• 한국토양비료학회지
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• 제45권6호
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• pp.1049-1057
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• 2012

본 연구는 메탄생성에 직접적으로 관여하는 혼합 메탄균과 셀롤로스 등의 고분자 물질의 가수분해 반응에 활성이 뛰어난 반추위 내 혐기성 섬유소분해균 중에서 대표적인 Fibrobacter succinogenes, Ruminococcus flavefaciens 및 Ruminococcus albus를 biochemical methane potential (BMP) 시험에 첨가하였을 때 메탄 발생에 미치는 영향을 조사하고자 수행되었다. BMP시험은 멸균증류수를 첨가한 control과 각각의 미생물 배양액을 첨가한 혼합 메탄균 첨가구 (M), F. succinogenes 첨가구 (FS) R. flavefaciens 첨가구 (RF), R. albus 첨가구 (RA) 및 RA+FS 혼합첨가구와 M+RA+FS 혼합 첨가구로 총 7개 처리구로 각 처리구별 3반복으로 진행되었다. 미생물 배양액의 첨가량은 식종액과 양돈슬러리에 1% (0.5 mL), 3% (1.5 mL) 및 5% (2.5 mL) 씩 첨가 하였다. BMP 시험을 위해 60일간 배양이 지속되었고 중온소화를 위해 $38^{\circ}C$의 배양기에서 수행되었다. 실험의 결과 총 바이오가스 발생량은 5% RF와 RA+FS가 대조구에 비하여 8.1 및 8.4%로 가스발생량이 유의적으로 높았다(p<0.05). 메탄발생량은 3% 미생물 배양액 첨가구 중 M+RA+FS를 제외하고 대조구에 비하여 증가시키는 경향을 보였으며, 5% 배양액을 첨가하였을 때는 대조구에 대하여 5%의 M, FS, RF, RA, RA+FS, 및 M+RA+FS RF가 각각 12.1, 12.6, 17.3, 13.7, 17.9 및 14.7%로 메탄가스발생량을 증가시켰다 (p<0.05). TS 및 VS 분해율은 가스발생량과는 관계없이 모든 처리구에서 미생물 배양액의 첨가량이 증가하더라도 차이가 없었다. BMP 종료시 배양액내 pH는 모든 처리구가 7.527~7.657의 범위로 메탄발효에 큰 영향을 주지 않았다. 결론적으로, 본 실험에서는 양돈 슬러리의 성분특성으로 인해 가수분해단계와 메탄생성단계 모두에 첨가한 미생물 배양액이 효과가 있었으나, 5% 첨가수준을 제외하고 낮은 첨가수준에서는 첨가효과가 나타나지는 않았다.

• 미생물학회지
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• 제34권3호
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• pp.91-95
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• 1998

한우의 반추위액에서 섬유소를 분해하는 균주 GPC-1, GPC-2, GNR-1, GNR-2, GNR-3를 선별하였다. 분리주 GPC-1과 GPC-2는 Gram 양성구균이며 편성혐기균으로서 Ruminococcus속으로 분류되었다. 분리주 GNR-1, GNR-2 및 GNR-3는 Gram 음성 간균이며 혐기균으로서 내생포자형성 유무에 따라 포자 형성균인 GNR-3는 Clostridium속으로 분류하고 포자를 형성하지 않은 균은 $H_2S$ 생성유무에 따라 Bacteroides 속과 Butyrivibrio 속으로 분류되었다. 종수준의 동정을 위하여 당분해능과 생화학적 조사를 통하여 GPC-1는 Ruminococcus albus로, GPC-2는 Ruminococcus flavefaciens로, GNR-1는 Bacteroides succinogenes로, GNR-2는 Butyrivibrio fibrisolvens로, GNR-3는 Clostridium cellobioparum로 각각 동정하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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• pp.211-218
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• 2016

