• Title/Summary/Keyword: inoculum

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In vitro Biohydrogenation of Linolenic and Linoleic Acids by Microorganisms of Rumen Fluid (반추위액의 미생물에 의한 In vitro 상에서의 리놀렌산과 리놀산의 Biohydrogenation)

  • Lee, S.W.;Chouinard, Yvan;Van, Binh N.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.985-1000
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    • 2005
  • In vitro anaerobic incubations of timothy (Phleum pretense L.) forage with bovine rumen fluid were conducted at 39℃ for 0, 3, 6, 9, 24, and 36 h in three trials to examine the biohy- drogenation of linolenic (C18:3) and linoleic acids (C18:2) and their bypass from the rumen. The objectives of the first trial was to study the effect of growth stage (stem elongation, early heading, late heading, and early flowering) and N-fertilization (0 and 120 kg N ha-1) on in vitro biohydrogenation of C18:2 and C18:3. The hydrogenable fraction, the effective disappearance and the bypass of C18:2 and C18:3 were high in timothy harvested at stem elongation, and decr- ease linearly with maturity. The N-fertilization increased the hydrogenable fraction of C18:3, the effective disappearance and the bypass of C18:2 and C18:3. However, the rate of disappearance of C18:2 and C18:3 were not affected by maturity and N-fertilization (P>0.1). In trial 2, the effect of timothy conservation method on in vitro C18:2 and C18:3 biohydrogenation was determined. Silage had the highest effective disappearance of C18:2 and C18:3, and grass hay had lowest one. The amounts of C18:2 and C18:3 biohydrogenated were higher in haylage and silage than in grass hay. Comparative to haylage timothy, the bypass of C18:3 was higher in fresh grass, wilted grass and grass hay. The bypass of C18:2 was higher in fresh grass and silage in comparison to grass hay and haylage. In trial 3, the effects of formic acid and Lactobacillus plantarum inoculum addition to timothy haylage and silage on C18:2 and C18:3 disappearance and bypass were studied. Haylage and silage additives had no effect (P>0.1) on effective disappearance and bypass of C18:2 and C18:3. The addition of formic acid increased the rate of biohydrogenation of C18:3 in haylage and silage, but it decreased the hydrogenable fraction of C18:2 in silage. The results of these three incubation trials show that the hydrogenable fraction and the bypass of C18:2 and C18:3 in timothy decreased with maturity and increased with N-fertilization. Higher amount of C18:2 and C18:3 were biohydrogenated in haylage and silage than in grass hay, and C18:3 ruminal disappearance was higher in fresh grass, wilted grass and grass hay than in haylage.

The Cd and Pb Accumulation in Various Tissues of Rooted Cuttings of Four Populus Species Inoculated with Ectomycorrhizal Fungi, Pisolithus tinctorius (모래밭버섯 균근균(菌根菌)으로 접종(接種)한 포플러 4개(個)개 수종(樹種) 삽목묘(揷木苗)의 체내(體內) 부위별(部位別) Cd과 Pb 축적(蓄積) 특성(特性))

  • Han, Sim Hee;Lee, Kyung Joon;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.90 no.4
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    • pp.495-504
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    • 2001
  • The objectives of this study were to understand the characteristics of Cd and Pb accumulation in various tissues of poplar species and the effects of ectomycorrhizal inoculation on the accumulation of above two heavy metals in the tissues. The mycelial inoculum of ectomycorrizal fungus, Pisolithus tinctorius was produced in peatmoss and vermiculite mixtures, and inoculated into potted soil with fresh cuttings of four species of Populus, P. alba ${\times}$ glandulosa (Pag). P. koreana ${\times}$ nigra var. italica (Pkn), P. nigra ${\times}$ maximowiczii(Pnm), and P. euramericana(Pe). The potted soils were added with 0, 30, and 80 ppm Cd, and 0, 50, and 300 ppm Pb. The cuttings were grown outdoors for about five months until the plants were harvested for measurement of growth, mycorrhizal infection, and metal contents in leaves, stems, and roots. The total dry weight of Pe treated with Cd and Pb was increased by mycorrhizal inoculation, while that of three other species was not affected by the inoculation. Cd was accumulated in the highest concentration in Pag and its concentration was increased by four times by mycorrhizal inoculation. The Pag accumulated Cd in the highest concentration in the leaves, while three other species accumulated Cd most in the roots. Pb was accumulated in the highest concentration in the roots of all the four species, while Pkn accumulated Pb in the leaves as much as in the roots. Without mycorrhizal inoculation Pe accumulated Pb most among the four species, while Pkn with mycorrhizal inoculation accumulated Pb two times more than in the same species without mycorrhizal inoculation. It was concluded that Pag was the most effective species among the four poplar species in Cd absorption from contaminated soil, and that Pe instead effectively absorbed Pb. Mycorrhizal inoculation increased the Cd accumulation in the tissues by four times in Pag and also increased Pb accumulation by two times in Pkn, with leaves being the major sites of metal accumulation. It may be possible to use these two poplar species in remedying the metal in the soil through the raking and removing the litter out of the contaminate site.

