• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.11
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• pp.1738-1746
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• 2018

Objective: The aim of the experiment was to assess the effect of increasing amounts of Leucaena leucocephala forage on dry matter intake (DMI), organic matter intake (OMI), enteric methane production, rumen fermentation pattern and protozoa population in cattle fed Pennisetum purpureum and housed in respiration chambers. Methods: Five crossbred heifers (Bos taurus${\times}$Bos indicus) (BW: $295{\pm}6kg$) were fed chopped P. purpureum grass and increasing levels of L. leucocephala (0%, 20%, 40%, 60%, and 80% of dry matter [DM]) in a $5{\times}5$ Latin square design. Results: The voluntary intake and methane production were measured for 23 h per day in respiration chambers; molar proportions of volatile fatty acids (VFAs) were determined at 6 h postprandial period. Molar concentration of VFAs in rumen liquor were similar (p>0.05) between treatments. However, methane production decreased linearly (p<0.005), recording a maximum reduction of up to ~61% with 80% of DM incorporation of L. leucocephala in the ration and no changes (p>0.05) in rumen protozoa population were found. Conclusion: Inclusion of 80% of L. leucocephala in the diet of heifers fed low-quality tropical forages has the capacity to reduce up to 61.3% enteric methane emission without affecting DMI, OMI, and protozoa population in rumen liquor.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.199-202
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• 2004

Cellulolytic bacterial strains have been isolated from the faeces of wild (blackbuck, Antilope cervicapra; nilgai, Baselophus tragocamelus chinkara, Gazella gazella spotted deer, Axis axis and hog deer, Cervus porcinus) and rumen liquor of domestic (sheep, Ovis aries) ruminants. Five best cellulose degrading bacterial isolates (Ruminococcus sp.) were used as microbial feed additive along with buffalo rumen liquor as inoculum to study their effect on digestibility of feed and enzyme production in in vitro conditions. The bacterial isolate from chinkara (CHI-2) showed the highest per cent apparent dry matter (DM) digestibility ($35.40{\pm}0.60$), true dry matter digestibility ($40.80{\pm}0.69$) and NDF ($26.38{\pm}0.83$) digestibility (p<0.05) compared to control ($32.73{\pm}0.56$, $36.64{\pm}0.71$ and $21.16{\pm}0.89$, respectively) and other isolates at 24 h of incubation with lignocellulosic feeds (wheat straw and wheat bran, 80:20). The same isolate also exhibited the highest activities of fibre degrading enzymes like carboxymethylcellulase, xylanase, ${\beta}$-glucosidase and acetyl esterase. The bacterial isolate from chinkara (Gazella gazella) appears to have a potential to be used as feed additive in the diet of ruminants for improving utilization of nutrients from lignocellulosic feeds.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.1
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• pp.38-41
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• 2005

An experiment was conducted on 3 male rumen fistulated adult buffaloes fed on wheaten straw and concentrate mixture in a Latin square design to study the impact of niacin supplementation on rumen metabolites. Three animals were fed wheaten straw+concentrate mixture (group I, control), wheaten straw+concentrate mixture+100 ppm niacin (group II), and wheaten straw +concentrate mixture+200 ppm niacin (group III). After 21 days feeding, rumen liquor was drawn for 3 consecutive days at different time intervals (0, 2, 4, 6 and 8 h) to study the various rumen metabolites i.e., rumen pH, ammonia-N, total-N, trichloroacetic acid precipitable-N, non-protein nitrogen, total volatile fatty acids, their fractions and number of protozoa. Mean pH values in strained rumen liquor (SRL) of animals in 3 groups were 6.64, 6.71 and 6.67, indicating no statistically significant difference. Results revealed a significant (p<0.01) increase in TVFA concentration among the supplemented groups (group II and III) in comparison to control group. Mean TVFA concentration (meq/dl) was 9.75, 10.97 and 11.44 in 3 groups respectively. The highest concentration of TVFA was observed at 4 h and minimum at 0 h in all the 3 groups. The percentage of acetic, propionic, butyric and isobutyric acid was statistically similar among the three groups. The mean ammonia-N concentration (mg/dl SRL) was significantly (p<0.01) lower in group II (16.38) and group III (15.42) than group I (18.14). Ammonia-N concentration was higher (p<0.01) at 4 h as compared to all the time intervals. The mean total-N concentration (mg/dl SRL) was higher (p<0.01) in group II (74.16) and group III (75.47) as compared to group I (62.04). Total-N concentration was higher (p<0.01) at 4 h as compared to other time intervals and lowest value was recorded at 0 h.Concentration of TCA-ppt-N (mg/dl SRL) was significantly (p<0.01) lower in control group as compared to niacin supplemented groups. Mean value of NPN (mg/dl SRL) was significantly (p<0.01) lower in group III (23.21) as compared to group I (25.71), whereas groups I and II, and groups II and III were similar to each other. Total protozoa number (${\times}10^4$/ml SRL) ranged from 18.06 to 27.41 in group I, 20.89 to 38.44 in group II and 27.61 to 39.45 in group III. The mean protozoa number was significantly (p<0.01) higher in SRL of group II (27.60) and III (30.59) as compared to group I (22.48). It can be concluded from the study that supplementation of niacin in the diet of buffaloes had improved the rumen fermentation by decreasing the concentration of ammonia-N and increasing protein synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1057-1066
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• 2007

