• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.200-207
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• 2007

In vitro rumen incubation studies were conducted to determine effects of initial pH on bacterial attachment and fiber digestion. Ruminal fluid pH was adjusted to 5.7, 6.2 and 6.7, and three major fibrolytic bacteria attached to rice straw in the mixed culture were quantified with real-time PCR. The numbers of attached and unattached Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminocococcus albus were lower (p<0.05) at initial pH of 5.7 without significant difference between those at higher initial pH. Lowering incubation media pH to 5.7 also increased bacterial numbers detached from substrate regardless of bacterial species. Dry matter digestibility, gas accumulation and total VFA production were pH-dependent. Unlike bacterial attachment, maintaining an initial pH of 6.7 increased digestion over initial pH of 6.2. After 48 h in vitro rumen fermentation, average increases in DM digestion, gas accumulation, and total VFA production at initial pH of 6.2 and 6.7 were 2.8 and 4.4, 2.0 and 3.0, and 1.2 and 1.6 times those at initial pH of 5.7, respectively. The lag time to reach above 2% DM digestibility at low initial pH was taken more times (8 h) than at high and middle initial pH (4 h). Current data clearly indicate that ruminal pH is one of the important determinants of fiber digestion, which is modulated via the effect on bacterial attachment to fiber substrates.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.12
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• pp.1625-1632
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• 2009

Four lactating Holstein Friesian crossbred cows, with an average initial weight of 450 kg, 48${\pm}$12 days in milk and initial milk yield of 18 kg/h/d, were randomly arranged according to a 2${\times}$2 factorial arrangement in a 4${\times}$4 in Latin square design with 21-d period to investigate the effects of type of total mixed ration (TMR) and type of whole cottonseed (WCS) on intake, digestibility and milk production. The dietary treatments were i) TMR and WCS supplementation at 0.5 kg/h/d, ii) TMR and cracked WCS (cWCS) supplementation at 0.5 kg/h/d, iii) fermented TMR (FTMR) and WCS supplementation at 0.5 kg/h/d, and iv) FTMR and cWCS supplementation at 0.5 kg/h/d. Voluntary feed intake was 15.9, 15.2, 15.4 and 15.6 kg DM/d in dietary treatment 1, 2, 3 and 4, respectively. Digestibility of DM, OM, CP, EE, NDF and ADF were not significantly different among dietary treatments. Ruminal pH, $NH_{3}-N$ and volatile fatty acids in the rumen were also not significantly different among type of TMR or type of WCS. Blood urea-N concentration was not significantly different among dietary treatments. Ruminal bacteria population tended to increase but ruminal protozoa population tended to decrease with supplementation of cWCS, but they were not affected by FTMR. Milk yield and 3.5% FCM were not statistically different among treatments (16.6, 16.2, 17.0, 16.3 kg/d and 18.0, 18.6, 19.9 and 19.0 kg/d, respectively). Milk composition was not significantly different among dietary treatments. However, unsaturated fatty acids in milk fat in cows fed FTMR were lower (p<0.05) than in cows fed TMR. In conclusion, fermentation is a conceivable method to improve the quality of TMR for long-time storage and the cracking method is suitable to release the fat from cottonseed for enhancing fatty acid deposition in milk. Thus, the combination of FTMR and cWCS supplementation would be an alternative strategy to improve performance of lactating cows.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.1
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• pp.48-53
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• 2001

An in vitro study was conducted to examine the effect of sources and the addition levels of carbohydrates on fermentation characteristics, bacterial growth, and hydrogenation of linoleic acid ($C_{18:2}$) by mixed ruminal bacteria. Starch and cellobiose were added to the 200 ml non-selective basal media at the levels of 0.20 and 0.35% (w/v), respectively. Linoleic acid (66.8~79.6 mg) in the absorbed form into the pieces of nylon cloth was also added to the media of 5 treatments including control which was not added with carbohydrate. Three mls of rumen fluid strained through 12 layers of cheese cloth were added to each medium, and were incubated anaerobically in the shaking incubator of $39^{\circ}C$ for 24 hours. During 24 h incubation the pH in incubation media of all treatments was maintained above 6.6 by the addition of sodium bicarbonate. The pH and ammonia concentration of incubation media were not clearly influenced by the sources and addition levels of carbohydrates while additions of carbohydrates increased (p<0.0001) VFA concentration at the 24 h incubation. Molar proportions of acetate were reduced (p<0.0004) while those of propionate were increased (p<0.0006) by the addition of carbohydrates. But the differences in concentration and molar proportions of the VFA were small between the sources or the addition levels. Bacterial growth was faster (p<0.0004) in the starch added treatments than in the cellobiose added ones and control, but no differences were found between addition levels. Increased (p<0.0487) hydrogenation was observed from the starch added treatments compared to the cellobiose added ones, but there was no difference between addition levels.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.1
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• pp.7-14
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• 2016

