentage of subclinical microorganisms from outside into the udder. Age
mastitis incidence. In the first lactation period, cows factor and degree of milk production of cow also
with teat length of 4.75 cm experience subclinical affects the incidence of mastitis. The older the cow and
mastitis incidence as high as 45%. Percentage of the higher its milk production, the more loose the teat
subclinical mastitis incidence as high as 48% occur in sphincter is. Teat with a loose sphincter will enable the
cows with teat length of 6.5 cm and has entered the cow to be infected with microorganism. The more a
third lactation period. Cows that has entered the fifth cow produces milk; therefore the longer time is needed
and sixth lactation period with teat length of 7.6 cm and for the sphincter to close completely [2].
8 cm has a 60% and 70% chance of being affected by subclinical mastitis. From this study it is obtained that
3.3 Correlation between Teat Length and Lactation teat length and lactation period can be used as Period with Degree of Incidence of Subclinical diagnosis indicator of subclinical mastitis and can also Mastitis
be used as a predisposed factor from subclinical Subclinical mastitis causes loss milk production
mastitis incidence.
Fig. 2 Correlation between lactation period and teat length with subclinical mastitis incidence in KPSBU Lembang Bandung Regency (r = 0.972, P < 0.05).
6 Teat Length and Lactation Period as A Predisposition Factor of Subclinical
Mastitis in Dairy Cattle in Bandung, Indonesia
This study is consistent with [20] that stated higher ke-4. Terjemahan: Srigandono B. Yogyakarta (ID): UGM Pr.
lactation period and longer teat length will cause the [7] Saragih, B. 2000. Kumpulan Pemikiran: Agribisnis
degree of incidence of subclinical mastitis to increase. Berbasis Peternakan. Bogor (ID): Pustaka Wirausaha This is due to weakness of ligament in maintaining the
Muda.
anatomical shape and the closing of the teat sphincter [8] Alluwaimi, A. M. 2004. The Cytokines of Bovine which need longer time. Sudarwanto M. [1] stated that Mammary Gland: Prospect for Diagnosis and Therapy. Research in Veterinary Science (in Pr).
time needed for the teat sphincter to close or restore of [9] BIF (Beef Improvement Federation). 2011. BIF teat width after milking is 10-15 sec. The more time the
Guidelines for Unifrom Beef Improvement Program. 8th sphincter needs to close, causes more amounts of
ed. Beef Improvement Federation. Athens (GR): Georgia microorganisms form the environment to enter the Univ. [10] Prihadi, S. 1997. Dasar Ilmu Ternak Perah. Yogyakarta
udder. One of the preventive measures to suppress the (ID): Fakultas Peternakan Universitas Gajah Mada. incidence of subclinical mastitis is teat dipping. Teat
[11] Sudono, A. 1999. Ilmu Produksi Ternak Perah. Diktat dipping is dipping of teats after milking into a liquid
Kuliah Fakultas Peternakan IPB, Bogor. Jurusan that contains disinfectant. Disinfectant used is Ilmu-ilmu Sosial Ekonomi Pertanian. Bogor (ID):
Fakultas Pertanian IPB.
iodophore with a 1% iodine concentration [15]. [12] Sudarwanto M. 1998. Pereaksi IPB-1 sebagai pereaksi
alternatif untuk mendeteksi mastitis subklinis. Bogor
4. Conclusions
(ID): Fakultas Kedokteran Hewan Institut Pertanian Lactation period and length of udder is a Bogor. [13] Agresti, A., and Finlay, B. 2009. Statistical Methods for
predisposing factor towards the level of incidence of the Social Science. New Jersey (US): Pearson E. subclinical mastitis in dairy cattle. Highest incidence
[14] Farmer, W. S., and Chrestman, G. 2006. Dairy Cattle of clinical mastitis occurred to dairy cattle with
Judging. Mississipi (US): Mississipi State University. average teat length of 7.5 cm and was on the third and [15] Falvey, L., and Chantalakhana, C. 1999. Smallholder Dairying in the Tropics. Bibliography: Milking, chapter
forth lactation period. 16. Nairobi. Kenya (KE): Internasional Livestock
Research Institute.
