• Title/Summary/Keyword: Screening of resistant strains

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Rapid Detection of Cadmium-Resistant Plant Growth Promotory Rhizobacteria: A Perspective of ELISA and QCM-Based Immunosensor

  • Agrawal, Ruchi;Satlewal, Alok;Chaudhary, Manav;Verma, Amit;Singh, Rachna;Verma, A.K.;Kumar, Rajesh;Singh, K.P.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.849-855
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    • 2012
  • Plant growth-promoting rhizobacteria (PGPR) pseudomonads have a large number of lipopolysaccharides on the cell surface, which induces immune responses. Cd-resistant PGPR prevalent at the Cd-affected sites under biophytostabilization was monitored. Transmissiom electron microscopy was used to the study the behavior of tolerance of PGPR to cadmium level and its effect on pseudomonad strains (Z9, S2, KNP2, CRPF, and NBRI). An immunosensor was developed by immobilizing antibody (anti-Z9 or anti-S2) against selected PGPR on a piezoelectric quartz crystal microbalance (QCM). Immunosensors were found to supplement the inherent specificity of antigen-antibody reactions with the high sensitivity of a physical transducer. On comparison of the efficiency of detection with ELISA, the spectrophotometric technique, the developed immunosensor was found to be more sensitive, fast, and reliable even after regeneration for several times. Thus, the immunosensor may be used for future detection of PGPR strains after automation of the screening process.

Description of 17 unrecorded bacterial species isolated from freshwater showing antibiotic resistance in Korea

  • Baek, Kiwoon;Kim, Eui-Jin;Han, Ji-Hye;Choi, Ahyoung
    • Korean Journal of Environmental Biology
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    • v.38 no.2
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    • pp.289-298
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    • 2020
  • As part of the research program "2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library" freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.

Semi-Rational Screening of Probiotics from the Fecal Flora of Healthy Adults against DSS-Induced Colitis Mice by Enhancing Anti-Inflammatory Activity and Modulating the Gut Microbiota

  • Wang, Weiwei;Xing, Wentao;Wei, Sichen;Gao, Qiaoying;Wei, Xinliang;Shi, Liang;Kong, Yu;Su, Zhenhua
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1478-1487
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    • 2019
  • Ulcerative colitis (UC), a chronic inflammatory bowel disease, substantially impacts patients' health-related quality of life. In this study, an effective strategy for discovering high-efficiency probiotics has been developed. First, in order to survive in the conditions of the stomach and intestine, high bile salt-resistant and strong acid-resistant strains were screened out from the fecal flora of healthy adults. Next, the probiotic candidates were rescreened by examining the induction ability of IL-10 (anti-inflammatory factor) production in dextran sodium sulfate (DSS)-induced colitis mice, and Lactobacillus sakei 07 (L07) was identified and selected as probiotic P. In the end, fourteen bifidobacterium strains isolated from stools of healthy males were examined for their antimicrobial activity. Bifidobacterium bifidum B10 (73.75% inhibition rate) was selected as probiotic B. Moreover, the colonic IL-6 and $TNF-{\alpha}$ expression of the DSS-induced colitis mice treated with L. sakei 07 (L07) - B. bifidum B10 combination (PB) significantly decreased and the IL-10 expression was up-regulated by PB compared to the DSS group. Furthermore, Bacteroidetes and Actinobacteria decreased and Firmicutes increased in the DSS group mice, significantly. More interestingly, the intestinal flora biodiversity of DSS colitis mice was increased by PB. Of those, the level of B. bifidum increased significantly. The Bacteriodetes/Firmicutes (B/F) ratio increased, and the concentration of homocysteine and LPS in plasma was down-regulated by PB in the DSS-induced colitis mice. Upon administration of PB, the intestinal permeability of the the DSS-induced colitis mice was decreased by approximately 2.01-fold. This method is expected to be used in high-throughput screening of the probiotics against colitis. In addition, the L. sakei 07 - B. bifidum B10 combination holds potential in UC remission by immunomodulatory and gut microbiota modulation.

Isolation of methicillin-resistant Staphylococcus aureus from a Shih-Tzu dog with canine distemper virus infection

  • Pak, Son-il;Hwang, Cheol-yong;Youn, Hwa-young;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.376-382
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    • 1999
  • A methicillin-resistant Staphylococcus aureus (MRSA) isolate was recovered from a 9-month-old female Shih-Tzu dog with canine distemper virus infection. We performed in vitro antimicrobial susceptibility test to determine the most effective antimicrobial drug against the isolate and thus, to emphasize its potential clinical importance in animal practices. Isolate was confirmed MRSA by oxacillin agar screening test. The isolate was fully resistant to all $\beta$-lactam antibiotics and was susceptible to glycopeptides. Of the other antibiotics, mupirocin, TMP/SMZ (trimethoprim-sulfamethoxazole), and chloramphenicol showed inhibitory effect at the concentration of 4x MIC. The MICs ranged 0.25->$128{\mu}g/ml$, and MBCs ranged 0.5->$128{\mu}g/ml$. The combined TMP/SMZ with cefamandole or novobiocin showed synergistic effect, whereas the combination of novobiocin plus cefamandole or teicoplanin resulted in antagonistic effects. Although MRSA in animals so far has been reported in the geographically limited countries, at least theoretically, it could be occurred in the future more frequently through either human or animal origin. The use of this combination may be of value in this situation. As with all antimicrobial agents, inappropriate or unnecessarily prolonged therapy may contribute to the emergence of resistance strains and loss of efficacy.

