Characteristics of Alcaligenes sp. LS2T Heterotrophic and Aerobic Ammonium Removal for Potential Livestock’s Wastewater Treatment - repository civitas UGM
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A.11.2 CULTIVATION AND CONSERVATION OF INDONESIAN NATIVE FISH (Rasbora lateristriata)
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•.····• .•.·····• .······• .·73
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A.I.1INTRA-SPECIFIC VARIATION OF THE M ITOCHONDRIAL
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KALOLO
LESSON 1831 (PISCES: GOBIIDAE) FROM BOGOW ONTO
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A.1.2 A SIM PLE OPTIM ISED DNA EXTRACTION M ETHOD FOR M ACROALGAE FROM COASTS OF GUNUNGKIDUL,
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76
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A.1.4INDONESIAN
SUSTAINABLE FISHERIES M ANAGEM ENT
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REGIONALIZATION
OF ASEAN
ECONOM IC
A B S T R A C T S
COM M UNITY
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·····························
•.····•.··78
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B.I.1 DIFFERENCES IN THE REARING SYSTEM TO BALI CADLE GASTROINTESTINAL
IN PRAFI DISTRICT M ANOKW ARI REGENCY W EST PAPUA PROVINCE
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INFESTATION
·····························
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B.1.2 TRYPANOSOM IASIS
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PRODUCTION IN INDONESiA···· •.····•.······················
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B.I.3 DETECTION THE PRESENCE OF BOVINE VIRAL DIARRHEA VIRUS PERSISTENT INFECTION
IN LOW REPRODUCTIVE
PERFORM ANCE DAIRY CADLE HERD IN CENTRAL JAVA, INDONESiA································································
•.··•.•.•.•.· ·•.·•.··82
B.l1.1FASCIOLOSIS IN CENTRAL JAVA: A CONTINUOUS THREAT ....................•.........•.........................················································83
B.II.2DETECTION OF TREM BOLONE ACETATE IN BEEF M USCLE AND LIVER USING HIGH PERFORM ANCE LIQUID
CHROM ATOGRAPHY
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B.l1.3 ANTIBIOTIC RESISTANCE IN ESCHERICHIA COLI ISOLATED FROM BROILER ASSOCIATED W ITH
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•.•.·····• .·•.··•.·86
B.l1.5RAPID DETECTION OF Verocytotoxigenic Escherichia coli (VTEC) BACTERIA BY POLYM ERASE
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B.II.6 INVESTIGATION OF LEAD CONTAM INATION
IN BADUNG RIVER USING FISHES AS BIOINDICA TORS ...•..................... 88
8.11.7 HAEM OPROTOZOA
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F .l11,2M A N A G E M E N T A N D R E P R O D U C T IV E
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··
260
261
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· ·..· 264
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265
266
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·
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·..· 267
268
A CASE STUDY OF
·
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A D O P T IO N O N IN T E G R A T E D P A LM O IL A N D C A T T LE F A R M IN G IN
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IN S T R U M E N T
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· ·
··
·
F .V .10 S U M M A R y
· 259
IN IN D O N E S IA
F .V .6 T H E P E R F O R M A N C E O F M ILK P R O D U C T IO N , T O T A L M ILK R E V E N U E A N D R E P R O D U C T IO N IN D IC A T O R S
D A IR Y S M A LLH O LD E R S IN D A E R A H IS T IM E W A Y O G Y A K A R T A A N D E A S T JA V A P R O V IN C E , IN D O N E S IA ·
F .V .7 P E R C E P T IO N O F B E E F C A T T LE F A R M E R S T O W A R D C R O P W A S T E A S A N IM A L F E E D
F .V .9 T H E R A T E O F T E C H N O LO G Y
JA M B I P R O V iN C E ..· ·..·..·
257
B U C K F E D R U M IN A LLY
A N D S O C IO E C O N O M IC
·
·
·
··
AN APPROACH
· ··
MODEL
GOAT
D IS T R IC T , B R E B E S R E G E N C Y ,
F .V .3 A N A LY S IS O F T H E IM P LE M E N T A T IO N
OF ETT AW A CROSSBRED GOAT M ANURE
K U LO N P R O G O Y O G Y A K A R T A IN D O N E S IA
F .V .s D E V E LO P M E N T O F M IC R O F IN A N C E IN S T IT U T IO N S
F A R M IN G S U S T A IN A B ILIT Y : A P ILO T S T U D Y
· · · 254
C A T T LE K E P T B Y
B R E E D IN G C E N T E R IN
·
F .lII.7 R E P R O D U C T IV E P E R F O R M A N C E O F JA B R E S C O W A T K E T A N G G U N A N
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F .V .2 M A R K E T S E G M E N T A T IO N O F G O A T M ILK C O N S U M E R S :
B E H A V IO R A L V A R IA B LE S ..·..· ·
·· ·
• ·..•..· · · ·
·
KACANG
···
sp. LS 2T
258
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TOW ARDS
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·
·
·
·
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P U R S LA N E
···
· 252
E LE C T R O N
2S 3
·
··
·..· 273
·..·..· ·..·..· ·..· 274
LIST OF POSTER SESSIONS .••.••.••..•..••.•••••••..•.•.•..••••••.•••••••••••••••••••••.••••••••••••••••• ••·•••••····276
ABSTRACTS OF POSTER SESSiONS· •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••···••••·•·••284
T he'·
1.
ID E N T IF IC A T IO N O F P O T E N T IA L F IS H E R Y C O M M O D IT Y
F O R S U P P O R T IN G S M E S O F P R O C E S S E D F IS H E R Y
P R O D U C T O F W E S T K A LIM A N T A N ,
IN D O N E S IA (A C A S E S T U D Y IN S A M B A S R E G E N C y)
·
·
·
2.
IN T E S T IN A L N E U R O T R O P IS M
3.
H O S T P LA N T S O F W E A V E R A N T (O E C O P H Y LLA
U N IV E R S IT A S G A D JA H M A D A C A M P U S ·
4.
E T lH N O B O T A N IC A L
U G M International
O F N E W C A S T LE
D IS E A S E V IR U S F IE LD IS O LA T E S IN LA Y E R C H IC K E N
S M A R A G D IN A )
·
·
·
·
S T U D Y O F S A C R E D F O R E S T O F B U LU P IT U ,
C onference
on T ropical
A griculture
2016
IN G R E E N A R E A O F
·
·
·
·..·
·
KEBUMEN,
·..·
C E N T R A L JA V A
·
·
·
·
·
285
· 286
·..· 287
288
1st International Conference on Tropical Agriculture (ICTA) 2016
Characteristics of Alcaligenes sp. LS2T Heterotrophic and
Aerobic Ammonium Removal for Potential Livestock’s
Wastewater Treatment
A.R. Azkarahmana, Y. Erwantoa, W. Hadisaputroa, L.M. Yusiatia, and N.A.
