• Title/Summary/Keyword: In-vitro Fermentation

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Comparison of in vitro ruminal fermentation between different originated corn grains

  • Kim, Do Hyung;Park, Ha Young;Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.541-548
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    • 2017
  • This study was conducted to compare in vitro rumen fermentation characteristics among corn grains imported from America, Brazil, Argentina and Ukraine A and Ukraine B. Two Holstein steers, each surgically fitted with a ruminal cannula, consuming total mixed ration were used as rumen fluid donors. In vitro rumen fermentation experiments were performed in a completely random design which included a control (no corn) and treatments with 3.0 g of corn from different geographical origins, i.e., America, Brazil, Argentina, and Ukraine A and Ukraine B, respectively. Ruminal pH, ammonia-N, volatile fatty acid (VFA) and total gas production were measured at 0, 1, 3, 6, 12, 24 and 48 h post-incubation, respectively. No differences (p > 0.05) in mean ruminal pH appeared among the treated groups, however, ruminal pH patterns differed; i.e. corn treated groups had dramatically lower pH compared with control during the entire incubation period. Similarly, no different patterns between the groups in ammonia-N (p > 0.05) appeared until 6 h post-incubation. Unexpectedly, higher ammonia-N concentration for control than that for the corn treated groups appeared after 12 h post-incubation despite that for all groups increased. Total VFA was similar between the groups until 6 h post-incubation, but VFA after 12 h post-incubation was different (p < 0.05), i.e. VFA for corn from Argentina, Ukraine A, Ukraine B, and Brazil were comparatively higher than for America. Overall, data in this study showed that the corns of different origins may have different feed values to ruminants despite having similar chemical compositions.

Effect of Lactic Acid Bacteria Treatment on Nutritive Value and In Vitro Ruminal Fermentation of Italian Ryegrass (Lolium multiflorum L.) Silage

  • Lee, Kihwan;Marbun, Tabita Dameria;Kim, Suyeon;Song, Jaeyong;Kwon, Chan Ho;Yoon, Duhak;Kang, Jungsun;Lee, Chanho;Cho, Sangbuem;Kim, Eun Joong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.182-189
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    • 2020
  • This study was conducted to evaluate the effect of lactic acid bacteria (LAB) inoculation to domestically-cultivated Italian ryegrass (IRG) on silage fermentation and in vitro ruminal fermentation. There were six treatments based on the LAB inoculants: 1) no addition of LAB (negative control: NC), additions of 2) commercially-available LAB (positive control: PC), 3) Lactobacillus plantarum (LPL), 4) L. paracasei (LPA), 5) L. acidophilus (LA), and 6) L. pentosus (LPT). All treatments were inoculated at a concentration of 106 CFU/g and ensiled for 3, 7, 21, and 42 days in triplicate and analyzed for nutritive values when ensiling was terminated. Day 42 silage from all treatments were also examined for in vitro ruminal fermentation. After 42 days, LAB-inoculated silages had higher (P<0.05) lactic acid concentration compared to the NC. In terms of nutritive values, the silages treated with LPA, LA, and LPT showed higher (P<0.05) crude protein and lower (P<0.05) neutral detergent fiber and acid detergent fiber content compared to the rest of the treatment. In vitro ruminal dry matter degradability was not affected by LAB addition. However, LAB-treated IRG had shown higher (P<0.05) ammonia-N compared with that of the NC. LPA had shown the highest (P<0.05) volatile fatty acid concentration among the LAB examined. In conclusion, the addition of a single strain of LAB appeared to produce a quality IRG silage compared with the NC and the PC. Among the strains examined, LPA seemed to be superior to the others.

