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d Mat Noor, Agriculture Engineering, Malaysia ul Park, Myong Ji University, Korea

evineni, Manipal University, India d Ben Haj Frej, POST University, USA

F. M. Monteiro, ULHT - Universidade Lusofona, Port ABU OJHA, Soil Scientist,Nepal Agricultural Research Bhatiya,BHK collage of Engineering, India

IMIRE,Agriculture Extension Officer Ministry of Agric ,Government of Nepal, Nepal

ayeswara,Department of Zoology, Sahyadri Science C ), Shivamogga-577203, Karnataka state,India

Swamy,Senior Research Fellow (SRF),Biopesticide La technology,(IIHR), Hessarghatta lake post, Bangalore Serdaroğlu,Engineering Faculty,Food Engineering De

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International journal of Agriculture and Environmental Research

Promote your research with IJAER| www.ijaer.in

Volume 1, Issue 1 , Nov 2015

Article

No. Title & Authors name Page download

1

ANTIMICROBIAL AND CYTOTOXICITY OF Ag AND ZnO NANOPARTICLES

Nabeel K.AL-Ani, Ayad,M.Ali,Sarah Ibrahim,Farooq I.Mohammad,Salah M.Al-chalabi

01-11view

2

POTENCY OF BACTERIA ISOLATED FROM BALI COLON WASTE AS LIGNOCELLULOSE SUBSTRATES DEGRADER

Mudita, I M., I G. Mahardika, I N. Sujaya, I. B. G. Partama

12-23view

3

NUTRITIVE EVALUATION OF SOME BROWSE TREES FRUITS (SEED AND PODS) AS DRY SEASON SUPPLEMENT FOR LIVESTOCK IN ARID AND SEMI-ARID LANDS OF SUDAN

Izeldin A. Babiker, Mohamed S.A. Abdulla

24-31view

4

THE EFFECT OF FEEDING THREE PEARL MILLET CULTIVARS ON PERFORMANCE OF LAYING HENS AND THYROID GLAND HISTOLOGY

Amal Eltayeb Mahmoud

32-41view

5

GROWTH OF SESAME AS INFLUENCED BY VARIETIES AND PLANT EXTRACTS ON THE CONTROL OF Cercospora sp. IN ARDO-KOLA AND GASSOL, TARABA STATE, NIGERIA

Tunwari, B.A., Nahunnaro, H.

42-64view

6

MINI REVIEW: HYDROPHONIC GREENHOUSE- THE COMMON PROBLEMS AND SOLUTIONS Unal Senel, Ilkay Senel, Ridvan Yildirim, Mustafa Cemek, Ibrahim Isildak, Ismail Agir

65-78view

7

IN VIVO EVALUATION OF FUNGICIDES ON FUSARIUM EQUISETI FOLIAR BLIGHT OF JATROPHA CURCAS L.

Nasiru A M, Shehu K, Mani U, Yerima M B 79-87view

8

OXALATE LEVELS IN SELECTED AFRICAN INDIGENOUS VEGETABLE RECIPES FROM THE LAKE VICTORIA BASIN, KENYA.

Mr. Wakhanu A. John, Prof. Kimiywe Judith, Prof. Nyambaka Hudson

88- 105 view

9

EFFECTS OF BANANA BORER WEEVIL (COSMOPOLITES SORDIDUS) ON PLANTAIN ORCHARD REHABILITATED WITH PARED CORMS, POULTRY MANURE AND BIO-ACTIVE MULCH.

Oso, A.A. and Longe O.O.

