• Title/Summary/Keyword: In vitro fermentation

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Effect of Encapsulating Nitrate in Sesame Gum on In vitro Rumen Fermentation Parameters

  • Mamvura, Chiedza Isabel;Cho, Sangbuem;Mbiriri, David Tinotenda;Lee, Hong-Gu;Choi, Nag-Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.11
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    • pp.1577-1583
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    • 2014
  • Encapsulation is a method used to protect material from certain undesirable environments, for controlled release at a more favorable time and place. Animal productivity would be enhanced if feed additives are delivered to be utilized at their site of action, bypassing the rumen where they are likely to be degraded by microbial action. A novel method of encapsulation with sesame gum was used to coat nitrate, a known enteric methane mitigating agent, and tested for the effect on methane reduction and other in vitro fermentation parameters using rumen fluid from cannulated Hanwoo steers. Orchard grass was used as basal diet for fermentation. The treatments were matrix (1.1 g sesame gum+0.4 g sesame oil cake) only, encapsulated nitrate (matrix+nitrate [21 mM]), free nitrate (21 mM), and a control that contained no additive. Analyses of fermentation parameters were done at 0, 3, 6, 9, 12, 24, and 48 h time periods. In comparison to control, both free and encapsulated nitrate produced significantly reduced (p<0.01) methane (76% less) and also the total volatile fatty acids were reduced. A significantly higher (p<0.01) concentration of ammonia nitrogen was obtained with the encapsulated nitrate treatment (44%) compared to the free form (28%) and matrix only (20%) (p = 0.014). This might suggest slow release of encapsulated nitrate so that it is fully reduced to ammonia. Thus, this pioneering study found a significant reduction in methane production following the use of sesame gum encapsulated nitrate that shows the potential of a controlled release system in enhancing sustainability of ruminant production while reducing/eliminating the risk of nitrite toxicity.

Silage Fermentative Quality and Characteristics of Anthocyanin Stability in Anthocyanin-rich Corn (Zea mays L.)

  • Hosoda, Kenji;Eruden, Bayaru;Matsuyama, Hiroki;Shioya, Shigeru
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.528-533
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    • 2009
  • The fermentative quality and quantitative change in anthocyanin of anthocyanin-rich corn (Zea mays L.) during storage and in vitro ruminal fermentation were studied. The anthocyanin-rich corn silages in bag silo, drum silo and round bale had good fermentative qualities, such as low pH (5% DM) and butyric acid-free, and its quality was maintained for more than 370 d. The amount of anthocyanin in the anthocyanin-rich corn decreased after ensiling by about 45% (from 3.34 to 1.88 mg/g DM), but stayed constant after day 60. The in vitro incubation of the anthocyanin-rich corn with ruminal fluid revealed little degradation of anthocyanin. These results indicate that the anthocyanin had no negative effect on silage fermentation, and the anthocyanin-rich corn silage is utilizable for practical use as a feedstuff. Our results also demonstrate alteration of the anthocyanin content during storage, and show that anthocyanin-rich corn is a suitable antioxidant source for ruminants because of the high stability of the anthocyanin in ruminal fluid.

In vitro fermentation profiles of different soybean oligosaccharides and their effects on skatole production and cecal microbiota of broilers