Six male Gayal (Bos frontalis), approximately two years of age and with a mean live weight of $203{\pm}17$ kg ($mean{\pm}standard\;deviation$), were housed indoors in metabolism cages and fed bamboo (Sinarundinaria) leaves and twigs. After an adjustment period of 24 days of feeding the diet, samples of rumen liquor were obtained for analyses of bacteria in the liquor. The diversity of rumen bacteria was investigated by constructing a 16S rDNA clone library. A total of 147 clones, comprising nearly full length sequences (with a mean length of 1.5 kb) were sequenced and submitted to an on-line similarity search and phylogenetic analysis. Using the criterion of 97% or greater similarity with the sequences of known bacteria, 17 clones were identified as Ruminococcus albus, Butyrivibrio fibrosolvens, Quinella ovalis, Clostridium symbiosium, Succiniclasticum ruminis, Selenomonas ruminantium and Allisonella histaminiformans, respectively. A further 22 clones shared similarity ranging from 90-97% with known bacteria but the similarity in sequences for the remaining 109 clones was less than 90% of those of known bacteria. Using a phylogenetic analysis it was found that the majority of the clones identified (57.1%) were located in the low G+C subdivision, with most of the remainder (42.2% of clones) located in the Cytophage-Flexibacter-Bacteroides (CFB) phylum and one clone (0.7%) was identified as a Spirochaete. It was apparent that Gayal have a large and diverse range of bacteria in the rumen liquor which differ from those of cattle and other ruminants. This may explain the greater live weights of Gayal, compared to cattle, grazing in the harsh natural environments in which Gayal are located naturally.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.6
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• pp.812-817
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• 2012

Nitrate can serve as a terminal electron acceptor in place of carbon dioxide and inhibit methane emission in the rumen and nitrate reducing bacteria might help enhance the reduction of nitrate/nitrite, which depends on the type of feed offered to animals. In this study the effects of three levels of sodium nitrate (0, 5, 10 mM) on fermentation of three diets varying in their wheat straw to concentrate ratio (700:300, low concentrate, LC; 500:500, medium concentrate, MC and 300:700, high concentrate, HC diet) were investigated in vitro using buffalo rumen liquor as inoculum. Nitrate reducing bacteria, isolated from the rumen of buffalo were tested as a probiotic to study if it could help in enhancing methane inhibition in vitro. Inclusion of sodium nitrate at 5 or 10 mM reduced (p<0.01) methane production (9.56, 7.93 vs. 21.76 ml/g DM; 12.20, 10.42 vs. 25.76 ml/g DM; 15.49, 12.33 vs. 26.86 ml/g DM) in LC, MC and HC diets, respectively. Inclusion of nitrate at both 5 and 10 mM also reduced (p<0.01) gas production in all the diets, but in vitro true digestibility (IVTD) of feed reduced (p<0.05) only in LC and MC diets. In the medium at 10 mM sodium nitrate level, there was 0.76 to 1.18 mM of residual nitrate and nitrite (p<0.01) also accumulated. In an attempt to eliminate residual nitrate and nitrite in the medium, the nitrate reducing bacteria were isolated from buffalo adapted to nitrate feeding and introduced individually (3 ml containing 1.2 to $2.3{\times}10^6$ cfu/ml) into in vitro incubations containing the MC diet with 10 mM sodium nitrate. Addition of live culture of NRBB 57 resulted in complete removal of nitrate and nitrite from the medium with a further reduction in methane and no effect on IVTD compared to the control treatments containing nitrate with autoclaved cultures or nitrate without any culture. The data revealed that nitrate reducing bacteria can be used as probiotic to prevent the accumulation of nitrite when sodium nitrate is used to reduce in vitro methane emissions.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.7
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• pp.990-996
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• 2005

