• Title/Summary/Keyword: In vitro Rumen Fermentation

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Effects of wild or mutated inoculants on rye silage and its rumen fermentation indices

  • Paradhipta, Dimas Hand Vidya;Joo, Young Ho;Lee, Hyuk Jun;Lee, Seong Shin;Kwak, Youn Sig;Han, Ouk Kyu;Kim, Dong Hyeon;Kim, Sam Churl
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.949-956
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    • 2020
  • Objective: This study was conducted to confirm the effects of new inoculants producing-antifungal or esterase substances on rye silage and its rumen fermentation indices by comparing wild with mutated types. Methods: Rye harvested at dough stage was ensiled into 3 L mini bucket silo (1 kg) for 90 d in triplicate following: distilled water at 20 μL/g (CON); Lactobacillus brevis 100D8 (AT) and its inactivation of antifungal genes (AT-m) at 1.2×105 cfu/g, respectively; and Leuconostoc holzapfelii 5H4 (FD) and its inactivation of esterase genes (FD-est) at 1.0×105 cfu/g, respectively. After silo opened, silage was sub-sampled for the analysis of ensiling quality and its rumen fermentation indices. Results: Among the wild type inoculants (CON vs AT vs FD), FD inoculant had higher (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in rumen, while AT inoculant had higher (p<0.05) lactate, acetate, and lactic acid bacteria in silage. Silage pH and the potentially degradable fraction in rumen increased (p<0.05) by inactivation of antifungal activity (AT vs AT-m), but lactate, acetate, and lactic acid bacteria of silage decreased (p<0.05). In silage, acetate increased (p<0.05) by inactivation of esterase activity (FD vs FD-est) with decreases (p<0.05) of pH, ammonia-N, lactate, and yeast. Moreover, inactivation of esterase activity clearly decreased (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in the rumen. Conclusion: This study concluded that FD inoculant confirmed esterase activity on rye silage harvested at dough stage, while AT inoculant could not be confirmed with antifungal activity due to the absence of mold in all silages.

Effects of Polyurethane Coated Urea Supplement on In vitro Ruminal Fermentation, Ammonia Release Dynamics and Lactating Performance of Holstein Dairy Cows Fed a Steam-flaked Corn-based Diet

  • Xin, H.S.;Schaefer, D.M.;Liu, Q.P.;Axe, D.E.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.491-500
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    • 2010
  • Three experiments were conducted to investigate the effects of polyurethane coated urea on in vitro ruminal fermentation, ammonia release dynamics and lactating performance of Holstein dairy cows fed a steam-flaked corn-based diet. In Exp. 1, a dual-flow continuous culture was run to investigate the effect of polyurethane coated urea on nutrient digestibility, rumen fermentation parameters and microbial efficiency. Three treatment diets with isonitrogenous contents (13.0% CP) were prepared: i) feedgrade urea (FGU) diet; ii) polyurethane coated urea (PCU) diet; and iii) isolated soy protein (ISP) diet. Each of the diets consisted of 40% steam-flaked corn meal, 58.5% forages and 1.5% different sources of nitrogen. PCU and FGU diets had significantly lower digestibility of NDF and ADF (p<0.01) than the ISP diet. Nitrogen source had no significant effect (p = 0.62) on CP digestibility. The microbial efficiency (expressed as grams of microbial N/kg organic matter truly digested (OMTD)) in vitro of the PCU diet (13.0 g N/kg OMTD) was significantly higher than the FGU diet (11.3 g N/kg OMTD), but comparable with the ISP diet (14.7 g N/kg OMTD). Exp. 2, an in vitro ruminal fermentation experiment, was conducted to determine the ammonia release dynamics during an 8 h ruminal fermentation. Three treatment diets were based on steam-flaked corn diets commonly fed to lactating cows in China, in which FGU, PCU or soybean meal (SBM) was added to provide 10% of total dietary N. In vitro $NH_3-N$ concentrations were lower (p<0.05) for the PCU diet than the FGU diet, but similar to that for the SBM diet at all time points. In Exp. 3, a lactation trial was performed using 24 lactating Holstein cows to compare the lactating performance and blood urea nitrogen (BUN) concentrations when cows were fed PCU, FGU and SBM diets. Cows consuming the PCU diet had approximately 12.8% more (p = 0.02) dietary dry matter intake than those consuming the FGU diet. Cows fed the PCU diet had higher milk protein content (3.16% vs. 2.94%) and lower milk urea nitrogen (MUN) concentration (13.0 mg/dl vs. 14.4 mg/dl) than those fed the FGU diet. Blood urea nitrogen (BUN) concentration was significantly lower for cows fed the PCU (16.7 mg/dl) and SBM (16.4 mg/dl) diets than the FGU (18.7 mg/dl) diet. Cows fed the PCU diet had less surplus ruminal N than those fed the FGU diet and produced a comparable lactation performance to the SBM diet, suggesting that polyurethane coated urea can partially substitute soybean meal in the dairy cow diet without impairing lactation performance.