106- 115 view

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International Journal of Agriculture and Environmental Research

Volume:01,Issue:01 www.ijaer.in

www.ijaer.in Page 12

POTENCY OF BACTERIA ISOLATED FROM BALI COLON WASTE AS LIGNOCELLULOSE SUBSTRATES DEGRADER

Mudita, I M., I G. Mahardika, I N. Sujaya and I. B. G. Partama

Faculty of Animal Husbandry, Udayana University, Denpasar

ABSTRACT

A research has been carried out to evaluate the potency of bacteria isolated from bali cattle colon waste as lignocellulosic substrates degrader. Bacteria were isolated by Hungate selective media using combination of tannic acid (lignin), xylan, and Carboxy Methyl Cellulose/CMC (cellulose) as selective substrates. The potency of lignocellulolytic was identified based on the degradation of lignocellulosic substrates measurement using clear/diffusion zone width and lignocellulolytic enzyme activity. This study showed that lignocellulolytic bacteria isolated from bali cattle colon produce clear/diffusion zones in the width of wide 0.078 – 3.440 cm², 0.007 – 0.072 cm², 0.172 – 4.497 cm² and 0,392 – 5,864 cm²respectively for lignocellulose, tannic acid, cellulose and xylan substrates, while isolates coded BCC 7 LC, BCC 4 LC and BCC 12.1 LC produced higher clear/diffusion zones. Meanwhile for lignocellulolytic enzyme activity, bacteria isolates coded BCC 12.1 LC and BCC 6 LC has higher enzyme activity at tannic acid, CMC, and xylan substrates. It was concluded that bacteria isolates coded BCC 12.1 LC is a lignocellulosic substrates degrader.

Keywords: Bali Cattle Colon Waste, Clear Zone Width, Enzyme Activity, Lignocellulosic Substrates

INTRODUCTION

Utilization of agricultural wastes as feed is one of the national policy in the development of cattle livestock to achieve beef self-sufficiency. This step is more strategic for the livestock sector in Bali in support of the Bali Green and Clean Provence by the Provincial Government of Bali, Indonesian Country. Utilization of waste as feed will reduce the emission of pollutants in the air (Hegarty, 2001). However, research results of Mudita et al. (2008-2012) and Putri et al.

(2009) revealed the use of waste as feed without the application of processing technology will decrease the productivity of livestock and increase pollutant emissions due to high crude fiber content in particular lignocellulose compounds and very difficult to be utilized by livestock.

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International Journal of Agriculture and Environmental Research

Lignocellulose is composed of three polymers, including lignin, cellulose and hemicellulose (Howard et al., 2003; Perez et al., 2002). Degradation of its polymer completely will supply all potential nutrients contained in the agricultural waste feedstuffs. Lignocellulose can only be degraded by certain microbes, such as lignocellulolytic bacteria. So the effort to develop lignocellulolytic bacteria is very important in optimizing utility agricultural waste as feedstuffs. Bali cattle colon contents waste is one of the source of lignocelluloses degrading bacteria isolates.

The cattle colon contains many fiber degrading microbes (bacteria and fungi) such as lignolytic, cellulolytic, hemycellulolytic, amylolytic and proteolytic and various probiotics microbes (Wahyudi et al., 2010ab). Wahyudi et al. (2010) showed that the lignocellulolytic bacteria of the colon and caecum have the ability to degrade crude fiber higher than the rumen bacteria. The bacteria isolates capable of associating with fiber rich feed ingredients that are not degraded in the rumen. Wahyudi et al. (2010) have isolated three lignolytic bacteria, bacterial xylanolytic 111, and 262 of cellulolytic bacteria from the colon of buffalo with the highest enzyme production of 0.1971 molecular weight (mol).Based on the potential of lignocellulosic substrates degradation, bacteria isolated from bali cattle colon contents waste was made to explore its potential as a source of inoculum agro-waste degrader on the development of competitive and sustainable bali cattle livestock.

MATERIALS AND METHODS

Isolation of source and sample preparation

The bali cattle colon contents waste sample taken from cattle slaughter house in Kediri Tabanan, Bali Provence-Indonesian Country. The colon content sample was obtained from gut after bali cattle’s death by filtering into a prewarm (39^oC) termos flask (Lowe, 1986). Anaerobic conditions were created by inserting the bali cattle colon contents waste up to full and filling CO2

into the flask, and then covering it with asterile butyl rubber stopper. The sample was brought to the laboratory for further processing as sources of bacteria isolates. Dilution of sample has been conducted in series until 10^-7foid dilution was obtained with dilution medium formula No 14 Bryant and Burkey (Ogimoto and Imai, 1981).

