• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.877-884
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• 2004

Although gut microbial functions have been analyzed through cultivation of isolated microbes, molecular analysis without cultivation is becoming a popular approach in recent years. Gene cloning studies have partially revealed the mechanisms involved in fiber digestion of individual microbe. The molecular approach finally made it possible to analyze full genomes of the representative rumen cellulolytic bacteria Fibrobacter and Ruminococcus. The coming database may contain useful information such as regulation of gene expression relating to fiber digestion. Meanwhile, unculturable bacteria are still poorly characterized, even though they are main constituents of gut microbial ecosystem. The molecular analysis is essential to initiating the studies on these unculturable bacteria. The studies dealing with rumen and large intestine are revealing considerable complexity of the microbial ecosystems with many undescribed bacteria. These bacteria are being highlighted as possibly functional members contributing to feed digestion. Manipulation of gut bacteria and gut ecology for improving animal production is still at challenging stage. Bacteria newly introduced in the rumen, whether they are genetically modified or not, suffer from poor survival. In one of these attempts, Butyrivibrio fibrisolvens expressing a foreign dehalogenase was successfully established in sheep rumen to prevent fluoroacetate poisoning. This expands choice of forages in tropics, since many tropic plants are known to contain the toxic fluoroacetate. This example may promise the possible application of molecular breeding of gut bacteria to the host animals with significance in their health and nutrition. When inoculation strategies for such foreign bacteria are considered, it is obvious that we should have more detailed information of the gut microbial ecology.

• 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.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1280-1290
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• 2014

In order to obtain the cellulolytic bacterial consortia, sediments from Great Basin hot springs (Nevada, USA) were sampled and enriched with cellulosic biomass as the sole carbon source. The bacterial composition of the resulting anaerobic ethanol-producing celluloytic bacterial consortium, named SV79, was analyzed. With methods of the full-length 16S rRNA library-based analysis and denaturing gradient gel electrophoresis, 21 bacteria belonging to eight genera were detected from this consortium. Clones with closest relation to the genera Acetivibrio, Clostridium, Cellulosilyticum, Ruminococcus, and Sporomusa were predominant. The cellulase activities and ethanol productions of consortium SV79 using different agricultural residues (sugarcane bagasse and spent mushroom substrate) and energy crops (Spartina anglica, Miscanthus floridulus, and Pennisetum sinese Roxb) were studied. During cultivation, consortium SV79 produced the maximum filter paper activity (FPase, 9.41 U/ml), carboxymethylcellulase activity (CMCase, 6.35 U/ml), and xylanase activity (4.28 U/ml) with sugarcane bagasse, spent mushroom substrate, and S. anglica, respectively. The ethanol production using M. floridulus as substrate was up to 2.63 mM ethanol/g using gas chromatography analysis. It has high potential to be a new candidate for producing ethanol with cellulosic biomass under anoxic conditions in natural environments.

• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.277-287
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• 2018

Current reports suggest that obesity is a serious global health issue. Emerging evidence has predicted strong links between obesity and the human gut microbiota. However, only a few such studies have been conducted in Asia, and the gut microbiota of lean and obese adult Asians remains largely unexplored. Here, we investigated the potential relationship between gut microbiota, body massindex (BMI), and metabolic parameters in adults from Thailand, where obesity is increasing rapidly. Fecal and blood samples were collected from 42 volunteers who were allocated into lean, overweight, and obese groups. The fecal microbiota was examined by quantitative PCR analysis. Bacteroidetes, Firmicutes, and Staphylococcus spp. and methanogens were most abundant in lean volunteers. Overweight volunteers majorly harbored Christensenella minuta and Akkermansia muciniphila, ${\gamma}-Proteobacteria$, and bacteria belonging to the genus Ruminococcus. Methanogens and bacteria belonging to the phylum Bacteroidetes were negatively correlated with adiposity markers (BMI and waist circumference), but positive correlated with high-density lipoprotein, suggesting that they can be used as leanness markers. While some of our results agree with those of previous reports, results regarding the contributions of specific taxa to obesity were inconsistent. This is the first study to report the adult gut microbiota in Southeast Asian populations using molecular techniques and biochemical markers and provides a foundation for future studies in this field.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.531-537
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• 2017