The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.10
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• pp.1482-1486
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• 2003

In order to compare effects of feeding systems on rumen fermentation characteristics and nutrient digestion, steers were fed either total mixed ration (TMR) or separate concentrate-roughage ration (CR). Total tract digestibility of nutrients was higher in steers receiving TMR. Especially, DM, ADF and NDF in TMR were digested to a greater extent than those in CR. Rumen pH was not influenced by the feeding systems. Holstein steers on TMR had higher ruminal $NH_3$-N than those on CR. Feeding system did not alter VFA production but TMR feeding resulted in lower A/P ratio. TMR feeding tended to increase the number of bacteria and protozoa in the rumen fluid. Also steers fed TMR generally had higher fiber degrading enzyme activities, which might be the result of increased number of cellulolytic microbes in the rumen of animals on TMR. Our results indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen and in the total tract of steers.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.2
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• pp.211-218
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• 2016

This study evaluated the effect of Candida norvegensis (C. norvegensis) viable yeast culture on in vitro ruminal fermentation of oat straw. Ruminal fluid was mixed with buffer solution (1:2) and anaerobically incubated with or without yeast at $39^{\circ}C$ for 0, 4, 8, 16, and 24 h. A fully randomized design was used. There was a decrease in lactic acid (quadratic, p = 0.01), pH, (quadratic, p = 0.02), and yeasts counts (linear, p<0.01) across fermentation times. However, in vitro dry matter disappearance (IVDMD) and ammonia-N increased across fermentation times (quadratic; p<0.01 and p<0.02, respectively). Addition of yeast cells caused a decrease in pH values compared over all fermentation times (p<0.01), and lactic acid decreased at 12 h (p = 0.05). Meanwhile, yeast counts increased (p = 0.01) at 12 h. C. norvegensis increased ammonia-N at 4, 8, 12, and 24 h (p<0.01), and IVDMD of oat straw increased at 8, 12, and 24 h (p<0.01) of fermentation. Yeast cells increased acetate (p<0.01), propionate (p<0.03), and butyrate (p<0.03) at 8 h, while valeriate and isovaleriate increased at 8, 12, and 24 h (p<0.01). The yeast did not affect cellulolytic bacteria (p = 0.05), but cellulolytic fungi increased at 4 and 8 h (p<0.01), whereas production of methane decreased (p<0.01) at 8 h. It is concluded that addition of C. norvegensis to in vitro oat straw fermentation increased ruminal fermentation parameters as well as microbial growth with reduction of methane production. Additionally, yeast inoculum also improved IVDMD.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.42-48
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• 2009

An in vitro incubation study was conducted to investigate effects of 2-bromoethanesulfonic acid (BES) on ruminal fermentation characteristics and methanogen population. BES at the final concentration of 0, 1 and 5 mM with two different substrates having a different ratio of timothy and concentrate (100% timothy vs. 40% timothy-60% concentrate) was incubated for 0, 24, 48 and 72 h in a $39^{\circ}C$ incubator. Total DNA extracted from culture fluid was used as a template for real-time PCR to measure the population of methanogens. Four different primer sets were used for amplification of total bacteria, total methanogens, the order Methanobacteriales and the order Methanomicrobiales. BES reduced (p<0.01) total gas and methane production in a dose-dependent manner. BES at 5 mM inhibited methane production by more than 95% compared to the control. An interaction between substrate and level of BES in total gas and methane was detected (p<0.01). The decrease of methane production with increasing BES level was more pronounced on mixed substrate than on timothy alone. However, hydrogen production was increased by BES treatment (p<0.01). Total VFA concentration was not affected, but molar percentage of propionate and butyrate was increased and acetate to propionate ratio was reduced by BES treatment (p<0.01). BES did not affect the population density of total bacteria but reduced (p<0.01) the population of total methanogens, the order Methanobacteriales and the order Methanomicrobiales in a dose-dependent manner. The type of substrate did not influence the trend, although the magnitude of response was different between all-roughage and 40% roughage substrate.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.631-639
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• 2001