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.

Effects of Thymol, Eugenol and Malate on In vitro Rumen Microbial Fermentation

  • Kim, Do-Hyung;Kim, Kyoung-Hoon;Choi, Chang-Won;Hong, Seong-Koo;Seol, Yong-Joo;Kwon, Eung-Gi;Kim, Wan-Young;Nam, In-Sik;Lee, Sung-Sill;Oh, Young-Kyoon
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.511-520
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    • 2009
  • The purpose of this study was to investigate effects of increased levels of eugenol, thymol and malate on pH and the concentrations of VFA, lactate and ammonia-N during in vitro ruminal incubation. One Hanwoo beef steer (741 kg) fitted with a rumen cannula was used and fed 0.5 kg/day rice straw and 10 kg/day corn-based concentrate (ratio of concentrate to rice straw = 95 : 5 on DM basis). Three different doses of thymol, eugenol and malate were used. Treatments of the experiment were as follows: Treatments of thymol were control (1g D-glucose/40ml), T1 (1g D-glucose + 40 mg thymol/40 ml), T2 (1g D-glucose + 50 mg thymol/40 ml) and T3 (1g D-glucose + 60 mg thymol/40 ml). Treatments of eugenol were control (1g D-glucose/40 ml), E1 (1g D-glucose + 55 mg eugenol/40 ml), E2 (1g D-glucose + 65 mg eugenol/40 ml) and E3 (1g D-glucose + 75 mg eugenol/40 ml). Treatments of malate were control (1g D-glucose/40ml), M1 (1g D-glucose + 25 mg malate/40ml), M2 (1g D-glucose + 50 mg malate/40 ml) and M3 (1g D-glucose + 100 mg malate/40 ml). The results of this study showed that eugenol and thymol have improved stability of the ruminal fermentation by decreasing lactic acid concentration and increasing ruminal pH. However, it inhibited the production of total VFA, acetate and propionate. Malate also improved stability of the ruminal fermentation by decreasing lactic acid concentration and increasing ruminal pH, but it had a very little effect on ruminal lactate concentrations and pH. On the other hand, malate did not decrease the concentrations of total VFA, acetate and propionate. Therefore, at the low ruminal pH expected in high-concentrate diets, thymol, eugenol, and malate are potentially useful in Hanwoo finishing diets. Further studies are necessary for determining the effectiveness of these additives on in vivo rumen fermentation and animal performance in Hanwoo finishing steers.

