The present study was conducted to investigate the effects of feed types (powder vs. pellet) from potato by-product containing glycoalkaloid on rumen fermentation with different rumen pH (low vs. high) using in vitro technique. Rumen fluid was collected from two cannulated Hanwoo heifers before morning feeding (high rumen pH at 6.71) and 3 h after feeding (low rumen pH at 6.20). A 0.5 gram of powder or pellet from potato by-product was incubated in the rumen buffer for 48 h at 39℃. Four replications for each treatment were used along with two blanks. Pellet had higher (p<0.05) the immediately degradable fraction, the fractional degradation rate, and the lag phase than those of powder. However, feed type had no effects on the potentially degradable fraction and the total degradable fraction. In fermentation characteristics, powder had higher (p<0.05) concentrations of ammonia-N, total volatile fatty acid (VFA), and acetate than those of pellet. In addition, a high rumen pH had higher (p<0.05) concentrations of ammonia-N and acetate, but lower (p<0.05) total VFA concentration than those of a low rumen pH. The interaction effects between feed type and rumen pH were found on the total degradable fraction and concentrations of ammonia-N, total VFA, and acetate. The present study concluded that generally feed type had no effects on rumen degradation kinetics, but potato by-product in the powder form is recommended due to produce higher fermentation characteristics than in the pellet form.

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.

This study was conducted to research on the efficacy of chemical treatment as an effective method for reducing mycotoxin in rice straw silage. As a chemical treatment method, ammonia and sodium hydroxid were treated at 4% level of rice straws contaminated with mycotoxin, and the effects of silage storage on fungal toxin reduction, fermentation quality, and fiber digestion were evaluated. Aflatoxin B1, B2, G1, G2 and fumonisin B1, B2 as well as deoxynivalenol were not detected in all experimental groups, and ochratoxin A and zearalenone were detected. Ochratoxin A was detected lower in the chemical treatment than control (41.23 g / kg) (p<0.05). Zearalenone showed lower results in sodium hydroxide treatment (297.44 μg / kg) than control (600.33 μg / kg) and ammonia treatment (376.00 μg / kg) (p<0.05). The pH of rice straw silage was the lowest in ammonia treatment and the highest in sodium hydroxide treatment (p<0.05). The lactic acid contents of control and ammonia treatments were similar, but sodium hydroxide treatment was the lowest (p<0.05). Propionic acid was higher in the control than in the chemical treatments (p<0.05), and showed similar contents in the ammonia and sodium hydroxide treatment. Both the rumen microbial degradation rate of NDF and ADF showed the highest in sodium hydroxide treatment, followed by ammonia treatment, and the control showed the lowest level (p<0.05). Therefore, the results of this study are demonstrated to have a good effect on the treatment of ammonia and sodium hydroxide to reduce the mycotoxins and increase the rumen microbial degradation rate in the rice straw silage. Sodium hydroxide treatment was more effective in reducing mycotoxins and improving fiber degradation rate than ammonia treatment, but it is thought to have an inefficient effect on silage fermentation in rice straw silage.

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.

This study was performed to investigate the effect of heat-stressed environment on rumen microbial diversity in Holstein cows. Rectal temperature and respiration rate were measured and rumen fluid was collected under normal environment (NE; Temperature humidity index (THI)=64.6) and heat-stressed environment (HE; THI=87.2) from 10 Holstein cows (60±17.7 months, 717±64.4 kg) fed on the basis of dairy feeding management in National Institute of Animal Science. The rumen bacteria diversity was analyzed by using the Illumina HiSeq™ 4000 platform. The rectal temperature and respiratory rate were increased by 1.5ﾟC and 53 breaths/min in HE compared to that in NE, respectively. In this study, HE exposure induced significant changes of ruminal microbe. At phylum level, Fibrobacteres were increased in HE. At genus level, Ruminococcaceae bacterium P7 and YAD3003, Butyrivibrio sp. AE2032, Erysipelotrichaceae bacterium NK3D112, Bifidobacterium pseudolongum, Lachnospiraceae bacterium FE2018, XBB2008, and AC2029, Eubacterium celulosolvens, Clostridium hathewayi, and Butyrivibrio hungatei were decreased in HE, while Choristoneura murinana nucleopolyhedrovirus, Calothrix parasitica, Nostoc sp. KVJ20, Anabaena sp. ATCC 33047, Fibrobacter sp. UWB13 and sp. UWB5, Lachnospiraceae bacterium G41, and Xanthomonas arboricola were increased in HE. In conclusion, HE might have an effect to change the rumen microbial community in Holstein cows.