Solid Media and Isolation

Bacteria from samples were grown in selective solid media by Hungate method (Ogimoto and Imai, 1981) countaining 0,02g Monopotassium phosphate (KH2PO4); 0,03g Dipotassium phosphate (K2HPO4); 0,01g Magnesium Sulfate (MgSO4); 0,01g Calcium

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International Journal of Agriculture and Environmental Research

Tetrachloride (CaCl4); 0,10g Sodium Cloride (NaCl); 0,10g Ammonium Sulfate {(NH4)2SO4};

0,10ml Rezasurin 0,1% solution; 0,02g Cystein-HCl.H2O; 0,40g Sodium Carbonate (Na2CO3);

30,00 ml rumen liquid; 1,00g substrate; 70,00ml Aquadest and 1,8% Agar. Selective substrate used were tannic acid (as lignin source), xylan (as hemicellulose source) and Carboxy Methyl Cellulose/CMC (as cellulose source). All ingredients were mixed in erlenmeyer, pH was determined 6.8 and heated until all ingredients dissolved. The flask then transferred aseptically with oxygen-free CO2 gas displacing all air until red color faded was closed with rubber stopper, sealed and then sterilized in the autoclave at 121^oC for 15 minutes.

The dilution colon fluid in series 10^-7in tube was transferred aseptically to a petri disc with oxygen free CO2 gas displacing all air. Then medium in the enlenmeyer flask wasinoculated into the petri disc and closed. The culture was incubated at 39^oC for 5 – 7 days. The colonies thus grown on this medium were selected for the isolation

Isolation of Colonies

From these inoculated tube containing the selection medium, the individual colonies of lignocellulolytic bacteria were pricked. Bacteria isolates were carefully pricked using bent platinum-irridium needle. The bacteria were then transferred to plate agar medium anaerobically with gasses oxygen-free CO2. The plate was incubated at 39^oC for 5 – 7 days. The bacteria colonies that produces clear or diffusion zone were chosen for furification by repeated streaking.

Ability of Lignocellulosic Substrates Degradation

The ability of lignocellulosic substrates degradation is determined from clear or diffusion zone formed by bacteria isolates tested (Ogimoto and Imai, 1981). The substrate used: CMC for cellulose, xylan for hemicellulose, tannic acid for lignin and combination of 50% CMC, 30%

Xylan and 20% tannic acid for lignocellulose substrate. Each pure bacteria isolate (15μl) was inoculated by spot method using paper disc blank (0.6 cm) and placed on the selective medium (solid growth medium containing 1% substrate test) for experimentation (Subbarao, 1993).

Diffusion and clear zone wide were measured after 24 hours of anaerobic incubation.

Lignoselulase enzyme activity

Enzyme extract was collected from centrifuged liquid media culture in 12.000 rpm for 15 minutes at 4^oC. Extracts enzyme were tested in three kinds of substrates that contained 1% CMC powder/xylan/Tannic Acid in 50 mM acetate buffer pH 5.5. Each substrate liquid in buffer was taken (8 ml), added 1 ml enzymes source, and 1ml aquadest. The mixture then were shaken by shaking bath, enzyme activity was measured in 30, 60, 180 and 360 minutes durations.

Reduction sugar (glucose from CMC and xylose from xylan), or vanillin from tannic acid

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International Journal of Agriculture and Environmental Research

(lignin) produced from the reaction were the cellulase/xylanase or lignaseenzyme activities (Efiok, 1996). For sugar reduction:1 ml of sample was added to 3 ml DNS reagent and 1 ml aquadest (Miller, 1959), for vanillin: 1ml of sample was added to 4 ml methanol, then measured the absorbent by spectrophotometer in wavelength (λ) 508,5 nm for glucose, 509 nm for xilosa and 279 nm for vanillin. Lignase/cellulase/xylanase enzyme activities was estimated by using vanillin/glucose/xylose calibration curve (Adney and Baker, 2008; Ghose, 1987). One unit (U) of enzyme activity was deﬁned as 1 μmolof vanillin/glucose/xylose equivalent released per minuteunder standard assay condition (Irfan et al., 2012; Lo et al., 2009).

