• Asian-Australasian Journal of Animal Sciences
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• 제12권1호
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• pp.139-147
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• 1999

Peptides are formed in the rumen as the result of microbial proteinase activity. The predominant type of activity is cysteine ptoteinase, but others, such as serine proteinases, are also present. Many species of protozoa, bacteria and fungi are involved in ptoteolysis; large animal-to-animal variability is found when proteinase activities in different animals are compared. The peptides formed from proteolysis are broken down to amino acids by peptidases. Different peptides are broken down at different rates, depending on their chemical composition and particularly their N-terminal structure. Indeed, chemical addition to the N-terminus of small peptides, such as by acetylation, causes the peptides to become stable to breakdown by the rumen microbial population; the microorganisms do not appear to adapt to hydrolyse acetylated peptides even after several weeks exposure to dietary acetylated peptides, and the amino acids present in acetylated peptides are absorbed from the small intestine. The amino acids present in some acetylated peptides remain available in nutritional trials with rats, but the nutritive value of the whole amino acid mixture is decreased by acetylation. The genus Prevotella is responsible for most of the catabolic peptidase activity in the rumen, via its dipeptidyl peptidase activities, which release dipeptides rather than free amino acids from the N-terminus of oligopeptides. Studies with dipeptidyl peptidase mutants of Prevotella suggest that it may be possible to slow the rate of peptide hydrolysis by the mixed rumen microbial population by inhibiting dipeptidyl peptidase activity of Prevotella or the rate of peptide uptake by this genus. Peptides and amino acids also stimulate the growth of rumen microorganisms, and are necessary for optimal growth rates of many species growing on tapidly fermented substrates; in rich medium, most bacteria use pre-formed amino acids for more than 90% of their amino acid requirements. Cellulolytic species are exceptional in this respect, but they still incorporate about half of their cell N from pre-formed amino acids in rich medium. However, the extent to which bacteria use ammonia vs. peptides and amino acids for protein synthesis also depends on the concentrations of each, such that preformed amino acids and peptides are probably used to a much lesser extent in vivo than many in vitro experiments might suggest.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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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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• 제13권7호
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• pp.918-921
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• 2000

A comparative study on rumen bacterial and protozoal population and fungal zoospores in cattle (Brahman$\times$Native) and swamp buffalo (Bubalus bubalis) was conducted. Forty animals, twenty of each, with same sex and similar age which were raised under similar condition in the Northeast of Thailand, were used. Rumen digesta were sampled immediately post slaughtering for total microscopic counts of bacteria, protozoa and fungal zoospores. It was found that total bacterial population were higher in swamp buffalo that those in cattle (1.6 vs $1.36{\times}10^{8}cells/ml$) having more population of cocci, rods and ovals. Lower rumen protozoal pupulation in swamp buffalo with lower numbers of Holotrichs and Entodiniomorphs were found as compared to those in cattle. Significant higher fungal zoospore counts were in swamp buffalo than those in cattle being 7.30 and $3.78{\times}10^6$, respectively. Study under electron microscope, revealed Anaeromyces sp. with acuminate apex were more predominant in the rumen of swamp buffalo. With these findings, cattle and swamp buffaloes showing differences in rumen bacterial, protozoal population and fungal zoospore counts, offer new additional information as why swamp buffaloes exhibit conditionally better than cattle especially during long dry season without green grass.

• Asian-Australasian Journal of Animal Sciences
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• 제17권6호
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• pp.820-824
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• 2004

The study was to see the effect of administration of ruminal fungi on feed intake, growth rate, rumen fermentation and nutrient digestion of calves (Tharparkar$\times$Holstein-Friesian, average age: 10 months, average body weight: 130 kg). The 6 calves in first group were fed a mixture consisted of 50% wheat straw and 50% concentrate (Maize 62%, Groundnut cake 35%, Mineral mix. 2% and Common salt 1%) along with 1 kg green oats $animal^{-1}$ $day^{-1}$ while second group calves were fed the above-mentioned diet in addition to a dose of 160 ml ($10^{6}$ CFU/ml) fungal culture $calf^{-1}$ $week^{-1}$. The average dry matter intake per day was slightly lowered in fungal fed calves yet feed conversion ratio was higher. The average daily weight gain was significantly higher (15.37%) in fungal administered group as compared to control. The nutrient digestibility was increased for crude fibre, NDF and ADF with fungal administration. Digestible energy value of straw-based diet in terms of percent TDN also increased. The pH and $NH_{3}$-N were lower whereas TVFA, total-N, TCA-N and number of zoospores were higher in rumen liquor in fungal administered group.