This study evaluated the effect of Candida norvegensis (C. norvegensis) viable yeast culture on in vitro ruminal fermentation of oat straw. Ruminal fluid was mixed with buffer solution (1:2) and anaerobically incubated with or without yeast at $39^{\circ}C$ for 0, 4, 8, 16, and 24 h. A fully randomized design was used. There was a decrease in lactic acid (quadratic, p = 0.01), pH, (quadratic, p = 0.02), and yeasts counts (linear, p<0.01) across fermentation times. However, in vitro dry matter disappearance (IVDMD) and ammonia-N increased across fermentation times (quadratic; p<0.01 and p<0.02, respectively). Addition of yeast cells caused a decrease in pH values compared over all fermentation times (p<0.01), and lactic acid decreased at 12 h (p = 0.05). Meanwhile, yeast counts increased (p = 0.01) at 12 h. C. norvegensis increased ammonia-N at 4, 8, 12, and 24 h (p<0.01), and IVDMD of oat straw increased at 8, 12, and 24 h (p<0.01) of fermentation. Yeast cells increased acetate (p<0.01), propionate (p<0.03), and butyrate (p<0.03) at 8 h, while valeriate and isovaleriate increased at 8, 12, and 24 h (p<0.01). The yeast did not affect cellulolytic bacteria (p = 0.05), but cellulolytic fungi increased at 4 and 8 h (p<0.01), whereas production of methane decreased (p<0.01) at 8 h. It is concluded that addition of C. norvegensis to in vitro oat straw fermentation increased ruminal fermentation parameters as well as microbial growth with reduction of methane production. Additionally, yeast inoculum also improved IVDMD.

• 한국초지조사료학회지
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• 제38권2호
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• pp.106-111
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• 2018

본 연구는 고창증 발병으로 폐사된 저지종 젖소의 반추위액과 캐놀라가 장착한 저지종 젖소의 반추위액을 샘플링하여 반추위 내 미생물 균총의 급작스런 변화에 따른 반추위 발효와 고창증 발병간에 연관성을 알아보고자 진행되었다. 채취한 반추위액 샘플은 PowerMax Soil DNA Isolation Kit (MO BIO Inc., CA, USA)를 이용하여 DNA를 추출한 후 PicoGreen (Turner BioSystems, Inc., CA, USA)과 Nanodrop spectrophotometer (ND-1000, NanoDrop Technologies Inc., DE, USA)를 이용하여 260/280 nm와 260/230 nm 흡광도 값을 측정하였다. DNA를 정량분석하고 $MiSeq^{TM}$ platform (Illumina, CA, USA)을 이용한 장내 미생물 균총의 다양성 분석을 마크로젠(Macrogen Inc., Seoul)에 의뢰하여 실시하였다. 반추위액 내 미생물 균총 분석 결과, 전분 분해균의 경우 고창증 발병으로 인한 폐사우의 반추위 내에서 R. bromii가 우점 하였으며, R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들이 정상우 대비 36배 높았다. 반면 반추위 발효와 밀접한 관련이 있는 F. succinogenes, R. albus 및 R. flavefaciens 등과 같은 대표적인 섬유소 분해균 비율은 정상우에서 고창증 발병으로 인한 폐사우 대비 12배 높았다. 이와 같은 결과를 볼 때, 반추위 내 균총 가운데 R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들의 급격한 증가가 반추위 내 pH 저하 및 가스 생성 증가를 초래했고 이는 저지종 젖소의 고창증 발병으로 인한 폐사와 밀접한 연관이 있었을 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권7호
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• pp.952-960
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• 2013

The objective of this study was to investigate the effect of carbohydrate source and cottonseed meal level in the concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in swamp buffaloes. Four, 4-yr old rumen fistulated swamp buffaloes were randomly assigned to receive four dietary treatments according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. Factor A was carbohydrate source; cassava chip (CC) and CC+rice bran at a ratio 3:1 (CR3:1), and factor B was level of cottonseed meal (CM); 109 g CP/kg (LCM) and 328 g CP/kg (HCM) in isonitrogenous diets (490 g CP/kg). Buffaloes received urea-treated rice straw ad libitum and supplemented with 5 g concentrate/kg BW. It was found that carbohydrate source did not affect feed intake, nutrient intake, digested nutrients, nutrient digestibility, ammonia nitrogen concentration, fungi and bacterial populations, or microbial protein synthesis (p>0.05). Ruminal pH at 6 h after feeding and the population of protozoa at 4 h after feeding were higher when buffalo were fed with CC than in the CR3:1 treatment (p<0.05). Buffalo fed with HCM had a lower roughage intake, nutrient intake, population of total viable and cellulolytic bacteria and microbial nitrogen supply than the LCM fed group (p<0.05). However, nutrient digestibility, ruminal pH, ammonia concentration, population of protozoa and fungi, and efficiency of microbial protein synthesis were not affected by cottonseed meal levels (p>0.05). Based on this experiment, concentrate with a low level of cottonseed meal could be fed with cassava chips as an energy source in swamp buffalo receiving rice straw.