References
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Purnawarman, T., Latif, H., Soejoedono, R. R., and melalui program mastitis subklinis. Orasi Ilmiah. Bogor,
Pisestyani, H. 2009. Higiene Pangan. Bagian Kesehatan 22 Mei 1999.
Masyarakat Veteriner Departemen Ilmu Penyakit Hewan [2] Subronto. 2003. Ilmu Penyakit Ternak 1. Yogyakarta (ID):
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[3] DSSP (Data Sementara Statistik Peternakan). 2009. [17] Ikawati, A. 2011. Analisis kandungan protein dan lemak Produksi Susu Sapi Perah. [Internet]. [diacu 2013 April
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perah di Wonocolo Surabaya [skripsi]. Surabaya (ID): tabel=1&id_subyek=36.
Fakultas Kedokteran Hewan Universitas Airlangga. [4] Luthan, F. 2011. Pengembangan Agribisnis Persusuan di
[18] Siregar, S. 1995. Jenis Teknik Pemeliharaan dan Analisa Indonesia. Sumatera Barat (ID): Universitas Andalas.
Usaha Sapi Perah. Jakarta (ID): Penebar Swadaya. [5] DSSP (Data Sementara Statistik Peternakan). 2008.
[19] Lush, S. 1990. “Jenetic Progress trough Selection in a Produksi Susu Sapi Perah. [Internet]. [diacu 2013 April
Closed Herd of Indian Cattle.” J. Dairy Sci. 51: 1059-64. 17]. Tersedia dari: http://faterna.unand.ac.id/index.php?
[20] Sudono, A., Rosdiana, R. F., and Setiawan, B. S. 2003. tabel=1&id_subyek=36.
Beternak Sapi Perah Secara Intensif. Jakarta (ID): [6] Blakely, J., and Bade, D. H. 1991. Ilmu Peternakan. Ed
Agromedia Pustaka.
Journal of Life Sciences 10 (2016) 7-15
doi: 10.17265/1934-7391/2016.01.002
DAVID PUBLISHING
The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
Mehmet Ufuk Kasim and Rezzan Kasim Kocaeli University, Graduate School of Natural and Applied Sciences, Vocational School of Arslanbey, Kocaeli 41285,
Kartepe-Kocaeli/Turkey
Abstract: This study aimed to evaluate the effect of calcium and ascorbic acid treatments on the quality of carrot shreds during storage. Towards this aim, carrot shreds were dipped into a 5 L solution of 2 g/L ascorbic acid containing 1%, 3%, or 5% CaCl 2 (Ca + AA) for 3 min at room temperature (~20 °C). In case of the control group (control, C), samples were dipped into distilled water for the same time interval. Subsequent to treatment, carrot shreds were stored in a cold room at 5 ± 1 °C, 85-90% RH for a period of 11 days. Color values (L*, a* b*), whiteness index, saturation index, hue angle values, visual quality, firmness scores, bitterness scores, total soluble solids (TSS) and electrolyte leakage measurements were conducted at various sampling dates. The results from this study demonstrated that brightness of carrot shreds was augmented by calcium and ascorbic acid treatments irrespective of the dosage used. Whiteness index values for the 5% Ca + AA treated samples were observed to be low whereas saturation indices of 5% Ca + AA and 3% Ca + AA treated carrot shreds were higher as compared to other treatments. This study concludes that treatment with calcium at high doses improves the color quality of carrot shreds under storage conditions. Visual quality and firmness of carrot shreds was maintained till day 4 of storage, thereafter it declined as compared to the control group. Bitterness of carrot shreds was also observed to increase upon treatment with calcium and ascorbic acid. However, calcium treatment of the test carrot shreds was seen to decrease weight loss and cause an increase in the TSS under storage conditions.
Key words: Calcium, ascorbic acid, color, bitterness, quality.