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).

This study was aimed to determine the effects of hybrid and inoculant on in vitro rumen digestibility and rumen fermentation indices of corn silage. Two corn hybrids(Kwangpyeongok (KW) vs. Pioneer 1543(PI)) were ensiled for 100 days with two inoculants(L. plantarum(LP) vs. L. buchneri(LB)) applied at 1.2 × 105 cfu/g of fresh forage. Silages were incubated in rumen fluid medium at 39℃ for 72 hours. Both of KW and PI had no difference(p>0.05) on in vitro digestibility of dry matter(IVDMD) and neutral detergent fiber(IVNDFD), while application of LB produced higher(p<0.05) IVDMD and IVNDFD than LP. Hybrid and inoculant applications did not affect(p>0.05) total gas volume, ammonia-N, total VFA, and acetate concentrations in the rumen. Applied LB was higher(p<0.05) propionate concentration than LP, while hybrids had no effect(p>0.05). Furthermore, the interaction effect(p<0.05) was occurred in propionate concentration. Butyrate concentration did not affected(p>0.05) by hybrid and inoculant applications, but there was an interaction effect between hybrid and inoculant(p<0.05). Applications of hybrid and inoculant had effects(p<0.05) on iso-butyrate and valerate, without any interaction effects(p>0.05). This study concluded that application of KW and PI has no effects on in vitro rumen digestibility and rumen fermentation indices of corn silage, while application of LB increase the rumen digestibility in both of corn hybrids.

This study was conducted to investigate the relationship between changes of rumen microflora and bloat in Jersey cow. Jersey cows (control age: 42 months, control weight: 558kg; treatment age: 29 months, treatment weight 507kg) were fed on the basis of dairy feeding management at dairy science division in National Institute of Animal Science. The change of microbial population in rumen was analyzed by using next generation sequencing (NGS) technologies due to metabolic disease. The diversity of Ruminococcus bromii, Bifidobacterium pseudolongum, Bifidobacterium merycicum and Butyrivibrio fibrisolvens known as major starch fermenting bacteria was increased more than 36-fold in bloated Jersey, while cellulolytic bacteria community such as Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens was increased more than 12-fold in non-bloated Jersey. The proportion of bacteroidetes and firmicutes was 33.4% and 39.6% in non-bloated Jersey’s rumen, while bacteroidetes and firmicutes were 24.9% and 55.1% in bloated Jersey’s. In conclusion, the change of rumen microbial community, in particular the increase in starch fermenting bacteria, might have an effect to occur the bloat in Jersey cow.

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 research was conducted to investigate the effects of feeding high and low forage diets with different forage sources on rumen fermentation characteristics and blood parameters of Holstein cows during the dry period. Eight Holstein cows were completely randomized assigned to two groups and repeated measurement was utilized in the analysis. Cows in two treatments were fed with diets with high (F:C = 70:30, 70F; forage source: mixed-sowing whole crop barley and Italian ryegrass silage, BIRG) and low (F:C = 55:45, 55F; forage source: tall fescue hay, TF) forage level. Rumen fluid pH was higher in 70F group. Levels of acetic acid, propionic acid, and butyric acid showed a similar pattern: from the lowest value at 07:30 h to the highest at 10:30 h and then decreased in both groups. The ratio of acetic acid to propionic acid was significantly higher (p < 0.05) in 55F group at 09:30 and 10:30 h. Rumen fluid NH3-N concentrations were significantly higher (p < 0.05) in 70F group at 09:30 and 10:30 h. Blood urea nitrogen was significantly higher (p < 0.05) in 70F group. It was concluded that BIRG based diet with a high forage level had no adverse effects on rumen fermentation, some blood chemical parameters, and immune system in dry Holstein cows and could be used as a forage source instead of imported TF.