Fitriyantoa*
a
Faculty of Animal Science, Universitas Gadjah Mada, Jl. Fauna No.3 Kampus UGM Bulaksumur, Yogyakarta, Indonesia
Abstract
Alcaligenes spp. was found to have the ability to grow and remove ammonium in the medium through the nitrificationdenitrification process. This ability was also known as an ammonium treatment in organic wastewater including livestock’s
wastewater. In order to further understand its potential to remove ammonium in livestock’s wastewater; microbial growth,
ammonium, nitrite, and nitrate production of Alcaligenes sp. LS2T were investigated in phosphate medium at different
carbon/nitrogen ratios (C/N ratios) along with constant airflow. Ammonium sulfate was used as sole nitrogen source with
acetate and citrate as an organic carbon source in a separate medium. Results showed that Alcaligenes sp. LS2T could utilize
ammonium as sole nitrogen source which associated with acetate and citrate as carbon source at different C/N ratios,
resulting in ammonium removal, production of nitrite and nitrate at various concentrations in the medium. Highest
ammonium removal was found in the acetate C/N 28 medium where 94.44% of initial ammonium was removed.
Keywords: Alcaligenes sp. LS2T; ammonium removal; heterotrophic nitrification
Introduction
Development in livestock’s industry was followed by the intensive farming system, causing accumulation
of ammonium in livestock’s wastes area. Excess of ammonium was known to cause several environmental
problems, one of it is eutrophication. Therefore, ammonium removal in wastewater treatment became essential
to overcome this problem thus the sustainable farming system could be achieved. Conventional ammonium
removal in wastewater treatment plants consist of nitrification by autotrophic bacteria under aerobic conditions1,
and difficult to apply because autotrophic bacteria are vulnerable to the high concentration of ammonium and
organic matters with C/N ratio of 0-22. Thus, wastewater pretreatment must be done to reduce the C/N ratio3 or
diluting the wastewater4,5. Other researches have been done to observe potential of various heterotrophic
microbial which could live in the higher C/N ratio medium and able to remove ammonium on livestock’s wastes
like on dairy wastewater6, piggery wastewater7, and poultry manure8.
Alcaligenes spp. was known as a heterotrophic bacteria that could be used for removing ammonium in
wastewater9 through simultaneous heterotrophic nitrification and aerobic denitrification 1,10,11. Alcaligenes sp.
LS2T is one bacteria that could remove ammonium in the medium12, allegedly through heterotrophic
nitrification and aerobic denitrification process. In this research, we further observe the potential of Alcaligenes
sp. LS2T for livestock’s wastewater treatment. Investigation on its characteristics to remove ammonium at
different C/N ratios in the phosphate medium was done, nitrite and nitrate production which was also the result
of the nitrification-denitrification process was also determined.
Materials and Methods
Microorganisms and culture conditions
Alcaligenes sp. LS2T was collected from the soil around layer farm in Yogyakarta, Indonesia, then cultured
and stocked in agar medium in the Laboratory of Animal Skins, By-products, and Animal Wastes Technology,
Faculty of Animal Science, Universitas Gadjah Mada. The phosphate medium was prepared by dissolving the
following in 1 L of distilled water: (NH 4)2SO4 0.472 g, MgSO4·7H2O 0.05 g, K2HPO4 0.2 g, NaCl 0.12 g,
MnSO4·4H2O 0.01 g, and FeSO4 0.01 g. Sodium acetate and trisodium citrate were used as organic carbon
source separately in the medium, abbreviated as acetate medium and citrate medium, respectively. The growth
of Alcaligenes sp. LS2T in this experiments were performed in shaken Erlenmeyer flasks containing 500 ml
medium and 1% preculture of Alcaligenes sp. LS2T on a rotary shaker at 120 rpm with constant airflow.
Shaking culture experiment
In order to elucidate the influence of carbon to nitrogen molar ratio (C/N) on the nitrifying capacity, the
amount of N-ammonium was fixed at approximately 100 mg·L-1 while the amount of carbon was adjusted to the
appropriate C/N ratio which was 7, 14, 21 and 28. The ammonium removal ability was investigated in the
acetate medium and citrate medium separately. Samples were taken periodically to examine changes in growth,
concentrations of ammonium, nitrite, and nitrate.
Analytical methods
The growth of Alcaligenes sp. LS2T was monitored by measuring the optical density at 600 nm (OD 600 nm) of
the culture medium using a spectrophotometer (Shimadzu, Japan)11. Culture samples were centrifuged at 10.000
rpm for ammonium, nitrite and nitrate analysis. Ammonium concentration was analyzed by Nessler’s reagent
photometry method13. Nitrite concentration was determined by N-(1-naphtalene)-diaminoethane photometry
method13. Nitrate concentration was measured by using EPA14 modified colorimetric, brucine method. All tests
were conducted in triplicate.
Results
Growth characteristics
The growth of Alcaligenes sp. LS2T at different C/N ratios in the acetate and citrate medium in shaking
culture showed at Fig. 1. Growth curves were established by measurement of the OD value at 600 nm.
(A)
Optical density (OD600)
1.600
1.400
1.200
1.000
0.800
0.600
0.400
0.200
0.000
0
9
18
27
36
45
54
63
72
Optical density (OD600)
time (h)
1.600
1.400
1.200
1.000
0.800
0.600
0.400
0.200
0.000
(B)
0
9
18
27
36
45
54
63
72
time (h)
Fig. 1. The Growth of Alcaligenes sp. LS2T at different C/N ratios in the (A) acetate medium; (B) citrate medium. Closed square: C/N7;
open square: C/N 14; closed circle: C/N 21; open circle: C/N 28.
