• Title/Summary/Keyword: Rumen Microbiology

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Molecular cloning, expression and characterization of a novel feruloyl esterase enzyme from the symbionts of termite (Coptotermes formosanus) gut

  • Chandrasekharaiah, Matam;Thulasi, Appoothy;Bagath, M.;Kumar, Duvvuri Prasanna;Santosh, Sunil Singh;Palanivel, Chenniappan;Jose, Vazhakkala Lyju;Sampath, K.T.
    • BMB Reports
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    • v.44 no.1
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    • pp.52-57
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    • 2011
  • Termites play an important role in the degradation of dead plant materials and have acquired endogenous and symbiotic cellulose digestion capabilities. The feruloyl esterase enzyme (FAE) gene amplified from the metagenomic DNA of Coptotermes formosanus gut was cloned in the TA cloning vector and subcloned into a pET32a expression vector. The Ft3-7 gene has 84% sequence identity with Clostridium saccharolyticum and shows amino acid sequence identity with predicted xylanase/chitin deacetylase and endo-1,4-beta-xylanase. The sequence analysis reveals that probably Ft3-7 could be a new gene and that its molecular mass was 18.5 kDa. The activity of the recombinant enzyme (Ft3-7) produced in Escherichia coli (E.coli) was 21.4 U with substrate ethyl ferulate and its specific activity was 24.6 U/mg protein. The optimum pH and temperature for enzyme activity were 7.0 and $37^{\circ}C$, respectively. The substrate utilization preferences and sequence similarity of the Ft3-7 place it in the type-D sub-class of FAE.

Effect of Diet on Enzyme Profile, Biochemical Changes and In sacco Degradability of Feeds in the Rumen of Buffalo

  • Kamra, D.N.;Saha, Sudipto;Bhatt, Neeru;Chaudhary, L. C.;Agarwal, Neeta
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.374-379
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    • 2003
  • Four rumen fistulated Murrah buffaloes were used to study the effect of four diets differing in roughage to concentrate ratio on rumen biochemical changes, microbial enzyme profile and in sacco degradability of feed in a $4{\times}4$ Latin Square design. The animals were fed four diets consisting of 80:20, 70:30, 60:40 and 50:50 ratios of wheat straw and concentrate mixtures, respectively. Wheat straw and concentrate mixture were mixed with water (0.6 l/kg feed) and complete feed mixture was offered to the animals at 8:00 h and 16:00 h in two equal parts. The variation in pH of rumen liquor (difference of maximum and minimum during 0-8 h post feeding) increased with increasing level of concentrate mixture in the diet. There was no effect of diet composition on volatile fatty acids, total nitrogen and trichloro-acetic acid precipitable nitrogen in the rumen liquor, but ammonia nitrogen increased with increasing level of concentrate mixture in the ration. Major portions of all fibre degrading enzymes were present in the particulate material (PM) of the rumen contents, but protease was absent in PM fraction. The activities of micro-crystalline cellulase, acetyl esterase and protease increased with increase in the level of concentrate mixture, but the activities of other enzymes (carboxymethylcellulase, filter paper degrading activity, xylanase, $\beta$-glucosidase and $\beta$-xylosidase) were not affected. The in sacco degradability and effective degradability of feeds increased with increasing level of concentrate mixture in the ration.

Effect of Feeding Ficus infectoria Leaves on Rumen Microbial Profile and Nutrient Utilization in Goats

  • Singh, B.;Chaudhary, L.C.;Agarwal, N.;Kamra, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.6
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    • pp.810-817
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    • 2011
  • A feeding trial was conducted to study the effect of tannin rich Pakar (Ficus infectoria) leaves on microbial profile, rumen fermentation and nutrient utilization in goats. Eight goats divided in two groups were fed pakar leaves (experimental group) and green oats (control group) as sole roughage source along with a fixed quantity of concentrate mixture for a period of 3 months. Two metabolic trials of six days duration were conducted after 30 and 90 days of experimental feeding. The dry matter intake was significantly higher (p<0.05) and digestibility's of DM, OM, CP, EE, NDF and ADF were reduced in experimental as compared with the control group. The TDN intake was similar (236.52 vs. 240.39 g/d) in both the groups. All the animals were in positive nitrogen balance. The concentration of ammonia nitrogen, TVFA, lactic acid and activities of xylanase and protease were reduced in pakar leaves fed goats. The rumen microbial profile as obtained by MPN technique showed no change in total bacterial population but total fungi and cellulolytic bacteria were reduced (p<0.05), whereas, tannin degrading/tolerant bacteria increased with the feeding of pakar leaves. Real time PCR data revealed a decrease in Ruminococcus flavefaciens, an increase in methanogens and no change in the Fibrobacter succinogenes population by feeding of pakar leaves.