• Asian-Australasian Journal of Animal Sciences
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• 제11권5호
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• pp.510-521
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• 1998

The effects of two types of protein, soybean meal (SBM) and fish meal (FM); and two types of energy supplements, corn flour (CF) and paper pulp (PP), on intake of guinea grass (Panicum maximum), fibre digestion and microbial activities in four Merino rams with an average weight of $54.4{\pm}4.5kg$ were studied. Each animal was fitted with a ruminal cannula and a duodenal cannula at the proximal position. The animals were fed twice daily with chopped guinea grass (5 cm) ad libitum and one of the four dietary supplements: 170 g FM+268 g PP; 170 g FM+268 g CF; 200 g SBM+200 g PP or 200 g SBM+200 g CF. All the supplements were mixed with 100 g molasses. In sacco and in vivo digestibilities, digesta flow rates, fermentation and microbial population were studied in a $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement of dietary treatments. The effects of energy or protein sources were not significant on grass intake of sheep. The potential degradabilities of NDF and ADF were not significantly affected by any of the supplements. However, the energy and protein sources had significant efects on disappearance rate of NDF and ADF. The disappearance rate of both NDF and ADF were significantly (p < 0.05) higher in animals fed PP when compared to animals fed CF. Animals fed FM also showed significantly (p < 0.03) higher disappearance rate of ADF than those fed SBM. Animals fed PP showed better digestion in the rumen and total tract. Total flow of NDF and ADF through the duodenum was not significantly affected by the various supplements. The mean rumen pH values (5.8-6.1) were not significantly different among the four different diets. The concentration of rumen ammonia was significantly (p < 0.0001) higher in animals fed SBM (235-266.4 mg N/L) supplement than in animals fed FM (174.9-179.7 mg N/L), while total VFA concentration was not significantly affected by both energy and protein supplements. Mean values of total VFA ranged from 72.5-82.3 mM. Molar proportions of acetate, propionate and butyrate were typical of a roughage type fermentation. Molar proportion of acetate was significantly (p < 0.0001) higher in sheep fed PP when compared to sheep fed CF. Animals fed FM had higher total viable bacterial counts, while animals fed CF showed higher protozoal numbers. Proportions of cellulolytic bacteria were only slightly higher in animals fed SBM or PP.

• 동물자원연구
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• 제30권3호
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• pp.111-120
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• 2019

생물학적 평가 방법 중 in situ와 in vitro 평가 방법은 비용 및 시간을 절약할 수 있으며 동물 복지 측면에서 제약이 적어 주목되고 있는 방법이다. In vitro 실험에서 사용되는 사료 접종법으로 배양병에 직접 사료를 담는 dispersion 방법은 실제 반추위 발효 상황에 근접한 조건에서 실험할 수 있는 장점이 있지만, 시간과 노동력이 많이 소모된다. 반대로 사료 주머니를 이용한 방법은 실험 수행의 편이성이 있지만 첨가제 실험 수행의 제약이 있으며, 미생물 활성과 섬유소 소화율이 저하되는 단점이 있다. 본 실험은 dispersion 방법과 사료 주머니 방법을 조합하여 보다 편리한 방법으로 dispersion 에 가까운 높은 재현성의 발효 성상과 소화율 평가 가능할 것이라고 생각하여 in vitro 발효 성상과 미생물 조성을 비교하였다. Dispersion 방법과 nylon bag 을 조합한 실험에서 건물 소화율은 세척 과정에서 미생물 및 사료 입자의 유실로 과대평가될 수 있지만 이론적 최대 가스 발생량 V_max, 가스 발생량에 대한 분해 속도 상수 K_g 에서 유의적 차이가 나타나지 않으며, IVNDFD, pH, NH₃-N, total VFA 발생량 그리고 VFA 조성에서도 유의적 차이가 나지 않았다. Real-time PCR 을 이용한 미생물 조성 분석에서는 general bacteria의 절대량에서 유의적 차이가 나지 않았다. 상기의 결과를 종합해 볼 때, in vitro 실험에서 사료 주머니의 사용은 충분히 적용가능하며 일반적인 방법인 dispersion과 비교해 볼 때, 영양소 이용에 있어 반추미생물의 사료 접촉을 방해하지 않을 것으로 생각된다. 하지만 in vitro bach culture 방법은 배양조건에서 제약이 있어, 향후 연속 배양 방법과 in situ의 실험을 통해 본 실험의 실험결과에 대한 추가적인 규명이 필요할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권6호
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• pp.820-826
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• 2013

