• Asian-Australasian Journal of Animal Sciences
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• v.30 no.11
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• pp.1590-1597
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• 2017

Objective: This study aims to identify the relationship between odd- and branched-chain fatty acids (OBCFAs) and microbial nucleic acid bases in the rumen, and to establish a model to accurately predict microbial protein flow by using OBCFA. Methods: To develop the regression equations, data on the rumen contents of individual cows were obtained from 2 feeding experiments. In the first experiment, 3 rumen-fistulated dry dairy cows arranged in a $3{\times}3$ Latin square were fed diets of differing forage to concentration ratios (F:C). The second experiment consisted of 9 lactating Holstein dairy cows of similar body weights at the same stage of pregnancy. For each lactation stage, 3 cows with similar milk production were selected. The rumen contents were sampled at 4 time points of every two hours after morning feeding 6 h, and then to analyse the concentrations of OBCFA and microbial nucleic acid bases in the rumen samples. Results: The ruminal bacteria nucleic acid bases were significantly influenced by feeding diets of differing forge to concentration ratios and lactation stages of dairy cows (p<0.05). The concentrations of OBCFAs, especially odd-chain fatty acids and C15:0 isomers, strongly correlated with the microbial nucleic acid bases in the rumen (p<0.05). The equations of ruminal microbial nucleic acid bases established by ruminal OBCFAs contents showed a good predictive capacity, as indicated by reasonably low standard errors and high R-squared values. Conclusion: This finding suggests that the rumen OBCFA composition could be used as an internal marker of rumen microbial matter.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.144-154
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• 2008

The rumen microbial ecosystem produces methane as a result of anaerobic fermentation. Methanogenesis in the rumen is thought to represent a 2-12% loss of energy intake and is estimated to be about 15% of total atmospheric methane emissions. While methanogenesis in the rumen is conducted by methanogens, PCR-based techniques have recently detected many uncultured methanogens which have a broader phylogenetic range than cultured strains isolated from the rumen. Strategies for reduction of methane emissions from the rumen have been proposed. These include 1) control of components in feed, 2) application of feed additives and 3) biological control of rumen fermentation. In any case, although it could be possible that repression of hydrogen-producing reactions leads to abatement of methane production, repression of hydrogen-producing reactions means repression of the activity of rumen fermentation and leads to restrained digestibility of carbohydrates and suppression of microbial growth. Thus, in order to reduce the flow of hydrogen into methane production, hydrogen should be diverted into propionate production via lactate or fumarate.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1439-1444
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• 2002

An objective of this study was to determine the effects of increasing contents of rumen undegradable protein (RUP) from formalin treated soy bean (FSBM) on rumen functions. Four rumen canulated non-lactating cows were randomly allocated to total mixed rations (TMR) containing different proportions of soy bean meal (SBM) and FSBM. Of rumen fermentation characteristics, concentrations of ruminal fluid ammonia and molar proportions of isoacids decreased with increasing contents of RUP in diets (p<0.01). The animals on TMR containing only SBM gained less weight and had smaller rumen volume than those on TMR containing RUP from FSBM (p<0.05). Organic matter and neutral detergent fiber digestibility in sacco were not different (p>0.05). The density of protozoa particularly small Entodinium sp. in ruminal fluid was higher in animal fed TMR containing SBM:FSBM (34:66) and FSBM than those fed TMR containing SBM:FSBM (66:34) and SBM (p<0.01). Total viable count, and net microbial protein synthesis as indicated by purine derivatives in urine increased with increasing contents of RUP from FSBM (p<0.01). It can be concluded that a reduction in net microbial protein synthesis in the rumen with increasing contents of RUP in the diet can be due to the reduction of preformed protein available for microbial growth as well as an increased turnover rate of microbial cells by predatory activity of protozoa.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.3
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• pp.326-331
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• 2005