Enumeration and Recovery of Bacterial Isolates from Ruminants Fed with Different Dietary Regimes and Their Antibacterial Activity

  • Pattnaik, P.;Grover, Sunita;Batish, V.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.811-815
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    • 2001
  • The study evaluated different synthetic and semisynthetic media for maximal recovery of rumen bacteria and expression of their antibacterial activity. Rumen Glucose Cellobiose Agar (RGCA) medium was found to be the best for recovery of rumen bacteria. However, L-10 medium was the best for expression of antibacterial activity of ruminal isolates followed by Easy, M-10, RGCA and M-98-5 medium. The present study recommends the use of L-10 medium as the medium of choice for screening of antibacterial activity of ruminal isolates. Comparative evaluation of bacterial counts on different dietary regimes indicated significant difference between different growth media on a specific diet and between diets on specific growth media within a species. However, there is no overall significant difference between total bacterial counts obtained from rumen liquor of cattle and buffalo with respect to either the feeding regime or growth media. Feeding straw based diet to the animal is the best for high recovery of rumen bacteria.

EFFECTS OF CHEMICAL TREATMENTS OF BARLEY STRAW ON LEACHING, AND DIGESTIBILITY BY RUMEN FLUID AND CELLULOLYTIC BACTERIA

  • Kudo, H.;Cheng, K.J.;Rode, L.M.;Abdullah, N.;Ho, Y.W.;Hussain, H.Y.;Jalaludin, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.389-396
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    • 1994
  • Effects of chemical treatments on in sacco and in vitro digestibility of barley straw by rumen fluid and pure cultures of cellulolytic bacteria were studied to evaluate the pretreatment and to improve the poor quality feed. Chemicals were applied by dissolving them in water equivalent to 40% of the weight of the straw (dry matter basis). Pretreatment with 5% NaOH yielded the largest increase in sacco digestion followed by pretreatment with 2% $(NH_4)_2SO_3$, 2.6% $NH_4OH$, 1.6% $NaHSO_3$ and untreated straw (control). In sacco dry matter digestibility of straw treated with NaOH and $(NH_4)_2SO_3$ continued to increase as the concentration of chemical increased (1 to 7.5%), as it was the in vitro dry matter loss by leaching. Treatment of barley straw with 5% NaOH enhanced significantly (p < 0.01) in vitro digestibility by rumen fluid, Fibrobacter suceinogenes and Ruminococcus albus though the fermentation products by cellulolytic bacteria were low, whereas the treatment with 5% $(NH_4)_2SO_3$ inhibited in vitro digestibility by F. succinogenes and R. albus together with lower fermentation products. Dry matter loss by leaching and bacterial digestion from barley straw treated with NaOH and $(NH_4)_2SO_3$ suggested the effect of pretreatment with these chemicals were based on leaching, and the cellulolytic bacteria had little to do with digestion.

In vitro Methanogenesis, Microbial Profile and Fermentation of Green Forages with Buffalo Rumen Liquor as Influenced by 2-Bromoethanesulphonic Acid

  • Agarwal, Neeta;Kamra, D.N.;Chatterjee, P.N.;Kumar, Ravindra;Chaudhary, L.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.6
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    • pp.818-823
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    • 2008
  • The interaction of fibre degrading microbes and methanogens was studied using two forages, lucerne (Medicago sativa) hay and maize (Zea mays) hay, as substrate and 2-bromoethanesulphonic acid (BES) as an additive in an in vitro gas production test. Gas and methane production (ml/g dry matter) were significantly higher (p<0.05) on lucerne as compared to maize hay. Inclusion of BES in the incubation medium significantly suppressed methane emission irrespective of substrate. The population density of total bacteria, fungi, Ruminococcus flavefaciens and Fibrobacter succinogenes was higher, whereas that of methanogens was lower with maize hay as compared to lucerne as substrate. BES suppressed methanogen population by 7 fold on lucerene and by 8.5 fold on maize at 24 h incubation as estimated by real time-PCR. This suppression was accompanied by almost complete (>98% of control) inhibition of methanogenesis. The proportion of acetate decreased, whereas that of propionate increased significantly by inclusion of BES, resulting in narrowing of acetate to propionate ratio. In vitro true digestibility (IVTD) of lucerne was significantly higher as compared to maize but BES inclusion did not affect IVTD.

