• Title/Summary/Keyword: Screening of resistant strains

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Rapid in vivo Colonization Screening of Probiotic Bacteria Isolated from Human Infants using Caenorhabditis elegans Surrogate Host (Caenorhabditis elegans 생체대체모델을 이용한 한국 영유아분변 유래 프로바이오틱스 균주의 in vivo 장 우점능 검토)

  • Park, Miri;Jeong, Eun-Seon;Oh, Sangnam;Song, Min-Ho;Doo, Jae-Kyun;Jeong, Yong-Seob;Moon, Yong-Il;Kim, Younghoon
    • Food Science of Animal Resources
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    • v.33 no.4
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    • pp.522-530
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    • 2013
  • The ability of probiotics to adhere to the intestinal epithelium likely plays an important role in their colonization of the gastrointestinal tract. Here, we performed high-throughput screening (HTS) for suitable characteristics of potential probiotic bacteria using attachment and colonization ability through a C. elegans surrogate in vivo model. A total of 100 strains of lactic acid bacteria (LAB) isolated from infant feces were subjected to the colonization assay using C. elegans intestine. Based on colonization ability, we showed that nine isolates have a high attachment ability during whole experimental periods (up to 168 h), compared to Lactobacillus rhamnosus strain GG as a control. Also, through the use of an in vitro cell attachment model, nine isolates revealed highly binding activity to the mucus layer. Next, the selected 9 isolates were assayed for their survival ability when exposed to acidic and bile conditions as well as cholesterol reduction and the utilization of prebiotic substrates. As a result, the isolated nine strains were determined to be highly resistant to acid and bile conditions. In addition, they have significant activity for the reduction of cholesterol and utilization of several prebiotic substrates as a carbon source. Finally, the selected nine strains were identified by either L. rhamnosus or L. plantarum (4 strains for L. rhamnosus and 5 strains for L. plantarum, respectively). Taken together, we propose that the direct colonization of probiotics using C. elegans may be applicable to the rapid screening of valuable probiotic strains in vivo.

Isolation of Antibiotics Effective to Multidrug-Resistant Cancer Cells from Sorangium cellulosum(Myxobacteria). (점액세균 Sorangium cellulosum이 생산하는 약제내성 암세포의 증식억제물질)

  • 안종웅;이정옥
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.47-51
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    • 2004
  • Drug resistance is one of the most significant impediments to successful chemotherapy of cancer. Multidrug-resistance Is characterized by decreased cellular sensitivity to anticancer agents due to the overexpression of P-glycoprotein. By using adriamycin-resistance CL02 cancer cells, we undertook the screening fur agents which were effective to multidrug-resistant cancer cells from strains of the species Sorangium cellulosum isolated in our laboratory. Sorangium cellulose, cellulose-degrading myxobacteria have recently proved to be a rich source of novel anticancer agents. One of the significant examples is the promising anticancer agent epothilone. JW 1006 is the first strain of Sorangium cellulosum which was selected by us for the isolation of a metabolite by a biological screening because of a high cytotoxic activity against the CL02 cancer cells. Cytotoxicity-guided chromatographic fractionation of the culture broth led to the Isolation of two active principles, disorazole $A_1$ and $A_2$. They showed potent cytotoxicity against CL02 cancer cells with $IC_{50}$ values in the picomolar range, and were as active against drug-resistant cancer cells CL02 and CP70 as against the corresponding sensitive cells.

Fungicidal Activities of 51 Fruit-Derived Extracts in vivo against Six Phytopathogenic Fungi

  • Lee, Hoi-Seon;Lee, Seon-Woo;Cho, Kwang-Yun;Kim, Moo-Key;Ahn, Young-Joon
    • Journal of Applied Biological Chemistry
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    • v.44 no.3
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    • pp.147-153
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    • 2001
  • Methanol extracts from 51 fruits were tested for their fungicidal activities against six phytopathogenic fungi in a greenhouse. The efficacy varied with both the plant pathogen and fruit species used. At 10 and 5 mg/pot, methanol extracts of Poncirus trifoliata peel and seed gave over 80% control values against Pyricularia grisea, and strong fungicidal activities against Rhizoctonia solani were showed from the extracts of Citrus paradisi peel and Punica granatum leaf. In a test with Botrytis cinerea at 5 mg/pot, the extracts of C. sinensis seed and D. kaki leaf produced potent fungicidal activities, and the extracts of C. crenata peel and leaf, Ch. sinensis seed, P. trifoliata peel, and Z. jujuba leaf had strong fungicidal activities. At 5 mg/pot, strong fungicidal activities were produced in the extracts of P. trifoliata peel and seed against Phytophthora infestans and in the extracts of P. ussriensis var. macrostipes fruit and seed, C. crenata peel, C. crenata leaf, C. paradisi peel, P. trifoliata peel, P. granatum peel, and Z. jujuba leaf against Puccinia recondita. In a test with E. graminis, potent activities at 10 mg/pot were produced from the extracts of Ch. sinensis seed, C. sinensis seed, P. trifoliata leaf, P. ussriensis var. macrostipes fruit and seed, and Vitis vinifera seed. In the control effect of seven extracts against B. cinerea strains resistant to carbendazim, procymidone, and diethofencarb, extracts of C. crenata peel and leaf, Ch. sinensis seed, and P. trifoliata peel were highly effective against all strains of B. cinerea. Furthermore, potent fungicidal activities were produced from the extracts of C. sinensis seed and D. kaki leaf against the SSR, SRR, and RRS, and Z. jujuba leaf against the SSR and RRS strains. As a naturally occurring fungicide, these fruit-derived materials could be useful as new fungicidal products against phytopathogenic fungi.