In the acetate medium, the log phase of the bacterium was all similar started at 3 h. However, the
growth peak in the C/N 7 medium was relatively lower than any other medium, and the stationary phase starts
later than any other acetate medium, indicating that carbon source was deficient for cell synthesis. However, the
shorter stationary phase was found in the C/N 14 medium which lasts for 9 hours while the longest stationary
phase was found in the C/N 28 medium which lasts for 24 hours. In the citrate medium under same conditions,
the log phase of the bacteria was all similar started at 3 h. The growth peak in the citrate C/N 7 medium was also
relatively lower, and the stationary phase was also started later than any other citrate medium. The growth phase
trend in citrate medium was relatively same than in the acetate medium, with longest stationary phase was found
in the citrate C/N 21 and 28 medium which lasts for 21 hours and the shortest was found in the citrate C/N 14
medium which lasts for only 9 hours.
Heterotrophic nitrification
70
0.7
60
0.6
50
0.5
40
0.4
30
0.3
20
0.2
10
0.1
80
0.78
70
0.76
60
0.74
50
0.72
40
0.7
30
20
0.68
10
0.66
0
0
0 12 24 36 48 60 72 84 96
time (h)
time (h)
90
0.9
80
0.8
70
0.7
60
0.6
50
0.5
40
0.4
30
0.3
20
0.2
10
0.1
0
0
(C)
(B)
0.64
0 12 24 36 48 60 72 84 96
0.84
0.82
0.8
0.78
0.76
0.74
0.72
0.7
0.68
0.66
0.64
0.62
90
NH4+ and NO3- concentration (mg·L-1)
NH4+ and NO3- concentration (mg·L-1)
0
0.8
NO2- concentration (mg.L-1)
0.8
90
80
70
60
50
40
30
20
10
0
0 12 24 36 48 60 72 84 96
0 12 24 36 48 60 72 84 96
time (h)
time (h)
(D)
NO2- concentration (mg.L-1)
80
NH4+ and NO3- concentration (mg·L-1)
0.9 (A)
NO2- concentration (mg.L-1)
90
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
In order to investigate the correlation between microbial growth, ammonium removal, and release of products
during heterotrophic nitrification, Alcaligenes sp. LS2T was inoculated into acetate and citrate medium under
different C/N ratios. Cultures were sampled every 12 hours for analysis. Fig. 2 shows the changes by
nitrification (ammonium, nitrite, and nitrate) during the 96-h growth period.
In the acetate medium, highest ammonium removal after 96-h was found in C/N 28 medium, about 94.44%
of initial ammonium removed; followed by C/N 21, C/N 14 and C/N 7 which removed 93.49%, 93.12% and
93.04% of initial ammonium respectively. Furthermore, highest nitrite production was found in C/N 14 medium
(0.79 mg·L-1), followed by C/N 28 (0.69 mg·L-1), C/N 21 (0.68 mg·L-1) and C/N 7 (0.63 mg·L-1). However,
highest nitrate production was found in C/N 28 medium (35.53 mg·L-1), while in the C/N 7 medium was 22.59
mg·L-1; 23.31 in C/N 14 and 26.71 mg·L-1 in the C/N 21 medium.
Fig. 2. Nitrification products of Alcaligenes sp. LS2T in acetate medium at (A) C/N 7; (B) C/N 14; (c) C/N 21; (d) C/N 28. Closed square:
NH4+-N; closed circle: NO2--N; closed triangle: NO3--N.
100
0.4
80
0.3
60
0.2
40
0.1
0
0
0 12 24 36 48 60 72 84 96
100
0.5
80
0.4
60
0.3
40
0.2
20
0.1
0
time (h)
80
0.4
60
0.3
40
0.2
20
0.1
0
time (h)
(C)
0.7
100
NH4+ and NO3- concentration (mg·L-1)
0.5
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
100
0 12 24 36 48 60 72 84 96
0
0 12 24 36 48 60 72 84 96
time (h)
0
(B)
0.6
80
0.5
60
0.4
40
0.3
0.2
20
0.1
0
(D)
NO2- concentration (mg.L-1)
20
(A)
NO2- concentration (mg.L-1)
0.5
NH4+ and NO3- concentration (mg·L-1)
120
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
In the citrate medium after 96-h growth period, highest ammonium removal was found in the C/N 21
medium, with 89.98% of initial ammonium removed; followed by C/N 28 with 88.45%, C/N 14 with 86.36%
and C/N 7 with 83.49% of initial ammonium removed. Highest nitrite production was found in the C/N 7 and 14
medium which both produced 0.27 mg·L-1, while in the C/N 21 and 28 medium both produced 0.24 mg·L-1
nitrite. The C/N 7 medium was found to produce highest nitrate production, with 23.89 mg·L-1 of nitrate
produced after 96-h, followed in the C/N 21; C/N 14 and C/N 28 medium which produced 22.45 mg·L-1; 22.08
mg·L-1 and 20.13 mg·L-1 of nitrate respectively. Heterotrophic nitrification process in citrate medium under
different C/N ratios shown in Fig. 3 below.
0
0 12 24 36 48 60 72 84 96
time (h)
Fig. 3. Nitrification products of Alcaligenes sp. LS2T in citrate medium at (a) C/N 7; (b) C/N 14; (c) C/N 21; (d) C/N 28. Closed square:
NH4+-N; closed circle: NO2--N; closed triangle: NO3--N.
Discussion
The previous study done by Gutama12 showed that Alcaligenes sp. LS2T has the ability utilize ammonium as
a nitrogen source for metabolism. Various studies showed that some Alcaligenes spp. has the ability to perform
heterotrophic nitrification under aerobic condition1,10,11. In this study, we investigate the growth profiles of
Alcaligenes sp. LS2T at different C/N ratios in the acetate and citrate medium. The results indicate that log
phase by Alcaligenes sp. LS2T was found in 3-h in both acetate and citrate medium at all C/N ratios researched.
In both acetate and citrate medium with C/N 7 stationary phase was found later than in any other C/N ratio
medium. Furthermore, the growth was relatively lower this could be caused the lack of carbon source for
metabolism, resulting in less ammonium removal in the medium. Highest ammonium removal in acetate
medium was found in C/N 28, and for citrate, the medium was found in 21, both was known to achieve earlier
and longer stationary phase than in any other C/N in the respective medium. The correlation between earlier and
longer stationary phase with higher ammonium removal is because, in heterotrophic nitrification, the
assimilation ratio is as high as heterotrophic nitrification ratio 15,16, the assimilated nitrogen is associated with
cell growth, and the nitrogen removal rate is affected by cell doubling time 17.Therefore, microorganisms with
earlier and longer stationary phase can take ammonium quickly into the cell, resulting in more effective
ammonium removal18.