Effect of flaking on the digestibility of corn in ruminants

  • Kang, Hamin;Lee, Mingyung;Jeon, Seoyoung;Lee, Sang Moon;Lee, Ju Hwan;Seo, Seongwon
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1018-1033
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    • 2021
  • In this study, we aimed to assess the effect of flaking on the nutrient digestibility of corn grain in ruminants. In this regard, in vitro rumen fermentation, in situ rumen degradability, and in vivo metabolic experiments were performed. The automated gas production technique was used for the in vitro fermentation experiments. Six types of corn flakes with various degrees of gelatinization (32%, 41%, 48%, 66%, 86%, and 89%) were ground and incubated in rumen fluid to measure rumen fermentation characteristics and digestion rate. The in situ degradability of ground corn, whole corn, and corn flakes with 62% and 66% gelatinization was measured by incubation in the rumen of two cannulated Holstein cows. In vivo metabolic experiments were performed using 12 crossbred goats (29.8 ± 4.37 kg) using a 3 × 3 Latin square design. The dietary treatments consisted of ground corn and flaked corn with 48% or 62% gelatinization. In vitro experiments showed that as the degree of gelatinization increased, the digestion rate increased linearly, while the discrete lag time decreased linearly (p < 0.05). The effective rumen dry matter degradability, determined by in situ fermentation, was 37%p lower in corn flakes than ground corn, assuming a passage rate of 6%/h (p < 0.01), and there was no difference between the two flakes. In the in vivo experiment, there was no difference in dry matter intake, average daily gain, feed efficiency, and nitrogen utilization among the treatment groups (p > 0.05); however, the crude fat digestibility was lower for corn flakes than for ground corn (p < 0.05). To summarize, the rate of fermentation of corn flakes increased as the degree of gelatinization increased. However, non-ground corn flakes had lower rumen digestibility and did not improve in vivo apparent nutrient digestibility, compared with ground corn. In contrast to the assumption that flaked corn provides more energy to ruminant animals than ground corn, we conclude that the digestibility and energy value of corn flakes are lower than those of ground corn if mastication does not sufficiently reduce the particle size of corn flakes.

Effect of Prebiotics on Intestinal Microflora and Fermentation Products in Pig In Vitro Model

  • Kim, Dong-Woon;Chae, Su-Jin;Cho, Sung-Back;Hwang, Ok-Hwa;Lee, Hyun-Jeong;Chung, Wan-Tae;Park, Jun-Cheal;Kim, In-Cheul;Kim, In-Ho
    • Journal of Animal Science and Technology
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    • v.52 no.3
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    • pp.199-204
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    • 2010
  • The objective of this study was to evaluate the effect of the different types and levels of prebiotics on intestinal microflora and fermentation products in the in vitro fermentation model. The prebiotcs used in this study were IMO (iso-malto oligosaccharide), CI (partially digested chicory-inulin), RA (raffinose) and CD (cyclodextrin). Experimental diet for growing pigs was predigested by digestive enzymes and this hydrolyzed diet was mixed with buffer solution containing 5% fresh swine feces. Then, the mixture was fermented with or without prebiotics at the concentrations of 0.5 and 1.0% for 24 h. Samples were taken at 24 h, and viable count of micoflora, gas, pH, volatile organic compounds and short-chain fatty acids were determined. The viable count of Enterobacteriaceae was significantly decreased (p<0.001) in all treatments added with prebiotics in comparison to control without prebiotics. However, the increase of lactic acid bacteria was observed in the prebiotics treatment. Gas production increased as the level of prebiotics increased. The pH values in the fermentation fluid decreased in a dose-dependent manner with increasing the concentration of prebiotics. The fermentation with prebiotics resulted in the reduction of malodorous compounds such as ammonia, hydrogen sulfide, indole and skatole. The increase in short-chain fatty acid (SCFA) production was observed in the treatments with prebiotics. In conclusion, the results of this study demonstrated that the fermentation with prebiotics was effective in reducing the formation of malodorous compounds and increasing lactic acid bacteria and SCFA. These effects depended on the concentration of prebiotics. Moreover, further study is needed to determine whether the in vitro efficacy on the reduction of malodorous compounds and increase of SCFA would also be observed in animals.

