In order to isolate thermophilic compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated SJ21 was selected by agar diffusion method. The strain SJ21 was identified as members of the Bacillus lincheniformis by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain SJ21 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain SJ21 was classified within the genus Bacillus, for which the name Bacillus sp. SJ21 is proposed. The cellulase and xylanase activity of Bacillus sp. SJ21 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. SJ21.
This study was conducted to evaluate effects of mixed microbes addition on physico-chemical, fermentative and microbial parameters of sawdust-based spent mushroom substrates(SMS). The SMS was inoculated with mixed microbes(Enterobacter ludwigii, Bacillus cereus, 2 strains of Bacillus subtillis, Saccharomyces cerevisiae and Lactobacillus plantarum) at 1% level(wet basis) and anaerobically fermented during the different periods(up to 8th week). Compared with the SMS before ensiling, the ensiled one had higher CP, NDF and ADF percentages and lower DM and NFC percentages. However, levels of change were very low. The in situ ruminal disappearance of SMS DM and NDF decreased with the ensiling period prolonged. For fermentative parameters, pH reduced and lactic acid contents increased after ensiling, compared with those after ensiling. At 8th week of ensiling, pH increased and lactic acid contents reduced again, compared with those at 4th week of ensiling; however, the silage still showed favorable fermentation status. Lactic acid bacteria counts did not change throughout 8 weeks of ensiling. Counts of total microbes and yeast reduced after 4th week of ensiling period. Counts of lactic acid bacteria and yeast at 8th week of ensiling were in the levels of 108cfu/g. These results indicate that anaerobic fermentation with microbial addition could be an effective way for the long term(8 weeks) storage of the SMS.
Kim, Y.I.;Jun, S.H.;Yang, S.Y.;Huh, J.W;Kwak, W.S.
Journal of Animal Science and Technology
/
v.49
no.5
/
pp.667-676
/
2007
This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.
Lee, Seul Bi;Kim, Jong Won;Bae, Sung Mun;Hwang, Yeon Hyeon;Lee, Heung-Su;Lee, Byeong Jeong;Hong, Kwang Pyo;Park, Chung Gyoo
Korean journal of applied entomology
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v.57
no.2
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pp.97-104
/
2018
The larva of the white-spotted flower chafer, Protaetia brevitarsis seulensis (Coleoptera: Cetoniidae), is known to have important medicinal properties, such as anti-cancer activity. However, the consumer market for the larvae is depressed because of its high production cost which needs to be reduced. This study was carried out to evaluate two spent mushroom substrates as food source for chafer larvae to improve the productivity and reduce production cost. The larvae were fed with spent substrates of king oyster mushroom (Pleurotus eryngii) (P-SMS) and shiitake mushroom (Lentinula edodes) (L-SMS), with a control of fermented oak sawdust. Effects of the diets on the development of P. brevitarsis seulensis and contents of minor nutrient components in the larvae produced were examined. In the P-SMS diet, the larval rearing period was shorter at 16.2 days, while the rate of larval weight gain and the cocoon weight were higher at 156.3% and 4.1 g, respectively, than that in the L-SMS diet. The pupation rate was 100% and the adult emergence was higher at 93.3% in the P-SMS diet than in the L-SMS diet with no difference to the control diet. In the P-SMS diet, the total nitrogen content during rearing was higher at 10.28% and the minor nutrient component, particularly Fe (145.8 mg/kg), was higher than that in other diets. The oviposition preference of adults showed no significant difference among the diets. Economic analysis showed that P-SMS costs approximately 667,960 Won less per 100 kg of larvae produced, compared to that of the control diet.
