A continuous culture study was conducted to determine the impact of ruminal degradable soy protein (S-RDP) level and dilution rate (D) on growth of ruminal non-structural carbohydrate-fermenting microbes. Corn starch, urea and isolated soy protein (ISP) were used to formulate three diets with S-RDP levels of 0, 35 and 70% of total dietary CP. Two Ds were 0.03 and $0.06h^{-1}$ of the fermenter volume in a single-effluent continuous culture system. As S-RDP levels increased, digestibilities of dietary dry matter (DM), organic matter (OM) and crude protein (CP) linearly (p=0.001) decreased, whereas digestion of dietary starch linearly (p=0.001) increased. Increasing D from 0.03 to $0.06h^{-1}$ resulted in decreased digestibilities of dietary DM and OM, but had no effect on digestibilities of dietary starch (p=0.77) and CP (p=0.103). Fermenter pH, the concentration of volatile fatty acids (VFA) and daily VFA production were unaffected (p=0.159-0.517) by S-RDP levels. Molar percentages of acetate, propionate and butyrate were greatly affected by S-RDP levels (p=0.016-0.091), but unaffected by D (p=0.331-0.442). With increasing S-RDP levels and D, daily bacterial counts, daily microbial N production (DMNP) and microbial efficiency (MOEFF; grams of microbial N produced per kilogram of OM truly digested) were enhanced (p=0.001). The increased microbial efficiency with increasing S-RDP levels is probably the result of peptides or amino acids that served as a stimulus for optimal protein synthesis. The quantity of ruminal degradable protein from soy proteins required for optimum protein synthesis of non-structural carbohydrate-fermenting microbes appears to be equivalent to 9.5% of dietary fermented OM.
Anaerobic rumen microorganisms mainly bacteria, protozoa and fungi degrade ligno-cellulosic feeds consumed by the ruminants. The ruminants in developing countries are predominantly maintained on low grade roughage and grazing on degraded range land resulting in their poor nutrient utilization and productivity. Hence, manipulation of rumen fermentation was tried during last two decades to optimize ruminal fermentation for improving nutrient utilization and productivity of the animals. Modification of rumen microbial composition and their activity was attempted by using chemical additives those selectively effect rumen microbes, introduction of naturally occurring or genetically modified foreign microbes into the rumen and genetically manipulation of existing microbes in the rumen ecosystem. Accordingly, rumen protozoa were eliminated by defaunation for reducing ruminal methane production and increasing protein outflow in the intestine, resulting in improve growth and feed conversion efficiency of the animals. Further, Interspecies trans-inoculation of rumen microbes was also successfully used for annulment of dietary toxic factor. Additionally, probiotics of bacterial and yeast origin have been used in animal feeding to stabilize rumen fermentation, reduced incidence of diarrhoea and thus improving growth and feed conversion efficiency of young stalk. It is envisaged that genetic manipulation of rumen microorganisms has enormous research potential in developing countries. In view of feed resource availability more emphasis has to be given for manipulating rumen fermentation to increase cellulolytic activity for efficient utilization of low grade roughage.
Three sheep fitted with cannulas in the rumen and the caecum were used in a $3{\times}3$ Latin square design to study the changes in ruminal and caecal microbial populations and their metabolite composition with ammoniated rice straw feeding. The 3 diets contained either 80% untreated rice straw (UTS) or ammoniated rice straw (ATS) and 20% formula feed. These were a control diet (C), a urea supplemented diet (U) containing urea at 1.1% and an ammoniated rice straw diet (AT). Data were analyzed by analysis of variance and means separated by the Student Neumann Kuel's multiple comparison. AT feeding increased ruminal bacterial counts, in particular cellulolytic bacterial counts (p < 0.05) which were 1.8, 2.4 and 7.0 (${\times}10^6/ml$ ruminal fluid) for C, U and AT, respectively. There was an increasing tendency (p < 0.10) in ruminal fungal population with U; values were 2.0, 5.2, 3.1 (${\times}10^3/ml$ ruminal fluid) for C, U and AT, respectively. Ruminal protozoa counts were not significantly (p > 0.05) altered with diets. Caecal total viable bacterial count with AT was about thrice the value with C. Total VFA concentration in the rumen was significantly increased (p < 0.025) (7.7 mmol/dl for C and 8.2 mmol/dl for AT) and correspondingly, pH lowered when AT was fed. Sheep on AT tended to produce less acetate and more butyrate in the rumen without significance (p > 0.05). Similar to the rumen, total VFA concentrations of 4.4, 3.8 and 5.2 mmol/dl were detected, respectively, for C, U and AT. Caecal ammonia-nitrogen concentrations were about six-fold of that in the rumen, though there were no differences (p > 0.05) among treatments.