RESULTS AND DISCUSSION

Ability of lignocellulosic compounds degradation

The results showed that the lignocellulolytic bacteria isolates were isolated from the bali cattle colon waste has the ability to return high enough to degrade lignocellulosic compounds shown with resultant wide clear zone large enough that 0.078 - 3.440 cm², 0.007 -0.072 cm², 0.172 - 4.497 cm² and 0.392 - 5.864 cm²respectively for on lignocellulosic, tannic acid, CMC and xylan substrates (Table 1). These showed its bacteria isolates as true lignocellulolytic bacteria (lignocellulose degrading bacteria). These isolates were isolated using the combination of lignin (tannic acid), cellulose (carboxy methyl cellulose/CMC) and xylan as selected substrates in bacteria isolated media. Every strain of bacteria isolates needed a spesific substrate as an energy source for growing (Howard et al., 2003; Perez et al., 2002). This suggests that in the colon of Bali cattle are various types of bacteria that have the ability to degrade lignocellulose compounds is high enough (Kamra, 2005) and capable of associating with fiber rich feed ingredients that are not degraded in the rumen (Wahyudi et al., 2010). Kamra (2005) stated that ruminants in tropic area such as Indonesia has higher fiber degrading bacteria than other animal farming, especially local ruminants (bali cattle).

Table 1. Width of clear zone (cm²) obtained from 15 μl Bacteria isolateson variety substrates

No Bacteria Isolates Lignocellulose Lignin CMC Xylan

1 BCC 1 LC 0,725ab¹ 0,007a 1,185b 2,178c

2 BCC 2 LC 2,803c 0,043abc 4,136cd 5,560ef

3 BCC 3 LC 0,885ab 0,027ab 1,335b 1,625b

4 BCC 4 LC 3,357c 0,045bc 4,206cd 5,864f

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International Journal of Agriculture and Environmental Research

5 BCC 5 LC 0,793ab 0,030ab 1,263b 2,253c

6 BCC 6 LC 0,978b 0,030ab 1,516b 2,339c

7 BCC 7 LC 3,440c 0,072c 4,497d 5,042d

8 BCC 11 LC 0,837ab 0,028ab 1,338b 1,858bc

9 BCC 12.1 LC 3,130c 0,044abc 3,901c 5,759f

10 BCC 13.1 LC 0,078a 0,030ab 0,172a 0,392a

SEM² 0,170 0,010 0,080 0,100

Notes: ¹⁾Means in the same column with different letter differ significantly (P<0,05), ²⁾SEM = Standard error of the treatmens and means

Herbivores digestive tract, specifically ruminants in Indonesia such as bali cattle, in general contain high lignocellulose feed, so that lignocellulolytic bacteria isolates were expected exist. This study proved that colon waste bali cattle possess lignocellulolytic bacteria. Several previous study showed that lignocellulolytic bacteria was exist in rumen and goat’s feces has used as rumen microbes replacement (Utomo et al., 2006). Another study showed that horse’s cecum and colon, and elephant have microbes composition such as rumen. (Ulrey et al., 1997;

Wahyudi et al., 2010). According to Table 1, it is proved that the lignocellulolytic bacteria isolates could be found from bali cattle colon waste.

