• 제목/요약/키워드: In vitro gas production

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Fermentation of Environmental Friend Total Mixed Ration and Alteration of Rumen Fermentation Characteristics (환경친화적 섬유질 배합사료의 발효와 반추위 발효특성 변화)

  • Ryu, Chae-Hwa;Park, Myung-Sun;Park, Chul;Choi, Nag-Jin;Cho, Sang-Buem
    • Korean Journal of Organic Agriculture
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    • 제25권2호
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    • pp.461-473
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    • 2017
  • Total mixed ration (TMR) including concentrate diet and roughage together have been used for the ruminant animal. Relatively high concentrations of moisture and water soluble carbohydrate are representative feature of TMR. Those moisture and water can also provide a niche for bacterial growth. Therefore, a possible fermentation of TMR induced by micro-organism is generally accepted. The present study hypothesized that different lactic acid bacteria could alter fermentation of TMR and subsequently rumen fermentation. Three lactic acid bacteria, Lactobacillus paracasei (A), L. plantarum (B) and L. parabuchneri (C), were employed and 7 treatments under full factorial design were compared with control without inoculation. TMR for dairy cow was used. Significant alterations by treatments were detected at lactic acid and butyric acid contents in TMR (p<0.05). Treatment AC (mixture of A and C) and BC (mixture of B and C) showed great lactate production. Great butyrate production was found at treatment C. At in vitro rumen fermentation, treatments B, C and AB (mixture of A and B) showed significantly great total gas production (p<0.05). All treatments except treatments B and AB, showed less dry matter digestibility, significantly (p<0.05). Total volatile fatty acid production at treatment AC was significantly greater than others (p<0.05). In individual volatile fatty acid production, treatment AB and AC showed great acetate and propionate productions, significantly (p<0.05). This study investigated correlation between organic acid production in TMR and rumen volatile fatty acid production. And it was found that butyric acid in TMR had significant negative correlation with acetate, propionate, total volatile fatty acid, AP ratio and dry matter digestibility.

Cytokines Stimulate Lung Epithelial Cells to Release Nitric Oxide

  • Robbins, Richard A.;Kwon, O-Jung
    • Tuberculosis and Respiratory Diseases
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    • 제42권4호
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    • pp.447-454
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    • 1995
  • Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

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Broken rice in a fermented total mixed ration improves carcass and marbling quality in fattened beef cattle

  • Kotupan, Salisa;Sommart, Kritapon
    • Animal Bioscience
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    • 제34권8호
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    • pp.1331-1341
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    • 2021
  • Objective: This study aimed to determine the effects of replacing cassava chips with broken rice in a fermented total mixed ration diet on silage quality, feed intake, ruminal fermentation, growth performance, and carcass characteristics in the final phase of fattening beef cattle. Methods: Eighteen Charolais-Thai native crossbred steers (average initial body weight: 609.4±46 kg; average age 31.6 mo) were subjected to three ad libitum dietary regimes and were maintained in individual pens for 90 d before slaughter. The experimental design was a randomized complete block design by initial age and body weight with six replicates. The dietary regimens used different proportions of broken rice (0%, 16%, and 32% [w/w] of dry matter [DM]) instead of cassava chips in a fermented total mixed ration. All dietary treatments were evaluated for in vitro gas production and tested in in vivo feeding trials. Results: The in vitro experiments indicated that organic matter from broken rice was significantly more digestible than that from a cassava-based diet (p<0.05). Silage quality, nutrient intake, ruminal fermentation characteristics, carcass fat thickness, and marbling score substantially differed among treatments. The ruminal total volatile fatty acids, propionate concentration, dietary protein intake, and digestibility increased linearly (p<0.05) with broken rice, whereas acetate concentration and the acetate:propionate ratio decreased linearly (p<0.05) with broken rice (added up to 32 g/kg DM). Broken rice did not influence plasma metabolite levels or growth performance (p>0.05). However, the marbling score increased, and the carcass characteristics improved with broken rice. Conclusion: Substitution of cassava chips with broken rice in beef cattle diets may improve fattened beef carcass quality because broken rice increases rumen fermentation, fatty acid biosynthesis, and metabolic energy supply.

