• Journal of Ginseng Research
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• 제22권4호
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• pp.304-309
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• 1998

Incubation condition affecting the chlamydospore formation and isolation from mycelia and conidia of Cylindrocarpon destructanse (isolate ACY-9701), isolated from the root rot lesion of the American ginseng (Panax quinquefolium) was investigated. Chlamydospores were formed from mycilia but not from conidia on the Czapek-Dox agar without carbon or nitrogen source after 20 days incubation at 2$0^{\circ}C$. In the medium added with nitrogen and carbon sources, immatured chlamy-dospore-like cells were formed from microconidia and mycelia as well. Immatured chlamydospore-like cells were formed from mycelia as well as microconidia In corn, kidney bean, and pea root extracts after 20 days incubation at 20"C, while typical chlamydospores were formed from both of them in the root extract of Panax quinquefolium. The 3.6 log chlamydospore/mm" was converted from microconidia in the medium, which was equal to 2.5% conidia formed. Under the light condition (251.1 pmol/m" sec, 12 hrs dark and light cycle), 4.2 log/mm" of chlamydospores were converted from interracially or terminal cells of macroconidia, which was 4.0% of macroconidia produced on Potato dextrose agar (PDA). When mycelia and microconidia were stored at -7$0^{\circ}C$ for 32 days and incubated on PDA after thawing at room temperature to isolate chlamydospores from them, microconidia and mycelia were still alive. Meanwhile, microconidial lysis was found after heating them at 32$^{\circ}C$ for 7 days, but the chlamydospores converted from macroconidia were not lysed up to 13 days at 32"C. to 13 days at 32"C.ot;C.

• 한국치위생학회지
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• 제4권2호
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• pp.153-163
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• 2004

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from normal inhabitants of children 's oral cavity, which inhibited the the production of halitosis by anaerobic bacteria. The authors identified the isolates by the lest using API 50 CHL medium kit. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively. 3. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. 4. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 5. When two isolates were tested with API 50 CHL medium kit, those were identified as Lactobaciallius salivarius and Lactobacillus delbrueckii subsp. lactis.

• 생명과학회지
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• 제22권6호
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• pp.723-729
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• 2012

난배양성 미생물의 배양과 보다 다양한 미생물의 분리를 위하여 Filter plate microbial trap (FPMT)을 개발하였고, 이 장치를 사용하여 Greenland의 토양에서 세균을 분리하였다. FPMT는 배지와 하단부의 membrane filter(0.45 ${\mu}m$ pore size)로 구성되어 있다. 시료에 포함된 세균과 화학물질이 FPMT 내로 이동하고, 세균이 증식하는 원리이다. 그 결과, FPMT를 이용한 새로운 in situ 배양방법을 통해 통상적인 Petri dish 배양방법보다 더 많은 종과 기존의 방법으로는 배양할 수 없는 세균을 분리하였다. 이 장치는 앞으로 다양한 환경에서 난배양성 미생물을 분리하는데 유용함을 확인하였다.

• 환경생물
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• 제19권1호
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• pp.87-92
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• 2001

For the biological treatment of industrial wastewater containing high concentration of phenol, isolation and characterization of phenol - degrading bacterium were carried out. A bacterial strain P2 capable of degrading phenol was isolated from contaminated soils by enrichment culture technique and identified as the genus Rhodococcus by morphological, cultural, biochemical characteristics, and Biolog system. The optimal medium composition and cultural conditions for the growth and degradation of phenol by Rhodococcus sp. P2 were 0.1% of (NH$_4$)$_2$SO$_4$, 0.2% of KH$_2$PO$_4$, 0.25% of Na$_2$HPO$_4$ㆍ12$H_2O$, 0.2% of MgSO$_4$ㆍ7$H_2O$, and 0.008% of CaC1$_2$ㆍ2$H_2O$ along with initial pH 8.5 at 3$0^{\circ}C$. Rhodococcus sp. P2 could grow with phenol as the sole carbon source up to 1,800 ppm in batch cultures, but did not grow in medium containing above 2,000 ppm of phenol. When 800 ppm phenol was given in the optimal media, Rhodococcus sp. P2 completely degraded it within 24 h. Meanwhile, 1,800 ppm of phenol was degraded within 9 days. Rhodococcus sp. P2 could utilize toluene, n-hexane, xylene and benzene as sole carbon source .

• The Plant Pathology Journal
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• 제16권4호
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• pp.236-241
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• 2000

A selective agar medium was developed and tested for the isolation of Acidovorax avenae subsp. avenae, the causal bacterial pathogen of bacterial brown stripe, from rice seeds. The new selective agar medium, designated sorbitol pyroglutamic acid agar (SPA) medium, contained 0.5 g of $K_2$HPO$_4$, 3.0 g of Na$_2$HPO$_4$, 2.0 g of D-sorbitol, 0.2 g of L-pyroglutamic acid, 10.0 $m\ell$ of tween 80, 40.0 mg of victoria blue B, 15.0 g of agar, 150.0 mg of ampicillin and 25.0 mg of vancomycin per litter. Colonies of A. avenae subsp. avenae on SPA medium were smooth, round, convex, shiny, blue and 1.5-2.0 mm in diameter 4 days after incubation at 28$^{\circ}C$. Blue colored colony having dark blue zone was typical type of A. avenae subsp. avenae colonies on the medium. Mean recovery of 8 isolates of A. avenae subsp. avenae on the selective SPA medium was 95.8% in comparison to that on KB medium. The saprophytic bacteria were reduced to 97.9% on SPA medium compared to those on KB medium. Most of other rice seedborne bacteria as well as reported pathogenic bacteria were failed to grow on SPA medium. This medium was highly selective for recovering A. avenae subsp. avenae from rice seed samples, and it could be used to enhance the recovery of this bacterium from rice seed samples, which may be contaminated with large numbers of competing microorganisms.