Two experiments were carried out concerning the effects of urea-molasses cake (UMC) and its separate components as supplements on rumen environment, in sacco feed degradability and intake of swamp buffaloes fed rice straw, grasses or a mixture of grasses and rice straw. Experiment 1 was a change-over design with 4 animals and 6 treatments. The buffaloes were fed rice straw ad libitum, and the experimental treatments were: no supplementation (R); 700 g of the complete urea-molasses cake (RUMC); 53.2 g urea (RU); 276 g rice bran and 52.5 coconut meal (RRC); 26.6 g salt, 26.6 g bone meal and 2.1 g trace minerals (RMi); and 25 g molasses (RMo). Experiment 2 was a Latin square design with four diets and four animals. The treatments were: rice straw ad libitum and mixed grass (RG) at 2.5 g dry matter per kg live weight (LW); RG plus 700 g urea-molasses cake (RGUMC); mixed grass ad libitum (G); and G plus 700 g cake (GUMC). In both experiments the supplements were fed once daily. In Exp. 1 although the rumen pH was significantly different (p<0.05) among diets, it varied only from 6.90 to 7.06. The ruminal ammonia was also significantly (p<0.05) different among the diets with RUMC significantly higher than R. Total bacterial and protozoal counts were significantly (p<0.05) higher for the RUMC, RU, RMo and RRC diets. Total feed and rice straw intakes were highest for RUMC (p<0.05) and lowest for the RMi and RMo diets, but in sacco degradability of four different roughages were not significantly different among diets. In Exp. 2, rumen pHs of the diets differed significantly and (p<0.01) ranged from 7.04 - 7.19. Ruminal $NH_3-N$ concentrations (mg/100 ml) were also significantly different (p<0.05), and higher for the RGUMC, G and GUMC diets. The total counts of bacteria and protozoa were significantly (p<0.05) higher for the RGUMC, G and GUMC diets. The total feed intake and roughage intake were significantly (p<0.05) higher for the RGUMC, G and GUMC diets compared to the RG diet. Correspondingly, LW changes also differed among treatments (p=0.06). It was concluded that there were significant increases in rumen $NH_3-N$ concentration, microbial populations and feed intake in the buffaloes by UMC supplementation, whereas the significant difference in in sacco DM degradation was not found by any type of supplementation. There seemed to be a need of a combination of urea, molasses, minerals and other protein nitrogen sources to enhance rice straw intake. Adding grass to the rice straw diet at 0.25% LW (DM) should also be considered to maintain buffalo rumen function and production with UMC supplementation, when rice straw is the main roughage.

• Journal of Animal Science and Technology
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• v.50 no.6
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• pp.831-838
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• 2008

This study was conducted to evaluate effects of mixed microbes addition on physico-chemical, fermentative and microbial parameters of sawdust-based spent mushroom substrates(SMS). The SMS was inoculated with mixed microbes(Enterobacter ludwigii, Bacillus cereus, 2 strains of Bacillus subtillis, Saccharomyces cerevisiae and Lactobacillus plantarum) at 1% level(wet basis) and anaerobically fermented during the different periods(up to 8th week). Compared with the SMS before ensiling, the ensiled one had higher CP, NDF and ADF percentages and lower DM and NFC percentages. However, levels of change were very low. The in situ ruminal disappearance of SMS DM and NDF decreased with the ensiling period prolonged. For fermentative parameters, pH reduced and lactic acid contents increased after ensiling, compared with those after ensiling. At 8th week of ensiling, pH increased and lactic acid contents reduced again, compared with those at 4th week of ensiling; however, the silage still showed favorable fermentation status. Lactic acid bacteria counts did not change throughout 8 weeks of ensiling. Counts of total microbes and yeast reduced after 4th week of ensiling period. Counts of lactic acid bacteria and yeast at 8th week of ensiling were in the levels of 108cfu/g. These results indicate that anaerobic fermentation with microbial addition could be an effective way for the long term(8 weeks) storage of the SMS.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.66-74
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• 2012

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.