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Sarsaponin Effects on Ruminal Fermentation and Microbes, Methane Production, Digestibility and Blood Metabolites in Steers

  • Lila, Zeenat Ara;Mohammed, Nazimuddin;Kanda, Shuhei;Kurihara, Mitsunori;Itabashi, Hisao
    • 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.

Effects of short-term fasting on in vivo rumen microbiota and in vitro rumen fermentation characteristics

  • Kim, Jong Nam;Song, Jaeyong;Kim, Eun Joong;Chang, Jongsoo;Kim, Chang-Hyun;Seo, Seongwon;Chang, Moon Baek;Bae, Gui-Seck
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.776-782
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    • 2019
  • Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

Effects of Flavonoid-rich Plant Extracts on In vitro Ruminal Methanogenesis, Microbial Populations and Fermentation Characteristics

  • Kim, Eun T.;Guan, Le Luo;Lee, Shin J.;Lee, Sang M.;Lee, Sang S.;Lee, Il D.;Lee, Su K.;Lee, Sung S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.4
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    • pp.530-537
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    • 2015
  • The objective of this study was to evaluate the in vitro effects of flavonoid-rich plant extracts (PE) on ruminal fermentation characteristics and methane emission by studying their effectiveness for methanogenesis in the rumen. A fistulated Holstein cow was used as a donor of rumen fluid. The PE (Punica granatum, Betula schmidtii, Ginkgo biloba, Camellia japonica, and Cudrania tricuspidata) known to have high concentrations of flavonoid were added to an in vitro fermentation incubated with rumen fluid. Total gas production and microbial growth with all PE was higher than that of the control at 24 h incubation, while the methane emission was significantly lower (p<0.05) than that of the control. The decrease in methane accumulation relative to the control was 47.6%, 39.6%, 46.7%, 47.9%, and 48.8% for Punica, Betula, Ginkgo, Camellia, and Cudrania treatments, respectively. Ciliate populations were reduced by more than 60% in flavonoid-rich PE treatments. The Fibrobacter succinogenes diversity in all added flavonoid-rich PE was shown to increase, while the Ruminoccocus albus and R. flavefaciens populations in all PE decreased as compared with the control. In particular, the F. succinogenes community with the addition of Birch extract increased to a greater extent than that of others. In conclusion, the results of this study showed that flavonoid-rich PE decreased ruminal methane emission without adversely affecting ruminal fermentation characteristics in vitro in 24 h incubation time, suggesting that the flavonoid-rich PE have potential possibility as bio-active regulator for ruminants.

In vitro ruminal fermentation of fenugreek (Trigonella foenum-graecum L.) produced less methane than that of alfalfa (Medicago sativa)

  • Niu, Huaxin;Xu, Zhongjun;Yang, Hee Eun;McAllister, Tim A;Acharya, Surya;Wang, Yuxi
    • Animal Bioscience
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    • v.34 no.4
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    • pp.584-593
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    • 2021
  • Objective: The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods: Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results: Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p<0.05) in vitro DM and neutral detergent fibre disappearance, but this difference was not observed between the mid-bloom Alf and 15-wk FG. Regardless plant maturity, in vitro ruminal fermentation of FG produced less (p<0.001) CH4 either on DM incubated or on DM disappeared basis than that of Alf during 48-h incubation. In vitro ruminal fermentation of FG yielded similar amount of total volatile fatty acids with higher (p<0.05) propionate percentage as compared to fermentation of Alf irrespective of plant maturity. Microbial protein synthesis was greater (p<0.001) with 11-wk FG than early bloom Alf as substrate and 16S RNA gene copies of total bacteria was higher (p<0.01) with 15-wk FG than mid-bloom Alf as substrate. Compared to mid-bloom Alf, 15-wk FG had lower (p<0.05 to 0.001) amount of 16S RNA methanogen gene copies in the whole culture during 48-h incubation. Conclusion: In comparison to Alf, FG emerges as a high quality forage that can not only improve rumen fermentation in vitro, but can also remarkably mitigate CH4 emissions likely due to being rich in saponins.