Fermentation quality and in vitro methane production of sorghum silage prepared with cellulase and lactic acid bacteria

  • Khota, Waroon;Pholsen, Suradej;Higgs, David;Cai, Yimin
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1568-1574
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    • 2017
  • Objective: The effects of lactic acid bacteria (LAB) and cellulase enzyme on fermentation quality, microorganism population, chemical composition and in vitro gas production of sorghum silages were studied. Methods: Commercial inoculant Lactobacillus plantarum Chikuso 1 (CH), local selected strain Lactobacillus casei (L. casei) TH 14 and Acremonium cellulase (AC) were used as additives in sorghum silage preparation. Results: Prior to ensiling Sorghum contained $10^4LAB$ and $10^6cfu/g$ fresh matter coliform bacteria. The chemical compositions of sorghum was 26.6% dry matter (DM), 5.2% crude protein (CP), and 69.7% DM for neutral detergent fiber. At 30 days of fermentation after ensiling, the LAB counts increased to a dominant population; the coliform bacteria and molds decreased to below detectable level. All sorghum silages were good quality with a low pH (<3.5) and high lactic acid content (>66.9 g/kg DM). When silage was inoculated with TH14, the pH value was significantly (p<0.05) lower and the CP content significantly (p<0.05) higher compared to control, CH and AC-treatments. The ratio of in vitro methane production to total gas production and DM in TH 14 and TH 14+AC treatments were significantly (p<0.05) reduced compared with other treatments while in vitro dry matter digestibility and gas production did not differ among treatments. Conclusion: The results confirmed that L. casei TH14 could improve sorghum silage fermentation, inhibit protein degradation and decrease methane production.

Lactic acid bacteria strains selected from fermented total mixed rations improve ensiling and in vitro rumen fermentation characteristics of corn stover silage

  • Huang, Kailang;Chen, Hongwei;Liu, Yalu;Hong, Qihua;Yang, Bin;Wang, Jiakun
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1379-1389
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    • 2022
  • Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×106 cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

Rumen Microbial Population in the In vitro Fermentation of Different Ratios of Forage and Concentrate in the Presence of Whole Lerak (Sapindus rarak) Fruit Extract

  • Suharti, Sri;Astuti, Dewi Apri;Wina, Elizabeth;Toharmat, Toto
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.8
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    • pp.1086-1091
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    • 2011
  • This experiment was designed to investigate the effect of lerak extract on the dynamic of rumen microbes in the in vitro fermentation of diet with different ratios of forage and concentrate. In vitro fermentation was conducted according to the method of Tilley and Terry (1963). The design of experiment was a factorial block design with 2 factors. The first factor was the ratio of forage and concentrate (90:10, 80:20, and 70:30 w/w) and the second factor was the level of lerak extract (0, 0.6, and 0.8 mg/ml). Total volatile fatty acid (VFA) concentration, proportional VFA and NH3 concentration were measured at 4 h incubation. Protozoal numbers in the buffered rumen fluid after 4 and 24 h of incubation were counted under a microscope. Bacterial DNAs of buffered rumen fluid were isolated from incubated samples after 24 h of incubation using a QiaAmp kit. Total bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Prevotella ruminicola were quantified using real time polymerase chain reaction (PCR). Lerak extract markedly reduced protozoal numbers in buffered rumen fluid of all diets after 24 h of incubation. Total bacteria did not change with lerak extract addition. While no difference in F. succinogenes was found, there was a slight increase in R. albus number and a significant enhancement in P. ruminicola number by increasing the level of lerak extract in all diets. Propionate concentration significantly increased in the presence of lerak extract at level 0.8 mg/ml. It was concluded that the addition of lerak extract could modify rumen fermentation and had positive effects on rumen microbes.

Effect of Temperature, pH and Salt Concnetration on formation of N-nitrosamines during Kimchi Fermentation (김치숙성중 숙성온도, pH 및 소금 농도가 니트로스아민의 생성에 미치는 영향)

