• Title/Summary/Keyword: In Vitro Fermentation

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Comparative analysis of silage fermentation and in vitro digestibility of tropical grass prepared with Acremonium and Tricoderma species producing cellulases

  • Khota, Waroon;Pholsen, Suradej;Higgs, David;Cai, Yimin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1913-1922
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    • 2018
  • Objective: To find out ways of improving fermentation quality of silage, the comparative analysis of fermentation characteristics and in vitro digestibility of tropical grasses silage applied with cellulases produced from Acremonium or Tricoderma species were studied in Thailand. Methods: Fresh and wilted Guinea grass and Napier grass silages were prepared with cellulases from Acremonium (AC) or Trichoderma (TC) at 0.0025%, 0.005%, and 0.01% on a fresh matter (FM), and their fermentation quality, chemical composition and in vitro digestibility were analyzed. Results: All silages of fresh Napier grass were good quality with lower pH, butyric acid, and ammonia nitrogen, but higher lactic acid content than wilted Napier grass and Guinea grass silage. Silages treated with AC 0.01% had the best result in terms of fermentation quality. They also had higher in vitro dry matter digestibility and in vitro organic matter digestibility at 6 and 48 h after incubation than other silages. Silages treated with lower levels at 0.005% or 0.0025% of AC and all levels of TC did not improve silage fermentation. Conclusion: The AC could improve silage fermentation and in vitro degradation of Guinea grass and Napier grass silages, and the suitable addition ration is 0.01% (73.5 U) of FM for tropical silage preparation.

Effects of wilting and additives on the ensiling quality and in vitro rumen fermentation characteristics of sudangrass silage

  • Wan, Jiang Chun;Xie, Kai Yun;Wang, Yu Xiang;Liu, Li;Yu, Zhu;Wang, Bing
    • Animal Bioscience
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    • v.34 no.1
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    • pp.56-65
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    • 2021
  • Objective: This study was conducted to investigate the effects of molasses and Lactobacillus plantarum on the ensiling quality and in vitro rumen fermentation of sudangrass silage prepared with or without wilting. Methods: The ensiling experiment, measured with 3 replicates, was carried out according to a 2×4 (wilted stages×additives) factorial treatment structure. Dry matter of the fresh (210 g/kg fresh matter) or wilted (305 g/kg fresh matter) sudangrass were ensiled (packed into 5.0-L plastic jars) without additive (control) or with molasses (M), Lactobacillus plantarum (LP), or molasses + Lactobacillus plantarum (M+LP). After 60 days of ensiling, the silages were analyzed for the chemical, fermentation, and in vitro characteristics. Results: After 60 days of ensiling, the fermentation parameters were affected by wilted, the additives and the interactions of wilted with the additives (p<0.05). The M+LP treatment at wilted had higher lactic acid levels and V-score (p<0.05) but lower pH values and butyric acid concentrations than the other treatments. In comparison with sudangrass before ensiling, after ensiling had lower dry matter and higher non-fibrous carbohydrate. The in vitro gas production, in vitro dry matter digestibility, in vitro crude protein digestibility, and in vitro acid fiber detergent digestibility changed under the effects of the additives. Significant interactions were observed between wilted and the additives in terms of in vitro gas production at 48 h, asymptotic gas production, gas production rate, half time, and the average gas production rate. The total volatile fatty acid levels in the additive treatments were higher than those in the control. Conclusion: Wilting and supplementation with molasses and Lactobacillus plantarum had the ability to improve the ensiling quality and in vitro nutrient digestibility of sudangrass silage. The M+LP treatment at wilted exhibited the strongest positive effects on silage quality and in vitro ruminal fermentation characteristics.

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.

Effect of γ-aminobutyric acid producing bacteria on in vitro rumen fermentation, growth performance, and meat quality of Hanwoo steers

  • Mamuad, Lovelia L.;Kim, Seon Ho;Ku, Min Jung;Lee, Sang Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.7
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    • pp.1087-1095
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    • 2020
  • Objective: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)-producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. Methods: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. Results: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH3-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. Conclusion: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH3-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers.