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.

본 연구는 약용실물 수피 추출물을 이용하여 in vitro 반추위 발효성상 및 반추위 메탄 발생에 미치 는 영향을 알아보기 위해 수행되었다. 공시재료는 무궁화, 음나무, 팽나무, 두충 및 후박나무 추출물을 사용하였다. McDougall’s buffer와 반추위액을 2:1 비율로 혼합한 발효액을 티모시 0.3g과 식물 추출 물이 담긴 50mL serum bottle에 혐기상태로 20mL 분주한 뒤, 39℃에서 3, 6, 9, 12, 24, 48 및 72시 간 동안 발효하였다. 약용식물 수피 추출물을 첨가하였을 때 반추위 발효 성상(pH, 건물소화율, 암모니 아태 질소, 미생물 성장량)에는 영향을 미치지 않았다. 배양 6시간대에 두충 추출물 첨가구에서 acetate의 농도가 대조구에 비해 유의적(p<0.05)으로 낮고, 메탄의 기질인 이산화탄소의 발생량이 대조 구에 비해 유의적(p<0.05)으로 낮아 메탄 저감 효과를 기대하였으나 메탄 저감효과는 없었다. 메탄생성 박테리아인 Methanogen archaea의 경우 무궁화 추출물 첨가구를 제외한 전 추출물 첨가구에서 대조구 에 비해 유의적(p<0.05)으로 높았으나 메탄발생량에는 유의적(p<0.05)인 차이가 없었다. 추후 첨가 농 도를 달리하거나 flavonoid 함량을 측정하는 실험이 필요한 것으로 사료되며, 메탄생성박테리아를 감소 시키는 연구도 추가적으로 필요할 것으로 생각된다

This study was conducted to investigate the effects of antiprotozoal plant extracts on in vitro ruminal fermentation and methane emission. Ruminal contents were collected from two ruminally-fistulated steers (Average BW: 450±30 kg) consuming 60% Timothy hay and 40% concentrate with free access to water and mineral block. Plant extracts, Lonicera japonica, Torreya nucifera, Zanthoxylum schinifolium, Poncirus trifoliate and Citrus junos, which have been known to have antiprotozoal and antiparasitic effects were added (5% of substrate) to the in vitro ruminal fermentation bottles containing the mixture of rumen fluid and McDougall’s buffer. The McDougall buffer (1000 ml) and ruminal inoculums (500 ml) were combined and 20 ml added to the 50 ml fermentation vessels containing 300 mg of substrate (DM basis). The fermentation vessels were placed in an incubator at 39°C for 3, 6, 9, 12, 24, 48 and 72 h with shaking. Plant extracts addition was decreased (p<0.05) acetate:propionate compared with control except with T. nucifera addition. Total gas production was increased (p<0.05), whereas methane emission was decreased (p<0.05) by all plant extracts compared with control at 12 h incubation. Methane emission at 24 h incubation was decreased for Lonicera japonica, Torreya nucifera, and Zanthoxylum schinifolium compared with control as well. In addition, methanogenic archaea was decreased (p<0.05) by all plant extracts at the 12 h incubation. These results indicate that the plant extracts may decrease methane emission by inhibiting methanogenesis without the decrease of total gas production in 12 h in vitro ruminal fermentation. Therefore, plant extracts which have the antiprotozoal effect may be promising feed additives for ruminants as a methane inhibitor