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.599-602
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• 1997

Fluctuation of fungal zoospores on agar strips were observed in the rumen of sheep fed three different levels of dietary concentrate, timothy hay: concentrate = 3:0 (AF diet), timothy hay: concentrate = 2:1 (MC diet), timothy hay : concentrate = 1:2 (HC diet) respectively. The number of zoospores on the strip was drastically decreased after morning feed with AF diet. The number was the highest at 0 h ($1.34{\times}10^2/cm^2$), then declined to $2.0{\times}10^3/cm^2$ at 9 h after feeding. In the rumen of animals fed MC diet, the number of zoospores decreased with time after feeding, although the decrement was slower than that with AF diet. During 0-3 h after feeding, number of zoospores was $1.6{\times}10^4/cm^2$. Although the number slightly decreased at 6 and 9 h, relatively high levels were maintained. It seems that the inducers for zoospore-release were maintained at relatively high concentration throughout incubation period. The fluctuation pattern of number of germinated zoospores was different in the rumen of animals fed HC diet from those of AF and MC diets. The number of zoospores was constantly maintained at lower level ($1.0{\times}10^3/cm^2$) than the other diets. For MC diet, continuous high number of germinated zoospores may be due to the continuous release of zoospores by hemes in timothy hay and concentrate feed, and by unknown mechanisms. Unlike AF diet which promoted relatively rapid decline of zoosporogenesis, supplementation of concentrate feed to the timothy hay did not promote such rapid decline of zoosporogenesis. It was suggested that release of inducers for zoosporogenesis from concentrate feed persisted longer time than from timothy hay. HC diet promoted the lowest zoospore production, suggested the lowest fungal population size in this experiment. These results show that an appropriate amount of concentrate may support fungal growth and stimulate zoosporogenesis in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• 제21권6호
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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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• 제20권2호
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• pp.283-294
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• 2007

Conventional culture-based methods of enumerating rumen microorganisms (bacteria, archaea, protozoa, and fungi) are being rapidly replaced by nucleic acid-based techniques which can be used to characterise complex microbial communities without incubation. The foundation of these techniques is 16S/18S rDNA sequence analysis which has provided a phylogenetically based classification scheme for enumeration and identification of microbial community members. While these analyses are very informative for determining the composition of the microbial community and monitoring changes in population size, they can only infer function based on these observations. The next step in functional analysis of the ecosystem is to measure how specific and, or, predominant members of the ecosystem are operating and interacting with other groups. It is also apparent that techniques which optimise the analysis of complex microbial communities rather than the detection of single organisms will need to address the issues of high throughput analysis using many primers/probes in a single sample. Nearly all the molecular ecological techniques are dependant upon the efficient extraction of high quality DNA/RNA representing the diversity of ruminal microbial communities. Recent reviews and technical manuals written on the subject of molecular microbial ecology of animals provide a broad perspective of the variety of techniques available and their potential application in the field of animal science which is beyond the scope of this treatise. This paper will focus on nucleic acid based molecular methods which have recently been developed for studying major functional groups (cellulolytic bacteria, protozoa, fungi and methanogens) of microorganisms that are important in nutritional studies, as well as, novel methods for studying microbial diversity and function from a genomics perspective.