Effects of Momordica charantia Saponins on In vitro Ruminal Fermentation and Microbial Population

  • Kang, Jinhe;Zeng, Bo;Tang, Shaoxun;Wang, Min;Han, Xuefeng;Zhou, Chuanshe;Yan, Qiongxian;He, Zhixiong;Liu, Jinfu;Tan, Zhiliang
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권4호
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    • pp.500-508
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    • 2016
  • This study was conducted to investigate the effects of Momordica charantia saponin (MCS) on ruminal fermentation of maize stover and abundance of selected microbial populations in vitro. Five levels of MCS supplements (0, 0.01, 0.06, 0.30, 0.60 mg/mL) were tested. The pH, $NH_3-N$, and volatile fatty acid were measured at 6, 24, 48 h of in vitro mixed incubation fluids, whilst the selected microbial populations were determined at 6 and 24 h. The high dose of MCS increased the initial fractional rate of degradation at t-value = 0 ($FRD_0$) and the fractional rate of gas production (k), but decreased the theoretical maximum of gas production ($V_F$) and the half-life ($t_{0.5}$) compared with the control. The $NH_3-N$ concentration reached the lowest concentration with 0.01 mg MCS/mL at 6 h. The MSC inclusion increased (p<0.001) the molar proportion of butyrate, isovalerate at 24 h and 48 h, and the molar proportion of acetate at 24 h, but then decreased (p<0.05) them at 48 h. The molar proportion of valerate was increased (p<0.05) at 24 h. The acetate to propionate ratio (A/P; linear, p<0.01) was increased at 24 h, but reached the least value at the level of 0.30 mg/mL MCS. The MCS inclusion decreased (p<0.05) the molar proportion of propionate at 24 h and then increased it at 48 h. The concentration of total volatile fatty acid was decreased (p<0.001) at 24 h, but reached the greatest concentration at the level of 0.01 mg/mL and the least concentration at the level of 0.60 mg/mL. The relative abundance of Ruminococcus albus was increased at 6 h and 24 h, and the relative abundance of Fibrobacter succinogenes was the lowest (p<0.05) at 0.60 mg/mL at 6 h and 24 h. The relative abundance of Butyrivibrio fibrisolvens and fungus reached the greatest value (p<0.05) at low doses of MCS inclusion and the least value (p<0.05) at 0.60 mg/mL at 24 h. The present results demonstrates that a high level of MCS quickly inhibits in vitro fermentation of maize stover, while MCS at low doses has the ability to modulate the ruminal fermentation pattern by regulating the number of functional rumen microbes including cellulolytic bacteria and fungi populations, and may have potential as a feed additive applied in the diets of ruminants.

Can Moringa oleifera Be Used as a Protein Supplement for Ruminants?