The gastrointestinal (GI) tract of ruminants contains diverse microbes that ferment various feeds ingested by animals to produce various fermentation products, such as volatile fatty acids. Fermentation products can affect animal performance, health, and well-being. Within the GI microbes, the ruminal microbes are highly diverse, greatly contribute to fermentation, and are the most important in ruminant nutrition. Although traditional cultivation methods provided knowledge of the metabolism of GI microbes, most of the GI microbes could not be cultured on standard culture media. By contrast, amplicon sequencing of 16S rRNA genes can be used to detect unculturable microbes. Using this approach, ruminant nutritionists and microbiologists have conducted a plethora of nutritional studies, many including dietary interventions, to improve fermentation efficiency and nutrient utilization, which has greatly expanded knowledge of the GI microbiota. This review addresses the GI content sampling method, 16S rRNA gene amplicon sequencing, and bioinformatics analysis and then discusses recent studies on the various factors, such as diet, breed, gender, animal performance, and heat stress, that influence the GI microbiota and thereby ruminant nutrition.
Hwang, I.H.;Kim, H.D.;Shim, S.S.;Lee, Sang S.;Ha, J.K.
Asian-Australasian Journal of Animal Sciences
/
v.14
no.4
/
pp.501-506
/
2001
This experiment was conducted to evaluate the effects of supplemental unsaturated fatty acids (UFA) on fermentation characteristics, especially on gas production, cellulose degradation and volatile fatty acid (VFA) concentration by mixed ruminal microorganisms. In order to attain this objective, unsaturated fatty acids including oleic acid (C 18:1), linoleic acid (C18:2) and arachidonic acid (C22:4) were added at varying level. Mixed ruminal microbes used in this experiment were obtained from the rumen of a cannulated Holstein cow. Medium pH values after 7 d incubation were significantly affected by type and level of unsaturated fatty acids (p<0.01). All of UFA inhibited total gas production, and especially treatment of arachidonic acid at the levels of 0.01% gave the lowest gas. production after 7 d incubation (p<0.01). Comparison of the population of protozoa revealed that UFA did not have any significant effect on the total protozoa number. The addition of UFA did not effect dry matter degradation. Volatile fatty acid (VFA) composition of the culture was influenced little by UFA, although the considerable amount of iso-type VFA were detected in UFA supplemented incubations. The ratio of acetic acids to propionic acids, however, was lower than control in all the treatments after 7 d incubation (p<0.01).
Ruminant animals develop a diverse and sophisticated microbial ecosystem for digesting fibrous feedstuffs. Plant cell walls are complex and their structures are not fully understood, but it is generally believed that the chemical properties of some plant cell wall compounds and the cross-linked three-dimensional matrix of polysaccharides, lignin and phenolic compounds limit digestion of cell wall polysaccharides by ruminal microbes. Three adaptive strategies have been identified in the ruminal ecosystem for degrading plant cell walls: production of the full slate of enzymes required to cleave the numerous bonds within cell walls; attachment and colonization of feed particles; and synergetic interactions among ruminal species. Nonetheless, digestion of fibrous feeds remains incomplete, and numerous research attempts have been made to increase this extent of digestion. Exogenous fibrolytic enzymes (EFE) have been used successfully in monogastric animal production for some time. The possibility of adapting EFE as feed additives for ruminants is under intensive study. To date, animal responses to EFE supplements have varied greatly due to differences in enzyme source, application method, and types of diets and livestock. Currently available information suggests delivery of EFE by applying them to feed offers the best chance to increase ruminal digestion. The general tendency of EFE to increase rate, but not extent, of fibre digestion indicates that the products currently on the market for ruminants may not be introducing novel enzyme activities into the rumen. Recent research suggests that cleavage of esterified linkages (e.g., acetylesterase, ferulic acid esterase) within the plant cell wall matrix may be the key to increasing the extent of cell wall digestion in the rumen. Thus, a crucial ingredient in an effective enzyme additive for ruminants may be an as yet undetermined esterase that may not be included, quantified or listed in the majority of available enzyme preparations. Identifying these pivotal enzyme(s) and using biotechnology to enhance their production is necessary for long term improvements in feed digestion using EFE. Pretreating fibrous feeds with alkali in addition to EFE also shows promise for improving the efficacy of enzyme supplements.