1. Introduction fresh-cut vegetables is minimal processing such that the produce retains fresh-like texture, color, flavor,
Fresh-cut vegetables are vegetables that are and safe-to-use quality. However, injuries that occur available in a ready-to-use format. They are during processes such as peeling, slicing, cutting, minimally-processed plant products that are peeled, shredding, etc. result in stress at the tissue cellular, trimmed and/or cut prior to being packaged in a subcellular and biochemical levels leading to several way that retains freshness whilst being convenient undesirable changes in the vegetables during the to the end user. Lettuce and pre-prepared salads are course of storage and transportation [2]. the most common types of fresh-cut vegetables In the case of fresh-cut carrots, the most significant available commercially, although fresh-cut carrots, problem faced is surface whitening. It is a tomatoes, broccoli, cauliflower, and cabbage can also phenomenon that arises as a result of dehydration and
be found [1]. lignin synthesis. Several treatments, such as In recent years, Turkey has witnessed an increase in application of edible coatings [3], treatment with citric the demand and availability of fresh-cut vegetables as acid [4, 5] or ascorbic acid [6] are available to prevent well as fruits; the examples include pre-washed and
the whitening.
trimmed spinach, sliced carrots, leeks, apples, etc. Results from previous studies have indicated that The basic premise for obtaining high quality treatment of carrot shreds with ascorbic acid is
successful in preventing the appearance of surface Corresponding author: Rezzan Kasim, Dr., associate
whitening. However, as this treatment results in professor, research field: postharvest technology of vegetables.
8 The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
softening of the shreds, the application of a firming containing 2 g/L ascorbic acid for 3 min. agent has been suggested for maintaining the crispness
(4) C: The control group samples were dipped in [6]. Calcium treatments that use either calcium
distilled water for 3 min.
chloride (CaCl 2 ) or calcium lactate have been shown All treatments were carried out at room temperature to be effective in maintaining the firmness of several
(~20 °C). Treated carrot shreds were dried by first fresh-cut fruits and vegetables during storage [7]. It is
using a salad spinner (2 min, room temperature) so as also known that treatment with Ca 2+ has the potential
to remove excessive surface solution and then at room to maintain the textural qualities of carrot for as long
temperature (15 min).
as up to 10 days of storage [2]. As softening and other
2.2 Packaging and Storage Condition undesirable textural changes in fresh-cut products are
related to their tissue calcium levels, application of The samples of shredded carrots (100 g) were calcium salts (calcium-chloride, -carbonate, -lactate,
placed in covered plastic boxes 110 × 110 × 50 mm in -propionate, -pectate, etc.) to fruits and vegetables,
size. Triplicates of each treatment were stored for 11 such as pears, strawberries, kiwis, shredded carrot,
days at 5 ± 1 °C with relative humidity of 85-90%. honeydew melon discs, nectarines, peaches and
2.3 Color Measurements
melons, helps in retaining tissue firmness [8]. Calcium, in a 1% CaCl 2 formulation, and ascorbic acid dips
Color measurements (L*, a* and b* values) were have been employed as firming agents that aid in
performed using a chroma meter CR-400 (Konica extending the postharvest shelf life of sliced pears and
Minolta Inc. Osaka, Japan) with an illuminant D65 strawberries that have been stored in a controlled
with 8 mm aperture. The instrument was calibrated atmosphere [9].
with a white reference tile (L* = 97.52, a* = -5.06, b* The objective of this study was to determine the
= 3.57) prior to measurements. The L* (0 = black, 100 effect of calcium chloride and ascorbic acid treatments
= white), a* (+red, green) and b* (+yellow, –blue) on the quality parameters of shredded carrots.
color coordinates were determined as per the CIELAB
2. Materials and Methods
coordinate color space system.
Whiteness index [WI, Eq. (1)], saturation index [SI,
2.1 Plant Material and Sample Preparation Eq. (2)] and hue angle [H, Eq. (3)] were calculated Carrots were obtained from the Kocaeli Wholesale
using L*, a* and b* values that were computed as Distribution Center. They were transported described below; these values were used to compare immediately to the laboratory, thoroughly washed,
the color changes of the test samples with that of the peeled, trimmed of tap root and stem plate prior to
control (fresh-cut carrot shreds) [10]. preparation. A grate was used to prepare carrot shreds
WI 100 100 L a b (1) (about 5 mm wide, 40 long, and 2 mm thick).
(2) Processed carrots (100 g for each replicate) were
(3) dipped into the following calcium and ascorbic acid
arctan
2.4 Visual Quality, Firmness and Bitterness solutions:
Assessments (1) 1% Ca + AA: 5 L solution of 1% CaCl 2
containing 2 g/L ascorbic acid for 3 min. Visual quality was evaluated by grading the freshness, (2) 3% Ca + AA: 5 L solution of 3% CaCl 2 appearance, color, uniformity and brightness of the containing 2 g/L ascorbic acid for 3 min.
test samples on a five-point Likert scale: 5, excellent; (3) 5% Ca + AA: 5 L solution of 5% CaCl 2 4, good quality; 3, fair quality; 2, poor quality; 1,
The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
extremely poor quality.