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Inheritance of Resistance to Phytophthora capsici by Inoculums in Korean Hot Pepper (고추 역병균의 접종원에 따른 역병 저항성의 유전 양식)

  • Soh, Jaewoo;Han, Kyung-Sook;Lee, Sung-Chan;Lee, Jung-Sup
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.317-323
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    • 2012
  • The study aims to identify the pathogenicity of Phytophthora. capsici isolates in major pepper-producing areas in Korea and the inherit genetic pattern of phytophthora blight resistance by inocula. With five kinds of testing materials including 'Kataguma (Sakata Korea)' peppers, a disease-susceptible material, '#308', a phytophthora blight resistance material, 'CM334', and their $F_1$ and $F_2$, respective isolates of P. capsici obtained from Icheon, Eumseong, Buan, Imsil and Yeongyang regions together with six kinds of peppers' inoculum including PA-159 (KACC No.40482) received from Korean Agricultural Culture Collection were used for inoculation. The disease-susceptible material '#308', the resistant material 'CM334' and the non-segregating generation of $F_1$ represented 4.94-5.00, 1.00-1.07, and 1.01-1.08 phytophthora blight incidence respectively in the group comparison by isolate. This result means that the phytophthora blight resistance was clearly distinguished among testing materials in the group comparison by P. capsici isolate. Moreover, $F_2$ segregating generation showed 1.79-2.31 phytophthora blight incidence which turned out to be identical in the group comparison by the six isolates of P. capsici isolate and with similarity between both the resistant and susceptible materials. Thus, the result proved that using the six isolates of P. capsici tested as inocula was suitable to investigate the phytophthora blight resistance. When it comes to group comparison of $F_2$ segregation generation, however, isolates were divided with PA-159 isolate being the center: a group consisting of isolates from Icheon, Buan, and Imsil and a group consisting of Yeongyang and Eumseong isolates with higher pathogenicity. The expected segregation ratio of the phytophthora blight resistance in $F_2$ generation by isolate was analyzed. PA-159 isolate showed 3:1 or 9:3:3:1, indicating that one to two genes were involved. On the other hand, results also proved that there is an interaction of genes since both Eumseong and Yeongyang isolates showed a segregation ratio of 11:5 while the Icheon isolate represented 12:3:1.

Fermentation Properties and Increased Health Functionality of Kimchi by Kimchi Lactic Acid Bacteria Starters (김치 유산균 Starter를 이용한 김치의 발효 특성 및 기능성 증진 효과)

  • Bong, Yeon-Ju;Jeong, Ji-Kang;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1717-1726
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    • 2013
  • Fermentation characteristics and health functionalities of kimchi by inoculating kimchi lactic acid bacteria (LAB) starters were studied. We manufactured single LAB starter kimchi (Lactobacillus plantarum pnuK, Lactobacillus plantarum 3099K, Leuconostoc mesenteroides pnuK), mixed LAB starter kimchi (Lb. plantarum pnu/Leu. mesenteroides pnuK, Lb. plantarum 3099/Leu. mesenteroides pnuK) with inoculum size of $10^6$ CFU/g, as well as naturally fermented kimchi (NK), and fermented them for 6 days at $15^{\circ}C$. The pH and acidity of the early phase of fermentation were not different, but kimchi with the starters showed rapid changes in the pH and acidity from 2 days of fermentation. As the fermentation progressed, the level of total aerobic bacteria and Lactobacillus sp. increased similarly with or without Lb. plantarum (LP) inoculation. However, the level of Leuconostoc sp. was high in kimchi inoculated with Leuconostoc sp. starter. In the sensory evaluation test, kimchi with starters received higher overall acceptability scores than those of NK; mixed starter added kimchi earned the highest score. In DPPH and hydroxyl radical scavenging activity, kimchi with the starters exhibited higher activity than that of NK. In the MTT assay of HCT-116 and HT-29 human colon cancer cells, NK showed inhibition rates of 63.4 and 51.9%, but LPpnuK achieved 77.1 and 68.8%, respectively. This study showed that inoculating starters in kimchi increased in vitro antioxidant and anticancer activities, and single starter (LP) added kimchi revealed higher functionality than the kimchi with mixed starter. Kimchis with the starters effectively up-regulated the gene expressions of the pro-apoptotic gene of Bax, but down-regulated Bcl-2. They promoted expressions of p53 and p21, and suppressed expressions of inflammation-related genes, iNOS and COX-2, compared with NK. Taken together, it is expected that using starters may help manufacture kimchi with improved sensory quality and health functionality.