Lignocellulose degrading bacteria (lignocellulolytic bacteria) isolates from bali cattle colon waste has degrading lignocellulosic substrates shown of producing a wide clear zone of 0.078 to 3.440 cm²on lignocellulose substrates (mixture substrates) and individual substrates of 0.007 - 0.072 cm², 0.172 – 4.497 cm², 0.392 -5.864 cm²respectively for lignin (tannic acid), CMC (cellulose), and xylan (hemicellulose) substrates. The bacteria isolates coded BCC2LC, BCC4LC, BCC7LC and BCC12.1LC with the higher clear zone wide and significantly different (P<0.05) compared to the other isolates. This shows the great potential of these bacteria isolates to degrade lignocellulosic compounds such as feed based on agricultural wasteand supply all potential nutrients contained in the substrates/feed.

Bacteria isolates coded BCC 7 LC with the highest wide of clear zone on lignocellulose, lignin, and cellulose substrates (3.440 cm², 0.072 cm² and 4.497 cm²) and significantly different (P<0.05) compared to the other isolates. While on the xylan substrate, bacteria isolate coded BCC 4 LC and BCC 12.1 LC with the highest wide of clear zone and significantly different (P<0.05) compared to the other isolates (5.864 cm² vs 1.685 – 5.560 cm²). These show its bacteria isolates (isolates BCC7LC, BCC4 LC and/or BCC 12.1 LC) able to degrade lignocellulose, lignin, cellulose and xylan to simple compounts and supply all potential nutrients contained in the materials/feeds. Lignocellulose, lignin and cellulose will degrade to simples sugars, energy (heat), CO2, H20 and the others compounts with action of thegroup lignocellulolytic enzyme such

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International Journal of Agriculture and Environmental Research

as lignase, cellulase and xylanase (Howard et al., 2003). At Table 2 also shows its bacteria isolate have higher lignocellulolytic enzyme activity so wide of clear zone produced its isolate are high. This shows the great potential of bacteria isolates to be applied as degrader of lignocellulosic compounts such as agriculture waste and potential benefit in optimizing the development of livestock based on agricultural waste

Lignocellulolytic Enzyme Activity

The results of study showed lignocellulose degrading bacteria from bali cattle colon waste has high lignocellulolytic enzyme activities i.elignase, cellulase, and xylanase enzyme activities (Table 2). These data proved that bacteria isolates has high lignocellulose degradation to simple coumponent such as vanillin, glucose, and/or xylose. This statement is strengthen by Wahyudi et al (2010) studies that the lignocellulolytic bacteria of the colon and/or caecum have the ability fiber to degrade higher than the rumen bacteria. The bacteria isolates capable of associating with fiber rich feed ingredients that are not degraded in the rumen.

The higher lignase enzyme activity of lignocellulolytic bacteria isolates from bali cattle colon waste found from bacteria isolate coded BCC 12.1 LC, BCC 6 LC, BCC 4 LC The bacteria isolates has high lignase activity 0.0045 – 0.0563 IU/ml, 0.0010 – 0.0427 IU/ml, 0.0050 – 0.0196 IU/ml and 0.0067 – 0.0105 IU/ml respectively after contact with the substrates for 30, 60, 180 and 360 minutes. The isolates coded BCC 12.1 LC and BCC 6 LC have shown higher (P<0.05) lignase enzyme activity than the other isolates except observations at 30 min; of the other bacteria isolates BCC 4 LC coded has the highest lignase enzyme activity similar to that of the bacteria isolate coded with BCC 12.1 LC (Table 2A). This show that the bacteria isolates from bali cattle colon waste capable produce lignase enzyme such as phenol oxidase/laccase, lignin peroxidases/Li-P, and/or manganese peroxidade/Mn-P to breakdown of lignin compounds from lignocellulose material (Howard et al., 2003). This statement is strengthen by Wahyudi et al (2010) studies that the lignocellulolytic bacteria of the colon and/or caecum from herbivores animal has produce lignase enzyme and has higher lignase enzyme activity compare with bacteria rumen. In generally, lignase enzyme activities of bali cattle colon waste bacteria isolate not too high. The enzyme activity this study similarly to the lignase enzyme activity of bacteria isolates from Wahyudi et al (2010) research is 0.02 – 0.04 µmol for bacteria isolate from buffalo, horse gastro intestinal tract and elephant dung. These numbers might be due lignin is the most recalcitrant to degradation whereas cellulose or hemicellulose/xylanose, because of its highly ordered crystalline structure and more resistant to hydrolysis (Howard et al., 2003).