The effect of diets varying in level and source of nitrogen (N) and fermentable organic matter on dynamic characteristics of microbial populations in rumen liquor and their impact on substrate fermentation in vitro was studied. The diets tested were straw alone, straw+concentrate mixture and straw+urea molasses mineral block (UMMB) lick. The same diets were taken as substrates and tested on each inoculum collected from the diets. Diet had no effect on the amino acid (AA) composition of either bacteria or protozoa. Differences among the diets in intake, source of N and OM affected bacterial and protozoal characteristics in the rumen. Upper asymptote of gas production (Y$\alpha$) had a higher correlation with bacterial pool size and production rate than with protozoal pool size and production rate. Among the parameters of the gas production model, Y$\alpha$ and lag time in total gas has showed significant (p<0.01) correlation with bacterial characteristics. Though the rate constant of gas production significantly differed (p<0.01) between diet and type of straw, it was least influenced by the microbial characteristics. The regression coefficient of diet and type of straw for Y$\alpha$ indicated that the effect of diet on Y$\alpha$ was threefold higher than that of the straw. As microbial characteristics showed higher correlation with Y$\alpha$, and diet had more influence on the microbial characteristics, gas production on a straw diet could be used effectively to understand the microbial characteristics.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.665-670
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• 2002

The expediency of replacing cost prohibitive and often inaccessible traditional protein supplements prompted the monitoring of hematological parameters was carried out in female goats at 0, 30, 60 and 90 days post feeding. Rumen environment was (3), respectively fed supplements containing either a leaf meal mixture (LMTM) of Leucaena leucocephala-Morus alba-Tectona grandis (2:1:1) or traditional protein supplements groundnut cake (GNC) or soybean meal (SBM) and wheat straw as basal diet. The periodic monitoring of hematological parameters was carried out in female goats at 0, 30, 60 and 90 days post feeding. Rumen environment was studied in bucks in a $3{\times}3$ switch over design. Rumen liquor was collected at 0, 2, 4, 6 and 8 h post feeding after 4 weeks of feeding. The goats fed on LMTM or GNC had similar dry matter intake (g/kg $W^{0.75}$), which was significantly (p<0.05) higher than SBM. Except for packed cell volume (PCV), none of the blood biochemical constituents (Hemoglobin, serum glucose, total protein, serum albumin (A) and globulin(G), A:G ratio, alkaline phosphatase, transaminases) varied significantly due to replacement of 50% dietary protein by LMTM throughout the experiment. GNC group had significantly higher level of PCV than other treatments. However, the level of serum total protein (p<0.01) tended to increase from 60th day onwards irrespective of dietary treatments. The average rumen pH was significantly higher (p<0.001) on SBM followed by LMTM and GNC, respectively. Total volatile fatty acid (TVFA) production was comparable in goats given LMTM or GNC supplements, the corresponding values were significantly different (p<0.001) when compared with SBM. The ammonical-N, total-N and TCA-precipitable-N (mg/100 ml SRL) did not differ significantly among dietary treatments. It may be concluded that supplementing wheat straw with LMTM based concentrate had no adverse effect on voluntary intake, blood biochemical profile and rumen fermentation pattern of the goats.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.5
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• pp.728-734
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• 1999

Thirty new born crossbred (Bos taurus${\times}$Bos indicus) calves, divided randomly in a $3{\times}2$ factorial design, were fed calf starters containing one of three protein sources i.e., groundnut cake (GN), cottonseed meal (CS) and meat and bone meal (MB) along with either raw (M) or gelatinized maize (MG) for 90d. Milk was fed upto 56d of age. Green oats and respective calf starters were offered from 14d of age onwards ad lib. Clinical profile of serum suggested significantly (p<0.05) higher albumin and lower alanine aminotransferase activity due to CS feeding. Alklaine phosphatase activity varied significantly (p<0.05) among dietary treatments showing interaction between protein and starch sources. Inclusion of gelatinized maize resulted in significantly higher concentration of serum globulin (p<0.05) and alkaline phosphatase activity (p<0.01). reduced (p<0.05) ruminal pH was accompanied by a significant decrease (p<0.01) in $NH_3-N$ concentration in the strained rumen liquor (SRL) of MG fed calves. Ruminal amylase activity was lower (p<0.05) on MG diets. Alanine aminotransferase activity in the rumen exhibited a significant (p<0.01) interaction between protein and starch sources. While feeding of CS significantly (p<0.01) reduced alanine aminotransferase activity, inclusion of thermally processed maize reduced (p<0.01) both aspartate and alanine aminotransferase activities in the rumen. The overall blood picture was similar among treatments, whereas rumen metabolites especially enzyme activities, seems to be altered with source of degradable protein an starch.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1606-1613
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• 2015

This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.