2.8 Statistical Analysis
Firmness of the carrot shreds was scored as a Experiments were conducted in a completely subjective variable; the perceived hardness or softness randomized design with a minimum of three experienced when carrot shreds were taken between replications per treatment per sampling date. The two fingers and pressure was applied and was graded resultant data were analyzed by application of the on a five-point Likert scale: 5, very firm; 4, firm; 3, ANOVA test and differences between mean values partially firm/soft; 2, soft; 1, very soft (not usable). were determined using Duncan’s multiple range test. Bitterness of the carrot shreds was also scored on a The results were regarded as significant when P < five-point Likert scale: 5, no bitterness; 4, slightly
0.05 and P < 0.001.
bitter; 3 bitter; 2, very bitter; 1, extremely bitter (not consumable).
3. Results and Discussions
The judging panel for sensory evaluation was
3.1 L* values and Whiteness Index composed of nine food-science students enrolled at
the university. All the students had prior classroom L* values of treated carrot shreds were observed to training and experience in the sensory evaluation of
have increased irrespective of the type of treatment food items.
applied. The highest value was observed on day 4 for shreds treated with 5% Ca + AA (59.757), which was
2.5 Electrolyte Leakage Measurement followed by those treated with 3% Ca + AA (57.790),
Electrolyte leakage (EL) was measured in the test 1% Ca + AA (57.003) and C (control, 56.287). The carrot shreds. Distilled water was used for washing as
difference between the treatments was statistically well as immersion of test sample shreds and
significant (P < 0.05). Post day 4, L* values of conductivity was measured 2 h after immersion. Total
samples were observed to be changing whilst in electrolyte conductivity of the carrot shreds was
storage: Initially a decrease was observed (day 8) measured after they had been frozen and thawed. EL
subsequent to which L* values started increasing was calculated as percentage of the conductivity after
again (Fig. 1).
2 h [11]. The whiteness index of carrot shreds stored after treatment with 3% Ca + AA and 5% Ca + AA was
2.6 Loss of Weight significantly lower than that of shreds treated with 1%
The weight of the triplicate samples was recorded Ca + AA as well as the control samples (P < 0.05). on the day of harvest and after the designated
Previous studies have determined that treatment with sampling dates. The loss in weight was calculated
ascorbic acid has a positive effect on the brightness of using the following formula:
carrot shreds [6] as well as carrot cubes [10]. weight loss (%) = (W i –W s /W i ) × 100; where W i =
Congruent with these findings, the present study initial weight; W s = weight at sampling period.
observed that during the first four days of storage, treatments that combined ascorbic acid with CaCl 2 (at
2.7 Total Soluble Solids (TSS) all doses) were observed to increase the brightness of
For each of the test replicates, TSS was determined carrot shreds as compared to the control group. for two parallel using an Atago DR-A1 digital
Although, L* values of samples were observed to be refractometer (Atago Co. Ltd., Japan). The experiment
decreasing by day 8, brightness of carrot shreds were was conducted at 20 °C and the results were expressed
maintained when treated with 3% Ca + AA as well as as percent value.
5% Ca + AA. Therefore, it can be concluded that
10 The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
Storage Duration (Days)
Fig. 1 L* and whiteness index (WI) values of carrot shreds. 1%: 1% CaCl 2 and 2 ppm ascorbic acid (AA); 3%: 3% CaCl 2
and 2 ppm AA; 5%: 5% CaCl 2 and 2 ppm AA; C: Control.
treating carrot shreds with a combination of calcium day 1 and day 11 of storage. During storage, it was chloride and ascorbic acid was effective in seen that treatment with a combination of ascorbic maintaining their brightness.