At four different C/N ratios in both media, nitrite, and nitrate were formed by Alcaligenes sp. LS2T during
heterotrophic ammonium removal process. The experimental results showed that the formation of nitrate is
higher than nitrite. Nitrite and nitrate concentration in this research demonstrate a whole time nitrification
process until 96-h, these results correspond to present study that indicates heterotrophic nitrite/nitrate production
is linked with growth, supports that heterotrophic nitrification can take place during entire growing phase 19,20.
Nitrate as dominant product in the process of heterotrophic nitrification by Alcaligenes sp. LS2T is similar with
A. faecalis strain NR10, Achromobacter sp.GAD3 and Comamonas sp. GAD421. While other strains like A.
faecalis C1611 and Bacillus methylotrophius strain L722 had nitrite as the dominant product over nitrate in the
process of heterotrophic nitrification.
Carbon dependency has been shown in this research. A possible reason is that ammonium is oxidized by a
sequence involved in some way with the metabolism of a certain or specific carbon source that could provide
the acceptor molecules for the synthesis of intermediate organic nitrogen compounds 23. Results of this research
also showed that even if the C/N ratio is as high as 28, which is too high for autotrophic nitrifying bacteria,
Alcaligenes sp. LS2T still exhibits nitrification ability. This results showed Alcaligenes sp. LS2T potential in
livestock’s wastewater treatment like in piggery wastewater which C/N ratio was around 5-207 or even chicken
manure which C/N ratio was around 824.
Conclusion
Alcaligenes sp. LS2T was capable of removing ammonium in the medium through nitrification process,
utilizing ammonium in the medium for cell synthesize and producing nitrite and nitrate with organic carbon
sources. The highest ammonium removal was found in the acetate medium at C/N 28; this indicates that
Alcaligenes sp. LS2T could perform heterotrophic nitrification under relatively high C/N ratio. Results of this
research suggest deeper research on the ammonium removal ability of Alcaligenes sp. LS2T which is in the
production of hydroxylamine and nitrogen gas and its application on various livestock’s wastewater, so that
clearer nitrogen pathway in this process could be conducted.
Acknowledgement
This research was supported by Program Penelitian Unggulan UGM 2016 DPKM Management contract
number 690/UN1-P.III/LT/DIT-LIT/2016. The authors also thank Laboratory of Animal Skins, By-products, and
Animal Wastes Technology, Faculty of Animal Science, Universitas Gadjah Mada for providing research needs.
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LESSON 1831 (PISCES: GOBIIDAE) FROM BOGOW ONTO
ESTUARY, YOGYAKARTA, INDONESiA •.•.··•.•.•.•.•.·•.•.·•.•.•.·•.· 75
A.1.2 A SIM PLE OPTIM ISED DNA EXTRACTION M ETHOD FOR M ACROALGAE FROM COASTS OF GUNUNGKIDUL,
YOGYAKARTA, USING CTAB-BASED M ETHOD·················································
...............•........•............•.•..•.•..•.•.•.•.•..•.•..•.•...•.•....•.
76
A.l.3 SASI:TRADITIONAL
CONSERVATION TO PROTECT POPULATION OF M ANGROVE CRABS
(SCYLLA SERRATA FORSKAL 1775) IN M ANGROVE AREA OF LEKSULA VILLAGE OF BURU SELATAN DiSTRICT· •.· 77
A.1.4INDONESIAN
SUSTAINABLE FISHERIES M ANAGEM ENT
IN GLOBALIZATIONAND
REGIONALIZATION
OF ASEAN
ECONOM IC
A B S T R A C T S
COM M UNITY
O F
(AEC) 2016··················································
S U S T A IN A B L E
A N IM A L
H E A L T H
·····························
•.····•.··78
•..............•••...............•.....•............••.
B.I.1 DIFFERENCES IN THE REARING SYSTEM TO BALI CADLE GASTROINTESTINAL
IN PRAFI DISTRICT M ANOKW ARI REGENCY W EST PAPUA PROVINCE
HELM INTHS
7 9
INFESTATION
·····························
•.·80
B.1.2 TRYPANOSOM IASIS
AND SURRA: VECTOR, DETECTION AND ERADICATION TO ENSURE LIVESTOCK
PRODUCTION IN INDONESiA···· •.····•.······················
•.····• .·•.··•.·•.....•......•.........•.........................................................................................81
B.I.3 DETECTION THE PRESENCE OF BOVINE VIRAL DIARRHEA VIRUS PERSISTENT INFECTION
IN LOW REPRODUCTIVE
PERFORM ANCE DAIRY CADLE HERD IN CENTRAL JAVA, INDONESiA································································
•.··•.•.•.•.· ·•.·•.··82
B.l1.1FASCIOLOSIS IN CENTRAL JAVA: A CONTINUOUS THREAT ....................•.........•.........................················································83
B.II.2DETECTION OF TREM BOLONE ACETATE IN BEEF M USCLE AND LIVER USING HIGH PERFORM ANCE LIQUID
CHROM ATOGRAPHY
M ETHOD ..............................................................•.....•.........................................·················································84
B.l1.3 ANTIBIOTIC RESISTANCE IN ESCHERICHIA COLI ISOLATED FROM BROILER ASSOCIATED W ITH
ENROFLOXACINE AND OXYTETRACYCLINE TREATM ENT ........•...•........•..•.•............•....................·················································85
B.l1.4SEROLOGICAL AND M OLECULAR BIOLOGICAL DIAGNOSIS FOR LEPTOSPiROSiS··········································
•.•.·····• .·•.··•.·86
B.l1.5RAPID DETECTION OF Verocytotoxigenic Escherichia coli (VTEC) BACTERIA BY POLYM ERASE