Development of an Efficient Screening Method for Resistance of Chili Pepper Plants to Meloidogyne incognita (뿌리혹선충 Meloidogyne incognita에 대한 저항성 고추를 선발하기 위한 효율적인 검정법 확립)

  • Hwang, Sung Min;Jang, Kyoung Soo;Choi, Yong Ho;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.282-293
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    • 2016
  • Root-knot nematodes (Meloidogyne spp.) are major plant pathogens that cause reductions in yield and quality of several solanaceous crops, including pepper (Capsicum spp.). These losses can be averted through planting of resistant cultivars. Plants are defined as resistant when they suppress nematode reproduction. In this study, the resistance degrees of 102 commercial cultivars of chili pepper (Capsicum annuum) to a root-knot nematode, Meloidogyne incognita, were evaluated by comparing the number of egg masses on their roots to those of 'PR huimangchan', a highly susceptible cultivar that exhibited the most egg masses of the chili pepper cultivars evaluated. Among these cultivars, forty-four (43.1%) showed resistance to M. incognita and eighteen (17.6%) were moderately resistant. The other cultivars (39.3%) were determined to be susceptible. For further study, six chili pepper cultivars (i.e., Gangryeokjosenggeon, Shinsegae, Muhanjilju, PR Bulrocho, PR Huimangchan, and Jjang) with different levels of resistance to the nematode were selected. Changes in resistance of the six cultivars under several conditions, such as inoculum concentration, plant growth stage, and cultivation period after transplanting were investigated. We found that an efficient screening method for resistance of chili pepper to M. incognita is to transplant the chili pepper seedlings 7 days before inoculation, to inoculate 28-day-old plants with M. incognita by loading 5,000 eggs per plant into the pot of soil, to cultivate the plants in a greenhouse ($25{\pm}5^{\circ}C$) for 45-60 days, to measure the number of egg masses on roots of the seedlings, and then to determine the resistance response of the plants by comparing the number of egg masses on the roots with a reference-susceptible cultivar 'PR huimangchan'.

A New Screening Method for the Selection of Calla Lily Zantedeschia aethiopica Cultivars Resistant to Calla Lily Soft Rot (칼라의 무름병 저항성 품종 선발을 위한 검정 방법 개발)

  • Joung, Hyang Young;Choi, Mok Pil;Han, Kyung Sook;Kim, Su;Goo, Dae Hoe;Kang, Yun Im;Choi, Youn Jung;Park, Sang Kun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.366-370
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    • 2013
  • This experiment was carried out to establish the screening methods for the selection of Zantedeschia aethiopica cultivars resistant to calla lily soft rot caused by Pectobacterium carotovorum subsp. carotovorum. A center of leaf and petiole tissues of four Z. aethiopica cultivars were wounded and inoculated with three different inoculum concentrations, $1{\times}10^7cfu/mL$, $1{\times}10^8cfu/mL$, and $1{\times}10^9cfu/mL$, of EccNHRI-21 isolate. And they were evaluated at 4, 18, and 26 hours after inoculation. The lesion of maceration was developed well in the leaf tissues inoculated with $1{\times}10^9cfu/mL$ concentration. And evaluation of resistance in 18 hours after inoculation was correlated with field resistance positively. Using this method, ten Z. aethiopica commercial cultivars and four wild types were screened. 'Crowbrough' and 'White Cutie' were selected as highly resistant genotypes and 'Mont Blanc' and 'Silky White' as resistant genotypes. 'Wedding March' and 'Kiwi Blush' were evaluated as moderate resistant and the others including 'Childsiana' were susceptible. And all of four wild types were evaluated as more than moderate resistant. In spite of control through cultural, biological and chemical methods, the use of resistant cultivars is most efficient to overcome calla lily soft rot. This newly developed screening method was helpful for breeding new varieties resistant to calla lily soft rot.

Distribution and Favorable Conditions for Mycelial Growth of Cordyceps pruinosa in Korea (한국에 자생하는 붉은자루동충하초(Cordyceps pruinosa)의 분포와 균사생장에 적합한 조건)

  • Shin, Jae-Chul;Shrestha, Bhushan;Lee, Won-Ho;Park, Young-Jin;Kim, Soo-Young;Jeong, Gwang-Ryel;Kim, Ho-Kyung;Kim, Tae-Woong;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.79-88
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    • 2004
  • Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.

Development of Eimeriu tenezla in MDEK cell culture with a note on enhancing effeet of preincubation with chicken spleen cells (MDBK 세포 배양에서 Eimeria tenella 발육 상황 및 닭 비장세포에 의한 발육 항진 효과)

  • 채종일;이순형
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.87-100
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    • 1989
  • Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that 2. tenezla first penetrate into the mucosal intraepithelial Iymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope labelled uracil ($^3H-uracil$) . Third, the E. tenella sporozoites viability was assayed after preincubation of them with thicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (I) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schisogonic cycle of E. tenella in 3~4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merogoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48~60 hours, and decreased thereafter. The uptake amount of $^3H-uracil$ depended not only upon the inoculum sixte of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.

