• Applied Biological Chemistry
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• v.17 no.2
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• pp.138-142
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• 1974

Biosynthesis of glycolipid from glucosamine and $C_{13:0}$ fatty acid was attempted using an enzyme preparation which was extracted from Selenomonas ruminantium grown in lactic acid medium by means of ultrasonication, and with cofactors of ATP, Co A, $Mg^{++}$, and UTP. The results are summarized as follows: 1. The rate of synthesis of glycolipid from $^{14}C-glucosamine$ and tridecyl CoA by an enzyme from Selenomonas ruminantium was promoted by the presence of co-factors, ATP, Mg, and UTP. 2. The rate of incorporation of $^{14}C-glucosamine$ into glycolipid by the centrifugal fraction of bacterial preparation was the highest in the 105,000 g supernatant fraction, indicating about twice the enzyme activity as the 6,000 rpm supernatant fraction. 3. The biosynthesis of glycolipid from $^{14}C-glucosamine$ and tridecyl-CoA by the crude enzyme of the 105,000 g supernatant fraction of Selenomonas ruminantium cell preparation proceeded the most part of it in 30 minutes and completed in an hour.

• Asian-Australasian Journal of Animal Sciences
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• v.2 no.3
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• pp.460-461
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• 1989

• Applied Biological Chemistry
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• v.17 no.2
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• pp.125-137
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• 1974

The chemical structure of glycolipid of Selenomonas ruminantium cell wall was to be elucidated. The bacterial cells were treated in hot TCA and the glycolipid fractions were extracted by the solvent $CHCl_3\;:\;CH_3OH$ (1 : 3). The extracted glycolipids fraction was further separated by acetone extraction. The acetone soluble fraction was named as the spot A-compound. The acetone insoluble but ether soluble fraction was named as the spot B-compound. These two compounds were examined for elucidation of their chemical structure. The results were as follows: 1. The IR spectral analysis showed that O-acyl and N-acyl fatty acids were linked to glucosamine moiety in the spot A-compound. However in the spot B-compound in addition to O and N-acyl acids phosphorus was shown to be attached to glucosamine. 2. It was recognized by gas liquid chromatography that spot A compound contained beta-OH $C_{13:0}$ fatty acid in predominance in addition to the fatty acid with beta-OH $C_{9:0}$, whereas the spot B compound was composed of the predominant fatty acid of beta-OH $C_{13:0}$ with small amount of beta-OH $C_{9:0}$. 3. According to the paper chromatographic analysis of hydrazinolysis products of the spot A compound, a compound of a similar Rf value as the chitobiose was recognized, which indicated a structure of two molecules glucosamine condensed. The low Rf value of the hydrazinolysis product of the spot B-compound confirmed the presence of phosphorus attached to glucosamine. 4. The appearance of arabinose resulting from. ninhydrin decomposition of the acid hydrolyzate of the spot A compound indicated that the amino group is attached to $C_2$ of glucosamine. 5. The amount of glucosamine in the N-acetylated spot A compound decreased in half of the original content by the treatment. with $NaBH_4$, indicating that there are two molecules of glucosamines in the spot A compound. The presence of 1, 6-linkage between two molecules of glucosamine was suggested by the Morgan-Elson reaction and confirmed by the periodate decomposition test. 6. By the action of ${\beta}-N-acetyl$ glucosaminidase the N-acetylated spot A compound was completely decomposed into N-acetyl glucosamine, whereas the spot B compound was not. This indicated the spot A compound has a beta-linkage. 7. When phosphodiesterase or phosphomonoesterase acted on $^{32}P-labeled$ spot B compound, $^{32}P$ was not released by phosphodiesterase, but completely released by phosphomonoesterase. This indicated that one phosphorus is linked to glucosamine moiety. 8. The spot A compound is assumed to have the following chemical structure: That is glucosaminyl, ${\beta}-1$, 6-glucosamine to which O-acyl and N-acyl fatty acids are linked, of which the predominant fatty acid is beta-OH $C_{13:0}$ fatty acid in addition to beta-OH $C_{9:0}$ fatty acid 9. The spot B compound is likely to have the linkage of $glucosaminyl-{\beta}-1$, 6-glucosamine to which phosphorus is linked in monoester linkage. Furthermore both O-acyl and N-acyl fatty acids contained beta-OH $C_{13:0}$ fatty acid predominantly in addition to beta-OH $C_{9:0}$ fatty acid.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.11
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• pp.1552-1559
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• 2011

