• Title/Summary/Keyword: In vitro gas production

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Effects of Tween 80 on In Vitro Fermentation of Silages and Interactive Effects of Tween 80, Monensin and Exogenous Fibrolytic Enzymes on Growth Performance by Feedlot Cattle

  • Wang, Y.;McAllister, T.A.;Baah, J.;Wilde, R.;Beauchemin, K.A.;Rode, L.M.;Shelford, J.A.;Kamande, G.M.;Cheng, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.968-978
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    • 2003
  • The effects of monensin, Tween 80 and exogenous fibrolytic enzymes on ruminal fermentation and animal performance were studied in vitro and in vivo. In Expt 1, the effects of the surfactant Tween 80 (0.2% wt/wt, DM basis) on ruminal fermentation of alfalfa, corn and orchardgrass silages were investigated using in vitro gas production techniques. Tween 80 did not affect (p>0.05) cumulative gas production at 24 h, but it reduced (p<0.05) the lag in fermentation of all three silages. With corn silage and orchardgrass silage, gas production rates and concentrations of total volatile fatty acids (VFA) were increased (p<0.05) by Tween 80; with alfalfa silage, they were reduced (p<0.05). Tween 80 increased (p<0.05) the proportion of propionate in total VFA, and reduced (p<0.05) acetate to propionate ratios (A:P) with all three silages. In Expt 2, exogenous fibrolytic enzymes (E; at 0, 37.5 or 75 g/tonne DM), monensin (M; at 0 or 25 ppm and Tween 80 (T; at 0 or 2 L/tonne DM) were added alone or in combination to backgrounding and finishing diets fed to 320 crossbred steers in a feeding trial with a $3{\times}2{\times}$2 factorial arrangement of treatments. The backgrounding and finishing diets contained barley grain and barley silage in ratios of 57.8:42.2 and 93.5:6.5 (DM basis), respectively. Added alone, none of the additives affected DM intake (p>0.1) in the backgrounding or in the finishing period, but interactive $M{\times}T$ effects were observed in the finishing period (p=0.02) and overall (p=0.04). In the finishing period, T without M tended to reduce DM intake (p=0.11), but T with M increased (p=0.05) DM intake. Monensin increased average daily gain (ADG) during backgrounding (p=0.07) and finishing (p=0.01), and this ionophore also improved overall feed efficiency (p=0.02). Warm carcass weight was increased (p<0.001) by M, but dressing percentage was reduced (p=0.07). In the backgrounding period, T increased ADG by 7% (p=0.06). Enzymes increased (p=0.07) ADG by 5 and 6% (low and high application rates, respectively) during backgrounding, but did not affect (p>0.10) ADG during finishing, or overall feed efficiency. Whereas T enhanced the positive effects of M on ADG during backgrounding (p=0.04) and overall (p=0.05), it had no impact (p>0.1) on the effects of E. Interactions between M and T suggest that the surfactant may have potential for enhancing the positive effects of monensin on beef production, but this requires further research.

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.

Effects of Cynanchum Wilfordii Extract on In vitro Ruminal Fermentation Characteristics and Methane Production (백하수오 추출물이 In vitro 반추위 발효성상 및 메탄가스 생성에 미치는 영향)

  • Yang, Seung-Hak;Lim, Joung-Soo;Kim, Byul;Hwang, Ok-Hwa;Cho, Sung-Back;Choi, Dong-Yoon;Choi, Seok-Geun;Hwang, Seong-Gu
    • Journal of Animal Environmental Science
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    • v.19 no.2
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    • pp.155-162
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    • 2013
  • The objective of this study is to investigate the effects of Cynanchum wilfordii (CW) on cell viability, anti-oxidant activity, volatile fatty acid (VFA) production and methane gas production. Collected rumen fluid incubated with CW powder (1% w/v) for 12 and 24 hours were analyzed for pH, VFAs and methane. Alamar blue assay showed no significant difference on the viability of 3T3-L1 and C2C12 cells treated with CW for 24 hours. TBARS data showed a dose dependent increase on the antioxidant activity of CW. VFAs increased in the CW-treated groups compared to the control group. In addition, propionate increased more than other VFAs by the treatment with CW. There was a significant decrease in methane gas production in batch culture treated with CW in 12hrs. In conclusion, it was suggested that Cynanchum wilfordii could manipulate rumen fermentation considered by increasing VFA production and inhibition of methanogenesis.