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International Journal of Agriculture and Environmental Research

Table 2. Enzyme Activies from lignocellulolytic bacteria isolates from Bali Cattle colon content waste

Bacteria Isolate

Enzyme Activities (IU/ml)

t30 t60 t180 t360

A.Lignase enzyme activities¹⁾

BCC 1 LC 0.0045a⁴ 0.0289bc 0.0121c 0.0067a

BCC 2 LC 0.0435bc 0.0366bc 0.0163d 0.0085b

BCC 3 LC 0.0116a 0.0328bc 0.0050a 0.0087bc

BCC 4 LC 0.0563d 0.0401bc 0.0110c 0.0071a

BCC 5 LC 0.0130a 0.0305bc 0.0075b 0.0080ab

BCC 6 LC 0.0507cd 0.0439c 0.0196e 0.0105d

BCC 7 LC 0.0381b 0.0010a 0.0174d 0.0099cd

BCC 11 LC 0.0057a 0.0260b 0.0050a 0.0087bc

BCC 12.1 LC 0.0563d 0.0427c 0.0196e 0.0105d

BCC 13.1 LC 0.0130a 0.0051a 0.0073b 0.0074ab

SEM⁵ 0,0021 0.0033 0.0004 0.003

B.Cellulase enzyme activities²⁾

BCC 1 LC 0.0493bc 0.0283bc 0,0108bcd 0.0127ab

BCC 2 LC 0.0448b 0.0238b 0.0093ab 0.0108a

BCC 3 LC 0.0503bc 0.0317cd 0.0105bc 0.0109a

BCC 4 LC 0.0682d 0.0309cd 0.0118cd 0.0138bcd

BCC 5 LC 0.0426b 0.0325cd 0.0123cde 0.0141bcd

BCC 6 LC 0.0720d 0.0398e 0.0141e 0.0154cd

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International Journal of Agriculture and Environmental Research

BCC 7 LC 0.0186a 0.0162a 0.0082a 0.0104a

BCC 11 LC 0.0214a 0.0140a 0.0087ab 0.0105a

BCC 12.1 LC 0.0716d 0.0398e 0.0142e 0.0157d

BCC 13.1 LC 0.0626cd 0.0359de 0.0129de 0.0133bc

SEM 0,0029 0,0014 0.0004 0.0005

C.Xylanase enzyme activities³⁾

BCC 1 LC 7.8734ab 9.7222abc 3.7282abc 1.9070a

BCC 2 LC 11.1111abc 11.5066bc 4.7216bcd 2.3485abcd

BCC 3 LC 8.0206ab 7.7134abc 2.5018a 1.9438a

BCC 4 LC 6.4018a 3.8355a 2.9065ab 2.0419ab

BCC 5 LC 11.6630abc 8.6553abc 4.2924abcd 2.3853abcd

BCC 6 LC 20.3826c 13.3830c 5.5923cd 2.9433d

BCC 7 LC 6.9904ab 6.3742ab 4.7952bcd 2.7349cd

BCC 11 LC 17.3657bc 8.7104abc 3.2009ab 1.9316a

BCC 12.1 LC 21.3392c 13.3462c 5.0711bcd 2.6613bcd

BCC 13.1 LC 16.9242abc 12.0438bc 6.4202d 2.2115abc

SEM 2.1088 1.2789 0.4389 0.1244

Notes: 1) Lignaseanalysis using tannic acid substrates, 2) Cellulase (Endo-glucanase)analysis using carboxy methyl cellulose/CMC powder substrates, 3) Xylanaseanalysis using Xylanose substrates, 4)Mean in the same colom with different letter differ significantly (P<0,05).

