본 연구의 목표는 S. boulardii 유래 세포벽 추출물 (CWSB)의 첨가가 반추위 발효에 미치는 영향을 평가하는 것이다. 본 연구의 in vitro 발효 실험은 CWSB의 첨가용 량이 다른 2개의 실험으로 구성되었다. 실험 1에서는 대 조구(CON, 시험사료)와 CWSB0.05(시험사료+CWSB 0.5g/kg 첨가), CWSB0.075(시험사료+CWSB 0.75g/kg 첨 가)의 처리구로 구성되었고, 실험 2의 처리구는 대조구 (CON, 시험사료)와 CWSB0.5(시험사료+ CWSB 5g/kg 첨 가), CWSB1.0(시험사료+CWSB 10g/kg 첨가)의 처리구로 구성되었다. In vitro 배양 이후 3, 6, 12, 24, 48h 동안 발 생한 가스 발생량 및 48h 배양 이후 발효 성상을 분석하 였다. 실험 1에서는 CWSB0.05 처리구에서 유의적으로 높 았던 Vmax(p<0.01)를 제외한 모든 반추위 발효성상에서 처 리구간 유의적 차이가 관찰되지 않았고, 이에 따라 사료 내 0.5%, 1%의 고용량 첨가 수준을 설정한 실험 2를 수행 하였다. 실험 2에서의 CWSB1.0 처리구는 대조구에 비해 총 VFA 발생량이 유의적으로 높았으나 전체 배양시간 내 가스 발생량 및 건물 소화율, 섬유소 소화율은 대조구와 유의적인 차이를 나타내지 않았다. 본 연구 결과를 종합 해 볼 때, CWSB의 첨가수준은 0.5% 이상에서 반추위 발 효 개선효과를 나타낼 것으로 추정된다.

반추위 미생물체 단백질의 아미노산 조성이 한우의 에 너지원 사료에 따라 차이가 있는지를 조사하고자 본 연구 가 수행되었다. 본 실험은 국내 한우 비육우 사육에 주로 이용되는 에너지원 사료인 옥수수(T1), 생미강(T2), 소맥 (T3), 소맥피(T4)를 이용하여 in vitro 반추위 발효실험을 진행하여 발효 24시간 후 배양액의 발효 성상, 미생물체 단백질 합성량(MPS), 미생물의 아미노산 조성 및 미생물 균총 변화를 분석하였다. 배양액의 pH는 T4가 6.02로 다 른 처리구들에 비해서 가장 높았으며, T1은 4.84로 가장 낮았다(p<0.05). Total VFA 농도는 T2가 96.80mM로 다른 처리구들에 비해서 가장 높았고, T1이 53.35mM로 가장 낮았다(p<0.05). 미생물체 단백질 합성량은 T3가 222.50g/L로 다른 처리구들에 비해서 높았으며(p<0.05), T1과 T4는 유의적인 차이는 없었지만 T1이 가장 낮았다. DGGE band 분석결과 모든 처리구는 57.5% 이하의 유사도를 나타내었으며, 급여되는 사료에 따라서 미생물 군집 이 다르게 나타났다. 미생물 내 총 아미노산 함량은 T3와 T4가 다른 처리구보다 높았다(p<0.05). 미생물의 아미노산 조성 중 대부분의 필수 아미노산은 T4가 다른 처리구들에 비해서 낮았다. 하지만 methionine은 다른 필수 아미노산 과 다르게 T4에서 가장 높았다(p<0.05). 곡류 사료 위주의 급여 시 결핍되기 쉬운 lysine은 모든 처리구에서 미생물 의 아미노산 조성이 8.05~8.98%의 범위에 있었다. 따라서, 본 연구에서 반추 가축 급여 에너지 사료원에 따른 반추 위 내 미생물 발효특성과 미생물 군집의 차이가 미생물의 아미노산 조성을 변화시킨다는 것을 확인하였다.