acid and calcium, irrespective of dose, resulted in a Previously Rico et al., (2007) have reported that in
reduction in h* values. This conclusion was arrived at instances of a colorimeter being used to analyze color,
because significantly higher values were found in increases in luminosity can be correlated with the
control samples compared to those that were treated development of whiteness in the test samples. In this
with Ca + AA (P < 0.05). Saturation index (SI) values study, however, the whiteness index (WI, Fig. 1)
of samples subjected to 3% Ca + AA and 5% Ca + values of high calcium treated samples (3% and 5%
AA were higher than those treated with 1% Ca + AA Ca + AA) were lower than those of control and 1% Ca
and control (Fig. 2; P < 0.05). On day 4, the highest + AA treatments. Therefore, on account of the
value of SI (53.756) was found in fruits treated with decrease in WI values of the samples, it can be
3% Ca + AA. By contrast, the lowest value was concluded that treatment with a high dose of calcium
observed in carrot shreds treated with 1% Ca + AA prevented the whitening of samples. Similar results
(44.303). However, from 4th day of storage till the were reported for ascorbic acid treatments of carrots
conclusion of the study, carrot shreds in the control [6, 10]. Interestingly, in case of carrot shreds calcium
group had the lowest measures of SI values. treatments alone did not impact white tissue formation
The main physiological effect of fresh-cut
and WI values were observed to increase [2]. processing in the case of carrots is surface whitening However, ascorbic acid alone was effective in
which results from a combination of dehydration and inhibiting white color formation on surface of carrots
lignin formation. This leads to significant loss of [6]. Therefore, as a result of this study, it can be
quality. In this study, the orange color of carrot shreds concluded that the combined use of calcium and
was observed to be maintained by treatment with ascorbic acid enhances the color quality and also
calcium and ascorbic acid. This was especially true prevents whitening of carrot shreds. Similar results
when samples of the control group were compared were found [12] in case of nectarine halves.
with those that received high dose treatments. In the case of minimally processed cabbage, results clearly
3.2 Hue Angle and Saturation Index Values demonstrated that while treatment with ascorbic acid
Fig. 2 shows the hue angle (h*) values measured at did not lead to significant differences between test and
The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
tion inde 48 a
hue angle 59
0 4 8 11 0 4 8 11 Storage Duration (Days)
Fig. 2 Hue angle (h*) and saturation index (SI) values of carrot shreds. 1%: 1% CaCl 2 and 2 ppm ascorbic acid (AA); 3%:
3% CaCl 2 and 2 ppm AA; 5%: 5% CaCl 2 and 2 ppm AA; C: Control.
control samples with respect to color or general + AA and 5% Ca + AA (Fig. 3). Also, statistically
significant differences amongst the treatments were resulted in consistent maintenance of high quality with
appearance, treatment with 2% CaCl 2 at 20 °C
observed at day 4 and 11 of storage (P < 0.05 and P < less intense browning and the best general appearance
0.001). Fresh-cut vegetables that maintain a firm, [13]. These results were also confirmed in the present
crunchy texture are highly desirable because study.
consumers associate such textures with freshness and wholesomeness of produce [14]. The development of
3.3 Visual Quality, Firmness and Bitterness Scores of such undesirable textural changes in minimally Carrot Shreds processed products can be reduced by the application
Visual quality scores of test samples, regardless of of calcium salts (calcium-chloride, -carbonate, -lactate, type of treatment, were observed to be decreased by
-propionate, -pectate, etc) because the rate of softening day 4. Subsequently, scores increased until the 8th day
was directly related to the reduction of calcium levels of storage after which they continued decreasing till
in fruit tissues [11]. Studies have shown that application the end of the storage (Fig. 3). However, the
of Ca salts to pears, strawberries, kiwifruits, shredded appearance of samples treated with calcium and
carrots, honeydew discs, nectarines, peaches and melons ascorbic acid was superior as compared to control
helps in retaining tissue firmness [15]. Firmness scores during the first eight days of storage with the
of treated carrot shreds across all Ca + AA doses were difference between treatments assuming statistical
lower than that obtained for control samples during significance at day 4 (P < 0.05). Therefore it can be
storage, and differences amongst the different
said that as compared to the control samples, the treatments were observed to be statistically significant visual quality scores of test carrot shreds were highest
at day 4 (P < 0.05) and day 11 (P < 0.001). Also, during the first 4 days of the storage, but subsequent
results clearly showed that the firmness scores of to that the effectiveness of the treatments decreased
samples treated with 1% CaCl 2 were higher than that such that by the end of the storage period the visual
of other calcium treatments leading to the conclusion quality scores of the Ca + AA treated samples were
that CaCl 2 treatments were not effective in improving much below that of the control group.
