• Asian-Australasian Journal of Animal Sciences
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• v.26 no.10
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• pp.1424-1436
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• 2013

The objective of this study was to investigate the effect of banana flower powder (BAFLOP) supplementation on gas production kinetics and rumen fermentation efficiency in in vitro incubation with different ratios of roughage to concentrate in swamp buffalo and cattle rumen fluid. Two male, rumen fistulated dairy steers and swamp buffaloes were used as rumen fluid donors. The treatments were arranged according to a $2{\times}2{\times}3$ factorial arrangement in a Completely randomized design by using two ratios of roughage to concentrate (R:C; 75:25 and 25:75) and 3 levels of BAFLOP supplementation (0, 2 and 4% of dietary substrate) into two different kinds of rumen fluid (beef cattle and swamp buffalo). Under this investigation, the results revealed that the rumen ecology was affected by R:C ratio. The pH declined as a result of using high concentrate ratio; however, supplementation of BAFLOP could buffer the pH which led to an improvement of ruminal efficiency. BAFLOP supplementation affected acetic acid (C2) when the proportion of concentrate was increased. However, there were no effect on total volatile fatty acid (TVFA) and butyric acid (C4) by BAFLOP supplementation. The microbial community was affected by BAFLOP supplementation, especially the bacterial population. As revealed by real-time PCR, the populations of F. succinogenes and R. albus were reduced by the high concentrate treatments while that of R. flavafaciens were increased. The populations of three dominant cellulolytic bacteria were enhanced by BAFLOP supplementation, especially on high concentrate diet. BAFLOP supplementation did not influence the ammonia nitrogen ($NH_3$-N) concentration, while R:C did. In addition, the in vitro digestibility was improved by either R:C or BAFLOP supplementation. The BAFLOP supplementation showed an effect on gas production kinetics, except for the gas production rate constant for the insoluble fraction (c), while treatments with high concentrate ratio resulted in the highest values. In addition, BAFLOP tended to increase gas production. Based on this study, it could be concluded that R:C had an effect on rumen ecology both in buffalo and cattle rumen fluid and hence, BAFLOP could be used as a rumen buffering agent for enhancing rumen ecology fed on high concentrate diet. It is recommended that level of BAFLOP supplementation should be at 2 to 4% of total dry matter of substrate. However, in vivo trials should be subsequently conducted to investigate the effect of BAFLOP in high concentrate diets on rumen ecology as well as ruminant production.

• Journal of Animal Science and Technology
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• v.51 no.1
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• pp.45-52
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• 2009

This experiment was conducted to observe the effects of anaerobic cellulolytic bacteria culture (Ruminococcus flavefaciens H-20 and Fibrobactor succinogenes H-23) on in vivo ruminal fermentation characteristics in Hanwoo heifers. Four ruminally cannulated Hanwoo heifers ($221\pm7.5kg$) receiving a basal diet containing 3 kg of mixture hay (tall fescue and ochardgrass) and 2 kg of concentrate per day were in a $4\times4$ Latin square with 21-day periods. Treatments were the basal diet without the culture additive (control), the basal diet plus 50 ml/day of bacteria culture of H-20 and H-23 (1%), 150 ml/day of H-20 and H-23 (3%), and 250 ml/day of H-20 and H-23 (5%). In the whole experimental periods, ruminal pH did not differ between treatments. However, the concentration of ruminal ammonia-N was increased in the 3% treatment relative to control and the 1% treatment at 1 hr post-feeding (p<0.05). Avicelase and CMCase (carboxymethyl cellulase) activities in rumen fluid showed no significant difference among treatments. However, xylanase activity was higher in the 5% (119.49, xylose ${\mu}mol$/ml/min) than the 3% treatment (71.02, xylose ${\mu}mol$/ml/min) at 0 hr post-feeding (p<0.05). Concentrations of ruminal total VFA, acetate, propionate and valerate were unaffected by treatments, while butyrate was higher in the 3% treatment (24.48 mM) than control (15.71 mM) at 1 hr post-feeding (p<0.05). Results indicate that minimum 3% inclusion of cellulolytic bacteria cultures improved ruminal fermentation, especially ammonia-N concentration and butyric acid production.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.12
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• pp.1746-1751
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• 2005