  • Zhu, Xin;Xu, Miao;Liu, Haiying;Yang, Guiqin
    • Animal Bioscience
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    • v.35 no.8
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    • pp.1195-1204
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    • 2022
  • Objective: The objective of this study was to investigate the in vitro fermentation profiles of different soybean oligosaccharides (SBOs) and their effects on skatole production and cecal microbiota of broilers. Methods: Five SBOs with varying main component contents were fermented using an in vitro batch incubation inoculated with broiler cecal microbiota. Gas production was recorded automatically, skatole, indole and short-chain fatty acids (SCFAs) were determined using high-performance liquid chromatography, and microbial changes were analyzed using 16S DNA gene sequencing. Results: The addition of SBOs increased (p<0.05) gas production, suggesting bacterial growth-stimulating activities. In addition, the concentrations of indole were significantly (p<0.05) decreased after SBO supplementation, and SBO III, with higher sucrose and stachyose contents, decreased (p<0.05) the skatole level. Our results also revealed that the fermentation of SBOs by cecal microbiota produced (p<0.05) SCFAs, which were dominated by propionic acid, butyrate acid and lactic acid compared to the control. In addition, SBO III increased (p<0.05) the abundance of Firmicutes and Subdoligranulum and decreased that of Bacteroides. Conclusion: These results suggest that SBOs with higher sucrose and stachyose contents are promising prebiotics in modulating gut microbiota and reducing odor emission in broilers.

Dietary lysophospholipids supplementation inhibited the activity of lipolytic bacteria in forage with high oil diet: an in vitro study

  • Kim, Hanbeen;Kim, Byeongwoo;Cho, Seongkeun;Kwon, Inhyuk;Seo, Jakyeom
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1590-1598
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    • 2020
  • Objective: The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods: Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH3-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results: There was no significant change in IVDMD, pH, NH3-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and <0.001, respectively). The LPL supplementation tended to increase the total bacteria in a linear manner (p = 0.089). There were significant decreases in the relative proportions of cellulolytic (Fibrobacter succinogenes and Ruminococcus albus) and lipolytic (Anaerovibrio lipolytica and Butyrivibrio proteoclasticus) bacteria with increasing levels of LPL supplementation (Lin, p = 0.028, 0.006, 0.003, and 0.003, respectively). Conclusion: The LPL supplementation had antimicrobial effects on several cellulolytic and lipolytic bacteria, with no significant difference in nutrient degradability (DM and neutral detergent fiber) and general bacterial counts, suggesting that LPL supplementation might increase the enzymatic activity of rumen bacteria. Therefore, LPL supplementation may be more effective as an antimicrobial agent rather than as an emulsifier in the rumen.

Roughage Energy and Degradability Estimation with Aspergillus oryzae Inclusion Using Daisy In vitro Fermentation

  • Chen, C.R.;Yu, B.;Chiou, P.W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.53-62
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    • 2004
  • The aim of this study was to predict the energy value and dynamic degradation of roughage in Taiwan using the $Daisy^{(R)}$. in vitro fermentation method to provide information on one of the very important nutrients for ration formulation. The second objective was to study the effects of Aspergillus oryzae (AFE) inclusion on nutrient utilization. Three ruminal fistulated dry dairy cows were used for rumen fluid and fifteen conventional forages used in dairy cattle were collected around this island. The degradability of these feedstuffs with and without AFE ($Amaferm^{(R)}$.) treatment was measured using the $Daisy^{(R)}$. in vitro method. The roughage energy values, including TDN and NEL, were calculated according to Robinson (2000). Results from the 30 h in vitro neutral detergent fiber (NDF) degradability and predicted energy evaluations showed that alfalfa (among the forages) contained the highest degradability and energy values, Bermuda straw having the lowest. Peanut vines and corn silage contained higher energy values and the lowest value found in Pangola and Napier grasses among the locally produced forages. Pangola and Napier grasses had lower values than most imported forages except Bermuda straw. Among the by-products, wheat middling contained the highest NDF degradability, while rice bran contained the richest energy value due to its high oil content. From the dynamic dry matter (DM), organic matter (OM), acid detergent fiber (ADF) and neutral detergent fiber (NDF) degradation, corn silage contained the highest effective degradation among the local forages; wheat middling (among the by-products) degraded the fastest in DM, OM, ADF and NDF and showed the highest effective degradability. AFE inclusion was inconsistent among the forages. Alfalfa hay showed significantly increased 30 h NDF degradability and energy values, Pangola hay, Napier grass and brewer's grains showed decreased degradability and energy values. AFE inclusion increased the DM, OM and NDF degradation rate in most forage, but only increased the DM degradation rate in sorghum distiller's grains, the OM degradation rate in bean curd pomace and the NDF and ADF degradation rates in soy pomace (among the by-products).