  • Kakengi, A.M.V.;Shem, M.N.;Sarwatt, S.V.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권1호
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    • pp.42-47
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    • 2005
  • The possibility of using Moringa oleifera as a ruminant protein supplement was investigated by comparison between nutritive and anti-nutritive value of its different morphological parts with that of conventionally used Leucaena leucocephala leaf meal (LL). Parameters determined were chemical composition, rumen degradable protein (RDP), acid detergent insoluble protein (ADIP), pepsin soluble protein (PESP), non-protein nitrogen (NPN) total soluble protein (TSP) and protein potentially digested in the intestine (PDI). Total phenols (TP) and total extractable tannins (TET) were also evaluated as anti-nutritive factors. In vitro gas production characteristics were measured and organic matter digestibility (OMD) was estimated basing on 24 h-gas production. Crude protein content ranged from 265-308 g/kg DM in M. oleifera leaves (MOL) and seed cake (MOC) respectively. Leucaena leucocephala and Moringa oleifera soft twigs and leaves (MOLSTL) had CP content of 236 and 195 g/kg DM while Moringa oleifera soft twigs alone (MOST) and Moringa oleifera bucks (MOB) had 160, 114 and 69.3 g/kg DM respectively. RDP was highest in (MOC) (181 g/kg DM) followed by (MOL) (177 g/kg DM) and was lowest in MOB (40 g/kg DM). The proportion of the protein that was not available to the animal (ADIP) was (p<0.05) higher in MOL and MOC (72 and 73 g/kg DM) respectively and lowest in LL (29 g/kg DM). The PDI was high in LL (74 g/kg DM) followed by MOC (55 g/kg DM) then MOL (16 g/kg DM). PESP was highest (p<0.05) in MOC followed by MOL then LL (273, 200 and 163 g/kg DM respectively). MOC exhibited highest NPN content (116 g/kg DM) and was lowest in MOB (18 g/kg DM) (p<0.05). Highly (p<0.05) TSP was observed in MOC and MOL (308 and 265 g/kg DM respectively) followed by LL (236 g/kg DM). MOL had negligible TET (20 g/kg DM) when compared with about 70 g/kg DM in LL. Highly (p<0.05) b and a+b values were observed for MOLSTL (602 and 691 g/kg DM respectively) followed by MOL (490 and 538 g/kg DM). Highest c value was observed in MOSTL followed by MOC and MOL (0.064, 0.056 and 0.053 rate/hour) respectively. OMD was highest (p<0.05) for MOSTL followed by MOC and then MOL (579, 579 and 562 g/kg DM respectively). LL exhibited lower (p<0.05) OMD (467 g/kg DM). It was concluded from this study that the high crude protein content in MOL and MOLST could be well utilized by ruminant animals and increase animal performance however, high proportion of unavailable protein to the lower gut of animals and high rumen degradable protein due to negligible tannin content render it a relatively poor protein supplement for ruminants. MOC can be a best alternative protein supplement to leaves and leaves and soft twigs for ruminants.

Effects of Methylcellulose on Cellulolytic Bacteria Attachment and Rice Straw Degradation in the In vitro Rumen Fermentation

  • Sung, Ha Guyn;Kim, Min Ji;Upadhaya, Santi Devi;Ha, Jong K.;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • 제26권9호
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    • pp.1276-1281
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    • 2013
  • An in vitro experiment was conducted to evaluate the effect of methylcellulose on the attachment of major cellulolytic bacteria on rice straw and its digestibility. Rice straw was incubated with ruminal mixture with or without 0.1% methylcellulose (MC). The attachment of F. succinogenes, R. flavefaciens and R. albus populations on rice straw was measured using real-time PCR with specific primer sets. Methylcellulose at the level of 0.1% decreased the attachment of all three major cellulolytic bacteria. In particular, MC treatment reduced (p<0.05) attachment of F. succinogenes on rice straw after 10 min of incubation while a significant reduction (p<0.05) in attachment was not observed until 4 h incubation in the case of R. flavefaciens and R. albus. This result indicated F. succinogenes responded to MC more sensitively and earlier than R. flavefaciens and R. albus. Dry matter digestibility of rice straw was subsequently inhibited by 0.1% MC, and there was a significant difference between control and MC treatment (p<0.05). Incubated cultures containing MC had higher pH and lower gas production than controls. Current data clearly indicated that the attachment of F. succinogenes, R. flavefaciens and R. albus on rice straw was inhibited by MC, which apparently reduced rice straw digestion.