Effect of feeding of blend of essential oils on methane production, growth, and nutrient utilization in growing buffaloes

  • Yatoo, M.A.;Chaudhary, L.C.;Agarwal, N.;Chaturvedi, V.B.;Kamra, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.5
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    • pp.672-676
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    • 2018
  • Objective: An experiment was conducted to study the effect of a blend of essential oils (BEO) on enteric methane emission and growth performance of buffaloes (Bubalus bubalis). Methods: Twenty one growing male buffaloes (average body weight of $279{\pm}9.3kg$) were divided in to three groups. The animals of all the three groups were fed on a ration consisting of wheat straw and concentrate mixture targeting 500 g daily live weight gain. The three dietary groups were; Group 1, control without additive; Group 2 and 3, supplemented with BEO at 0.15 and 0.30 mL/kg of dry matter intake (DMI), respectively. Results: During six months feeding trial, the intake and digestibility of dry matter and nutrients (organic matter, crude protein, ether extract, neutral detergent fibre, and acid detergent fibre) were similar in all the groups. The average body weight gain was tended to improve (p = 0.084) in Group 2 and Group 3 as compared to control animals. Feeding of BEO did not affect feed conversion efficiency of the animals. The calves of all the three groups were in positive nitrogen balance with no difference in nitrogen metabolism. During respiration chamber studies the methane production (L/kg DMI and L/kg digestible dry matter intake was significantly (p<0.001) lower in Group 2 and Group 3 as compared to control animals. Conclusion: The results indicated that the BEO tested in the present study have shown potential to reduce enteric methane production without compromising the nutrient utilization and animal performance and could be further explored for its use as feed additive to mitigate enteric methane production in livestock.

Fumarate Reductase-Producing Enterococci Reduce Methane Production in Rumen Fermentation In Vitro

  • Kim, Seon-Ho;Mamuad, Lovelia L.;Kim, Dong-Woon;Kim, Soo-Ki;Lee, Sang-Suk
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.558-566
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    • 2016
  • Biotic agents such as fumarate-reducing bacteria can be used for controlling methane (CH4) production in the rumen. Fumarate-reducing bacteria convert fumarate to succinate by fumarate reductase, ultimately leading to the production of propionate. Fumarate-reducing bacteria in the genus Enterococcus were isolated from rumen fluid samples from slaughtered Korean native goats. The enterococci were identified as Enterococcus faecalis SROD5 and E. faecium SROD by phylogenetic analyses of 16S rRNA gene sequences. The fumarate reductase activities of the SROD5 and SROD strains were 42.13 and 37.05 mM NADH oxidized/min/mg of cellular nitrogen (N), respectively. Supplementation of rumen fermentation in vitro with the SROD5 and SROD strains produced significantly higher propionate, butyrate, and total volatile fatty acid (VFA) concentrations than controls at 12 h; VFA concentrations tended to increase after 24 h of incubation. The generated CH4 concentration was significantly lower in the SROD5 and SROD treatment groups after 24 h of incubation. These findings indicate that E. faecium SROD has potential as a direct-fed microbial additive for increasing total VFAs while decreasing CH4 production in rumen fermentation in vitro.

Comparative Study between Swamp Buffalo and Native Cattle in Feed Digestibility and Potential Transfer of Buffalo Rumen Digesta into Cattle

  • Wanapat, M.;Nontaso, N.;Yuangklang, C.;Wora-anu, S.;Ngarmsang, A.;Wachirapakorn, C.;Rowlinson, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.4
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    • pp.504-510
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    • 2003
  • Rumen ecology plays an important role in the fermentation process and in providing end-products for ruminants. These studies were carried out to investigate variations in rumen factors namely pH, $NH_3-N$ and microorganisms in cattle and swamp buffaloes. Furthermore, studies on diurnal patterns of rumen fermentation and the effect of rumen digesta transfer from buffalo to cattle was conducted. Based on these studies, diurnal fermentation patterns in both cattle and buffaloes were revealed. It was found that rumen NH3-N was a major limiting factor. Rumen digesta transfer from buffalo to cattle from buffalo to cattle was achievable. Monitoring rumen digesta for 14d after transfer showed an improved rumen ecology in cattle as compared to that of original cattle and buffalo. It is probable that buffalo rumen digesta could be transferred. However, further research should be undertaken in these regards in order to improve rumen ecology especially for buffalo-based rumen.