Evaluation of the nutritional value of locally produced forage in Korea using chemical analysis and in vitro ruminal fermentation

  • Ki, Kwang Seok;Park, Su Bum;Lim, Dong Hyun;Seo, Seongwon
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.3
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    • pp.355-362
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    • 2017
  • Objective: The use of locally produced forage (LPF) in cattle production has economic and environmental advantages over imported forage. The objective of this study was to characterize the nutritional value of LPF commonly used in Korea. Differences in ruminal fermentation characteristics were also examined for the LPF species commonly produced from two major production regions: Chungcheong and Jeolla. Methods: Ten LPF (five from each of the two regions) and six of the most widely used imported forages originating from North America were obtained at least three times throughout a year. Each forage species was pooled and analyzed for nutrient content using detailed chemical analysis. Ruminal fermentation characteristics were also determined by in vitro anaerobic incubations using strained rumen fluid for 0, 3, 6, 12, 24, and 48 h. At each incubation time, total gas, pH, ammonia, volatile fatty acid (VFA) concentrations, and neutral detergent fiber digestibility were measured. By fitting an exponential model, gas production kinetics were obtained. Results: Significant differences were found in the non-fiber carbohydrate (NFC) content among the forage species and the regions (p<0.01). No nutrient, other than NFC, showed significant differences among the regions. Crude protein, NFC, and acid detergent lignin significantly differed by forage species. The amount of acid detergent insoluble protein tended to differ among the forages. The forages produced in Chungcheong had a higher amount of NFC than that in Jeolla (p<0.05). There were differences in ruminal fermentation of LPF between the two regions and interactions between regions and forage species were also significant (p<0.05). The pH following a 48-h ruminal fermentation was lower in the forages from Chungcheong than from Jeolla (p<0.01), and total VFA concentration was higher in Chungcheong than in Jeolla (p = 0.05). This implies that fermentation was more active with the forages from Chungcheong than from Jeolla. Analysis of gas production profiles showed the rate of fermentation differed among forage species (p<0.05). Conclusion: The results of the present study showed that the nutritional values of some LPF (i.e., corn silage and Italian ryegrass) are comparable to those of imported forages widely used in Korea. This study also indicated that the nutritional value of LPF differs by origin, as well as by forage species. Detailed analyses of nutrient composition and digestion kinetics of LPF should be routinely employed to evaluate the correct nutritional value of LPF and to increase their use in the field.

Ruminal pH pattern, fermentation characteristics and related bacteria in response to dietary live yeast (Saccharomyces cerevisiae) supplementation in beef cattle

  • Zhang, Xiangfei;Dong, Xianwen;Wanapat, Metha;Shah, Ali Mujtaba;Luo, Xiaolin;Peng, Quanhui;Kang, Kun;Hu, Rui;Guan, Jiuqiang;Wang, Zhisheng
    • Animal Bioscience
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    • v.35 no.2
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    • pp.184-195
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    • 2022
  • Objective: In this study we aimed to evaluate the effect of dietary live yeast supplementation on ruminal pH pattern, fermentation characteristics and associated bacteria in beef cattle. Methods: This work comprised of in vitro and in vivo experiments. In vitro fermentation was conducted by incubating 0%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15% active dried yeast (Saccharomyces cerevisiae, ADY) with total mixed ration substrate to determine its dose effect. According to in vitro results, 0.1% ADY inclusion level was assigned in in vivo study for continuously monitoring ruminal fermentation characteristics and microbes. Six ruminally cannulated steers were randomly assigned to 2 treatments (Control and ADY supplementation) as two-period crossover design (30-day). Blood samples were harvested before-feeding and rumen fluid was sampled at 0, 3, 6, 9, and 12 h post-feeding on 30 d. Results: After 24 h in vitro fermentation, pH and gas production were increased at 0.1% ADY where ammonia nitrogen and microbial crude protein also displayed lowest and peak values, respectively. Acetate, butyrate and total volatile fatty acids concentrations heightened with increasing ADY doses and plateaued at high levels, while acetate to propionate ratio was decreased accordingly. In in vivo study, ruminal pH was increased with ADY supplementation that also elevated acetate and propionate. Conversely, ADY reduced lactate level by dampening Streptococcus bovis and inducing greater Selenomonas ruminantium and Megasphaera elsdenii populations involved in lactate utilization. The serum urea nitrogen decreased, whereas glucose, albumin and total protein concentrations were increased with ADY supplementation. Conclusion: The results demonstrated dietary ADY improved ruminal fermentation dose-dependently. The ruminal lactate reduction through modification of lactate metabolic bacteria could be an important reason for rumen pH stabilization induced by ADY. ADY supplementation offered a complementary probiotics strategy in improving gluconeogenesis and nitrogen metabolism of beef cattle, potentially resulted from optimized rumen pH and fermentation.