CHAIN REACTION (PCR) M ETHOD AGAINST ISOLATES INOCULATED IN BGLB·····································································87
B.II.6 INVESTIGATION OF LEAD CONTAM INATION
IN BADUNG RIVER USING FISHES AS BIOINDICA TORS ...•..................... 88
8.11.7 HAEM OPROTOZOA
A B S T R A C T
O F
INFECTIONS
S U S T A IN A B L E
OF DOM ESTIC
BIRDS IN HILLY AREAS OF BANGLADESH
T R O P IC A L
F O R E S T R y
·························
•.·············
•. 89
· 9 0
cnu
ASSESSM ENT OF LANDSLIDE RISK AND DISASTER M ANAGEM ENT
IN W ATUGEDE CATCHM ENT,
GUNUNGKIDUL
DISTRICT .........................................................................•....................•............•.•.•.· ·•.····• .•.·•.•.·•.•.·•.·•.·•.·•.·•.···················91
-----------------T h e
1 "U G M
In te rn a tio C
na
oln fe re nocnT
e ro p icA
a lg ric u ltu2re
016
T
F .V 13 E S T A B LIS H IN G Q U A IL P O P U LA T IO N S
F O R LE S S B IA S E D E X P E R IM E N T A L
F .V 1.4R E S T R lalO N E N Z Y M E M A P P IN G O F C Y T O C H R O M E
IN T H E W O R LD ..·
·..·
·
·
·
· · · ·· ·
F .V I,5 C O M P A R A T IV E S T U D Y O F B O D Y M E A S U R E M E N T S
F .V I,6T H E E F F E C T O F B IR T H T Y P E O N P H E N O T Y P IC
F .V I.7T H E M IC R O R N A -A M P K G E N E C A N D ID A T E
LE U T E IN IZ A T IO N : A N IN S ILIC O S T U D y
R E S U LT S
B GENE ON ANGUS
·
240
IN D IF F E R E N T S T R A IN O F IN D O N E S IA N
PERFORM ANCE
LO C A L D U C K S
O F S A P U D I S H E E P IN S A P U D IIS LA N D ,
· 241
MADURA
242
IN R E G U LA T E LIP ID A C C U M U LA T IO N
O F B O V IN E G R A N U LO S A C E LL
·· ·· · ··
·
·
· ··· ··
·..............................•.................................. 243
F .V I,8IM P A C T O F B U LL'S B R E E D A T C O N C E N T R A T IO N
O F P R O S T A G LA N D IN
S E M IN A LIS B Y IN V IT R O M E T H O D
··
· ·..· · · · ·
··· ·· ·· ··· ··
F .V I.9 A N IM A L W E LF A R E A S S T R E S S M A N A G E M E N T T O
IM P R O V E
C O LLE C T E D
F .V I.12 T H E M O T ILIT Y O F D E G R A M 'S S P E R M A T O Z O A
A N T IO X ID A N T A N D D IF F E R E N T E X T E N D E R
F R O M V E S IC U LA R
244
B E E F C A T T LE R E P R O D U C T IO N
F .V 1.10S P E R M Q U A LIT Y O F B A LI C A T T LE A F T E R G IV E N C R U D E T A N IN S U P P LE M E N T
F .V I.11 B LO O D C H E M IS T R Y P A R A M E T E R S O F A D U LT K A C A N G G O A T S
·
245
IN T H E S E M E N
246
247
B E F O R E A N D A F T E R F R E E Z IN G W IT H A D D IT IO N
·
F .IV .1 P O T E N T IA L O F M A R M E LO S IN B IO A C T IV E C O M P O U N D
IN A E G LE M A R M E LO S
B A C T E R IA L U R E A S E F O R R E D U C E F O R M IN G A M M O N IA
(N H 3) IN E X C R E T A
T O IN H IB IT T H E G R O W T H
FROM FARM TO FORK THREATEN
HUMAN
· · 248
OF
249
F .IV .2 T H E A D D IT IO N E F F E C T O F F E R M E N T E D A E G LE M A R M E LO S F R U IT A N D B A M B O O
C A m E F E C E S S LU R R Y T O T H E R E D U C T IO N O F A M M O N IA
G A S E M IS S IO N
F .IV .3 M E A T C O N T A M IN A T IO N
239
G R A D E C A T T LE A N D O T H E R C A T T LE
SHOOTS
IN
2S 0
H E A LT H
251
F .lV .4 E F F E C T SO F A D M IN IS T R A T IO N
O F C O M B IN A T IO N
O F P A LM K E R N E L M E A L, K A T U K LE A F A N D B E T E L N U T
P O W D E R O N S E N S O R Y V A LU E O F M E A T IN T H E G O A T
F .lV .s Q U A N T IT A T IV E E LE M E N T A L A N A LY S IS
M iC R O S C O P E
O F V A R IO U S
F .lV .6 P H Y S IC A L Q U A LIT Y O F E G G O F LA Y IN G
R IC H IN A LP H A -LIN O LE N IC A C ID
H E N S F E D D IE T S C O N T A IN IN G
F .1I1.1H E T E R O T R O P H IC A N D A E R O B IC A M M O N IU M
F .l11,2M A N A G E M E N T A N D R E P R O D U C T IV E
F A R M E R S IN A T R A D IT IO N A L S yS T E M
F .1I1,3A R T IF IC IA LIN S E M IN A T IO N
P O U LT R Y E G G S H E LL C U T IC LE U S IN G S C A N N IN G
REMOVAL
IN W A S T E W A T E R
B Y A lcaligenes
Y E A R LIN G
F .l11.6A N IM A L W E LF A R E A S S E S S M E N T S Y S T E M F O R S U S T A IN A B LE
C A T T LE P R O D U C T IO N
E N V IR O N M E N T A L
2S 5
2S 6
F .V .4 R O LE O F F E M A LE M E M B E R S IN S M A LLH O LD E R
F A R M E R S ' F A M ILY
T H E V ILLA G E A T T H E U P LA N D A R E A , Y O G Y A K A R T A
IN D O N E S IA
··
260
261
D E V E LO P M E N T
IN
··
··
·
··
·
U S IN G D E M O G R A P H IC
·..·
·
263
TREATM ENT
· ····
F O R G O A T F A R M IN G
IN T H E D IS T R IC T
·
·
··· ·
· ·..· 264
A C T IV IT IE S
IN
265
266
TO M ODEL
··
·•
S M A LLH O LD E R S :
·
·..·
·
ON
·..· 267
268
A CASE STUDY OF
·
·
·..· 269
A D O P T IO N O N IN T E G R A T E D P A LM O IL A N D C A T T LE F A R M IN G IN
· ·..·
·..·..·
·
·.....................................................•............................................. ·
·
·..·
·..·..·..·..· ·..·..·
· 262
AND
F O R F A R M E R S G R O U P T O S U P P O R T B E E F C A T T LE
F .v.8 U S IN G C A U S A L LO O P D IA G R A M A S A Q U A LIT A T IV E
IN S T R U M E N T
G O A T F A R M IN G IN G U M E LA R - B A N Y U M A S ..· ·
· ·
··
·
F .V .10 S U M M A R y
· 259
IN IN D O N E S IA
F .V .6 T H E P E R F O R M A N C E O F M ILK P R O D U C T IO N , T O T A L M ILK R E V E N U E A N D R E P R O D U C T IO N IN D IC A T O R S
D A IR Y S M A LLH O LD E R S IN D A E R A H IS T IM E W A Y O G Y A K A R T A A N D E A S T JA V A P R O V IN C E , IN D O N E S IA ·