The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39°C for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybeansupplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

본 연구는 조사료 원료인 마늘 껍질의 사료적 가치와 메탄 발생량을 조사하기 위해 in vitro 발효 실험을 수행하였다. Garlic husk를 제외하고 국내에서 널리 사용되는 4가지 조사료 원료(Oat hay, Annual ryegrass, Timothy, Tall fescue)를 실험의 처리구로 구성하였다. In vitro 48 시간 발효 후 건물 소화율, 섬유소 소화율, 암모니아태 질소, 휘발성 지방산, pH, 메탄 발생량을 평가하였다. 가스 발생량은 3, 6, 9, 12, 24, 36 및 48 시간에 각각 측정되었다. 최종 가스 발생량에서 Oat hay가 유의적으로 가장 높았다(p<0.01). Garlic husk는 처리구 중 가장 높은 메탄 발생량을 나타내었다(p<0.01). Garlic husk의 건물 소화율은 Oat hay보다 유의적으로 낮았지만 Annual ryegrass, Timothy, Tall fescue보다 높았다(p<0.01). 섬유소 소화율은 Annual ryegrass에서 가장 낮았다(p<0.01). 총 휘발성 지방산 생성량은 Oat hay에서 가장 높았고(p<0.01), Garlic husk, Timothy, Tall fescue는 유사하였다. 본 연구의 in vitro 발효 성상과 메탄 발생량을 고려할 때, Garlic husk는 국내 주요 조사료를 대체할 영양적 가치를 포함하고 있으나 메탄 저감 효과는 미미할 것으로 추정된다.

This in vitro study investigated the enhancement of rumen bacterial adhesion on a substrate to increase the digestibility of rice straw in Hanwoo cattle. The rice straw was pretreated with enzymes, probiotics, or ammonia, and the effects on the enhancement of bacterial adhesion and fiber degradation were analyzed using in vitro rumen fermentation. Enzyme treatment included spraying of cellulase and xylanase at 40 and 120 U per g of rice straw, respectively; Saccharomyces cerevisiae culture of 1.0×107 CFU was sprayed as a probiotic treatment per gram of rice straw; ammonia was sprayed at 0.3% per gram of rice straw. Following enzyme treatment, Fibrobacter succinogenes formed a higher adhesion colony than the control group (7.26±0.03 and 8.43±0.20) after 6h and 12h of in vitro culture (p<0.05), respectively. Attachment of Ruminococcus flavefaciens also increased following enzyme treatment (p<0.05) after 6 and 12 h compared to that of the control (5.18±0.06 and 6.60±0.15); and R. albus attachment showed a significant increase compared to that of the control (5.94±0.15) after 6 h of incubation (p<0.05). Probiotic treatment increased attachment of F. succinogenes in comparison with untreated rice straw after 6 h and 12 h of fermentation (p<0.05); R. flavefaciens attachment showed an increase only after 6 h of culture (p<0.05); R. albus was not affected. Attachment of F. succinogenes, R. flavefaciens, and R. albus increased with ammonia treatment after 6 h and 12 h (p<0.05). Dry matter digestibility was higher after the enzyme treatment (3.45±0.21 and 7.04±0.09) than in the control group(1.85±0.08 and 3.94±0.04) after 6 and 12 h of in vitro culture (p<0.05), respectively. It was also higher than that of untreated rice straw after probiotic and ammonia treatments (p<0.05). There was an increase in the enhancement levels of bacterial adhesion depending on the type of fibrolytic bacteria following enzyme, probiotic, and ammonia treatment. These treatments improved digestibility. This enhancement is considered to be greater following enzyme and ammonia treatments than with probiotics.

The objective of this study was to evaluate effects of feeding methods on in vitro ruminal fermentation, total gas and methane production in Hanwoo steers. Six Hanwoo steers fitted with rumen cannula (430 ± 21 kg of body weight) were randomly assigned to one of three feeding systems: 1) feeding forage 1 hour after concentrate, 2) feeding concentrate 1 hour after forage, 3) feeding mixed ration. Rumen fluid sampled from each animals was incubated 24 hours with maize or timothy substrates in in vitro. Ruminal pH was increased in feeding method 2 or maize substrate than that of other methods or timothy substrate (P < 0.001). The production of total volatile fatty acid, acetate, propionate, butyrate, and valerate were increased when steers fed diets using feeding method 1 or rumen fluid was incubated with maize substrate (P < 0.001). Increased production of total gas and methane was observed in feeding method 1 and maize substrate compared to those of other methods or timothy substrate (P < 0.001). Due to the inconsistent results between ruminal fermentation and gas production in this study, further research is required to estimate effects of feeding method on enteric fermentation and gas production in in vivo.