Effects of lactic acid bacteria and molasses on fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro ruminal fermentation of rice straw silage

  • Zhao, Jie;Dong, Zhihao;Li, Junfeng;Chen, Lei;Bai, Yunfeng;Jia, Yushan;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.783-791
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    • 2019
  • Objective: This study was to evaluate the fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro gas production of rice straw ensiled with lactic acid bacteria and molasses. Methods: Fresh rice straw was ensiled in 1-L laboratory silos with no additive control (C), Lactobacillus plantarum (L), molasses (M) and molasses+Lactobacillus plantarum (ML) for 6, 15, 30, and 60 days. After storage, the silages were subjected to microbial and chemical analyses as well as the further in vitro fermentation trial. Results: All additives increased lactic acid concentration, and reduced pH, dry matter (DM) loss and structural carbohydrate content relative to the control (p<0.05). The highest organic acid and residual sugar contents and lignocellulose reduction were observed in ML silage. L silage had the highest V-score with 88.10 followed by ML silage. L and ML silage improved in vitro DM digestibility as compared with other treatments, while in vitro neutral detergent fibre degradability (IVNDFD) was increased in M and ML silage (p<0.05). M silage significantly (p<0.05) increased propionic acid (PA) content and decreased butyric acid content and acetic acid/PA as well as 72-h cumulative gas production. Conclusion: The application of ML was effective for improving both the fermentation quality and in vitro digestibility of rice straw silage. Inclusion with molasses to rice straw could reduce in vitro ruminal gas production.

Effects of Lactic Acid Bacteria Inoculant on Fermentation Quality and in vitro Rumen Fermentation of Total Mixed Ration

  • Choi, Yeon Jae;Lee, Sang Suk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.3
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    • pp.132-140
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    • 2019
  • Fermented total mixed ration (TMR) is a novel feed for ruminants in South Korea. The purpose of this study was to evaluate the effects of lactic acid bacteria (LAB) on the quality of TMR and in vitro ruminal fermentation. Strains of three LAB spp. (Lactobacillus plantarum, L. brevis, L. mucosae) were used in fermentation of TMR. Inoculations with the three LAB spp. lowered pH and increased concentrations of lactic acid, acetic acid, and total organic acid compared to non-LAB inoculated control (only addition of an equivalent amount of water) (p<0.05). Bacterial composition indicated that aerobic bacteria and LAB were higher. However, E. coli were lower in the fermented TMR than those in the control treatment (p<0.05). Among the treatments, L. brevis treatment had the highest concentration of total organic acid without fungus detection. Gas production, pH, and ammonia-nitrogen during ruminal in vitro incubation did not differ throughout incubation. However, ruminal total VFA concentration was higher (p<0.05) in the LAB spp. treatments than the control treatment at 48 hours. Overall, the use of L. brevis as an inoculant for fermentation of high moisture. TMR could inhibit fungi growth and promote lactic fermentation, and enhance digestion in the rumen.

Investigation of Dietary Lysophospholipid (LipidolTM) to Improve Nutrients Availability of Diet with In Vitro Rumen Microbial Fermentation Test

  • Cho, Sangbuem;Kim, Dong Hyun;Hwang, Il Hwan;Choi, Nag-Jin
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.3
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    • pp.206-212
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    • 2013
  • This study was conducted to investigate the effect of biological membrane transfer modifier, lysophospholipd (LPLs) on the parameters from in vitro rumen simulated fermentation. Commercially available LPLs product (Lipidol$^{TM}$) was supplemented into experimental diets which consisted of orchard grass and concentrate diet (60:40) in different levels (0.1%, 0.3% and 0.5%). Then in vitro rumen simulated fermentation was performed. Although, a declining trend of pH was found in treatments, all pH values were detected in a range relevant to normal rumen fermentation. Gas production, ammonia nitrogen and total VFA production were greatly influenced by the supplementation of LPLs. All parameters were increased along with increased levels of LPLs in diet. As a result, 0.1% of Lipidol$^{TM}$ is recommended based on the determined in vitro rumen fermentative parameters in this study.

Effects of Fermented Products by Formitella fraxinea and Sarcodon aspratus on In Vitro Ruminal Fermentation (In vitro 반추위 발효에 미치는 Formitella fraxinea와 Sarcodon aspratus 발효물질의 영향)