The objective of this study was to evaluate the effects of sarsaponin on methane production, ruminal fermentation, nutrient digestion and blood metabolites using three Holstein steers in a 3${\times}$3 Latin Square design. The steers were fed Sudangrass hay plus concentrate mixture at a ratio 1.5:1 twice daily, and sarsaponin (0, 0.5 and 1% of DM), which was given at 09:00 and 17:00 h daily by mixing with concentrate. Rumen samples were collected 0, 2, and 5 h after morning dosing. Ruminal pH was numerically decreased and numbers of protozoa were decreased linearly (p<0.01) by treatment. Ruminal ammonia-N was reduced (linear; p<0.05) and total VFA was increased (quadratic; p<0.05) at 2 and 5 h after sarsaponin dosing. The molar proportion of acetate was decreased (quadratic; p<0.05) and propionate was increased (linear; p<0.01) at all sampling times. Blood plasma glucose was increased and urea-N was decreased (linear; p<0.05) at 2 and 5 h after dosing. Methane was decreased by approximately 12.7% (linear; p<0.05). The apparent digestibility of DM and NDF were decreased (quadratic; p<0.05) and that of CP remained unchanged due to the sarsaponin. The numbers of cellulolytic bacteria were decreased (quadratic; p<0.05), while numbers of total viable bacteria remained unchanged due to the sarsaponin. These results show that sarsaponin can partially inhibit rumen methanogenesis in vivo and improve ruminal fermentation, which supports our previous in vitro results.

• Animal Bioscience
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• v.37 no.1
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• pp.74-83
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• 2024

Objective: Mitragyna speciosa Korth is traditionally used in Thailand. They have a high level of antioxidant capacities and bioactive compounds, the potential to modulate rumen fermentation and decrease methane production. The aim of the study was to investigate the different levels of microencapsulated-Mitragyna leaves extracts (MMLE) supplementation on nutrient degradability, rumen ecology, microbial dynamics, and methane production in an in vitro study. Methods: A completely randomized design was used to assign the experimental treatments, MMLE was supplemented at 0%, 4%, 6%, and 8% of the total dry matter (DM) substrate. Results: The addition of MMLE significantly increased in vitro dry matter degradability both at 12, 24, and 48 h, while ammonia-nitrogen (NH₃-N) concentration was improved with MMLE supplementation. The MMLE had the greatest propionate and total volatile fatty acid production when added with 6% of total DM substrate, while decreased the methane production (12, 24, and 48 h). Furthermore, the microbial population of cellulolytic bacteria and Butyrivibrio fibrisolvens were increased, whilst Methanobacteriales was decreased with MMLE feeding. Conclusion: The results indicated that MMLE could be a potential alternative plant-based bioactive compound supplement to be used as ruminant feed additives.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.6
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• pp.852-862
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• 2003