F .V .7 P E R C E P T IO N O F B E E F C A T T LE F A R M E R S T O W A R D C R O P W A S T E A S A N IM A L F E E D
F .V .9 T H E R A T E O F T E C H N O LO G Y
JA M B I P R O V iN C E ..· ·..·..·
257
B U C K F E D R U M IN A LLY
A N D S O C IO E C O N O M IC
·
·
·
··
AN APPROACH
· ··
MODEL
GOAT
D IS T R IC T , B R E B E S R E G E N C Y ,
F .V .3 A N A LY S IS O F T H E IM P LE M E N T A T IO N
OF ETT AW A CROSSBRED GOAT M ANURE
K U LO N P R O G O Y O G Y A K A R T A IN D O N E S IA
F .V .s D E V E LO P M E N T O F M IC R O F IN A N C E IN S T IT U T IO N S
F A R M IN G S U S T A IN A B ILIT Y : A P ILO T S T U D Y
· · · 254
C A T T LE K E P T B Y
B R E E D IN G C E N T E R IN
·
F .lII.7 R E P R O D U C T IV E P E R F O R M A N C E O F JA B R E S C O W A T K E T A N G G U N A N
C E N T R A L JA V A P R O V IN C E , IN D O N E S IA
F .V .2 M A R K E T S E G M E N T A T IO N O F G O A T M ILK C O N S U M E R S :
B E H A V IO R A L V A R IA B LE S ..·..· ·
·· ·
• ·..•..· · · ·
·
KACANG
···
sp. LS 2T
258
O F LO C A L C O W S A T V ILLA G E
TOW ARDS
oleracea)
·
·
G R A D E D O E S U S IN G F R O Z E N S E M E N S O F G E M B R O N G
F .III.4 E D IB LE P O R T IO N O F C A R C A S S A N D O F F A LS O F IN D O N E S IA N
U N D E G R A D A B LE P R O T E IN
F .V .1 B E E F C A T T LE F A R M E R S A T T IT U D E
Y O G Y A K A R T A P R O V IN C E
(P ortulaca
··· ···· ··
P E R F O R M A N C E S O F A C E H C O W S , LO C A L IN D O N E S IA N
··· ·
·
·
·
·
···
ON THE ETAW AH
F .lll.s P O S T P A R T U M E S T R U S V A R IA T IO N
T H E Y O G Y A K A R T A P R O V IN C E
P U R S LA N E
···
· 252
E LE C T R O N
2S 3
·
··
·..· 273
·..·..· ·..·..· ·..· 274
LIST OF POSTER SESSIONS .••.••.••..•..••.•••••••..•.•.•..••••••.•••••••••••••••••••••.••••••••••••••••• ••·•••••····276
ABSTRACTS OF POSTER SESSiONS· •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••···••••·•·••284
T he'·
1.
ID E N T IF IC A T IO N O F P O T E N T IA L F IS H E R Y C O M M O D IT Y
F O R S U P P O R T IN G S M E S O F P R O C E S S E D F IS H E R Y
P R O D U C T O F W E S T K A LIM A N T A N ,
IN D O N E S IA (A C A S E S T U D Y IN S A M B A S R E G E N C y)
·
·
·
2.
IN T E S T IN A L N E U R O T R O P IS M
3.
H O S T P LA N T S O F W E A V E R A N T (O E C O P H Y LLA
U N IV E R S IT A S G A D JA H M A D A C A M P U S ·
4.
E T lH N O B O T A N IC A L
U G M International
O F N E W C A S T LE
D IS E A S E V IR U S F IE LD IS O LA T E S IN LA Y E R C H IC K E N
S M A R A G D IN A )
·
·
·
·
S T U D Y O F S A C R E D F O R E S T O F B U LU P IT U ,
C onference
on T ropical
A griculture
2016
IN G R E E N A R E A O F
·
·
·
·..·
·
KEBUMEN,
·..·
C E N T R A L JA V A
·
·
·
·
·
285
· 286
·..· 287
288
1st International Conference on Tropical Agriculture (ICTA) 2016
Characteristics of Alcaligenes sp. LS2T Heterotrophic and
Aerobic Ammonium Removal for Potential Livestock’s
Wastewater Treatment
A.R. Azkarahmana, Y. Erwantoa, W. Hadisaputroa, L.M. Yusiatia, and N.A.
Fitriyantoa*
a
Faculty of Animal Science, Universitas Gadjah Mada, Jl. Fauna No.3 Kampus UGM Bulaksumur, Yogyakarta, Indonesia
Abstract
Alcaligenes spp. was found to have the ability to grow and remove ammonium in the medium through the nitrificationdenitrification process. This ability was also known as an ammonium treatment in organic wastewater including livestock’s
wastewater. In order to further understand its potential to remove ammonium in livestock’s wastewater; microbial growth,
ammonium, nitrite, and nitrate production of Alcaligenes sp. LS2T were investigated in phosphate medium at different
carbon/nitrogen ratios (C/N ratios) along with constant airflow. Ammonium sulfate was used as sole nitrogen source with
acetate and citrate as an organic carbon source in a separate medium. Results showed that Alcaligenes sp. LS2T could utilize
ammonium as sole nitrogen source which associated with acetate and citrate as carbon source at different C/N ratios,
resulting in ammonium removal, production of nitrite and nitrate at various concentrations in the medium. Highest
ammonium removal was found in the acetate C/N 28 medium where 94.44% of initial ammonium was removed.
Keywords: Alcaligenes sp. LS2T; ammonium removal; heterotrophic nitrification
Introduction
Development in livestock’s industry was followed by the intensive farming system, causing accumulation
of ammonium in livestock’s wastes area. Excess of ammonium was known to cause several environmental
problems, one of it is eutrophication. Therefore, ammonium removal in wastewater treatment became essential
to overcome this problem thus the sustainable farming system could be achieved. Conventional ammonium
removal in wastewater treatment plants consist of nitrification by autotrophic bacteria under aerobic conditions1,
and difficult to apply because autotrophic bacteria are vulnerable to the high concentration of ammonium and
organic matters with C/N ratio of 0-22. Thus, wastewater pretreatment must be done to reduce the C/N ratio3 or
diluting the wastewater4,5. Other researches have been done to observe potential of various heterotrophic
microbial which could live in the higher C/N ratio medium and able to remove ammonium on livestock’s wastes
like on dairy wastewater6, piggery wastewater7, and poultry manure8.