Rumen fermentation, methane production, and microbial composition following in vitro evaluation of red ginseng byproduct as a protein source

  • Hamid, Muhammad Mahboob Ali;Moon, Joonbeom;Yoo, Daekyum;Kim, Hanbeen;Lee, Yoo Kyung;Song, Jaeyong;Seo, Jakyeom
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.801-811
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    • 2020
  • The main objective of this in vitro study was to evaluate red ginseng byproduct (RGP) as a protein resource and its effects on rumen fermentation characteristics, microflora, CO2, and CH4 production in ruminants. Four treatments for in vitro fermentation using buffered rumen fluid over a 48 h incubation period were used: 1, RGP; 2, corn gluten feed (CGF); 3, wheat gluten (WG); and 4, corn germ meal. In vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), in vitro crude protein digestibility (IVCPD), volatile fatty acids, pH, and ammonia nitrogen (NH3-N) were estimated after 48 h incubation. Gas production was investigated after 3, 6, 12, 24, 36 and 48 h. The CO2 and CH4 were evaluated after 12, 24, 36, and 48 h. A significant difference in total gas production and CO2 emissions was observed (p < 0.01) at all incubation times. CH4 production in RGP were higher (p < 0.05) than that in other treatments but a higher CH4 portion in the total gas production was observed in WG (p < 0.05) at 48 h incubation. The IVDMD, IVNDFD, and IVCPD of RGP was lower than those of other conventional ingredients (p < 0.01). The RGP had the lowest NH3-N value among the treatments (p < 0.01). The RGP also had the lowest total VFA concentration (p < 0.01), but presented the highest acetate proportion and acetate to propionate ratio among the treatments (both, p < 0.01). The abundance of Prevotella ruminicola was higher in RGP than in WG (p < 0.01), whereas RGP has lower methanogenic archaea (p < 0.01). In conclusion, based on the nutritive value, IVDMD, low NH3-N, and decreased methanogenic archaea, RGP inclusion as a protein source in ruminant diets can be an option in replacing conventional feed sources.

Effects of Coconut Materials on In vitro Ruminal Methanogenesis and Fermentation Characteristics

  • Kim, E.T.;Park, C.G.;Lim, D.H.;Kwon, E.G.;Ki, K.S.;Kim, S.B.;Moon, Y.H.;Shin, N.H.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.12
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    • pp.1721-1725
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    • 2014
  • The objective of this study was to evaluate the in vitro effects of coconut materials on ruminal methanogenesis and fermentation characteristics, in particular their effectiveness for mitigating ruminal methanogenesis. Fistulated Holstein cows were used as the donor of rumen fluid. Coconut materials were added to an in vitro fermentation incubated with rumen fluid-buffer mixture and timothy substrate for 24 h incubation. Total gas production, gas profiles, total volatile fatty acids (tVFAs) and the ruminal methanogens diversity were measured. Although gas profiles in added coconut oil and coconut powder were not significantly different, in vitro ruminal methane production was decreased with the level of reduction between 15% and 19% as compared to control, respectively. Coconut oil and coconut powder also inhibited gas production. The tVFAs concentration was increased by coconut materials, but was not affected significantly as compared to control. Acetate concentration was significantly lower (p<0.05), while propionate was significantly higher (p<0.05) by addition of the coconut materials than that of the control. The acetate:propionate ratio was significantly lowered with addition of coconut oil and coconut powder (p<0.05). The methanogens and ciliate-associated methanogens in all added coconut materials were shown to decrease as compared with control. This study showed that ciliate-associated methanogens diversity was reduced by more than 50% in both coconut oil and coconut powder treatments. In conclusion, these results indicate that coconut powder is a potential agent for decreasing in vitro ruminal methane production and as effective as coconut oil.