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Antimicrobial Effects of Essential Oils for Multidrug-Resistant Acinetobacter baumanii (다제내성 아시네토박터 바우마니의 에센셜 오일에 대한 항균효과)

  • Park, Chang-Eun;Kwon, Pil Seung
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.431-437
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    • 2018
  • Acinetobacter baumannii is categorized as a red alert pathogen that is increasingly associated with a high mortality rate in infected patients because of its resistance to extensive antibiotics. This study evaluated the antibacterial activities of some essential oils (tee tree, rosemary, and lavender oils) against 18 clinical isolates of multidrug-resistant A. baumannii (MRAB). The carbapenemase screening Hodge test showed that all 20 strains of A. baumannii were resistant to imipenem. The identification of multidrug-resistant microbes was carried out using the VITEK system. The antimicrobial activity of essential oils was tested by a disk diffusion method against MRAB. In the disk diffusion method, tea tree showed the largest increase in inhibition size compared to lavender oil, and rosemary had no antibacterial effect. These results proved the antimicrobial effect of multidrug resistance A. baumannii. Tee tree oil would be a useful alternative natural product for the treatment and prevention of most common human pathogens and MRAB infections. This is expected to be used as an antimicrobial agent, such as hand disinfectant using natural essential oil in the future.

Screening of Resistant Chinese Cabbage Cultivar against Clubroot (Plasmodiophora brassicae) for Cultivation in Highland (고랭지 배추재배를 위한 뿌리혹병 저항성 품종선발)

  • Yoon, Cheol-Soo;Jung, Eun-Kyoung;Lee, Sang-Jun;Zhang, Yan;Lee, Jae-Eun;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.59-65
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    • 2010
  • This study was conducted to select the cultivars and strains which are suitable in cultivation of the Chinese cabbage having good horticultural characteristics and the resistance to clubroot caused by Plasmodiophora brassicae, which is a widespread disease that causes serious problems in many Brassica growing areas. Thirty four cultivars and nineteen strains of the Chinese cabbage were planted in the contaminated fields in highland and investigated the wilting symptom due to the initial disease occurrence and high temperature. Disease occurrence rate and severity of clubroot and the horticultural characteristics were also examined. According to the results, Bulamplus, Kangryeokyeoleumbaechu among cultivars of Chinese cabbage, SP-05, SP-02, CR DB 50, and CR DB 29 among strains of Chinese cabbage were resistant to hot weather by showing delayed above-ground symptom (wilting) of clubroot in 30 days after transplanting. In case of below-ground clubroot rate, Daetong, super CR Kimi 85 and super CR Hiroki among the cultivars of Chinese cabbage were resistant to clubroot in 60 days after transplanting. When fresh weight of Chinese cabbage among cultivars were compared, super CR Hiroki of over 2,500 g was the highest among other cultivars. Although the resistant and susceptible reactions of Chinese cabbage cultivars and strains against clubroot disease were somewhat different from the above-ground symptoms (wilting) of clubroot in 30 days and below-ground symptoms of clubroot in 60 days, we recommend that super CR Hiroki, CR Kimi 85, CR Janggun among cultivars of Chinese cabbage and CR DB 50 and CR DB 29 among strains of Chinese cabbage will be more suitable in cultivation of Chinese cabbage in highland.

Screening rhizobacteria for biological control of root rot and Phytophthora blight on glnseng.

  • Bae, Yeoung-Seuk;Park, Kyungseok;Kim, Choong-Hoe
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.103.2-104
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    • 2003
  • Ginseng (Panax ginseng) is one of the most widely cultivated medicinal herbs in Korea. During 3 or 5 years cultivation of ginseng, yield losses can reach as high as 30-60% due to numerous diseases in Korea. Among 106 Bacillus strains isolated from various plant internal roots, we selected three promising biocontrol agents by screening against root rot caused by Cylindrocarpon destructan in a greenhouse. Preinoculation of selected isolates to seed or one-year-old root resulted in stimulation of shoot and/or root growth of seedlings, and control of root rot in infested soils with Cylindronrpon destructans (P=0.05). Furthermore, drenching of selected isolates on seedling-growing pots reduced the incidence of Phytophthora blight when the seedlings were challenged with zoospores of Phytophthora cactorum (P=0.05). However, isolates B1141 and B1142 did not show any antifungal activity against various soilborne pathogens while B1146 did in vitro. Our results provide an insight that rhizobacteria can induce resistance against various plant diseases on ginseng even if any resistant breeds have been unknown on ginseng yet.