the texture of carrot shreds during storage. But, as per According to the firmness scores, the texture of the
the weight loss results obtained in this study (Fig. 4), shredded carrots was retained best in the control group
the recorded weight loss of the 3% Ca + AA and 5% followed by samples treated with 1% Ca + AA, 3% Ca
Ca + AA treatments were lower than those obtained
12 The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
Storage Duration (Days)
C ittern 2 K
Storage Duration (Days) Storage Duration (Days)
Fig. 3 Visual quality, firmness and bitterness scores of carrot shreds. 1%: 1% CaCl 2 and 2 ppm ascorbic acid (AA); 3%: 3%
CaCl 2 and 2 ppm AA; 5%: 5% CaCl 2 and 2 ppm AA; C: Control.
for 1% Ca + AA and control. Hence, the high firmness present study corroborate the above mentioned results. values of samples in control and 1% Ca + AA
3.4 Weight Loss
treatments can be potentially explained as a byproduct of water loss.
Weight loss of all the treated samples was observed
Bitterness of carrot shreds increased upon to increase during storage (Fig. 4). The highest weight increasing CaCl
2 dose (Fig.3) with the highest (least
bitter) values being obtained by 1% Ca + AA
C 0.5 1%
treatment (4.33) followed by 3% Ca + AA (2.0) and 5%
Ca + AA as recorded on day 4, and these results
continued during the storage. Differences amongst the
treatments were statistically significant at the level of
P < 0.001 (4 and 8 days of storage) and P < 0.05 (11th
Weight losses
day of storage). Studies [16] determined that 0.1
exogenous administration of CaCl 2 in form of a
0 4 8 solution can reduce browning as well flesh softening 11
Storage Duration (Days)
in case of zucchini squash slices. However, CaCl 2 ,
Fig. 4 Weight losses of carrot shreds. 1%: 1% CaCl 2 and
when used in high concentrations (> 0.5%), has been
2 ppm ascorbic acid (AA); 3%: 3% CaCl 2 and 2 ppm AA;
known to cause a detectable off-flavor. The results of
5%: 5% CaCl 2 and 2 ppm AA; C: Control.
The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
loss was observed in the control group (0.18 and 0.29)
and followed by the 1% Ca + AA (0.10 and 0.26), 5%
Ca + AA (0.09 and 0.18) and 3% Ca + AA (0.008 and
0.14) groups as noted on day 4 and 8. Also,
statistically significant differences were observed
between the various treatment groups while in storage.
TSS
Therefore it can be concluded that CaCl 2 —ascorbic
acid treatments have a significant effect on weight loss
especially at higher doses.
Peel or skin is a very important barrier against 0 4 8 11 desiccation and loss of turgor. Several fruits and
Storage Duration (Days)
vegetables have a protective waxy coating that makes
Fig. 5 Total soluble solids (TSS) of carrot shreds. 1%: 1%
them highly resistant to water loss. Mechanical injury
CaCl 2 and 2 ppm ascorbic acid (AA); 3%: 3% CaCl 2 and 2
ppm AA; 5%: 5% CaCl 2 to the skin brought about by peeling, cutting, slicing, and 2 ppm AA; C: Control.
shredding, etc. makes fresh cut products highly ranged from 3% to 7%. Subsequent to day 4, TSS of susceptible to weight loss as the protective peel is no
the control group was observed to steadily decrease longer intact [11, 15]. In the present study, water lost
till the end of the storage period whereas in case of the by carrot shreds was reduced when treated with a
Ca treated carrots it was observed to increase. It was combination of calcium and ascorbic acid. Izumi and
also seen that TSS of samples treated with a Watada [2] have previously reported that Ca has no
combination of Ca and AA was higher than that of observable effect on weight loss in case of carrot
control and that the higher values were observed in slices and sticks but is effective in preventing the
correlation with high doses of CaCl 2 treatment during same in case of carrot shreds. Their results also proved
storage. Differences in TSS values amongst the that carrot shreds have almost two and three times
various treatments were statistically significant (P < more Ca content as compared to sticks and slices
respectively. Additionally, Ca has widely been The edible portion of carrot contains about 10% reported to play an important role in preserving the
carbohydrate with the soluble carbohydrate structural integrity and mechanical strength of cell
composition ranging from 6.6 to 7.7 g per 100 g [14]. walls [9]. The basis for the reduced weight loss
In the present study, the initial TSS content of carrot observed in case of carrots shreds treated with Ca +
shreds was observed to be 8%. This value was seen to AA in the present study, can be accounted for by the
decrease across treatments under storage conditions. Ca absorbed by the samples under test.