Reducing CH4 Emissions from Grazing Ruminants in New Zealand: Challenges and Opportunities

  • Clark, H.;Kelliher, F.;Pinares-Patino, C.
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권2호
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    • pp.295-302
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    • 2011
  • Almost half of New Zealand's greenhouse gas emissions arise from agriculture and enteric methane ($CH_4 $) emissions arising from ruminant animals constitute 30% of total $CO_2$-e emissions. Enteric $CH_4$ emissions have increased by 9% since 1990. Extensive research has been undertaken to develop reliable methods for measuring enteric $CH_4$ emissions. New Zealand studies using the SF6 tracer technique suggest that on average this technique yields similar values to the 'gold' standard of calorimetry, but with a larger variance. National inventory estimates based on results obtained using the $SF_6$ technique will therefore overestimate the uncertainty. Mitigating emissions can be achieved by changing feed type but there are practical and cost barriers to the use of alternative feeds. Forages containing condensed tannins do reduce emissions but are agronomically inferior to the forages currently used. Rumen additives have shown some success in-vitro but results from in-vivo trials with both monensin and fumaric acid have been disappointing. The development of methods for directly manipulating rumen microorganisms are at an early stage and work to develop vaccines that can inhibit methanogenesis has yielded mixed results. The successful identification of sheep with contrasting $CH_4$ yields raises the possibility that, in the long term, a breeding approach to $CH_4$ mitigation is feasible.

In vitro Glutathione Production using Mixed Cells in an Aerated Slurry Bioreactor (혼합세포를 이용한 Aerated Slurry Bioreactor에서의 in vitro Glutathione 생산)

  • Go, Seong-Yeong;Gu, Yun-Mo
    • KSBB Journal
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    • 제14권4호
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    • pp.445-451
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    • 1999
  • Glutathione production was carried out using mixed cells of E. coli TG1/pDG7 $\alpha$ and bakers yeast in an Aerated Slurry Bioreactor. Glutathione-producing enzymes were stable for 34 hours, yielding 4.6 mM glutathione in suspension reaction. Glutahione production with high density mixed cells was studied as a function of flow rate in an Aereated Slurry Bioreactor. Glutathione concentration was higher than that in suspension reaction for 32 hours at the substrate feeding rate of 5.2 mL/hr with cell recycle in continuous Aerated Slurry Bioreactor. It was for 42 hours at 2.6 mL/hr and 22 hours at 5.2 mL/hr without cell recycle. Glutahione productivity was 25.7 mg/g wet $cell{\cdot}hr$ at the substrate feeding rate of 10.4 mL/hr with cell recycle, but 5.28 mg/g wet $cell{\cdot}hr$ at 5.2 mL/hr and 1.65 mg/g wet $cell{\cdot}hr$ at 2.6 mL/hr without cell recycle. Effective production time increased from 25 to 45 hours, by using a surfactant, tween 80. As a purfing gas, nitrogen was tested instead of air to avoid a possible oxidizing effect on glutathione-producing enzymes, resulting in the increase of effective production time to 40 hours.

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Effect of Byproducts Supplementation by Partically Replacing Soybean Meal to a Total Mixed Ration on Rumen Fermentation Characteristics In Vitro (대두박 대체 부산물 위주의 TMR 사료가 반추위 내 미생물의 In Vitro 발효특성에 미치는 영향)