Kim, C.H.;Park, B.K.;Park, J.G.;Kim, H.S.;Sung, K.I.;Shin, J.S.;Ohh, S.J.
Journal of Animal Science and Technology
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v.47
no.5
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pp.759-768
/
2005
The study was designed to estimate the in vitro rumen by-pass rate of both chromium methionine chelate as an organic supplement and $ClCl_3$ as an inorganic supplement. Rumen by-pass rates of the supplements were evaluted by comparing ruminal metabolites in rumen fluid and Cr and methionine contents in the body of ruminal microorganism. For in vitro digestion examination, basic nutrients for ruminal microbes were supplied with 7g(DM) of feed, 2g of rice straw, and 2g of corn silage per each incubation jar. Three treatments including Control(no supplementation of Cr), T1(1000ppb supplementation of $ClCl_3$) and T2(chromium methionine chelate supplementation equivalent to 1000ppb of Cr content) were prepared with five replications per each treatment. pH of T2 was lower than that of Control and T1 regardless of incubation time. Ammonia content was higher in T2 than in Control and T1 during first 6 hours of incubation. However, the ammonia content in Control was remained low after 6 hours. Total volatile fatty acids(VFA) content in control was increased constantly as incubation time was extended. Therefore, VFA content in T1 and T2 were significantly lower (P<0.05) than those of Control. Dry matter recovery rate by ruminal microorganism was the lowest in T1, however ruminal microbial population was increased most efficiently in T2 during 12 hours of in vitro incubation. Cr concentrations in the body of ruminal microbes were not different(P>0.05) between Control and T2, but it was significantly high in T1(P<0.05). Contents of methionine and cystine in ruminal microbes also were not different between Control and T2(P>0.05), but it was relatively low in T1. Based on the above results, the chromium methionine chelate was believed to by-pass rumen and could remain intact until it reaches small intestine compared to inorganic chromium. This results implies that chromium methionine chelate could be more effective to function in the small intestine of ruminant animals.
Ando, S.;Khan, R.I.;Takahasi, J.;Gamo, Y.;Morikawa, R.;Nishiguchi, Y.;Hayasaka, K.
Asian-Australasian Journal of Animal Sciences
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v.17
no.1
/
pp.68-72
/
2004
The effects of the addition of yeast on in vitro roughage degradability and methane production were investigated in order to clarify the effects of yeast on the rumen microbes and to establish methods of rumen manipulation. Three roughages (whole crop corn, rice straw and Italian ryegrass) were incubated for 3, 6, 12 and 24 h with or without dried beer yeast following the method described by Tilley and Terry. Using the same method, these roughages were incubated with or without yeast extract, albumin or purified DNA. In vitro methane production was measured with or without dried beer yeast at 12 and 24 h. The degradability of yeast was found to be 57 and 80% at 12 and 24 h, respectively. The rate of degradation of fraction b was 6.16%/h. There was a significant increase in roughage degradability at 6 h (p<0.05), 12 h (p<0.05) and 24 h (p<0.01) by dried yeast addition. The degradability of all three roughages was higher in the samples treated with yeast extract than in the no addition samples except in the case of rice straw incubated for 12 h. Nevertheless, the magnitude of increment was smaller with the addition of yeast extract than without the addition of yeast. With the addition of purified DNA, there were significant increases in roughage degradability at 6 h (p<0.01), 12 h (p<0.01) and 24 h (p<0.05); however, higher degradability values were detected in the samples to which albumin was added, particularly at 6 h. If the degradability values of the no addition samples with those of samples containing yeast, yeast extract, DNA and albumin were compared, the largest difference was found in the samples to which yeast was added, although it is worth noting that higher values were observed in the yeast extract samples than in the DNA or albumin samples, with the exception of the case of rice straw incubated for 24 h. Methane production was significantly increased at both 12 and 24 h incubation. The increment of roughage degradation and methane production brought about by the addition of dried beer yeast to the samples was thought to be due to the activation of rumen microbes. Water soluble fraction of yeast also seemed to play a role in ruminal microbe activation. The increment of degradability is thought to be partially due to the addition of crude protein or nucleic acid but it is expected that other factors play a greater role. And those factors may responsible for the different effects of individual yeast on ruminal microbes.