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Effect of the Pretreatment by Thermal Hydrolysis on Biochemical Methane Potential of Piggery Sludge (열가수분해 전처리가 양돈 슬러지의 메탄생산퍼텐셜에 미치는 영향)

  • Kim, Seung-Hwan;Kim, Ho;Kim, Chang-Hyun;Yoon, Young-Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.524-531
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    • 2012
  • The objective of this study was to investigate the organic solubilization (SCOD) and improvement of methane production for pig slurry by thermal hydrolysis. A sludge cake was pretreated by thermal hydrolysis at different reaction temperatures (200, 220, 250, $270^{\circ}C$). Ultimate methane potential (Bu) was determined at several substrate and inoculum (S/I) ratios (1:9, 3:7, 5:5, 7:3 in volume ratio) by biochemical methane potential (BMP) assay for 73 days. Pig slurry SCOD were obtained with 98.4~98.9% at the reaction temperature of $200{\sim}270^{\circ}C$. Theoretical methane potentials ($B_{th}$) of thermal hydrolysates at the reaction temperature of $200^{\circ}C$, $220^{\circ}C$, $250^{\circ}C$, $270^{\circ}C$ were 0.631, 0.634, 0.705, $0.748Nm^3\;kg^{-1}-VS_{added}$, respectively. $B_u$ of $200^{\circ}C$ thermal hydrolysate were decreased from $0.197Nm^3\;kg^{-1}-VS_{added}$ to $0.111Nm^3\;kg^{-1}-VS_{added}$ with the changes of S/I ratio from 1:9 to 7:3, and also $B_u$ of different thermal hydrolysates ($220^{\circ}C$, $250^{\circ}C$, $270^{\circ}C$) showed same tendency to $B_u$ of $200^{\circ}C$ thermal hydrolysate according to the changes of S/I ratio. Anaerobic biodegradability ($B_u/B_{th}$) of $200^{\circ}C$ thermal hydrolysate at different S/I ratios was decreased from 32.2% for S/I ratio of 1:9 to 17.6% for S/I ratio of 7:3. $B_u/B_{th}$ of $220^{\circ}C$, $250^{\circ}C$, and $270^{\circ}C$ thermal hydrolysat were decreased from 36.4% to 9.6%, from 31.3% to 0.8%, and from 26.6% to 0.8%, respectively, with the S/I ratio change, respectively. In this study, the rise of thermal reaction temperature caused the decrease of anaerobic digestibility and methane production while organic materials of pig slurry were more solubilized.

Effects of Supplementation of Mixed Methanogens and Rumen Cellulolytic Bacteria on Biochemical Methane Potential (혼합 메탄균과 반추위 섬유소 분해균 첨가가 메탄발생에 미치는 영향)

  • Kim, Ji-Ae;Yoon, Young-Man;Kim, Chang-Hyun
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.515-523
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    • 2012
  • The study investigated the biochemical methane potential (BMP) assay of cellulose supplementing with mixed methanogens and cellulolytic bacteria to improve anaerobic digestion for methane production. For the BMP assay, 7 different microbial supplementation groups were consisted of the cultures of mixed methanogens (M), Fibrobacter succinogenes (FS), Ruminococcus flavefaciensn (RF), R. albus (RA), RA+FS and M+RA+FS including control. The cultures were added in the batch reactors with the increasing dose levels of 1% (0.5 mL), 3% (1.5 mL) and 5% (2.5 mL). Incubation for the BMP assay was carried out for 40 days at $38^{\circ}C$ and anaerobic digestate obtained from an anaerobic digester with pig slurry as inoculum was used. In results, 5% FS increased total biogas and methane production up to 10.4~22.7% and 17.4~27.5%, respectively, compared to other groups (p<0.05). Total solid (TS) digestion efficiency showed a similar trend to the total biogas and methane productions. Generally the TS digestion efficiency of the FS group was higher than that of other groups showing at the highest value of 64.2% in the 5% FS group. Volatile solid (VS) digestion efficiencies of 68.4 and 71.0% in the 5% FS and the 5% RF were higher than other groups. After incubation, pH values in all treatment groups were over 6.4 indicating that methanogensis was not inhibited during the incubation. In conclusion, the results indicated that the hydrolysis stage for methane production in anaerobic batch reactors was the late-limiting stage compared with the methanogenesis stage, and especially, as the supplementation levels of F. succinogenes supplementation increased, the methane production was increased in the BMP assay compared with other microbial culture addition.