Alcaligenes spp. was known as a heterotrophic bacteria that could be used for removing ammonium in
wastewater9 through simultaneous heterotrophic nitrification and aerobic denitrification 1,10,11. Alcaligenes sp.
LS2T is one bacteria that could remove ammonium in the medium12, allegedly through heterotrophic
nitrification and aerobic denitrification process. In this research, we further observe the potential of Alcaligenes
sp. LS2T for livestock’s wastewater treatment. Investigation on its characteristics to remove ammonium at
different C/N ratios in the phosphate medium was done, nitrite and nitrate production which was also the result
of the nitrification-denitrification process was also determined.
Materials and Methods
Microorganisms and culture conditions
Alcaligenes sp. LS2T was collected from the soil around layer farm in Yogyakarta, Indonesia, then cultured
and stocked in agar medium in the Laboratory of Animal Skins, By-products, and Animal Wastes Technology,
Faculty of Animal Science, Universitas Gadjah Mada. The phosphate medium was prepared by dissolving the
following in 1 L of distilled water: (NH 4)2SO4 0.472 g, MgSO4·7H2O 0.05 g, K2HPO4 0.2 g, NaCl 0.12 g,
MnSO4·4H2O 0.01 g, and FeSO4 0.01 g. Sodium acetate and trisodium citrate were used as organic carbon
source separately in the medium, abbreviated as acetate medium and citrate medium, respectively. The growth
of Alcaligenes sp. LS2T in this experiments were performed in shaken Erlenmeyer flasks containing 500 ml
medium and 1% preculture of Alcaligenes sp. LS2T on a rotary shaker at 120 rpm with constant airflow.
Shaking culture experiment
In order to elucidate the influence of carbon to nitrogen molar ratio (C/N) on the nitrifying capacity, the
amount of N-ammonium was fixed at approximately 100 mg·L-1 while the amount of carbon was adjusted to the
appropriate C/N ratio which was 7, 14, 21 and 28. The ammonium removal ability was investigated in the
acetate medium and citrate medium separately. Samples were taken periodically to examine changes in growth,
concentrations of ammonium, nitrite, and nitrate.
Analytical methods
The growth of Alcaligenes sp. LS2T was monitored by measuring the optical density at 600 nm (OD 600 nm) of
the culture medium using a spectrophotometer (Shimadzu, Japan)11. Culture samples were centrifuged at 10.000
rpm for ammonium, nitrite and nitrate analysis. Ammonium concentration was analyzed by Nessler’s reagent
photometry method13. Nitrite concentration was determined by N-(1-naphtalene)-diaminoethane photometry
method13. Nitrate concentration was measured by using EPA14 modified colorimetric, brucine method. All tests
were conducted in triplicate.
Results
Growth characteristics
The growth of Alcaligenes sp. LS2T at different C/N ratios in the acetate and citrate medium in shaking
culture showed at Fig. 1. Growth curves were established by measurement of the OD value at 600 nm.
(A)
Optical density (OD600)
1.600
1.400
1.200
1.000
0.800
0.600
0.400
0.200
0.000
0
9
18
27
36
45
54
63
72
Optical density (OD600)
time (h)
1.600
1.400
1.200
1.000
0.800
0.600
0.400
0.200
0.000
(B)
0
9
18
27
36
45
54
63
72
time (h)
Fig. 1. The Growth of Alcaligenes sp. LS2T at different C/N ratios in the (A) acetate medium; (B) citrate medium. Closed square: C/N7;
open square: C/N 14; closed circle: C/N 21; open circle: C/N 28.
In the acetate medium, the log phase of the bacterium was all similar started at 3 h. However, the
growth peak in the C/N 7 medium was relatively lower than any other medium, and the stationary phase starts
later than any other acetate medium, indicating that carbon source was deficient for cell synthesis. However, the
shorter stationary phase was found in the C/N 14 medium which lasts for 9 hours while the longest stationary
phase was found in the C/N 28 medium which lasts for 24 hours. In the citrate medium under same conditions,
the log phase of the bacteria was all similar started at 3 h. The growth peak in the citrate C/N 7 medium was also
relatively lower, and the stationary phase was also started later than any other citrate medium. The growth phase
trend in citrate medium was relatively same than in the acetate medium, with longest stationary phase was found
in the citrate C/N 21 and 28 medium which lasts for 21 hours and the shortest was found in the citrate C/N 14
medium which lasts for only 9 hours.
Heterotrophic nitrification
70
0.7
60
0.6
50
0.5
40
0.4
30
0.3
20
0.2
10
0.1
80
0.78
70
0.76
60
0.74
50
0.72
40
0.7
30
20
0.68
10
0.66
0
0
0 12 24 36 48 60 72 84 96
time (h)
time (h)
90
0.9
80
0.8
70
0.7
60
0.6
50
0.5
40
0.4
30
0.3
20
0.2
10
0.1
0
0
(C)
(B)
0.64
0 12 24 36 48 60 72 84 96
0.84
0.82
0.8
0.78
0.76
0.74
0.72
0.7
0.68
0.66
0.64
0.62
90
NH4+ and NO3- concentration (mg·L-1)
NH4+ and NO3- concentration (mg·L-1)
0
0.8
NO2- concentration (mg.L-1)
0.8
90
80
70
60
50
40
30
20
10
0
0 12 24 36 48 60 72 84 96
0 12 24 36 48 60 72 84 96
time (h)
time (h)
(D)
NO2- concentration (mg.L-1)
80
NH4+ and NO3- concentration (mg·L-1)
0.9 (A)
NO2- concentration (mg.L-1)
90
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
In order to investigate the correlation between microbial growth, ammonium removal, and release of products
during heterotrophic nitrification, Alcaligenes sp. LS2T was inoculated into acetate and citrate medium under
different C/N ratios. Cultures were sampled every 12 hours for analysis. Fig. 2 shows the changes by
nitrification (ammonium, nitrite, and nitrate) during the 96-h growth period.