The Effect of Alcohol Fermented Feedstuff Made of Byproducts on In vitro Fermentation Characteristics and NDF Disappearance in the Rumen (조사료 대용 산업부산물을 이용한 알코올 발효사료가 In vitro 반추위내 발효특성 및 NDF 분해율에 미치는 영향)

  • Kim, Byong-Wan;Lin, Guang-Zhe;Park, Byung-Ki;Kim, Jong-Duk;Shin, Jong-Suh
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.4
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    • pp.323-334
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    • 2007
  • An in vitro study was conducted to examine the influence of alcohol-fermented feedstuff formulated with byproducts on the fermentation characteristics and NDF disappearance in the rumen. Dietary treatments were either a soybean curd-based alcohol-fermented feedstuff (AFS) or brewery grain-based alcohol-fermented feedstuff (AFB). The AFS and AFB are composed of 50% commercial beef cattle feed, 50% soybean curd dreg, 5% molasses and 0.5% yeast and 25% commercial beef cattle feed, 25% brewery grain, 25% soybean curd dreg, 25% corn grit, 5% molasses and 0.5% yeast, respectively. The change of ammonia, pH alcohol, volatile fatty acids, and NDF disappearance were measured at 0, 2, 4, 8 and 12 hr after in vitro incubation in the rumen. After 2 hr incubation, higher ammonia concentrations were resulted in AFS (12.47 mg/dl) and AFB (12.85 mg/dl) compared to control (11.84 mg/dl) (p<0.05). Ruminal pH of AFS and AFB were significantly higher than control during 1 to 6 hr fermentation, but the pH of AFS and AFB were decreased after 6 hr. At 12 hr fermentation, the alcohol concentration of AFS and AFB was significantly increased by 43.9% and 48.0%, respectively. The acetate concentration was rapidly decreased in control, while the concentration was slowly decreased in AFS and AFB with increasing the fermentation time. Lower concentrations propionate and butyrate were observed in AFS and AFB during every fermentation time (p<0.05). The NDF disappearance was significantly lower in AFS and AFB after 4 hr fermentation. These results suggest that alcohol fermented feedstuff can control the fermentation pattern in the rumen.

Influence of Monensin and Virginiamycin on In Vitro Ruminal Fermentation of Ammoniated Rice Straw

  • Kook, K.;Sun, S.S.;Yang, C.J.;Myung, K.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.544-547
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    • 1999
  • The object of this study was to determine the influence of monensin and virginiamycin (VM) on in vitro ruminal fermentation of rice straw or ammoniated rice straw. Rumen fluid was collected from 4 wethers fed 200 g of concentrate supplement with 400 g of untreated (U) or ammoniated (A) rice straw once daily for 28 days. Mixed ruminal microorganisms were incubated in anaerobic media that contained 20% (vol/vol) ruminal fluid and 0.3 g of either U or A rice straw. Monensin and/or VM, dissolved in ethanol, were added in centrifuge tubes at final concentrations of 0, 15, 30, 15+15 and 30+30 ppm of culture fluid. The addition of monensin and VM combination to A rice straw fermentation decreased (p<0.05) the acetate to propionate ratio, total VFA and lactate production, but increased (p<0.05) pH. Total gas production tended to be decreased by the addition of monensin plus VM. Antimicrobial agents decreased $NH_3$ N concentration and dry matter digestibility.

Effects of Moisture and a Saponin-based Surfactant during Barley Processing on Growth Performance and Carcass Quality of Feedlot Steers and on In vitro Ruminal Fermentation