Effects of Dietary Nitrogen Sources on Fiber Digestion and Ruminal Fluid Characteristics in Sheep Fed Wheat Straw

  • Tan, Z.-L.;Lu, D.-X.;Hu, M.;Niu, W.-Y.;Han, C.-Y.;Ren, X.-P.;Na, R.;Lin, S.-L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1374-1382
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    • 2001
  • Fifteen Inner Mongolian wethers with permanent ruminal and duodenal cannulas were used to study the effects of dietary rumen-undegradable protein (RUP) to rumen-degradable protein (RDP) ratios or protein sources on fiber digestion in the gastrointestinal tract and ruminal fluid characteristics. Fiber digestion and ruminal fermentation were not affected (p>0.05) by dietary RUP to RDP ratios (from 1.54 to 0.72). Soybean meal supplementation improved ruminal digestion. Fish meal supplementation increased (p<0.05) the ruminal degradability of fiber. The different RUP to RDP ratios (from 1.54 to 0.72) did not influence (p>0.05) ruminal fluid pH, but there were differences (p<0.05) in ruminal fluid $NH_3-N$ concentration because of urea replacement. Soybean meal as a dietary protein source decreased (p<0.05) ruminal fluid pH and increased (p<0.05 or p<0.01) $NH_3-N$, acetate, propionate and butyrate concentrations in the rumen. Fish meal as a dietary protein source decreased (p<0.05 or p<0.01) ruminal $NH_3-N$ and acetate concentrations and increased (p<0.05) ruminal propionate concentration. It can be concluded that dietary protein sources have more significant effect on fiber digestion and ruminal fermentation than different dietary RUP to RDP ratios, when the dietary crude protein requirements of growing sheep are satisfied.

Effect of Ruminal NH3-N Levels on Ruminal Fermentation, Purine Derivatives, Digestibility and Rice Straw Intake in Swamp Buffaloes

  • Wanapat, M.;Pimpa, O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.6
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    • pp.904-907
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    • 1999
  • The experiment was aimed at studying the effect of ruminal $NH_3-N$ levels on ruminal fermentation, microbial population, urinary purine derivative excretion, digestibility and rice straw intake in swamp buffaloes. Five, 3 to 4 years old, rumen fistulated swamp buffaloes were randomly assigned according to a $5{\times}5$ Latin square design to rceive five different intraruminal infusions of $NH_4HCO_3$ (0, 150, 300, 450 and 600 g/d) on a continuous daily basis. Rice straw as a roughage was offered ad libitum while concentrate was given at 0.8% BW daily. The results were that as levels of $NH_4HCO_3$ increased, ruminal $NH_3-N$ concentrations increased from 7.1 to 34.4 mg%. The highest digestibility and voluntary straw intakes were found at 13.6 to 17.6 mg% ruminal $NH_3-N$ levels; straw intake was highest at 13.6 mg%. Total bacterial and protozoal counts linearly increased as the ruminal $NH_3-N$ increased and were highest at 17.6 mg%. Total urinary purine derivatives and allantoin excretion were highest (44.0, 37.4 mM/d) at 17.6 mg% ruminal $NH_3-N$. Highest total VFAs (115 mM) were obtained a 13.6 mg% ruminal $NH_3-N$ while blood urea nitrogen significantly increased as ruminal $NH_3-N$ increased. The results from this experiment suggest that optimum ruminal $NH_3-N$ in swamp buffaloes is higher than 13.6 mg%, for improving rumen ecology, microbial protein synthesis, digestibility and straw intake.