Effect of 2-Bromoethanesulfonic Acid on In vitro Fermentation Characteristics and Methanogen Population

  • Lee, S.Y.;Yang, S.H.;Lee, W.S.;Kim, H.S.;Shin, D.E.;Ha, Jong K.
    • 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.

Effects of Gelidium amansii extracts on in vitro ruminal fermentation characteristics, methanogenesis, and microbial populations

  • Lee, Shin Ja;Shin, Nyeon Hak;Jeong, Jin Suk;Kim, Eun Tae;Lee, Su Kyoung;Lee, Il Dong;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.71-79
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    • 2018
  • Objective: Gelidium amansii (Lamouroux) is a red alga belonging to the family Gelidaceae and is commonly found in the shallow coasts of many East Asian countries, including Korea, China, and Japan. G. amansii has traditionally been utilized as an edible alga, and has various biological activities. The objective of this study was to determine whether dietary supplementation of G. amansii could be useful for improving ruminal fermentation. Methods: As assessed by in vitro fermentation parameters such as pH, total gas, volatile fatty acid (VFA) production, gas profile (methane, carbon dioxide, hydrogen, and ammonia), and microbial growth rate was compared to a basal diet with timothy hay. Cannulated Holstein cows were used as rumen fluid donors and 15 mL rumen fluid: buffer (1:2) was incubated for up to 72 h with four treatments with three replicates. The treatments were: control (timothy only), basal diet with 1% G. amansii extract, basal diet with 3% G. amansii extract, and basal diet with 5% G. amansii extract. Results: Overall, the results of our study indicate that G. amansii supplementation is potentially useful for improving ruminant growth performance, via increased total gas and VFA production, but does come with some undesirable effects, such as increasing pH, ammonia concentration, and methane production. In particular, real-time polymerase chain reaction indicated that the methanogenic archaea and Fibrobacter succinogenes populations were significantly reduced, while the Ruminococcus flavefaciens populations were significantly increased at 24 h, when supplemented with G. amansii extracts as compared with controls. Conclusion: More research is required to elucidate what G. amansii supplementation can do to improve growth performance, and its effect on methane production in ruminants.

Effect of Disodium Fumarate on In vitro Rumen Fermentation of Different Substrates and Rumen Bacterial Communities as Revealed by Denaturing Gradient Gel Electrophoresis Analysis of 16S Ribosomal DNA

  • Mao, S.Y.;Zhang, G.;Zhu, W.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.543-549
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    • 2007
  • Two experiments were conducted to investigate the effects of disodium fumarate on the in vitro rumen fermentation profiles of different substrates and microbial communities. In experiment 1, nine diets (high-forage diet (forage:concentrate, e.g. F:C = 7:3, DM basis), medium-forage diet (F:C = 5:5, DM basis), low-forage diet(F:C = 1:9, DM basis), cracked corn, cracked wheat, soluble starch, tall elata (Festuca elata), perennial ryegrass and rice straw) were fermented in vitro by rumen microorganisms from local goats. The results showed that during 24 h incubations, for all substrates, disodium fumarate increased (p<0.05) the gas production, and tended to increase (p<0.10) the acetate, propionate and total VFA concentration and decrease the ratio of acetate to propionate, whereas no treatment effect was observed for the lactate concentration. The apparent DM loss for tall elata, perennial ryegrass and rice straw increased (p<0.05) with the addition of disodium fumarate. With the exception of tall elata, perennial ryegrass and rice straw, disodium fumarate addition increased the final pH (p<0.05) for all substrates. In experiment 2, three substrates (a high-forage diet, a medium-forage diet and a high concentrate diet) were fermented by mixed rumen microbes in vitro. A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique was applied to compare microbial DNA fingerprints between substrates at the end of 24 h incubation. The results showed that when Festuca elata was used as substrate, the control and disodium fumarate treatments had similar DGGE profiles, with their similarities higher than 96%. As the ratio of concentrate increased, however, the similarities in DGGE profiles decreased between the control and disodium fumarate treatment. Overall, these results suggest that disodium fumarate is effective in increasing the pH and gas production for the diets differing in forage: concentrate ratio, grain cereals and soluble starch, and in increasing dry matter loss for the forages (tall elata, perennial ryegrass and rice straw) in vitro, whereas its effect on changes of ruminal microbial community may largely depend on the general nature of the substrate.