A rumen simulated continuous culture (RSCC) system was used to study the influence of supplementation of the three different types of protein sources such as urea, casein and soy protein on rumen microbial synthesis in terms of rumen microbial synchronization. The urea treatment showed the highest pH value. Ammonia nitrogen concentration was rapidly increased after feeding and not significantly different in the urea treatment (13.53 mg/100 ml). Protozoa numbers were not significantly different for soy protein and casein treatment compared to urea treatments during incubation. The average concentration of total VFA (mMol) was not detected with significant difference among treatments, but iso-butyrate production showed the highest for soy protein treatment among treatments (p<0.001). The lowest concentration in total iso-acids (iso-butyrate and iso-valerate) production was observed in urea treatment. The soy protein treatment showed no significantly change in acetate/propionate. The amounts of dry matter (DM) out flow showed no significant difference among treatments. Organic matter (OM) flow was the highest for urea treatments and the lowest for casein treatment (p<0.03). The nitrogen flow for casein treatment was not significantly different from other treatments. The efficiency of microbial protein synthesis in terms of microbial nitrogen (MN) synthesis (g MN/kg ADOM) digested in the rumen was highest for casein treatment (58.53 g MN/kg ADOM) compared to soy protein and urea (p<0.05). This result suggests that rumen ammonia releasing rate may influence on microbial protein synthesis in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.3
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• pp.303-316
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• 1994

Microbial digestion of feed in the rumen involves a sequential attack culminating in the formation of fermentation products and microbial cells that can be utilized by the host animal. Most feeds are protected by a cuticular layer which is in effect a microbial barrier that must be penetrated or circumvented for digestion to proceed. Microorganisms gain access to digestible inner plant tissues through damage to the cuticle, or via natural cell openings (e.g., stomata) and commence digestion from within the feed particles. Primary colonizing bacteria adhere to specific substrates, divide to form sister cells and the resultant microcolonies release soluble substrates which attract additional microorganisms to the digestion site. These newly attracted microorganisms associate with primary colonizers to form complex multi-species consortia. Within the consortia, microorganisms combine their metabolic activities to produce the diversity of enzymes required to digest complex substrates (e.g., cellulose, starch, protein) which comprise plant tissues. Feed characteristics that inhibit the microbial processes of penetration, colonization and consortia formation can have a profound effect on the rate and extent of feed digestion in the rumen. Strategies such as feed processing or plant breeding which are aimed at manipulating feed digestion must be based on an understanding of these basic microbial processes and their concerted roles in feed digestion in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.3
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• pp.209-216
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• 1998

The paper presented here is aimed at increasing knowledge on purine metabolism in ruminants and hence the quantification of microbial cells entering the small intestine from urinaη excretion of purine derivatives. Nucleic acid metabolisms of micro-organisms in the rumen, digestion and absorption of nucleic acids entering the intestines, metabolisms of absorbed and endogenous purines involving de novo synthesis of nucleic acids in the ruminants host, and the relationship between absorbed and excreted purines are reviewed. Principal concerns about an amount of purine derivatives excreted in urine in relation to a change in purine-N: total-N ratios in rumen microbes that leave the rumen are discussed. The use of urinary excretion of purine derivatives as an indicator of the amount of microbial biomass leaving the rumen has to be done with some caution since it may be impossible to get a representative sample of microbes entering the intestine and thus yield estimates are relative rather than absolute.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.4
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• pp.351-362
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• 1998

Microbial growth efficiency in the rumen was studied in sheep given hourly, 31.25 g oaten chaff with either 0.31 and 0.88 g urea or 1.88 and 5.63 g casein (exp. 1) and 33.33 g oaten chaff with 1.04 casein or 0.3, 0.6 and 0.9 g urea or the mixture of the casein and urea (exp. 2). Concentrations of ruminal fluid ammonia increased with increasing nitrogenous supplements. Organic matter digestibility in sacco in the rumen was not different irrespective of N sources. Isoacids and valeric acid increased with increasing ingested casein but decreased with increasing urea intake. Peptide and amino acid pools in ruminal fluid increased with increasing ammonia concentrations (exp. 2) suggesting that proteolytic activity and transportation of peptides and amino acids across microbial membrane of rumen microbes may be regulated by the metabolite mechanism (intracellular amino acids and $NH_4{^+}$, respectively). Densities of total viable and cellulolytic bacteria in ruminal fluid increased with increasing ammonia levels but that of small Entodinia decreased. The density of fungal sporangia growth on oat leaf blades decreased with increasing ammonia concentrations but appeared to remain constant in the presence of casein. Efficiency of net microbial cell synthesis was 15-28% higher when ammonia concentrations increased from 100 to above 200 mg N/l regardless of N sources. In conclusion, supplementation of preformed protein had no effect on rumen digestion and microbial growth efficiency. This could not be accounted for its effect on ruminal fluid ammonia. Increased microbial growth efficiency with increasing ammonia levels may be due to a reduction in the turnover of microbial cells within the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.6
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• pp.661-672
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• 1998