Evaluation of nutritive value of chestnut hull for ruminant animals using in vitro rumen fermentation (밤 가공 부산물의 반추가축용 사료 가치 평가: in vitro 반추위 배양)

  • Jeong, Sin-Yong;Jo, Hyeon-Seon;Park, Gi-Su;Kang, Gil-Nam;Jo, Nam-Chul;Seo, Seongwon
    • Korean Journal of Agricultural Science
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    • v.39 no.3
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    • pp.335-340
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    • 2012
  • During the manufacturing process of chestnut, 50% of biomass is produced as chestnut shell (CS) or chestnut hull (CH), a forestry by-product. Due to its high fiber content and economic benefit, there is a possibility of using chestnut hull as a supplement for a ruminant diet. Few studies, however, have been conducted on evaluating nutritive value of chestnut hull for ruminant animals. The objective of this study were thus to analyze chemical composition of CS, a by-product after the first processing of chestnut, and CH, a by-product after the second processing, and access in vitro rumen fermentation characteristics of them. For the in vitro fermentation using strained rumen fluid obtained from a fistulated Hanwoo steer, commercial total mixed ration (TMR) for dairy goat was used as a basal diet and was replaced with different proportions of chestnut shell and hull. A total number of 13 treatments were carried out in this study: 100% TMR, 100% CS, 100% CH, a mix with 50% CS and 50% of CH (MIX), TMR replaced with 5%, 10%, or 15% of CS, CH, or MIX, respectively. For each treatment, in vitro dry matter digestibility (IVDMD) and pH after 48 hours of rumen fermentation were measured. Gas production at 6, 12, 24, 48 hours of incubation was also analyzed. Compared to CH, CS contains higher level of fiber (NDF, ADF, lignin) and consequently has a lower amount of non-fiber carbohydrate, but no difference was observed in the other nutrients (i.e. crude protein, crude fat, and ash). IVDMD was significantly (p<0.05) the highest in 100% CH (71.97%) and the lowest in 100% CS (42.80%). Addition of CH by replacing TMR did not affect IVDMD, while an increase in the proportion of CS tended to decrease IVDMD. The total gas production after 48 hours of incubation and the rate of gas production were also the highest in 100% CH and the lowest in 100% CS (P<0.05). Likewise, the pH after 48 hours of fermentation was significantly (p<0.05) the lowest in 100% CH (6.33) and the highest in 100% CS (6.50), and no significant difference in gas production was observed when TMR was replaced with CS or CH up to 15% (P>0.05). In conclusion, CH may successfully be used for a supplement in a ruminant diet. The nutritive value of CS is relative low, but can replace, if not 100%, low quality forage. This study provides valuable information about the nutritive value of CS and CH. An in vivo trials, however, is needed for conclusively accessing the nutritive value of CS and CH.

The Effects of Negative- and Positive- Charged Surfactants on In vitro DM Digestibility and the Growth of Ruminal Mixed Microorganisms (양(+) 이온성 및 음(-) 이온성 계면활성제 첨가가 반추위 혼합 미생물에 의한 In vitro 건물소화율 및 미생물 성장에 미치는 영향)