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Screening of Radio-resistant Lactic Acid Bacteria

  • Hwang, E-Nam;Kang, Sang-Mo;Kim, Jae-Kyung;Lee, Ju-Woon;Park, Jong-Heum
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.335-340
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    • 2013
  • This study screened for radio-resistant strains lactic acid bacteria (LAB) by evaluating their capability to survive exposure to ionizing radiation. Ten strains of LAB - Lactobacillus bulgaricus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus delbruekii, Lactococcus lactis, Streptococcus thermophilus, Bifidobacterium breve, and Pediocuccos pentosaceus - were selected and subcultuted twice. The LAB was then further cultured for 3 d at $37^{\circ}C$ to reach 7-10 Log colony-forming units (CFU)/mL prior to irradiation and immediately exposed to gamma rays or electron beams with absorbed doses of 0, 1, 2, 3, 4, 5, 6, 8, and 10 kGy. Gamma irradiation gradually decreased the number of the tested viable LAB, and the effect was irradiation dose dependent. A similar effect was found in electron beam-irradiated LAB. Radiation sensitivity of LAB was calculated as $D_{10}$ values, which ranged from 0.26 kGy to 0.9 kGy and 0.5 kGy to 1.44 kGy with exposure to gamma and electron beam irradiation, respectively, in all tested LAB. L. acidophilus was the most resistant to gamma and electron beam irradiation, with $D_{10}$ values of 0.9 kGy and 1.44 kGy, respectively. These results suggest that L. acidophilus might be suitable for the preparation of probiotics as direct-fed microbes for astronauts in extreme space environments.

Selection of High Tobramycin-Producing Mutants (Tobramycin 고생산성 변이주의 분리)

  • 나규흠;김학주;김기태;양중익;김계원
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.343-347
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    • 1991
  • An improved method for the selection of high tobramycin-producing mutants of Streptomyces tenebrarius ATCC 17920 was investigated. By the use of apramycin-containing media, low nebramycin-producing mutants were eliminated. Strain No. 23, resistant to apramycin and kanamycin B and sensitive to tobramycin, was isolated from soils, identified as Pseudomonas paucimobilis and used as a test organism for overlaying the mutants on agar plate. While inhibition zones were not shown when the parent strains were overlaid with soft agar containing the strain No. 23, clear zones were shown when high tobramycin-producing mutants were overlaid. Using this screening strategy, 58 mutants showing clear zones had been isolated. antibiotic activities of which were incresed to 3~8 fold compared to that of parent strain.

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Study on Inheritance of Potato virus X Resistance in Capsicum annuum

  • Shi, Jinxia;Choi, Do-Il;Kim, Byung-Dong;Kang, Byoung-Cheorl
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.433-438
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    • 2008
  • Potato virus X (PVX) resistance in potato is one of the best-characterized resistance models, however little is known in pepper. To evaluate the resistance to PVX in Capsicum annuum, a total of eleven pepper accessions were used for resistance screening against two PVX strains, USA and UK3. None of them were resistant against strain UK3, whereas four resistant genotypes were found against strain USA, three of which were further characterized. Two unlinked dominant genes were identified for both genotypes Bukang and Perennial; resistance in the genotype CV3 seemed to be conferred by two complementary dominant genes. These results demonstrated that the resistance to PVX in C. annuum is different from that in potato. This is the first report on genetic analysis of PVX resistance in C. annuum.

Screening and Partial Purification of Bacteriocins by Strains of Lactobacillus acidophilus Isolated from Human Origin (인체에서 분리된 Lactobacillus acidophilus가 생산하는 박테리오신의 선별과 정제)

  • Kim, Se-Heon;Kim, Yeong-Gyo;Gilliland, S.E.
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.21-26
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    • 1997
  • Lactobacillus acidophilus 223, 606, and NCFM-F among 21 isolated from fecal contents of humans demonstrated inhibitory activity attributed to bacteriocin(s). The bacteriocin(s) were heat stable and nondialyzable proteinous compounds and exhibited narrow inhibitory spectra of activity. Neither hydrogen peroxide nor pH were responsible for inhibitory action. All of the producer strains were resistant to their own bacteriocin(s). The bacteriocin(s) were purified by ammonium sulfate precipitation, gel chromatography and ion exchange chromatography for further characterization. The bacteriocin(s) of human origin exhibited similar characteristics.

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Screening and Characterization of Thermotolerant Alcohol-producing Yeast

  • Sohn, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.215-221
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    • 1994
  • Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

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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.