Real time PCR was used in this study to determine the effect of triticale dried distillers grains with solubles (TDDGS) as a replacement for grain or barley silage in finishing diets on the presence of six classical ruminal bacterial species (Succinivibrio dextrinosolvens, Selenomonas ruminantium, Streptococcus bovis, Megasphaera elsdenii, Prevotella ruminicola and Fibrobacter succinogenes) within the rumen contents of feedlot cattle. This study was divided into a step-wise adaptation experiment (112 days) that examined the effects of adaptation to diets containing increasing levels of TDDGS up to 30% (n = 4), a short-term experiment comparing animals (n = 16) fed control, 20%, 25% or 30% TDDGS diets over 28 days, and a rapid transition experiment (56 days) where animals (n = 4) were rapidly switched from a diet containing 30% TDDGS to a barley-based diet with no TDDGS. It was found that feeding TDDGS as replacement for barley grain (control vs. 20% TDDGS) decreased 16S rRNA copy numbers of starch-fermenting S. ruminantium and S. bovis (p<0.001 and p = 0.04, respectively), but did not alter 16S rRNA copy numbers of the other rumen bacteria. Furthermore, feeding TDDGS as a replacement barley silage (20% vs. 25% and 30% TDDGS) increased 16S rRNA copy numbers of S. ruminantium, M. elsdenii and F. succinogenes (p<0.001; p = 0.03 and p<0.001, respectively), but decreased (p<0.001) the 16S rRNA copy number of P. ruminicola. Upon removal of 30% TDDGS and return to the control diet, 16S rRNA copy numbers of S. ruminantium, M. elsdenii and F. succinogenes decreased (p = 0.01; p = 0.03 and p = 0.01, respectively), but S. dextrinosolvens and S. bovis increased (p = 0.04 and p = 0.009, respectively). The results suggest that replacement of TDDGS for grain reduces 16S rRNA copy numbers of starch-fermenting bacteria, whereas substitution for barley silage increases 16S rRNA copy numbers of bacteria involved in fibre digestion and the metabolism of lactic acid. This outcome supports the contention that the fibre in TDDGS is highly fermentable.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.672-676
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• 2003

Effect of a surfactant Tween 80 on the bacterial growth in the rumen was examined on the in vitro pure cultures of Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Prevotella ruminicola, Megasphaera elsidenni, Fibrobacta succinogenes, Ruminanococcus albus and Ruminococcus flavefaciens. Dry matter degradability (DMD), concentrations and compositions of volatile fatty acids (VFA), and the most probable number (MPN) of cellulolytic bacteria and total number of bacteria in the presence of Tween 80 were also examined on the in vitro rumen mixed culture either with barley grain or orchardgrass hay. The growth of S. bovis, S. ruminantium, B. fibrisolvens, P. ruminicola, M. elsidenni and F. succinogenes were significantly higher (p<0.05) at over 0.05% concentrations of Tween 80 than those of the control cultures, while was not changed with R. albus and R. flavefaciens. With rumen mixed culture the DMD of barley grain and orchardgrass hay was significantly higher (p<0.05) at a 0.2% concentration of Tween 80 than the control, being reflected in the significantly higher (p<0.05) VFA production (mmol $g^{-1}$DDM) with orchardgrass hay. The higher (p<0.05) ratio of propionate to acetate at a 0.2% concentration of Tween 80 was also observed with orchardgrass hay, showing a similar trend with barley grain. No changes in the total bacterial number and MPN of cellulolytic bacteria were observed.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.5
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• pp.708-714
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• 1999