  • Lee, S.J.;Shin, N.H.;Kim, W.Y.;Moon, Y.H.;Kim, H.S.;Ha, J.K.;Lee, S.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.647-656
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    • 2007
  • In order to investigate the effects of supplemental ionic surfactants in in vitro ruminal fermentation, N-Lauroylsarcosine sodium salt(N-LSS) and sodium dodecyl sulfate(SDS) for negative(-) ionic surfactant, and hexadecylpyridinium chloride monohydrate(HPCM) and hexadecyltrimethyl ammonium bromide(HTAB) for positive (+) ionic surfactant were supplemented by 0.05% and 0.1% into the Dehority’s artificial medium containing rice straw(1mm) as a substrate. In vitro DM digestibility, the growth of rumen mixed microbes, pH, cumulative gas production and SEM(Scanning Electron Microscopy) observation of microbial attachment on rice straw particle were investigated through the experiment composing 9 treatments (two supplemental levels of two positive ionic(+) surfactant, two supplemental levels of two negative(-) ionic surfactant) including the control. The sample collection was at 6, 12, 24, 48 and 72 h post fermentation with 3 replications per treatments. DM digestibility in treatments supplemented (+) or (-) surfactants almost stopped afterward 12 h fermentation, in vitro DM digestibility at 72 h post fermentation in the ionic surfactants was at half level of that of the control(P<0.05). Accumulative gas production in in vitro was less(P<0.05) with addition of ionic surfactants compared to the control. The amount of rumen mixed microbes recovered from in vitro incubation fluid pleateaued at 12 h post fermentation for the positive (+) ionic surfactants, but steadily increased as fermentation time elapsed for the control. Rumen microbial growth rate was significantly(P<0.05) low in the negative(-) ionic surfactant compared to the control. pH of the incubation fluid was ranged from 6.02 to 7.20, and was the highest in the negative(-) ionic surfactants, and was the lowest in the control(P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was less with addition of ionic surfactants than the control. In conclusion we could not found any positive effects of negative- and positive- charged surfactants on rumunal fermentation characteristics and rumen microbial growth rates.

Effect of Total Mixed Ration with Fermented Feed on Ruminal In vitro Fermentation, Growth Performance and Blood Characteristics of Hanwoo Steers

  • Kim, S.H.;Alam, M.J.;Gu, M.J.;Park, K.W.;Jeon, C.O.;Ha, Jong-K.;Cho, K.K.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.2
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    • pp.213-223
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    • 2012
  • In this study, two experiments were conducted to evaluate the total mixed ration with fermented feed (TMRF) and total mixed ration (TMR) by rumen in vitro fermentation and their effects on the growth performance and blood characteristics of Hanwoo steers. In experiment 1, three Hanwoo steers ($600{\pm}47$ kg), each permanently fitted with a ruminal cannula were used. In this experiment, three diets designated as T1, TMRF (18.4% fermented feed, tall fescue, mammoth wild rye forage and whole crop barley); T2, TMRF (17.7% fermented feed, rice straw and whole crop barley); and T3, TMR (rice straw, whole crop barley and probiotics, but no fermented feed), which were subjected to rumen in vitro fermentation for 48 h. The results demonstrated that DM disappearance rate gradually increased with advancing fermentation time, but T1 and T2 were higher than the T3 (p<0.05) from 3 h to 12 h, but insignificant (p>0.05) at 24 and 48 h. None of the specific VFAs were affected except for acetic and non volatile lactic acids, which were produced more in T2 than in T1 and T3 at 24 h and 48 h of incubation. A/P was lower in T1 and T2 than inT3 at 24 h (p<0.05) and 48 h (p>0.05) of incubation. These results confirmed that TMRF-related treatment shows a superior performance to that of TMR during the ruminal fermentation period. In experiment 2, the three diets in experiment 1 plus 1 more control diet (concentrates, probiotics and 2% rice straw of body weight) were fed to the 48 Hanwoo steers ($160{\pm}10$ kg) for a period of 168 d. The results demonstrated that the daily and total live weight gain and feed efficiency were higher (p<0.05) in the TMRF and TMR groups than in the control group. SGOT, SGPT and BUN (p<0.05) were reduced in TMRF relative to the control and TMR groups by 168 d which confirmed that TMRF shows better blood profiles than the TMR and control groups. Overall, these results appear to show that TMRF has better in vitro ruminal characteristics than those of TMR; growth performance and blood profiles were also found to be superior in TMRF than in the TMR and control groups. Thus, our findings suggest that TMRF-based feed supplies are favorable for Hanwoo cattle.