Objective: This experiment was conducted to explore the impact of diet on the ruminal microbial community in goats. Methods: Twelve goats were divided into two groups and fed complete feed (CF) or all forage (AF) diet. The total microbial DNAs in the rumen liquid were extracted. The V4 region of microbial 16S rRNA genes was amplified and sequenced using high-throughput. Information of sequences was mainly analyzed by QIIME 1.8.0. Results: The results showed that Bacteroidetes and Firmicutes were the most predominant microbial phyla in the rumen of all goats. At genus level, the abundance of fiber-digesting bacteria such as Ruminococcus and Lachnospiracea incertae sedis was significantly higher in AF than that in CF, while the levels of fat-degrading bacterium Anaerovibrio and protein-degrading bacterium Pseudomonas were opposite. The core shared genera, Prevotella and Butyrivibrio were widespread in the rumen of goats and no significant difference was observed in relative abundance between groups. Conclusion: We concluded that the richness of fiber-, protein-, and fat-digesting bacteria was affected by diet and tended to increase with the rise of their corresponding substrate contents in the ration; some bacteria shared by all goats maintained stable despite the difference in the ration, and they might be essential in maintaining the normal function of rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.495-504
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• 2017

Objective: Manipulating the fermentation to improve the performance of the ruminant has attracted the attention of both farmers and animal scientists. Propionate salt supplementation in the diet could disturb the concentration of propionate and total volatile fatty acids in the rumen. This study was conducted to evaluate the effect of calcium propionate supplementation on the ruminal bacterial community composition in finishing bulls. Methods: Eight finishing bulls were randomly assigned to control group (CONT) and calcium propionate supplementation (PROP) feeding group, with four head per group. The control group was fed normal the total mixed ration (TMR) finishing diet, and PROP group was fed TMR supplemented with 200 g/d calcium propionate. At the end of the 51-day feeding trial, all bulls were slaughtered and rumen fluid was collected from each of the animals. Results: Propionate supplementation had no influence the rumen fermentation parameters (p>0.05). Ruminal bacterial community composition was analyzed by sequencing of hypervariable V3 regions of the 16S rRNA gene. The most abundant phyla were the Firmicutes (60.68%) and Bacteroidetes (23.67%), followed by Tenericutes (4.95%) and TM7 (3.39%). The predominant genera included Succiniclasticum (9.43%), Butyrivibrio (3.74%), Ruminococcus (3.46%) and Prevotella (2.86%). Bacterial community composition in the two groups were highly similar, except the abundance of Tenericutes declined along with the calcium propionate supplementation (p = 0.0078). Conclusion: These data suggest that the ruminal bacterial community composition is nearly unchanged by propionate supplementation in finishing bulls.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.10
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• pp.1590-1598
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• 2020

Objective: The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods: Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH₃-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results: There was no significant change in IVDMD, pH, NH₃-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and <0.001, respectively). The LPL supplementation tended to increase the total bacteria in a linear manner (p = 0.089). There were significant decreases in the relative proportions of cellulolytic (Fibrobacter succinogenes and Ruminococcus albus) and lipolytic (Anaerovibrio lipolytica and Butyrivibrio proteoclasticus) bacteria with increasing levels of LPL supplementation (Lin, p = 0.028, 0.006, 0.003, and 0.003, respectively). Conclusion: The LPL supplementation had antimicrobial effects on several cellulolytic and lipolytic bacteria, with no significant difference in nutrient degradability (DM and neutral detergent fiber) and general bacterial counts, suggesting that LPL supplementation might increase the enzymatic activity of rumen bacteria. Therefore, LPL supplementation may be more effective as an antimicrobial agent rather than as an emulsifier in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.2
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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.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1086-1091
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• 2011

This experiment was designed to investigate the effect of lerak extract on the dynamic of rumen microbes in the in vitro fermentation of diet with different ratios of forage and concentrate. In vitro fermentation was conducted according to the method of Tilley and Terry (1963). The design of experiment was a factorial block design with 2 factors. The first factor was the ratio of forage and concentrate (90:10, 80:20, and 70:30 w/w) and the second factor was the level of lerak extract (0, 0.6, and 0.8 mg/ml). Total volatile fatty acid (VFA) concentration, proportional VFA and NH3 concentration were measured at 4 h incubation. Protozoal numbers in the buffered rumen fluid after 4 and 24 h of incubation were counted under a microscope. Bacterial DNAs of buffered rumen fluid were isolated from incubated samples after 24 h of incubation using a QiaAmp kit. Total bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Prevotella ruminicola were quantified using real time polymerase chain reaction (PCR). Lerak extract markedly reduced protozoal numbers in buffered rumen fluid of all diets after 24 h of incubation. Total bacteria did not change with lerak extract addition. While no difference in F. succinogenes was found, there was a slight increase in R. albus number and a significant enhancement in P. ruminicola number by increasing the level of lerak extract in all diets. Propionate concentration significantly increased in the presence of lerak extract at level 0.8 mg/ml. It was concluded that the addition of lerak extract could modify rumen fermentation and had positive effects on rumen microbes.

• Korean Journal of Organic Agriculture
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• v.20 no.3
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.