An experiment was conducted to study the effect of ionophore enriched cold processed mineral block supplemented with urea molasses on microbial growth and rumen fermentation. Twelve adult male crossbred cattle were divided into four groups on body weight basis. Animals were given wheat straw as a basal diet. The animals of group I and II were supplemented with concentrate mixture and animals of group III and IV were supplemented with cold processed urea molasses mineral block (UMMB). Thirty mg monensin/day/animal were supplemented to the animals of group II and 35 ppm monensin were incorporated in the UMMB supplemented to the animals of group IV. Dry matter (DM) intake did not differ significantly among groups. Mean rumen pH was higher in UMMB fed animals. Total volatile fatty acids (TVFA) concentration (mmole/L strained rumen liquor (SRL) in group III (113.19) was significantly (p<0.05) higher than those of group I (105.83) and II (108.74) but similar to group IV (109.34). TVFA production (mole/day) was similar in all the groups. The molar proportion of acetate was significantly (p<0.01) higher in the group I (59.56) than those of group II (51.73) and IV (55.91) but similar to group III (57.12). The molar proportion of propionate was significantly (p<0.01) higher in the monensin treated groups i.e. group II (38.38) and IV (36.26) than those of group I (27.78) and III (33.06). Butyrate molar percent was significantly (p<0.01) higher in group I (12.65) than those of group II (10.19), group III (9.83) and IV (7.84). The reduction of acetate and butyrate was due to UMMB and monensin resulted in lower A:P ratio. Average bacterial pool and bacterial production rate did not differ significantly among groups. Total N concentration (mg/100 ml SRL) was significantly (p<0.01) higher in the group I (55.30) and III (57.70) as compared to the group II (47.97) and IV (47.59). Ammonia-N concentration (mg/100 ml SRL) of group III (34.99) was significantly (p<0.01) higher than that of the group I (25.76) which was again significantly (p<0.01) higher than that of the group II (20.79) and IV (19.83) indicating slower release of ammonia due to monensin in diet. Total bacterial, cellulolytic, proteolytic bacterial and fungal count at 4 h post feeding did not differ significantly (p<0.05) among treatment groups. However, methanogenic bacterial count was significantly (p<0.01) higher in the group I (11.80) compared to the group II (8.43) which was significantly (p<0.01) higher than that of the group III (4.70) and IV (2.90). Average protozoal population was affected by both treatments. Thus feeding of UMMB and monensin in diet affected the rumen fermentation pattern towards propionate production, slower release of ammonia and reduction in methanogenic bacteria in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.607-615
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• 2018

Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1364-1373
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• 2012

Four crossbred (75% Holstein Friesian) lactating dairy cows, with an average live weight of $418{\pm}5$ kg and $36{\pm}10$ d in milk were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design to evaluate the effects of cassava hay (CH) and rice bran oil (RBO) on feed intake, nutrient digestibility, ruminal fermentation, milk yield, and milk composition. Factor A was non-supplementation or supplementation with CH in the concentrate. Factor B was supplementation with RBO at 0% or 4% in the concentrate mixture. The four dietary treatments were (T1) control (Concentrate with non-CH plus 0% RBO; C), (T2) Concentrate with CH plus 0% RBO (CH), (T3) Concentrate with non-CH plus 4% RBO (RBO), and (T4) Concentrate with CH plus 4% RBO (CHRBO). The cows were offered concentrate, at a ratio of concentrate to milk production of 1:2, and urea-lime treated rice straw was fed ad libitum. Urea-lime treated rice straw involved 2.5 g urea and 2.5 g $Ca(OH)_2$ (purchased as hydrated lime) in 100 ml water, the relevant volume of solution was sprayed onto a 100 g air-dry (91% DM) straw, and then covering the stack with a plastic sheet for a minimum of 10 d before feeding directly to animals. The CH based concentrate resulted in significantly higher roughage intake and total DM intake expressed as a percentage of BW (p<0.05). Ruminal pH, $NH_3$-N, BUN and total VFA did not differ among treatments, while RBO supplementation increased propionate, but decreased acetate concentration (p<0.05). Furthermore, the population of total ruminal bacteria was significantly lower on the RBO diet (p<0.05). In contrast, the total ruminal bacteria and cellulolytic bacteria on the CH diet were higher than on the other treatments. Supplementation with CH increased (p<0.05) F. succinogens and R. flavefaciens populations, whereas the populations of B. fibrisolvens and M. elsdenii were increased on the RBO diet. In addition, supplementation with CH and RBO had no effect on milk production and composition in dairy cows, while fatty acid composition of milk was influenced by RBO supplementation, and resulted in significantly lower (p<0.05) concentrations of both short-chain and medium-chain FA, and increased (p<0.05) the proportion of long-chain FA in milk fat, as well as significantly increased cis-9, trans-11 CLA and total CLA. In conclusion, RBO or CH exhibited specific effects on DMI, rumen fermentation, microbial population, milk yield and composition in lactating dairy cows, which were not interactions between CH and RBO in the diets. Feeding lactating dairy cows with RBO could improve fatty acid in milk fat by increasing cis-9, trans-11 CLA.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.1
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• pp.46-54
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• 2014