In vitro Metabolism of Stanozolol to 3'-Hydroxystanozolol in the Liver S-9 Fraction of Polychlorinated Biphenyl-treated Rats (Polychlorinated biphenyl 전처리한 횐쥐 간장의 S-9 분획에서 Stanozolol의 Hydroxylation 대사체의 생성)

  • 권오승;류재천
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.379-383
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    • 2000
  • Stanozolol (STZ, 17$\alpha$-methyl-17$\beta$-hydroxy-5$\alpha$-androstano-(3,2-c) pyrazole), an anabolic steroid, is an abused drug by body-builders or atheletes, as well as medicine for treatment of aplastic anemia and vascular thrombosis. In human volunteers, the major urinary metabolite of STZ was reported to be 3'-hydroxystanozolol that was identified by gas chromatography-mass selective detector (GC/MSD). The objective of this experiment is to investigate the in vitro metabolism of STZ in liver S-9 faction of polychlorinated biphenyl-induced rats. Reaction mixture including STZ as substrate and the S-9 faction was extracted with diethyl ether and quantified by the selected ion monitoring mode of GC/MSD. The selected concentration of substrate STZ is 100 nmole and the selected time for incubation in the reaction mixture was determined to 60 min. The amount of 3'-hydroxystanozolol produced was increased by about 6-fold in the reaction medium including the liver S-9 fraction of polychlorinated biphenyl-induced rats, compared to that of untreated rats. Inhibitors of cytochrome P450, SKF-525A and 7,8-benzoflavone, decreased the production of 3'-hydroxystanozolol by about 89~100% and 65~75%, respectively; In conclusion, hydroxylation of STZ into 3'-hydroxystanozolol is confirmed by GC/MSD and is catalyzed by cytochrome P450.

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Effect of storage time and the level of formic acid on fermentation characteristics, epiphytic microflora, carbohydrate components and in vitro digestibility of rice straw silage

  • Zhao, Jie;Wang, Siran;Dong, Zhihao;Li, Junfeng;Jia, Yushan;Shao, Tao
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1038-1048
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    • 2021
  • Objective: The study aimed to evaluate the effect of storage time and formic acid (FA) on fermentation characteristics, epiphytic microflora, carbohydrate components and in vitro digestibility of rice straw silage. Methods: Fresh rice straw was ensiled with four levels of FA (0%, 0.2%, 0.4%, and 0.6% of fresh weight) for 3, 6, 9, 15, 30, and 60 d. At each time point, the silos were opened and sampled for chemical and microbial analyses. Meanwhile, the fresh and 60-d ensiled rice straw were further subjected to in vitro analyses. Results: The results showed that 0.2% and 0.6% FA both produced well-preserved silages with low pH value and undetected butyric acid, whereas it was converse for 0.4% FA. The populations of enterobacteria, yeasts, moulds and aerobic bacteria were suppressed by 0.2% and 0.6% FA, resulting in lower dry matter loss, ammonia nitrogen and ethanol content (p<0.05). The increase of FA linearly (p<0.001) decreased neutral detergent fibre and hemicellulose, linearly (p<0.001) increased residual water soluble carbohydrate, glucose, fructose and xylose. The in vitro gas production of rice straw was decreased by ensilage but the initial gas production rate was increased, and further improved by FA application (p<0.05). No obvious difference of FA application on in vitro digestibility of dry matter, neutral detergent fibre, and acid detergent fibre was observed (p>0.05). Conclusion: The 0.2% FA application level promoted lactic acid fermentation while 0.6% FA restricted all microbial fermentation of rice straw silages. Rice straw ensiled with 0.2% FA or 0.6% FA improved its nutrient preservation without affecting digestion, with the 0.6% FA level best.