Fermentation of legume fodder tree leaves by rumen microorganisms was evaluated. The substrates were sun-dried, ground leaves. Gas and volatile fatty acid (VFAs) production were estimated. Using gas production as an index of fermentation at 12 h, the leaves tested ranked as follows; Chamaecytisus palmensis>Gliricidia sepium>Sebania sesban>Tephrosia bracteolate>Leucaena pallida>Vernonia amygdalina>Acacia sieberiana>Sesbania goetzei>Acacia angustissima. Using VFA production, the ranking was a follows; G. sepium>S. sesban>S. goetzei>L. pallida>C. palmensis/V. amygdalina>T. bracteolate> A. sieberiana>A. angustissima. Absolute gas or VFA production rates, were also used to rank the leaves. Extracts (70% acetone) of A. angustissima inhibited the growth of Ruminococcus albus 8, R. flavefaciens FD-1, Prevotella ruminicola D3ID and Streptococcus bovis JBI while the trowth of Selenomonas ruminantium D was depressed when 0.6 ml exracts were added. C. palmensis water extracts enhanced cellulose hydrolysis by R. flavefaciens FD-1. All extracts reduced celluloysis by R. albus 8. R. flavefaciens FD-1 hydrolyzed more (p<0.001) cellulose than R. albus 8.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.1
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• pp.50-57
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• 2015

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1433-1441
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• 2015

This study examined changes of rumen fermentation, ruminal bacteria biodiversity and abundance caused by nitrate addition with Ion Torrent sequencing and real-time polymerase chain reaction. Three rumen-fistulated steers were fed diets supplemented with 0%, 1%, and 2% nitrate (dry matter %) in succession. Nitrate supplementation linearly increased total volatile fatty acids and acetate concentration obviously (p = 0.02; p = 0.02; p<0.01), butyrate and isovalerate concentration numerically (p = 0.07). The alpha (p>0.05) and beta biodiversityof ruminal bacteria were not affected by nitrate. Nitrate increased typical efficient cellulolytic bacteria species (Ruminococcus flavefaciens, Ruminococcus ablus, and Fibrobacter succinogenes) (p<0.01; p = 0.06; p = 0.02). Ruminobactr, Sphaerochaeta, CF231, and BF311 genus were increased by 1% nitrate. Campylobacter fetus, Selenomonas ruminantium, and Mannheimia succiniciproducens were core nitrate reducing bacteria in steers and their abundance increased linearly along with nitrate addition level (p<0.01; p = 0.02; p = 0.04). Potential nitrate reducers in the rumen, Campylobacter genus and Cyanobacteria phyla were significantly increased by nitrate (p<0.01; p = 0.01).To the best of our knowledge, this was the first detailed view of changes in ruminal microbiota by nitrate. This finding would provide useful information on nitrate utilization and nitrate reducer exploration in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.6
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• pp.837-844
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• 2006

Four, lactating dairy cows were randomly assigned according to a $2{\times}2$ Factorial arrangement in a $4{\times}4$ Latin square design to study supplementation of urea level (U) at 2 and 4% and sodium dl-malate (M) at 10 and 20 g/hd/d in concentrate. The treatments were as follows U2M10, U2M20, U4M10 and U4M20, respectively. The cows were offered the treatment concentrate at a ratio to milk yield at 1:2.5 and urea-treated rice straw was fed ad libitum. The results have revealed that rumen fermentation and blood metabolites were similar for all treatments. The populations of protozoa and fungal zoospores were significantly different as affected by urea level and sodium dl-malate. In addition, the viable bacteria were similar for amylolytic and proteolytic bacteria. Cellulolytic bacteria were significantly affected by level of sodium dl-malate especially Selenomonas ruminantium and Megasphaera elsdenii while Butyrivibrio fibrisolvens was significantly affected by level of urea supplementation. In conclusion, the combined use of concentrate containing high level of cassava chip at 75% DM with urea at 4% in concentrate and sodium dl-malate at 20 g/hd/d with UTS as a roughage could improv rumen ecology and microbial protein synthesis efficiency in lactating dairy cows.

• 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.