In this study, the effect of forage sources in the total mixed ration (TMR) on in vitro goat rumen fermentation was investigated. Rice straw (RS), Italian ryegrass (IRG), timothy (TIM), and alfalfa (ALF) were used as forage sources. Each forage source was mixed with a commercial goat concentrate diet in the ratio of 1:1. Total 4 TMR were prepared. Rumen simulated in vitro fermentation using goat rumen fluid collected from the slaughterhouse was conducted until 72 th . For fermentation parameters, gas production (GP), volatile fatty acids (VFAs), and ammonia nitrogen (NH3-N) were examined. All assays were performed at 24 th , 48 th , and 72 th h of incubation individually. Contents of crude protein and non-fibrous carbohydrate were greater in the order of RS < IRG < TIM < ALF. Significant treatment effects were found in valerate and NH3-N at 24 th h of incubation (p<0.05). ALF showed the greatest contents of them and RS was the lowest. At 48 th incubation, a significant effect was detected at GP (p<0.05) and RS was greater than others. However, GP of RS was lower than others at 72 th . Significant effects on Total VFA, butyrate, and valerate productions were found at 72 th h of incubation (p<0.05). ALF showed the greatest production. Methane production from all treatments was not significantly different for each incubation time (p>0.05). The present study provided primary information on how goat rumen fermentation responds to different nutrient contents and forage sources of TMR. And the information could be used for the design or optimizing economical diet formulation for goats.

This study evaluated effects of the addition of garlic powder allicin (GPA) mixture on rumen fermentation with methane in Hanwoo steer. On in vitro trial, two experimental groups were used: control (without GPA) and treatment group (addition of 0.1% GPA mixed with the basal concentrate). Similar to in vitro trial, two experimental groups were used in vivo trial. Five Hanwoo steers (3 steers in one group and 2 in another group; average body weight = 500 ± 43 kg) were assigned by crossover design for 20 d consists of 15 d diet adaptation and 5 d data collection in each experimental period. Daily feed intake and enteric methane production were recorded by an automated head chamber system. The results of in vitro study showed that the GPA treatment group had higher acetic acid (24.30 vs 23.45 mmol/L) and butyric acid (16.55 vs 15.47 mmol/L) concentrations, but lower CH4 production (1.40 vs 2.71 mmol/ml) after 24 h of incubation compared to the control (p<0.05). Total gas, propionic acid, total volatile fatty acid (VFA), and acetic acid: propionic acid ratio were not affected by treatment after 24 h incubation. In the in vivo experiment, rumen pH and VFA were not significantly different between treatments (p>0.05), except acetic acid, which was significantly higher in GPA mixture group (60.97 vs 53.94 mM) than in the control group (p<0.05). Furthermore, no significant differences were recorded in CH4 production (g/d) and CH4 yield (g/kg DMI) between the two groups (p>0.05). In conclusion, the addition of 0.1% GPA mixture reduced CH4 proudcition on in vitro trial, but no effect on in vivo trial.

In in vitro fermentation studies, feed samples can either be included in the in vitro rumen medium using filter bags or can be directly dispersed. The objective of this study was to investigate the effects of different pore sizes of filter bags on the rumen fermentation characteristics in an in vitro system. Corn, soybean meal, and timothy were ground to pass through a 1.0-mm screen and were formulated in the ratio of 70:7:23 based on DM, respectively. The formulated experimental diet (2g/DM) was put in F57 filter bags and R510 nylon bags (Ankom®) which pore sizes were 25 and 50 μm, respectively. An in vitro study was conducted to determine the rumen fermentation characteristics for 3, 6, 12, 24, and 48 h and rumen microbial community at 48 h of incubation. A significantly higher production of gas was observed in the R510 bags than in F57 at all the incubation times (p<0.01). IVDMD (p<0.01) and IVNDFD (p<0.01) were significantly higher, whereas pH (p<0.01) and NH3-N (p<0.01) were lower when R510 bags were used. In the VFA composition, acetate and butyrate were significantly higher (p<0.01) in R510 bags, and propionate and total VFA concentration did not differ (p=0.55 and 0.25, respectively) between F57 and R510 bags. The log copy numbers of bacteria and protozoa did not differ (p=0.69 and 0.94, respectively) between F57 and R510 bags, whereas those of fungi were significantly higher in R510 than in F57 bags (p<0.01). Therefore, the use of R510 may reflect actual rumen fermentation characteristics more precisely than those of F57 because increased gas production, nutrient digestibility and acetate, butyrate proportion were founded in R510.