This study was conducted to investigate the effects of eucalyptus (E. Camaldulensis) crude oils (EuO) supplementation on voluntary feed intake and rumen fermentation characteristics in swamp buffaloes. Four rumen fistulated swamp buffaloes, body weight (BW) of $420{\pm}15.0$ kg, were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The dietary treatments were untreated rice straw (RS) without EuO (T1) and with EuO (T2) supplementation, and 3% urea-treated rice straw (UTRS) without EuO (T3) and with EuO (T4) supplementation. The EuO was supplemented at 2 mL/h/d in respective treatment. Experimental animals were kept in individual pens and concentrate mixture was offered at 3 g/kg BW while roughage was fed ad libitum. Total dry matter and roughage intake, and apparent digestibilites of organic matter and neutral detergent fiber were improved (p<0.01) by UTRS. There was no effect of EuO supplementation on feed intake and nutrient digestibility. Ruminal pH and temperature were not (p>0.05) affected by either roughage sources or EuO supplementation. However, buffaloes fed UTRS had higher ruminal ammonia nitrogen and blood urea nitrogen as compared with RS. Total volatile fatty acid and butyrate proportion were similar among treatments, whereas acetate was decreased and propionate molar proportion was increased by EuO supplementation. Feeding UTRS resulted in lower acetate and higher propionate concentration compared to RS. Moreover, supplementation of EuO reduced methane production especially in UTRS treatment. Protozoa populations were reduced by EuO supplementation while fungi zoospores remained the same. Total, amylolytic and cellulolytic bacterial populations were increased (p<0.01) by UTRS; However, EuO supplementation did not affect viable bacteria. Nitrogen intake and in feces were found higher in buffaloes fed UTRS. A positive nitrogen balance (absorption and retention) was in buffaloes fed UTRS. Supplementation of EuO did not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.4
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• pp.515-523
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• 2012

The study investigated the biochemical methane potential (BMP) assay of cellulose supplementing with mixed methanogens and cellulolytic bacteria to improve anaerobic digestion for methane production. For the BMP assay, 7 different microbial supplementation groups were consisted of the cultures of mixed methanogens (M), Fibrobacter succinogenes (FS), Ruminococcus flavefaciensn (RF), R. albus (RA), RA+FS and M+RA+FS including control. The cultures were added in the batch reactors with the increasing dose levels of 1% (0.5 mL), 3% (1.5 mL) and 5% (2.5 mL). Incubation for the BMP assay was carried out for 40 days at $38^{\circ}C$ and anaerobic digestate obtained from an anaerobic digester with pig slurry as inoculum was used. In results, 5% FS increased total biogas and methane production up to 10.4~22.7% and 17.4~27.5%, respectively, compared to other groups (p<0.05). Total solid (TS) digestion efficiency showed a similar trend to the total biogas and methane productions. Generally the TS digestion efficiency of the FS group was higher than that of other groups showing at the highest value of 64.2% in the 5% FS group. Volatile solid (VS) digestion efficiencies of 68.4 and 71.0% in the 5% FS and the 5% RF were higher than other groups. After incubation, pH values in all treatment groups were over 6.4 indicating that methanogensis was not inhibited during the incubation. In conclusion, the results indicated that the hydrolysis stage for methane production in anaerobic batch reactors was the late-limiting stage compared with the methanogenesis stage, and especially, as the supplementation levels of F. succinogenes supplementation increased, the methane production was increased in the BMP assay compared with other microbial culture addition.