• 생명과학회지
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• 제28권10호
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• pp.1244-1253
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• 2018

반추위 속에는 박테리아, 고세균, 프로토조아, 곰팡이 및 바이러스와 같은 다양한 미생물들이 편성의 혐기조건에서 공생하고 있다. 사료의 발효에 중요한 역할을 하고 있는 반추위 미생물은 위내 발효과정에서 에너지 손실에 영향을 주는 메탄의 발생을 제외하면 에너지와 단백질 대사에 필수적인 다양한 휘발성 지방산을 생산한다. 반추위내 미생물의 이용효율을 개선시키기 위해 사료배합비조절, 천연사료첨가제, 생균제첨가 등의 다양한 접근방법들이 사용되고 있다. 최근에 반추위 군집에 대한 메타유전체 또는 메타전사체와 같은 차세대 유전체 해독기술 또는 차세대 시퀀싱 기술의 적용으로 반추위 미생물의 다양성 및 기능에 대한 이해가 크게 증가하였다. 특히 메타단백질체와 메타대사체는 반추위 생태계의 복잡한 미생물네트워크에 대한 더 깊은 통찰력을 제공할 뿐만 아니라, 다양한 반추가축용 사료에 대한 반응을 제공함으로서 생산효율을 개선시키는데 기여하였다. 본 논문에서는 반추위내 사료의 발효와 이용을 향상시키기 위한 메타오믹스 기술, 즉, 메타유전체, 메타전사체, 메타단백질체 및 메타대사체의 최신 응용기술을 요약하고자 한다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권11호
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• pp.1583-1591
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• 2013

The objective of this study was to investigate microbial population in the rumen of dairy steers as influenced by supplementing with dietary condensed tannins and saponins and different roughage to concentrate ratios. Four, rumen fistulated dairy steers (Bos indicus) were used in a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The main factors were two roughage to concentrate ratios (R:C, 60:40 and 40:60) and two supplementations of rain tree pod meal (RPM) (0 and 60 g/kg of total DM intake). Chopped 30 g/kg urea treated rice straw was used as a roughage source. All animals received feed according to respective R:C ratios at 25 g/kg body weight. The RPM contained crude tannins and saponins at 84 and 143 g/kg of DM, respectively. It was found that ruminal pH decreased while ruminal temperature increased by a higher concentrate ratio (R:C 40:60) (p<0.05). In contrast, total bacterial, Ruminococus albus and viable proteolytic bacteria were not affected by dietary supplementation. Numbers of fungi, cellulolytic bacteria, Fibrobactor succinogenes and Ruminococus flavefaciens were higher while amylolytic bacteria was lower when steers were fed at 400 g/kg of concentrate. The population of Fibrobactor succinogenes, was found to be higher with RPM supplementation. In addition, the use of real-time PCR technique indicated that the population of protozoa and methanogens were decreased (p<0.05) with supplementation of RPM and with an increasing concentrate ratio. Supplementation of RPM and feeding different concentrate ratios resulted in changing the rumen microbes especially, when the animals were fed at 600 g/kg of concentrate and supplemented with RPM which significantly reduced the protozoa and methanogens population.

• Asian-Australasian Journal of Animal Sciences
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• 제5권2호
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• pp.323-327
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• 1992

An experiment was conducted to determine whether there were any apparent differences in the microbial population, colonization pattern and digestion of guinea grass in situ, between cattle and swamp buffalo. Percentage losses in dry matter (DM), nitrogen (N) and neutral detergent fibre (NDF) of guinea grass were significantly (p<0.01) higher when incubated in the rumen of buffalo than in cattle. Buffalo also showed significantly (p<0.05) faster degradation rates than cattle for each grass component (DM, N, DNF). Light microscopy and SEM examination of the incubated grass materials showed that there were no apparent differences in the pattern of bacterial and fungal invasion and colonization of the grass materials between cattle and buffalo. Attachment of bacteria and fungal zoospores on the grass fragments occurred at 15 min after rumen incubation. After 3 h of rumen incubation, dense population of bacteria was observed in the thin-walled mesophyll and parenchyma tissues, whereas root-like fungal rhizoids were observed in both thin-walled and thick-walled cells. By 6 h, eroded zones were apparent in the thin-walled tissues and in thick-walled tissues with profuse rhizoids. After 24. 48 and 72 h of rumen incubation, most thin-walled tissues were degraded leaving mostly the thick-walled tissues. The predominant bacteria were the curved rods resembling Butyrivibrio sp., the thick rods resembling Fibrobacter sp., the diplococcoids resumbling Ruminococcus sp. And spirochetes. Fungi were predominantly those with spherical or oval sporangia. Fusiform sporangia with acuminate apices which resembled Ruminomyces sp. Were of lesser occurrence. Few protozoa were found on the grass fragments at all incubation times.