Effects of Ruminal Infusion of Garlic Oil on Fermentation Dynamics, Fatty Acid Profile and Abundance of Bacteria Involved in Biohydrogenation in Rumen of Goats

  • Zhu, Zhi;Mao, Shengyong;Zhu, Weiyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.962-970
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    • 2012
  • This study aimed to investigate the effects of ruminal infusion of garlic oil (GO) on fermentation dynamics, fatty acid (FA) profile, and abundance of bacteria involved in biohydrogenation in the rumen. Six wethers fitted with ruminal fistula were assigned to two groups for cross-over design with a 14-d interval. Each 30-d experimental period consisted of a 27-d adaptation and a 3-d sample collection. Goats were fed a basal diet without (control) or with GO ruminal infusion (0.8 g/d). Ruminal contents collected before (0 h) and at 2, 4, 6, 8, and 10 h after morning feeding were used for fermentation analysis, and 0 h samples were further used for FA determination and DNA extraction. Garlic oil had no influence on dry matter intakes of concentrate and hay. During ruminal fermentation, GO had no effects on total VFA concentration and individual VFA molar proportions, whereas GO increased the concentrations of ammonia nitrogen and microbial crude protein (p<0.05). Compared with control, GO group took a longer time for total VFA concentration and propionate molar proportion to reach their respective maxima after morning feeding. The ratio of acetate to propionate in control reduced sharply after morning feeding, whereas it remained relatively stable in GO group. Fatty acid analysis showed that GO reduced saturated FA proportion (p<0.05), while increasing the proportions of C18, t11-18:1 (TVA), c9,t11-conjugated linoleic acid (c9,t11-CLA), t10,c12-CLA, and polyunsaturated FA (p<0.05). The values of TVA/(c9,t11-CLA+TVA) and C18:0/(TVA+C18:0) were reduced by GO (p<0.05). Real-time PCR showed that GO tended to reduce Butyrivibrio proteoclasticus abundance (p = 0.058), whereas GO had no effect on total abundance of the Butyrivibrio group bacteria. A low correlation was found between B. proteoclasticus abundance and C18:0/(TVA+C18:0) (p = 0.910). The changes of fermentation over time suggested a role of GO in delaying the fermentation process and maintaining a relatively modest change of ruminal environment. The inhibitory effects of GO on the final step of biohydrogenation may be related to its antibacterial activity against B. proteoclasticus and other unknown bacteria involved.

Effects of dietary cation and anion difference on eating, ruminal function and plasma leptin in goats under tropical condition

  • Nguyen, Thiet;Chanpongsang, Somchai;Chaiyabutr, Narongsak;Thammacharoen, Sumpun
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.941-948
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    • 2020
  • Objective: This study was carried out to determine the effects of elevated dietary cation and anion difference (DCAD) on dry matter intake (DMI) and ruminal fermentation pattern in lactating dairy goats under tropical conditions. Methods: Ten dairy goats were divided into two groups of five animals each. The groups received diets at different DCAD levels, either a control diet (22.81 mEq/100 g dry matter [DM], DCAD-23) or a DCAD-39 diet (39.08 mEq/100 g DM, DCAD-39). After parturition, DMI and water intake were recorded daily. Ruminal fluid and urine were collected, and nutrient digestibility measurements were carried out at 8th weeks postpartum (PP-8). Blood samples were collected at PP-4 and PP-8 to measure plasma leptin. Results: Dry matter intake/body weight (DMI/BW) at PP-8 of the animals fed the DCAD-39 diet was significantly higher than those fed with DCAD-23 diet (p<0.05). Animals fed with DCAD-39 consumed more water than those fed DCAD-23 over 24 h, particularly at night (p<0.05). Ruminal pH, acetate concentration, and urinary allantoin excretion increased with the DCAD-39 diet, whereas ruminal butyrate concentration was lower with the DCAD-39 diet. On the other hand, other ruminal parameters, such as total volatile fatty acid concentration, propionate molar proportion and acetate/propionate average ratio, were not affected by increased DCAD supplementation. Apparent digestibility was improved by increased DCAD supplementation. Plasma leptin concentration was higher with DCAD supplementation. Conclusion: When feeding goats with DCAD-39 under tropical conditions, an increase in DMI was associated with improved apparent digestibility of nutrients, ruminal fermentation and microbial protein synthesis. An increase in plasma leptin concentration could not explain the effect of high DCAD on DMI.