  • Bae, Gui Seck;Kim, Eun Joong;Song, Tae Ho;Song, Tae Hwa;Park, Tae Il;Choi, Nag Jin;Kwon, Chan Ho;Chang, Moon Baek
    • Journal of The Korean Society of Grassland and Forage Science
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    • 제34권2호
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    • pp.129-140
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    • 2014
  • This study was performed to evaluate the effects of replacing basic total mixed ration (TMR) with fermented soybean curd, Artemisia princeps Pampanini cv. Sajabal, and spent coffee grounds by-product on rumen microbial fermentation in vitro. Soybean in the basic TMR diet (control) was replaced by the following 9 treatments (3 replicates): maximum amounts of soybean curd (SC); fermented SC (FSC); 3, 5, and 10% FSC + fermented A. princeps Pampanini cv. Sajabal (1:1, DM basis, FSCS); and 3, 5, 10% FSC + fermented coffee meal (1:1, DM basis, FSCC) of soybean. FSC, FSCS, and FSCC were fermented using Lactobacillus acidophilus ATCC 496, Lactobacillus fermentum ATCC 1493, Lactobacillus plantarum KCTC 1048, and Lactobacillus casei IFO 3533. Replacing dairy cow TMR with FSC treatment led to a pH value of 6 after 8 h of incubation-the lowest value measured (p<0.05), and FSCS and FSCC treatments were higher than SC and FSC treatment after 6 h (p<0.05). Gas production was higher in response to 3% FSC and FSCC treatments than the control after 4-10 h. Dry matter digestibility was increased 0-12 h after FSC treatment (p<0.05) and was the highest after 24 h of 10% FSCS treatment. $NH_3-N$ concentration was the lowest after 24 h of FSC treatment (p<0.05). Microbial protein content increased in response to treatments that had been fermented by the Lactobacillus spp. compared to control and SC treatments (p<0.05). The total concentration of volatile fatty acids (VFAs) was increased after 6-12 h of FSC treatment (p<0.05), while the highest acetate proportion was observed 24 h after 5% and 10% FSCS treatments. The FSC of propionate proportion was increased for 0-10 h compared with among treatments (p<0.05). The highest acetate in the propionate ration was observed after 12 h of SC treatment and the lowest with FSCS 3% treatment after 24 h. Methane ($CH_4$) emulsion was lower with A. princeps Pampanini cv. Sajabal and spent coffee grounds treatments than with the control, SC, and FSC treatments. These experiments were designed to replace the by-products of dairy cow TMR with SC, FSC, FSCS, and FSCC to improve TMR quality. Condensed tannins contained in FSCS and FSCC treatments, which reduced $CH_4$ emulsion in vitro, decreased rumen microbial fermentation during the early incubation time. Therefore, future experiments are required to develop a rumen continuous culture system and an in vivo test to optimize the percentages of FSC, FSCS, and FSCC in the TMR diet of the dairy cows.

Effects of Feeding Heat Treated Protein and Mineral Complex on In Vitro Fermentation Characteristics, Milk Production and Composition of Holstein Dairy Cows (열처리 단백질-광물질 복합제제 첨가가 In Vitro 발효성상과 착유우의 유량 및 유성분에 미치는 영향)

  • Choi, N.J.;Bae, G.S.;Nam, K.P.;Chang, M.B.;Um, J.S.;Ko, J.Y.;Ha, J.K.
    • Journal of Animal Science and Technology
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    • 제44권5호
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    • pp.541-548
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    • 2002
  • This study, consisting of two experiments, was conducted to determine the effects of feeding heat treated protein and mineral complex (HPM) on milk production and composition, and ruminal fermentation of Holstein dairy cows. In in vitro experiment, HPM levels were 0, 0.2, 1 and 2%, and Timothy hay, which was substrate, was milled as 1 mm size, and the effects of HPM on pH, ammonia and VFA were analyzed after incubation times of 0, 6, 12, 24 and 48 h, respectively. The pH and ammonia production were not significantly different between treatments during the incubation. In addition, generally, total VFA and individual VFA were not affected by HPM on 0, 6 and 24 h. While, total VFA and individual VFA were increased in 0.2% and 1% of HPM supplemented treatments, but decreased in 2% of HPM treatment compared with control on 12 h. On 48 h, total VFA and individual VFA were increased in HPM treatments compared to control (P<0.05). However, A/P ratio was not affected by HPM supplementation. Gas production was higher in HPM treatment compared to control on 24 h (P<0.05) and 48 h (P<0.05). In lactating experiment, fourteen lactating Holstein cows were used for 4 months in a cross over experimental design. There were two treatments; no added HPM as a control and 0.2% of HPM added as a test treatment. Daily milk yield (P<0.001), 4% FCM (P<0.001), milk protein (P<0.05) and SNF (solid not fat; P<0.05) were increased in HPM treatment compared to control. While, milk fat, MUN (milk urea nitrogen) and SCC (somatic cell count) were not significantly different between treatments.