  • Kim, Yong-Kook
    • Korean Journal of Agricultural Science
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    • v.31 no.1
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    • pp.15-25
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    • 2004
  • In order to determine the effect of fermentation by the mycelia of fungal species, Formitella fraxinea and Sarcodon aspratus, on the in vitro dry matter digestibility and pH of mixtures with sawdust plus 20% wheat bran w/w, on dry matter basis to use as a feedstuff or an additive including fungal mycelium into a feedstuff. The mixtures were unfermented (UF) and fermented by Formitella fraxinea(FF) and Sarcodon aspratus(SA) for two weeks at $29^{\circ}C$ in a incubator. Fungal fermentation products were added to the basal diet to the level of 0, 1, 3 and 5%, w/w of diets each. The in vitro dry matter digestibilities, soluble sugar contents and pH of fermentation fluids were measured at 24, 48 and 72hr after fermentation begin. Neutral detergent fiber(NDF) contents in mixtures were lower for SA and UF(80.4 and 82.2%) than for FF(88.3%) (P<0.05). In vitro DM digestibility for 48h was higer for SA(21.2%) than for UF and FF(17.9 and 12.2%). The in vitro dry matter digestibility for 24hr was higher for diets added with FF 1% as 49.18%, and lower for diet added with FF 5%(43.07%) than basal diet(44.98%)(P<0.05), and tended to be higher for the diets added with fungal products. The pH of in vitro fermentation fluids for 24 and 48 hrs fermentation were lower for diets added with all FF and SA than for UF(P<0.05). However, those for 72 hrs fermentation were higher for SA 1%(6.74) than other diets(P<0.05). The soluble sugar concentration of in vitro fermentation fluid was not different between diets for 24 hr fermentation. However, those were higher for all additive diets than basal diet for 48 and 72 hrs fermentation(P<0.05). It could be concluded that dairy cow's diets added with fungal fermentation products have positive effects, and expected it will be more beneficial if more fungal mycelium was contained.

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Effect of Herbal Extracts Supplementation on Ruminal Methane Production and Fermentation Characteristics In vitro (한약재 추출물 첨가가 in vitro 반추위 발효 시 메탄생성 및 발효성상에 미치는 영향)

  • Lee, Shin-Ja;Lee, Sung-Sill;Moon, Yea-Hwang
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1315-1322
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    • 2011
  • This study was conducted to investigate the effects of several herbal extracts (obtusifolia, cinnamon, chinese pepper, licorice root) on the characteristics of rumen fermentation in vitro. Soybean meal was used as a substrate for fermentation in vitro. Herbal extracts were supplemented to media by 10% of the substrate. The substrates supplemented to Dehority artificial media with herbal extracts were fermented in 30ml serum bottles for 0, 3, 6, 9, 12 and 24 hr at $39^{\circ}C$. Cumulative gas production was significantly (p<0.05) greater in the herbal extract supplements than in the control, in the order of licorice root, chinese pepper, cinnamon and obtusifolia. Methane proportions of the herbal extracts were significantly (p<0.05) higher than that of the control. Licorice root extract supplementation resulted in the lowest methane proportion at 3 hr fermentation. Proportion of hydrogen was significantly (p<0.05) higher in the herbal extract supplements than in the control at 12 hr fermentation. Compared to the control, ammonia concentration in the licorice root was significantly higher at 3 hr fermentation, but lower at 12 hr fermentation (p<0.05). Based on these results, supplementation of the herbal extracts used in this experiment resulted in increased cumulative gas production and stimulating methane production in vitro rumen fermentation.

Effects of Rumen Protozoa of Brahman Heifers and Nitrate on Fermentation and In vitro Methane Production

  • Nguyen, S.H.;Li, L.;Hegarty, R.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.6
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    • pp.807-813
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    • 2016
  • Two experiments were conducted assessing the effects of presence or absence of rumen protozoa and dietary nitrate addition on rumen fermentation characteristics and in vitro methane production in Brahman heifers. The first experiment assessed changes in rumen fermentation pattern and in vitro methane production post-refaunation and the second experiment investigated whether addition of nitrate to the incubation would give rise to methane mitigation additional to that contributed by defaunation. Ten Brahman heifers were progressively adapted to a diet containing 4.5% coconut oil distillate for 18 d and then all heifers were defaunated using sodium 1-(2-sulfonatooxyethoxy) dodecane (Empicol). After 15 d, the heifers were given a second dose of Empicol. Fifteen days after the second dosing, all heifers were allocated to defaunated or refaunated groups by stratified randomisation, and the experiment commenced (d 0). On d 0, an oral dose of rumen fluid collected from unrelated faunated cattle was used to inoculate 5 heifers and form a refaunated group so that the effects of re-establishment of protozoa on fermentation characteristics could be investigated. Samples of rumen fluid collected from each animal using oesophageal intubation before feeding on d 0, 7, 14, and 21 were incubated for in vitro methane production. On d 35, 2% nitrate (as $NaNO_3$) was included in in vitro incubations to test for additivity of nitrate and absence of protozoa effects on fermentation and methane production. It was concluded that increasing protozoal numbers were associated with increased methane production in refaunated heifers 7, 14, and 21 d after refaunation. Methane production rate was significantly higher from refaunated heifers than from defaunated heifers 35 d after refaunation. Concentration and proportions of major volatile fatty acids, however, were not affected by protozoal treatments. There is scope for further reducing methane output through combining defaunation and dietary nitrate as the addition of nitrate in the defaunated heifers resulted in 86% reduction in methane production in vitro.