In the acetate medium, highest ammonium removal after 96-h was found in C/N 28 medium, about 94.44%
of initial ammonium removed; followed by C/N 21, C/N 14 and C/N 7 which removed 93.49%, 93.12% and
93.04% of initial ammonium respectively. Furthermore, highest nitrite production was found in C/N 14 medium
(0.79 mg·L-1), followed by C/N 28 (0.69 mg·L-1), C/N 21 (0.68 mg·L-1) and C/N 7 (0.63 mg·L-1). However,
highest nitrate production was found in C/N 28 medium (35.53 mg·L-1), while in the C/N 7 medium was 22.59
mg·L-1; 23.31 in C/N 14 and 26.71 mg·L-1 in the C/N 21 medium.
Fig. 2. Nitrification products of Alcaligenes sp. LS2T in acetate medium at (A) C/N 7; (B) C/N 14; (c) C/N 21; (d) C/N 28. Closed square:
NH4+-N; closed circle: NO2--N; closed triangle: NO3--N.
100
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80
0.3
60
0.2
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0.1
0
0
0 12 24 36 48 60 72 84 96
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0.5
80
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0.3
40
0.2
20
0.1
0
time (h)
80
0.4
60
0.3
40
0.2
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0.1
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time (h)
(C)
0.7
100
NH4+ and NO3- concentration (mg·L-1)
0.5
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
100
0 12 24 36 48 60 72 84 96
0
0 12 24 36 48 60 72 84 96
time (h)
0
(B)
0.6
80
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NO2- concentration (mg.L-1)
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0.5
NH4+ and NO3- concentration (mg·L-1)
120
NO2- concentration (mg.L-1)
NH4+ and NO3- concentration (mg·L-1)
In the citrate medium after 96-h growth period, highest ammonium removal was found in the C/N 21
medium, with 89.98% of initial ammonium removed; followed by C/N 28 with 88.45%, C/N 14 with 86.36%
and C/N 7 with 83.49% of initial ammonium removed. Highest nitrite production was found in the C/N 7 and 14
medium which both produced 0.27 mg·L-1, while in the C/N 21 and 28 medium both produced 0.24 mg·L-1
nitrite. The C/N 7 medium was found to produce highest nitrate production, with 23.89 mg·L-1 of nitrate
produced after 96-h, followed in the C/N 21; C/N 14 and C/N 28 medium which produced 22.45 mg·L-1; 22.08
mg·L-1 and 20.13 mg·L-1 of nitrate respectively. Heterotrophic nitrification process in citrate medium under
different C/N ratios shown in Fig. 3 below.
0
0 12 24 36 48 60 72 84 96
time (h)
Fig. 3. Nitrification products of Alcaligenes sp. LS2T in citrate medium at (a) C/N 7; (b) C/N 14; (c) C/N 21; (d) C/N 28. Closed square:
NH4+-N; closed circle: NO2--N; closed triangle: NO3--N.
Discussion
The previous study done by Gutama12 showed that Alcaligenes sp. LS2T has the ability utilize ammonium as
a nitrogen source for metabolism. Various studies showed that some Alcaligenes spp. has the ability to perform
heterotrophic nitrification under aerobic condition1,10,11. In this study, we investigate the growth profiles of
Alcaligenes sp. LS2T at different C/N ratios in the acetate and citrate medium. The results indicate that log
phase by Alcaligenes sp. LS2T was found in 3-h in both acetate and citrate medium at all C/N ratios researched.
In both acetate and citrate medium with C/N 7 stationary phase was found later than in any other C/N ratio
medium. Furthermore, the growth was relatively lower this could be caused the lack of carbon source for
metabolism, resulting in less ammonium removal in the medium. Highest ammonium removal in acetate
medium was found in C/N 28, and for citrate, the medium was found in 21, both was known to achieve earlier
and longer stationary phase than in any other C/N in the respective medium. The correlation between earlier and
longer stationary phase with higher ammonium removal is because, in heterotrophic nitrification, the
assimilation ratio is as high as heterotrophic nitrification ratio 15,16, the assimilated nitrogen is associated with
cell growth, and the nitrogen removal rate is affected by cell doubling time 17.Therefore, microorganisms with
earlier and longer stationary phase can take ammonium quickly into the cell, resulting in more effective
ammonium removal18.
At four different C/N ratios in both media, nitrite, and nitrate were formed by Alcaligenes sp. LS2T during
heterotrophic ammonium removal process. The experimental results showed that the formation of nitrate is
higher than nitrite. Nitrite and nitrate concentration in this research demonstrate a whole time nitrification
process until 96-h, these results correspond to present study that indicates heterotrophic nitrite/nitrate production
is linked with growth, supports that heterotrophic nitrification can take place during entire growing phase 19,20.
Nitrate as dominant product in the process of heterotrophic nitrification by Alcaligenes sp. LS2T is similar with
A. faecalis strain NR10, Achromobacter sp.GAD3 and Comamonas sp. GAD421. While other strains like A.
faecalis C1611 and Bacillus methylotrophius strain L722 had nitrite as the dominant product over nitrate in the
process of heterotrophic nitrification.
Carbon dependency has been shown in this research. A possible reason is that ammonium is oxidized by a
sequence involved in some way with the metabolism of a certain or specific carbon source that could provide
the acceptor molecules for the synthesis of intermediate organic nitrogen compounds 23. Results of this research
also showed that even if the C/N ratio is as high as 28, which is too high for autotrophic nitrifying bacteria,
Alcaligenes sp. LS2T still exhibits nitrification ability. This results showed Alcaligenes sp. LS2T potential in
livestock’s wastewater treatment like in piggery wastewater which C/N ratio was around 5-207 or even chicken
manure which C/N ratio was around 824.
Conclusion
Alcaligenes sp. LS2T was capable of removing ammonium in the medium through nitrification process,
utilizing ammonium in the medium for cell synthesize and producing nitrite and nitrate with organic carbon
sources. The highest ammonium removal was found in the acetate medium at C/N 28; this indicates that
Alcaligenes sp. LS2T could perform heterotrophic nitrification under relatively high C/N ratio. Results of this
research suggest deeper research on the ammonium removal ability of Alcaligenes sp. LS2T which is in the
production of hydroxylamine and nitrogen gas and its application on various livestock’s wastewater, so that
clearer nitrogen pathway in this process could be conducted.
Acknowledgement
This research was supported by Program Penelitian Unggulan UGM 2016 DPKM Management contract
number 690/UN1-P.III/LT/DIT-LIT/2016. The authors also thank Laboratory of Animal Skins, By-products, and
Animal Wastes Technology, Faculty of Animal Science, Universitas Gadjah Mada for providing research needs.
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