  • 김준환;장영상;신효선
    • Journal of Food Hygiene and Safety
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    • v.13 no.4
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    • pp.332-336
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    • 1998
  • The effect of ripening temperature, pH and salinity on the formation of Nnitrosamine (NA) during Kimchi fermentation and in vitro was studied, respectively. During Kimchi fermentation for six weeks at cold storage temperature ($4^{\circ}C$) and room temperature ($16{\pm}2^{\circ}C$), the contents of nitrite and dimethylamine (DMA) showed variation at room temperature but no variation at cold storage temperature. The maximum generation of nitrosodimethylamine (NDMA) resulted low content ($2.69\;\mu\textrm{g}/kg$) at cold storage temperature but started to increase after one week fermentation and reached to the 18-fold higher generation ($49.6\;\mu\textrm{g}/kg$) at room temperature. During Kimchi fermentation, no correlation was observed between the variation of nitrite and DMA content and the generation of NDMA. However, pH showed effective relation to NDMA generation such as the highest NDMA generation was obtained at lowest pH 4. During in vitro test, higher temperature and lower pH resulted more NDMA generation and generation amount was affected more by pH. Also, the salinity of Kimchi provided inhibitory effects on the formation of NDMA. NDMA was produced $5.86\;\mu\textrm{g}/kg$ at normal salinity (2.5%) but $90.9\;\mu\textrm{g}/kg$ at lower salinity (15%) after three week. The higher salinity showed lower formation of NDMA in vitro test, too.

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Effects of Non-ionic or Zwitterionic Surfactant on in vitro Digestibility of Rice Straw and Growth of Rumen Mixed Microorganisms. (비이온성 및 양쪽 이온성 계면활성제 첨가가 반추위 혼합 미생물의 성장과 볏짚의 in vitro 소화에 미치는 영향)

  • Lee, Shin-Ja;Kim, Wan-Young;Moon, Yea-Hwang;Kim, Hyeon-Shup;Kim, Kyoung-Hoon;Ha, Jong-Kyu;Lee, Sung-Sill
    • Journal of Life Science
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    • v.18 no.4
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    • pp.515-521
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    • 2008
  • This experiment was conducted to investigate effects of non-ionic or zwitterionic (+/-) surfactants on digestibility of rice straw, and changes of growth of rumen mixed microbes, pH, and gas production during in vitro fermentation. Also, during in vitro ruminal fermentation, microbial attachment on rice straw was investigated using scanning electron microscopy (SEM). Tween 80 or SOLFA-850 for non-ionic surfactant (NIS), and 3-(Dodecyldimethylammonio) propanesulfanate (DDAP) for zwitterionic surfactant (ZIS) was supplemented by 0.05% and 0.1% in Dehority's artificial medium containing Holtein rumen fluid, respectively, and the substrate for fermentation was rice straw passed through 1 mm screen. The experiment was composed of 7 treatments (two levels of two NISs, two levels of a ZIS) including the control, and 6, 12, 24, 48 and 72 hr of fermentation time with 3 replications per treatment. Treatment of Tween 80 increased in vitro DM digestibilities during 48 hr and 72 h post fermentations compared to the other treatments, whereas treatment of DDAP as a ZIS resulted in decreased DM digestibility than that of the control from 24 hr post fermentation (P<0.05). Gas production in vitro was greater (P<0.05) with addition of NIS than the control or ZIS, and increased as fermentation time elapsed. Rumen mixed microbial growth was greatest with addition of Tween 80 as NIS, and lowest when DDAP as ZIS was supplemented to the fermentation tube (P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was greater with addition of NIS, but was less with addition ZIS compared with the control. In conclusion we could not found any positive effects of ZIS surfactants on rumunal fermentation characteristics and rumen microbial growth rates.

Fermentation Properties and In vitro Anticancer Effect of Kimchi Prepared with Potato (감자를 첨가한 김치의 발효 특성 및 항암효과)

  • Chang, Sang-Keun
    • Korean journal of food and cookery science
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    • v.23 no.2 s.98
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    • pp.227-234
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    • 2007
  • Potato kimchi, fermentation was carried out at $10^{\circ}$C for 15 days using various ratios of potato to kimchi (2.5%, 5%, 10%). The samples were determined according to the fermentation time, pH, acidity and growth of lactic acid bacteria in potato kimchi. The addition ratio of potato to kimchi had little effect on the pH, acidity or growth of lactic acid bacteria in potato kimchi. In comparison to baechu kimchi and mul-kimchi, the pH, acidity and growth of lactic acid bacteria was better in potato kimchi than in the other kimchi samples. The in vitro anticancer effect of potato kimchi was investigated using human cancer cells, AGS human gastric adenocarcinoma cells and HT-29 human colon adenocarcinoma cells. MTT assay revealed that the methanol extract of potato kimchi showed the highest anticarcinogenic effects.