• Preventive Nutrition and Food Science
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• 제7권3호
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• pp.305-309
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• 2002

Raw soybean sprouts were tested for contamination with the following bacteria which have potential for pathogenesis or food spoilage : Salmonella spp., Escherichia coli O157:H7, Yersinia enterocolitica, Vibrio parahae-molyticus, Aeromonas hydrophila, Plesidomonas shigeloides, Pseudomonas aeruginosa, Staphylococcus aureus, Lis-teria monocytogenes, Bacillus cereus, Clostridium perfringens, Campylobacter jejuni, Erwinia spp., and Fusarium spp. Three of the above strains were isolated from the sprouts, and identified by morphological and biochemical methods including an API kit and ATB automated identification system. The isolate cultured in Cereus selective agar, a selective medium, was a Gram-positive, rod shaped, anaerobic spore former. The biochemical and culture tests revealed the following characteristics: catalase-positive, no growth on Simmon's citrate, NO₂ production and requirement of arginine for growth; the ATB automated identification system gave 99.8 % agreement for the identification of Bacillus cereus to the species level. The isolate cultured in Macconkey agar selective medium was Gram-negative, rod shaped and a gas former; the ATB-system gave 99.9% agreement for the identification of Aeromonas hydrophila to the species level. The isolate found in Pseudomonas isolation agar was Gram-negative, rod shaped, cytochrome oxidase-positive, a reducer of nitrates to nitrogen, and pyocyanin producer; the ATB-system gave 99.9 % agreement for the identification of Pseudomonas aeruginosa to the species level. These results indicate that the three bacteria species present in the soybean sprouts were Bacillus cereus, Aero-monas hydrophila, and Pseudomonas aeruginosa. Salmonella spp., Escherichia coli O157:H7, and Yersinia enter-ocolitica, which are associated with serious disease in humans, were not isolated from soybean sprouts examined in this study.

• 한국산림과학회지
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• 제74권1호
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• pp.29-36
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• 1986

이태리포푸라 I-214 (Populus euramericana cv. I-214)의 기내배양(器內培養)한 엽육조직(葉肉組織)에서 원형질체(原形質體) 분리(分離)에 미치는 몇가지 요인(要因)에 대(對)해 조사(調査), 검토(檢討)하였다. 기내(器內)에서 배양(培養)된 아(芽)를 다량(多量)으로 증식(增植)하기 위한 배지(培地)는 MS 기본배지(基本培地)에 $0.1mg/{\ell}$의 BAP를 첨가(添加)한 것이 가상 좋은 성적을 나타냈다. 엽(葉) 1g 당(當) $2.4{\times}10^6$개의 가장 높은 원형질체(原形質體) 분리(分離) 빈도를 나타낸 것은 Cellulase R-10 2 %, Macerozyme R-10 0.8 %, Hemicellulase 1.2 %, Driselase 2.0 %, Pectolyase Y-23 0.05 %에 DTT와 MES 완충액을 첨가(添加)한 후 삼투압 안정제로 0.6 M의 Mannitol을 넣고 pH를 5.6으로 조정한 효소용액(酵素溶液)이었다. CPW 용액(溶液)으로 세정(洗淨)한 후 0.6 M의 Sucrose 용액(溶液)에 처리(處理)한 것이 회수율(回收率) 51.8 %로 가장 높게 나타났다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권10호
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• pp.1407-1415
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• 2016

Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.

• Reproductive and Developmental Biology
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• 제33권3호
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• pp.133-137
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• 2009

Pluripotent embryonic stem (ES) cells isolated from inner cell mass (ICM) of blastocyst-stage embryos are capable of differentiating into various cell lineages and demonstrate germ-line transmission in experimentally produced chimeras. These cells have a great potential as tools for transgenic animal production, screening of newly-developed drugs, and cell therapy. Miniature pigs, selectively bred pigs for small size, offer several advantages over large breed pigs in biomedical research including human disease model and xenotransplantation. In the present study, factors affecting primary culture of somatic cell nuclear transfer blastocysts from miniature pigs for isolation of ES cells were investigated. Formation of primary colonies occurred only on STO cells in human ES medium. In contrast, no ICM outgrowth was observed on mouse embryonic fibroblasts (MEF) in porcine ES medium. Plating intact blastocysts and isolated ICM resulted in comparable attachment on feeder layer and primary colony formation. After subculture of ES-like colonies, two putative ES cell lines were isolated. Colonies of putative ES cells morphologically resembled murine ES cells. These cells were maintained in culture up to three passages, but lost by spontaneous differentiation. The present study demonstrates factors involved in the early stage of nuclear transfer ES cell isolation in miniature pigs. However, long-term maintenance and characterization of nuclear transfer ES cells in miniature pigs are remained to be done in further studies.

• Journal of Life Science
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• 제10권1호
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• pp.64-65
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• 2000

Insoluble phosphate-solubilizing bacteria are routinely screened by a plate assay method using Pikovskaya agar and a modified Pikovskaya medium. A modified Pikovskaya medium to improve the clarity of the yellow-colored halo has not necessarity improved the plate assay. Colonies of phosphate-solubilizing bacteria tested could be redily selected after 48 h of incubation by green-colored colony formation on plate in which bromcresol green(BCG) was included. Among them, two bacterial strains did not produce distinct yellow halos after 48 h of incubation. We suggest that the green colony formation on plate medium containing BCG is advantageous ofr rapid isolating phosphate-solubilizing bacteria directly from soil.