On cellulase enzyme activity, the lignocellulolytic bacteria isolates has cellulase activities 0.0186 – 0.0720 IU/ml, 0.0140 – 0.0398 IU/ml, 0.0082 – 0.0142 IU/ml, and 0.0104 – 0.0157 IU/ml respectively for 30, 60, 180 and 360 minutes after contact with the carboxy methyl cellulose substrate. The bacteria isolates coded BCC 12.1 LC and BCC 6 LC have shown higher (P<0.05) cellulase enzyme activity than the other isolates (Table 2B). This show that cellulases enzyme from bali cattle colon bacteria isolates responsible for the hydrolysis of cellulose through the breakdown of hydrogen bond in cellulose crystallin structure (α1,4 glukoside bond) from

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cellulose componds. CMC-ase (endoglucanase) are proposed to initiate attack randomly at multiple internalsites in the amorphous regions of the cellulose fibre (Howard et al., 2003; Perez et al., 2002).

Xylan is the most abundant hemicellulose, consist of xylosa polymer and other sugar with ß-1,4 bond and end side chainwith a-1,2 or a-1,3 bonds (Peres et al., 2002).Xylanase enzyme is one of the major hemicellulases which hydrolyse the β-1.4 bond in the xylan backbone yielding short xylooligomers which are further hydrolysed intosingle xylose units by β- xylosidase (Howard et al., 2003). Xylanase enzyme activities of lignocellulolytic bacteria isolates from bali cattle colon waste showed at Table 2C. This study showed that isolate coded BCC 6 LC, BCC 12.1 LC and BCC 13.1 LC has produce higher xylanase enzyme activity (P<0.05) compared to the other isolates on all time durations. This show its bacteria isolates capable better hydrolysis of xylanose to simple sugars ie xylose and the others sugars. Kamra (2005) stated that ruminants in tropic area such as Indonesia has higher fiber degrading bacteria than other animal farming.

In general, the activity of xylanase enzyme of bacterial isolates is quite high. This might be due to xylan/hemicellulose has a structure that is relatively more easily degraded than the other components of lignocellulose. The xylanase enzyme activity in this study is higher than the xylanase enzyme activity of rumen bacteria isolates Bali cattle is 1.449 to 17.1264 U/ml (Partama et al., 2012). This confirms that the isolated bacteria from the colon having enzyme activity were higher than bacteria isolates of rumen cattle. This statement is strengthen by Wahyudiet al (2010) studies that the lignocellulolytic bacteria of the colon and/or caecum have the ability fiber to degrade higher than the rumen bacteria.

Based on the enzyme activities value obtained, the bacteria isolates coded BCC 12.1 LC and BCC 6 LC have the higher quality and most potencial as lignocellulosic substrates degrader.

Lignoselulase high enzyme activity show a high ability of bacterial isolates to degrade lignocellulose compounds as well as a variety of fiber-rich feedstuffs to simple nutrients/compounds which easy to degrade or digestible for animal/livestock. It is feasible that the isolates most potential to be used as fermentor/degrader of material/feed resources rich in crude fiber and lignocellulose such as agriculture waste product.

CONCLUSION

1. Lignocellulolytic bacteria isolates coded BCC 7 LC, BCC 4 LC, BCC 12.1 LC, and BCC 2 LC has produced higher width of clear zone

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2. Lignocellulolytic bacteria isolates coded BCC 12.1 LC and BCC 6 LC higher lignocellulolytic enzyme activity

3. The lignocellulolytic bacteria isolates from bali cattle colon most potential to be used as degrader of feedstuff rich in lignocellulosic substrates such as agriculture waste product.

ACNOWLEDGEMENTS

The authors acknowledge to the Directorate General of Higher Education, National Education Department, Republic of Indonesia and Udayana University through Competitive Research Grant 2014 for fund support in this experimentation. Thanks are also due to the Laboratory of feed and nutrition animal, Faculty of Animal Husbandry Udayana University for assistance in isolation and characterization bacteria isolates analysis.

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