Two experiments were conducted to examine the effects of a range of concentrations of ruminal fluid ammonia ($NH_3$-N) on forage digestibility, microbial growth efficiency and the mix of microbial species. Urea was either continuously infused directly into the rumen of sheep fed 33.3 glh of oaten chaff (Exp. I) or sprayed onto the oaten chaff (750 g/d) given once daily (Exp. 2). Concentrations of $NH_3$-N increased with incremental addition of urea (p < 0.01). Volatile fatty acids (VFA) concentrations and 24 h in sacco organic matter digestibility in the rumen were higher when supplemental urea was given (p < 0.01). The (C2 + C4) : C3 VFA ratio was lower (p < 0.05) when $NH_3$-N was above 200 mgN/I. The fungal sporangia appearing on oat leaf blades were significantly higher when urea was supplemented, indicating that $NH_3$-N was a growthlimiting nutrient for fungi at levels of $NH_3$-N below 30 mgN/l. The density of protozoa was highest when $NH_3$-N concentrations were adjusted to 30 mgN/I for continuously fed ($4.4{\times}10^5/ml$) and to 168 mgN/1 for once daily feeding ($2.9{\times}10^5/ml$). Thereafter increasing concentrations of $NH_3$-N, were associated with a concomitant decline in protozoal densities. At the concentration of $NH_3$-N above 200 mgN/l, the density of protozoa was similar to the density of protozoa in ruminal fluid of the control sheep ($1.8{\times}10^5/ml$). The efficiency of net microbial protein synthesis in the rumen calculated from purine excretion was 17-47% higher when the level of $NH_3$-N was above 200 mgN/1. The possibilities are that 1) there is less bacterial cell lysis in the rumen because of the concomitant decrease in the protozoal pool and/or 2) microbial growth per se in the rumen is more efficient with increasing $NH_3$-N concentrations.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.1
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• pp.75-81
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• 2007

The object of this study was to determine the influence of supplementation of concentrate containing high levels of cassava chip on rumen ecology, microbial protein and digestibility of nutrients. Four, rumen fistulated crossbred beef steers with initial body weight of 400${\pm}$10 kg were randomly assigned according to a 4${\times}$4 Latin square design. The dietary treatments were concentrate cassava chip based offering at 0, 1, 2 and 3% BW with urea-treated rice straw fed ad libitum. It was found that ruminal pH was significantly decreased with increase of concentrate. Volatile fatty acids (VFA) concentration in the rumen was significantly different among treatments. In addition, a molar proportion of propionate was higher in supplemented groups at 2 and 3% BW (p<0.05), leading to significantly decreased acetate:propionate ratio. Furthermore, microbial N supply was significantly improved and was highest at 2% BW supplementation. The efficiency of rumen microbial-N synthesis based on organic matter (OM) truly digested in the rumen was highest in level of concentrate supplementation at 2% BW (80% of cassava chip in diets). Moreover, bacterial populations such as amylolytic bacteria was linearly increased, while cellulolytic bacteria was linearly decreased (p<0.01) when cattle received concentrate supplementation in all levels. The total protozoal counts were significantly increased, while fungal zoospores were dramatically decreased in cattle receiving increased levels of concentrate. In conclusion, cassava chip can be use as energy source at 80% in concentrate and supplementation of concentrate at 2% BW with urea-treated rice straw as roughage could improve rumen fermentation efficiency in beef cattle.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1725-1731
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• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.