Fermented total mixed ration (TMR) is a novel feed for ruminants in South Korea. The purpose of this study was to evaluate the effects of lactic acid bacteria (LAB) on the quality of TMR and in vitro ruminal fermentation. Strains of three LAB spp. (Lactobacillus plantarum, L. brevis, L. mucosae) were used in fermentation of TMR. Inoculations with the three LAB spp. lowered pH and increased concentrations of lactic acid, acetic acid, and total organic acid compared to non-LAB inoculated control (only addition of an equivalent amount of water) (p<0.05). Bacterial composition indicated that aerobic bacteria and LAB were higher. However, E. coli were lower in the fermented TMR than those in the control treatment (p<0.05). Among the treatments, L. brevis treatment had the highest concentration of total organic acid without fungus detection. Gas production, pH, and ammonia-nitrogen during ruminal in vitro incubation did not differ throughout incubation. However, ruminal total VFA concentration was higher (p<0.05) in the LAB spp. treatments than the control treatment at 48 hours. Overall, the use of L. brevis as an inoculant for fermentation of high moisture. TMR could inhibit fungi growth and promote lactic fermentation, and enhance digestion in the rumen.

The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 μm, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

본 시험은 항염에 효과가 있다고 알려진 식물 추출물의 첨가가 반추위 발효와 메탄 생성에 미치는 영 향을 알아보기 위해 in vitro 실험을 수행하였다. 항염과 항산화 효과가 있는 산뽕나무, 뽕나무, 예덕나 무, 오동나무, 방아풀과 은행나무 6종을 선발하였다. 반추위액과 McDougall buffer 혼합액 15mL과 티 모시 0.3g, 각각의 추출물을 기질의 5%로 넣고 39℃에서 3, 9, 12, 24, 48 그리고 72시간 배양하였다. 항염 효과가 있는 식물 추출물의 첨가는 반추위 발효 성상(pH, 건물소화율, Glucose 농도, 암모니아 농도, 단백질 농도, 미생물 성장량, 휘발성지방산)에는 영향을 미치지 않았다. 총 가스 발생량은 각기 다른 양상을 보였으며, 이산화탄소 발생량은 48시간대에서 예덕나무와 방아풀에서 대조구에 비해 유의 적(p<0.05)으로 높았다. 또한 메탄 발생량은 배양 초기에는 대조구보다 첨가구에서 유의적(p<0.05)으로 감소하였으나, 발효가 진행될수록 대조구에 비해 첨가구에서 더 많은 메탄이 발생하였다. Polyphenol과 flavonoid는 은행나무 추출물구에서 가장 높았다. 본 시험의 결과에서 항염에 효과가 있는 식물 추출물 을 in vitro 반추위 배양액에 첨가하였을 때, 반추위 발효에는 영향을 미치지 않았고, 메탄은 초기 발효 에 저감 효과가 있는 것으로 사료된다.

This study was to research the relationships between rice straw degradation and changes of fibrolytic bacteria population during the in vitro rumen fermentation. Dry matter(DM) digestion of rice straw and population of fibrolytic bacteria were measured at the 0. 4, 8, 12 and 48 hours during the incubation. The populations of F. succinogenes. R. albus and R. flavefaciens were defined as log copy number of 16S rDNA by technical method of Quantitative real-time PCR. Total population of F. succinogenes, R. flavefaciens and R. albus was sum of bactera attached on rice straw and suspended in medium. It's population was increased with incubation, reached top level of 29.0 Log copy No at the 24 hour and then decreased. In the meantime, DM digestion of rice straw showed the higher increasement from the 8 hour to the 24 hour than from the 0 hour to the 8 hour, and then a slowdown in increasing trend of digestibility. Attachments of F. succinogenes, R. flavefaciens and R. albus were detected immediately after start of in vitro rumen incubation. At the same time, the colonized bacterial share were respectively 34.5%, 84.4% and 67.9% in total population. All of them was reached the highest colonized bacterial share above 94.7% at the 4 hour incubation. However population of attached bacteria was shown the highest level at the 12 hour or the 24 hour incubation. Kinetics of colonization were formed area of top speed from the 12 hour to the 24 hour and respectively reached 10.33, 9.28 및 8.30 Log copy No/h/g DM at the 24 hour by F. succinogenes, R. flavefaciens and R. albus. The kinetics of rice straw degradation was formed top level of 0.95% DM/h at the 24 hour. The present results gave clear evidence that degradation of rice straw was increased with the development of total fibrolytic bacteria in process of rumen fermentation. Also, their attachment was largely occurred immediately after insertion of rice straw, the colonized bacteria was actively proliferated, and then degradation of rice straw was maximized.