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LOVASTATIN 고생산성 균주 개발 및 반응표면분석법 (RSM)에 의한 생산배지 최적화

  • Kim, Je-Gyeong;Kim, Gyeong-Hui;Kim, Hyeon-Suk;Lee, Sang-Jong;Jang, Yong-Geun;Jeong, Yeon-Ho;Jeong, Yong-Seop;Jeon, Gye-Taek
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.228-232
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    • 2003
  • Lovastatin produced by Aspergillus terreus via polyketide pathway is a secondary metabolite with high anti-hypercholesterolemic activity. In this paper we are going to present effective strain development strategies for lovastatin production by comparing the productivity of the mutants obtained through traditional rational screening process and protoplast fusion method. Mutants resistant against various antibiotics and/or antimetabolites showed significantly higher lovastatin productivity than the corresponding mother strains, demonstrating that rational screening method was very efficient in selecting high yielding producers. Recombinant fusants obtained using protoplast fusion between high producers were observed to have very different morphology and physiology as represented by the production and secretion of lovastatin, as well as cell growth pattern. In parallel with the strain development, optimization process for the production medium was carried out in order to find optimal concentrations of the medium components using such a powerful statistical method as response surface method (RSM). It was concluded that not only the optimum production medium but also good morphological characteristics of the high-yielding producers led to higher lovastatin production.

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Screening of Thermotolerant Yeast for Use as Microbial Feed Additive

  • Lee, Jae-Heung;Lim, Yoo-Beom;Koh, Jong-ho;Baig, Soon-Yong;Shin, Hyung-Tai
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.162-165
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    • 2002
  • With the objective of identifying the commercial potential of new direct-fed microbials, several temperature-tolerant strains were isolated from cane molasses at $39^{\circ}C$ and tested for their tolerance to pH, bile salts, and a mixture of volatile fatty acids (acetic acid:propionic acid:butyric acid=6.5:2.0:1.5). It was found that the isolated strain DY 252 grew very well up to pH 2.0 and was resistant to relatively high concentrations of bile salts. Among the strains tested, DY 252 was least inhibited by the addition of volatile fatty acids to the growth medium at $39^{\circ}C$. Accordingly, it would appear that strain DY 252, identified as yeast Issatchenkia orientalis, may be a potential candidate for use as a microbial feed additive.

Antibiotics produced by anaerobic fermentation of Streptococcus sp. An-21-1 isolated from domestic soil I. Screening and identification of anaerobic bacteria (국내토양에서 분리한 혐기성 세균 Streptococcus sp. An-21-1 이 생성하는 항생물질 I. 혐기성 세균의 선별과 동정)

  • Park, Seung-chun;Yun, Hyo-in;Oh, Tae-kwang
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.53-60
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    • 1993
  • Anaerobic bacteria are suggested to be potential source for new antibiotics. In order to search for antibiotics from domestic origin, we collected 800 soil samples across Korean locations and could isolate as many as 989 anaerobic strains. Among them 10, strains were found to have good producing capacity of antibiotics. An anaerobe was finally selected due to secreting antibiotics having high antimicrobial activity towards multiple resistant microorganism(E coli JM 83) transformed by genetic engineering technique. Its morphological, physiological and biochemical charateristics were investigated, together with antimicrobial spectrum therefrom. On antimicrobial spectrum study, substance secreted from this strain, had no activities to fungus and yeast. The selected strain showed G(+) and coccal shape, on Gram, staining and electron scanning microscopy, respectively. Biochemically this strain utilized glucose, fructose lactose, sucrose, but did not arabinose, cellulose, rhamnose, sorbitol, trehalose, mannitol. Catalase test showed negative property. Optimal growth temperature was $37^{\circ}C$. The results obtained above suggest this strain Streptococcus faecium subspp. and we named it Streptococcus sp. An-21-1.

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Identification and Sequence Analysis of RNA3 of a Resistance-Breaking Cucumber mosaic virus Isolate on Capsicum annuum

  • Lee Mi-Yeon;Lee Jang-Ha;Ahn Hong-Il;Yoon Ju-Yeon;Her Nam-Han;Choi Jang-Kyung;Choi Gug-Seon;Kim Do-Sun;Harn Chee-Hark;Ryu Ki-Hyun
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.265-270
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    • 2006
  • Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.