Interestingly, the maximum decrease in TSS values was observed in case of the control group where
3.5 Total Soluble Solids values fell from 1% on day 4 to below 1% on days 8
Fig. 5 shows total soluble solid (TSS) values for and 11. In contrast, TSS of calcium treated carrot shreds carrot shreds subjected to different treatments. TSS of
maintained constant high values especially in the case carrot shreds was observed to be decreased on day 4
of shreds treated with 5% calcium where values but this decrease was higher in the control group as
ranged from 7.1% to 7.7%. TSS of samples in 1% Ca compared to the calcium and ascorbic acid treated
+ AA and 3% Ca + AA treatment groups was also samples. In quantitative terms, TSS of the control
found to be high as compared to the control. Therefore group was 1.2%; for the Ca treated carrot shreds it
it can be concluded that calcium and ascorbic acid
The Effects of Calcium Chloride and Ascorbic Acid Treatment on Ready-to-use Carrot Shreds
14
Fig. 6 Electrolyte leakage of carrot shreds. 1%: 1% CaCl 2
and 2 ppm ascorbic acid (AA); 3%: 3% CaCl 2 and 2 ppm
AA; 5%: 5% CaCl 2 and 2 ppm AA; C: Control.
treatments prevent loss of TSS in case of carrot shreds especially when used in high doses.
3.6 Electrolyte Leakage On day 4, it was observed that electrolyte leakage
(EL) from carrot shreds in the control group and 1% Ca + AA treated group decreased, whereas in the 3% Ca + AA and 5% Ca + AA treated samples it was increased (Fig. 6). The EL values of the control samples continued to decline but an increase was noted in the 1% Ca + AA and 3% Ca + AA treated groups. Moreover, it showed variations between decrease and increase in 3% Ca + AA treatments during the storage. Differences in EL values among the treatments were statistically significant (P < 0.001). Leakage of electrolytes or cellular content is commonly used as an index for evaluating changes in membrane integrity arising due to ripening, stress damage or mechanical injury [7]. Electrolyte leakage is considered as an indirect measure of plant cell membrane damage [17]. In the present study, EL values for calcium and ascorbic acid treated samples were observed to be higher than that of the control
group. Therefore, it can be concluded that CaCl 2
treatments have no membrane stabilizing effect conferred by exogenous calcium ions.
4. Conclusions
This study aimed to determine the impact of
calcium and ascorbic acid on the quality of carrot shreds during storage. For this purpose, carrots were grated and treated with solutions containing varying doses of calcium along with 2 g/L ascorbic acid. The carrots were then stored for 11 days in a cold room at
5 ± 1 °C and 85-90% RH. According to the results obtained from this study, calcium was found to improve color quality and brightness while decreasing the development of whiteness on carrot shreds. As a cautionary note, however, it was observed that calcium, especially at higher doses, could cause bitterness of carrot shreds. Weight losses of carrot shreds treated with calcium and ascorbic acid was found to be higher than that of the control group; also firmness values of these test samples were low compared with the control. While calcium treatment was observed to improve the visual quality of the produce during the first eight days of storage, it was found to lose its efficacy after that. In addition, the calcium treatments showed no membrane stabilizing effect.
References
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[2] Izumi, H., and Watada, A. E. 1994. “Calcium Treatments
Affect Storage Quality of Shredded Carrots.” Journal of Food Science 59 (1): 106-9.
[3] Sargent, S. A., Brecht, J. K., Zoelner, J. J., Baldwin, E. A.,
and Campbell, C. A. 1994. “Edible Films Reduce Surface Drying of Peeled Carrots.” Proc. Fla. State Hort. Soc. 107: 245-7.
[4] Bolin, H. R., and Huxsoll, C. C. 1991. “Control of