Journal of Practical Agriculture & Fisheries Research
/
v.20
no.1
/
pp.35-47
/
2018
A edible mushroom, Clitocybe maxima (Lentinus giganteusis) commercially cultivated in China and Taiwan. However, the researches of cultivation and cultural characteristics were not reported in Korea. In this study, we conducted on cultural characteristics and artificial cultivation of C. maxima. Six isolates were collected from China(3 isolates, commercial strain), Taiwan(1 isolate, commercial strain) and Korea(2 isolates, wild type). C. maxima and L. giganteus collected in China and Taiwan, respectively, are the same in China and are estimated to be of the same species as cultured characteristics. The mycelial growth of the collected strains was not significantly different in agar medium but it showed the best growth in YPMG in liquid culture. Optimum temperature for mycelial growth and induction of fruit body were 25℃ and 30℃, respectively. In order to artificial cultivation of C. maxima, cultural characteristics and artificial cultivation were carried out using agricultural by-products and forestry by-products materials. Mycelial growth was suitable in rice straw, cottonwood sawdust, corncob and rice seed medium, and it was selected as a cultivation medium. The suitable medium for artificial cultivation of C. maxima was selected to mixed medium 2(compounding ratio(v/v): 55% of hardwood sawdust, 5% of cottonseed pellets, 10% of cottonseed, 15% of beet pulp, 15% of swollen rice husks). It took about 30 days to be able to harvest, it was faster than oyster mushrooms. The cultivation period was about 30days. A isolate, CMA-002 was not initiation to fruit body primordiuma on the used cultivation substrate. Other 5 isolates were initiate and development to fruit body on the substrate used in this study. The strain CMA-003 was initiated to be fruiting body by 8~10 days after induction of fruiting body in all of the substrates. Isolate CMA-003 was generate to a bundle fruit body. Other isolates, however, were form fruit body individually. The CMA-003 strain was likely highly recommendable strains for farming. The optimum conditions for the induction and growth of C. maxima fruit body were 25~30℃, 8 hr illumination per day with white fluorescent lamp, 90~95% relative humidity, and 1,500 ppm of CO2 concentration in a cultivation room.
Kim, Hye Soo;Park, Hyun-Young;Kwon, Hyun-Sook;Lee, Sang-Ho;Ha, Jun;Lee, Sang-Won;Cho, Soo-Jeong
Journal of Mushroom
/
v.17
no.4
/
pp.261-267
/
2019
The objective of this study was to evaluate the antioxidant activity of extracts of Protaetia brevitarsis larvae fed on fermented oak sawdust (FOS) or spent mushroom substrates (SMS, Pleurotus eryngii). Total polyphenol content was 32% higher in extracts of larvae fed on SMS (P. eryngii) (75.33±0.43 mg GAE/g) than in extracts of larvae fed on FOS (57.02±1.73 mg GAE/g). The flavonoid content of extracts of larvae grown on FOS and SMS (P. eryngii) was 24.6±0.28 mg/g and 25.4±0.75 mg/g, respectively. DPPH radical scavenging activity increased in an extract concentration-dependent manner, and the DPPH radical scavenging capacity of the extract of larvae produced on SMS (P. eryngii) was higher than that of the larvae produced on FOS. The reducing power of the larval extracts produced on FOS and SMS (P. eryngii) increased in an extract concentration-dependent manner, but there was no significant difference between them. The extract of larvae fed on SMS (P. eryngii) (66.55±0.99 uM TE/g) had a higher oxygen radical absorbance capacity (ORAC) than extracts of larvae grown on FOS (76.32±0.48 uM TE/g). The effect of larval extracts on cell proliferation was investigated using a WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) assay on RAW 264.7 cells. When cells were treated with larval extracts produced on FOS and SMS (P. eryngii) at concentrations of 0, 2, 4, 8, 16, 32, 40, and 64 mg/ml, RAW 264.7 cells proliferated at 90% or more. Therefore, larval extracts produced on FOS and SMS (P. eryngii) were not toxic to RAW 264.7 cells.
To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.
Mutual growth limitation was observation when the two antagonistic fungi was come in contact with each other. Brown line was formed 2day after contact with Trichoderma spp., and then, green spores formed overnight. The laccase activity of L. edodes was stimulated when this fungus wsa co-incubated with Trichoderma spp. for a few days in liquid media. In sawdust-rice bran nixtures, outstanding broun line developed when the two antagonistic fungi co-cultured. The pH of the substrates changed from 5.5 to 4.5 after overgrowth, suggesting a difference in the degradation ability and the preference of the two fungi for the lignocellulose material.
The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.
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