The objective of this study was to determine the effect of rain tree pod meal with palm oil supplementation on feed intake, digestibility, microbial protein synthesis and microbial populations in dairy cows. Four, multiparous early-lactation Holstein-Friesian crossbred (75%) lactating dairy cows with an initial body weight (BW) of $405{\pm}40$ kg and $36{\pm}8$ DIM were randomly assigned to receive dietary treatments according to a $4{\times}4$ Latin square design. The four dietary treatments were un-supplementation (control), supplementation with rain tree pod meal (RPM) at 60 g/kg, supplementation with palm oil (PO) at 20 g/kg, and supplementation with RPM at 60 g/kg and PO at 20 g/kg (RPO), of total dry matter intake. The cows were offered concentrates, at a ratio of concentrate to milk production of 1:2, and chopped 30 g/kg of urea treated rice straw was fed ad libitum. The RPM contained condensed tannins and crude saponins at 88 and 141 g/kg of DM, respectively. It was found that supplementation with RPM and/or PO to dairy cows diets did not show negative effects on feed intake and ruminal pH and BUN at any times of sampling (p>0.05). However, RPM supplementation resulted in lower crude protein digestibility, $NH_3$-N concentration and number of proteolytic bacteria. It resulted in greater allantoin absorption and microbial crude protein (p<0.05). In addition, dairy cows showed a higher efficiency of microbial N supply (EMNS) in both RPM and RPO treatments. Moreover, NDF digestibility and cellulolytic bacteria numbers were highest in RPO supplementation (p<0.05) while, supplementation with RPM and/or PO decreased the protozoa population in dairy cows. Based on this study, supplementation with RPM and/or PO in diets could improve fiber digestibility, microbial protein synthesis in terms of quantity and efficiency and microbial populations in dairy cows.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1635-1642
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• 2018

Objective: This study was conducted to evaluate the effects of Rhus succedanea extract addition on in vitro ruminal fermentation and microbial growth. Methods: Two ruminally-fistulated steers consuming 600 g/kg timothy- and 400 g/kg cracked corn-based concentrate with free access to water and mineral block were used as rumen fluid donors. In vitro batch fermentation, with timothy as a substrate, was conducted for up to 72 h, with Rhus succedanea extracts added to achieve final concentrations of 0, 10, 30, 50, 70, and 90 mg/L. Results: Effective dry matter (DM) degradability rate linearly decreased (p = 0.046) depending on extract dosing levels. Total gas production after 24 to 72 h incubation tended to decrease following extract addition, beginning with 50 mg/L starting dose (significance of quadratic effects: p = 0.006, p<0.001, and p = 0.008 for 24, 48, and 72 h, respectively). Methane production decreased depending on dosing levels following 24 h (p<0.05) and 48 h (p<0.005) incubations and was the lowest with the 50 mg/L dose. The Rhus succedanea extracts increased the abundance of Fibrobacter succinogenes (p<0.05) and Ruminococcus flavefaciens (p = 0.0597) and decreased the abundance of methanogenic archaea (p<0.05) following 24 h incubation. Conclusion: Rhus succedanea was shown to reduce methane production and increase cellulolytic bacteria without any signs of toxic effects and with a minor effect on DM degradability.

• 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.