Lee, S.S.;Shin, K.J.;Kim, W.Y.;Ha, J.K.;Han, In K.
Asian-Australasian Journal of Animal Sciences
/
v.12
no.6
/
pp.988-1001
/
1999
The rumen ecosystem is increasingly being recognized as a promising source of superior polysaccharide-degrading enzymes. They contain a wide array of novel enzymes at the levels of specific activities of 1,184, 1,069, 119, 390, 327 and $946{\mu}mol$ Reducing sugar release/min/mg protein for endoglucanase, xylanase, polygalactouronase, amylase, glucanase and arabinase, respectively. These enzymes are mainly located in the surface of rumen microbes. However, glycoside-degrading enzymes (e.g. glucosidase, fucosidase, xylosidase and arabinofuranosidase, etc.) are mainly located in the rumen fluid, when detected enzyme activities according to the ruminal compartments (e.g. enzymes in whole rumen contents, feed-associated enzymes, microbial cell-associated enzymes, and enzymes in the rumen fluid). Ruminal fungi are the primary contributors to high production of novel enzymes; the bacteria and protozoa also have important functions, but less central roles. The enzyme activities of bacteria, protozoa and fungi were detected 32.26, 19.21 and 47.60 mol glucose release/min/mL mediem for cellulose; 42.56, 14.96 and 64.93 mmol xylose release/min/mL medium after 48h incubation, respectively. The polysachharide-degrading enzyme activity of ruminal anaerobic fungi (e.g. Neocallimastix patriciarum and Piromyces communis, etc.) was much higher approximately 3~6 times than that of aerobic fungi (e.g. Tricoderma reesei, T. viridae and Aspergillus oryzae, etc.) used widely in industrial process. Therefore, the rumen ecosystem could be a growing source of novel enzymes having a tremendous potential for industrial applications.
Riasi, A.;Mesgaran, M. Danesh;Stern, M.D.;Moreno, M.J. Ruiz
Asian-Australasian Journal of Animal Sciences
/
v.25
no.5
/
pp.642-647
/
2012
Eight continuous culture fermenters were used in a completely randomized design to evaluate various nutritional values of Kochia (Kochia scoparia) compared with Atriplex (Atriplex dimorphostegia). Dried and pelleted samples (leaves and stems) provided substrate for metabolism by ruminal microbes maintained in a continuous culture fermentation system. Results indicated that there were no differences (p>0.05) in dry matter (DM) and crude protein (CP) digestibility between the two halophytic plants. Atriplex had higher (p<0.05) organic matter (OM) digestibility compared with Kochia. Neutral detergent fiber (aNDF) digestibility of Atriplex (411 g/kg) was higher (p<0.05) than that of Kochia (348 g/kg), however acid detergent fiber (ADF) digestibility was higher (p<0.05) in Kochia compared with Atriplex (406 vs. 234 g/kg). There were no differences (p>0.05) between the two halophytic plants in molar proportion of acetate and propionate, but the concentration of butyrate and valerate in Kochia were about two fold of Atriplex (p<0.05). When Kochia provided substrate to the microbes, protein synthesis was higher (p<0.05) compared with feeding Atriplex (5.96 vs. 4.85 g N/kg of OM truly digested). It was concluded that Kochia scoparia and Atriplex dimorphostegia had similar digestibility of DM and CP. It appears that these halophytic plants may not have enough digestible energy for high producing ruminants.
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