In vitro Fermentation of Rumen Microorganisms Cultured in Medium Supplemented with Bacterio-mineral Water (BMW) Produced from Bio-reacted Swine Manure

  • Kim, Chang-Hyun;Park, Joong Kook;Lee, Gi Yeong;Seo, In Joon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1435-1439
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    • 2005
  • Bacterio-mineral water (BMW) produced from manure has been known to exert a number of positive effects on animal production and odor control. An experiment was conducted to examine the effects of BMW produced from bio-reacted swine manure on in vitro gas production, cellulose degradation, microbial growth and fibrolytic enzyme activities of mixed rumen microorganisms. The five levels of 0, 0.001, 0.005, 0.01 and 1.0% BMW were supplemented into serum vials containing mixed rumen microorganisms. Incubations were carried out anaerobically at $39^{\circ}C$ without shaking for 0, 12, 24, 48, 72 and 96 h. There were no significant (p>0.05) differences among the treatments for the initial rate of gas production. At 72 h incubation, the gas production tended (p<0.1) to be increased by the 0.01 and 1.0% BMW treatments compared with control and the 0.001% BMW treatment. At the end of incubation (96 h), the sample supplemented with 0.01% BMW was higher (p<0.05) than control (0% BMW) in the gas production. The microbial growth rate was increased by all the BMW treatments, while 0.01% BMW was most effective in stimulating the growth rate. Although the addition of BMW on the filter paper DM degradation was not significantly influenced throughout the incubation period except the 48 h incubation, DM degradation tended to be increased by all BMW treatments compared with control. The addition of both 0.005 and 0.01% BMW highly increased (p<0.05) CMCase activity compared with control after 24 h and 48 h incubation, while at the 72 h incubation the 0.01% BMW addition only significantly increased (p<0.05). After 72 h incubation, the xylanase activity was significantly (p<0.05) increased with the addition of 1.0% BMW compared with the addition of 0.001 and 0.005% BMW, while at the other incubation times, the xylanase activity was not different among the treatments. In conclusion, the 0.01% BMW of supplementation level would be the suitable addition level to stimulate rumen fermentation increasing microbial growth and cellulose degradation.

Evaluation of Some Agri-industrial By-products Available in Samoa for Goats

  • Aregheore, E.M.;Abdulrazak, S.A.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.11
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    • pp.1593-1598
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    • 2003
  • Nutritional evaluation of some agro-industrial byproducts available in Samoa [dry brewers' grains (DBG), cocoa shell (CS), cocoa dust (CD) and desiccated coconut waste meal (DCWM)] available in Samoa was carried out using both the in vivo and in vitro techniques. In the in vivo study 24 Anglo-nubian goats were offered by-products with other feed ingredients to compound four different diets. The goats were randomly allocated to 4 diets on the basis of liveweight (18.7-0.3kg). The ADF content of the byproducts followed a similar trend to NDF. The byproducts have a high content of organic matter (91.0-95.4%). Gross energy (GE) content was higher in DCWM (25.1 MJ/kg DM), closely followed by CD (23.2 MJ/kg DM). Concentrate intake was significantly different (p<0.05) among the goats. Average daily live weight gains were 105, 92, 88 and 97 g/goat/day for DBG, CS, CD and DCWM, respectively. Daily live weight gains were higher (p<0.05) in the goats that received DBG, while the least gain was obtained in the goats that received CS byproduct diet. DM digestibility was significantly higher (p<0.05) in the goats on DBG diet than in the other goats. The least DM digestibility was obtained in the goats that received CD diet (p>0.05). CP digestibility followed a similar pattern to DM digestibility. The digestibility of NDF and ADF was influenced by the nature of the diets. The digestibility of OM and GE were best (p<0.05) in the goats that received DBG, DCWM and CS byproduct diets than in CD. Significant differences (p<0.05) among the byproducts were recorded for net gas production. Potential gas production (a+b) ranged from 7.064 to 42.17 ml. Organic matter digested (OMD) from gas production value at 24 h was higher in DBG (47.6 g/kg DM) and this was followed by DCWM (42.5 g/kg DM). The least OMD was obtained in CD (17.9 g/kg DM). A significant difference (p<0.05) in DM disappearance after 4, 8, 16, 24, 48 and 72 h was recorded. The potential and effective degradability varied significantly (p<0.05) from 85.95-99.6 g/kg DM and from 39.9-65.8%, respectively. The digestibility of the byproducts in both the in vivo and in in vitro techniques demonstrated that they are potential source of feed ingredients for ruminant livestock in Samoa and possibly in the other small Pacific Island countries. On the basis of their potential degradability the byproducts could be ranked in the following order:DCWM>DBG>CD>CS. In conclusion, the results obtained suggest that all the byproducts can contribute to ruminant livestock diets without adverse effects on feed intake, growth rate and apparent nutrient digestibility coefficients.