  • Wang, Y.;Gibb, D.;Greer, D.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1690-1698
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    • 2011
  • Feedlot and in vitro ruminal experiments were conducted to assess the effects of saponin-containing surfactant applied during tempering of barley grain on cattle growth performance and on ruminal fermentation. In the feedlot experiment, treatments with three barley grain/barley silage based diets were prepared using barley grain at 7.7% moisture (dry, D), after tempering to 18% moisture (M), or after tempering with a saponin-based surfactant included at 60 ml/t (MS). Each treatment was rolled at settings determined previously to yield optimally processed barley. A total of 180 newly weaned British${\times}$Charolais steers were fed three diets in 18 pens for a 63-d backgrounding period and 91-d finishing period to determine feed intake, growth rate and feed efficiency. Cattle were slaughtered at the end of the experiment to measure the carcass characteristics. Tempering reduced (p<0.001) volume weight and processing index, but processing characteristics were similar between MS and M. Tempering increased (p<0.05) growth during backgrounding only, compared with D, but did not affect feed intake in either phase. During backgrounding, feed efficiency was improved with tempering, but during finishing and overall this response was only observed with the surfactant. Tempering did not affect carcass weight, fat content or meat yield. Surfactant doubled the proportion of carcasses grading AAA. In the in vitro experiment, barley (500 mg; ground to <1.0 mm or steam-rolled) was incubated in buffered ruminal fluid (40 ml) without or with surfactant up to 20 ${\mu}l/g$ DM substrate for 24 h. Surfactant increased (p<0.05) apparent DM disappearance and starch digestibility but reduced productions of gas and the volatile fatty acid and acetate:propionate ratio, irrespective of barley particle size. Compared with feeding diets prepared with non-tempered barley, tempering with surfactant increased the feed efficiency of feedlot steers. This may have arisen from alteration in processing characteristics of barley grain by surfactant rather than its direct effect on rumen microbial fermentation.

Influence of Temperature and pH on Fermentation Pattern and Methane Production in the Rumen Simulating Fermenter (RUSITEC)

  • Bhatta, R.;Tajima, K.;Kurihara, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.376-380
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    • 2006
  • An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

Study on Rumen Cellulolytic Bacterial Attachment and Fermentation Dependent on Initial pH by cPCR (cPCR 기법을 이용한 초기배양 pH에 의한 반추위 섬유소 분해 박테리아의 부착 및 발효에 관한 연구)

  • Kim, M.S.;Sung, H.G.;Kim, H.J.;Lee, Sang-S.;Chang, J.S.;Ha, J.K.
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.615-624
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    • 2005
  • The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.

The Effect of Plant Extracts on In-vitro Ruminal Fermentation, Methanogenesis and Methane-related Microbes in the Rumen

  • Kim, E.T.;Min, K.S.;Kim, C.H.;Moon, Y.H.;Kim, S.C.;Lee, Sung-Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.517-522
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    • 2013
  • The effect on methanogens attached to the surface of rumen ciliate protozoa by the addition of plant extracts (pine needles and ginkgo leaves) was studied with particular reference to their effectiveness for decreasing methane emission. The plant extracts (pine needles and ginkgo leaves) were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, ciliated-associated methanogen, four different groups of methanogens and Fibrobacter succinogenes were quantified by using the real-time PCR. Gas profiles including methane, carbon dioxide and hydrogen, and runinal fermentation characteristics were observed in vitro. The methane emission from samples with an addition of individual juices from pine needles, ginkgo leaves and 70% ethanol extract from ginko leaves was significantly lower (p<0.05, 27.1, 28.1 and 28.1 vs 34.0 ml/g DM) than that of the control, respectively. Total VFAs in samples with an addition of any of the plant extracts were significantly lower than that of the control (p<0.05) as well. The order Methanococcales and the order Methanosarcinales were not detected by using PCR in any incubated mixtures. The ciliate-associated methanogens population decreased from 25% to 49% in the plant extacts as compared to control. We speculate that the supplementation of juice from pine needles and ginkgo leaves extract (70% ethanol extract) decreased the protozoa population resulting in a reduction of methane emission in the rumen and thus inhibiting methanogenesis. The order Methanobacteriales community was affected by addition of all plant extracts and decreased to less than the control, while the order Methanomicrobiales population showed an increase to more than that of the control. The F. succinogenes, the major fibrolytic microorganism, population in all added plant extracts was increased to greater than that of the control. In conclusion, pine needles and ginkgo leaves extracts appear to have properties that decrease methanogenesis by inhibiting protozoa species and may have a potential for use as additives for ruminants.