• 한국수산과학회지
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• 제43권2호
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• pp.169-173
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• 2010

This study evaluated the effects of gamma ray irradiation on the safety of dried seafood products. Dried salted squid, Engraulis japonica, Hizikia fusiformis, Mytilus coruscus, and Porphyra tenera were gamma-irradiated at doses of 0, 1, 3, and 5 kGy. The total bacterial populations were then enumerated on total plate count agar, and bacteria isolated from the samples were identified by 16S rDNA sequencing. In addition, the isolated strains were inoculated in the products to determine $D_{10}$ values. The total bacterial populations in the dried seafood products ranged from 3.40 to 6.59 log CFU/g, and those of yeasts and molds ranged from 2.21 to 4.56 log CFU/g. The sequence analysis identified Staphylococcus sp. as the most common species in the dried seafood products, except for dried P. tenera. The $D_{10}$ values of the contaminating bacteria were lower than 0.7 kGy, except for Deinococcus sp., which had a value of 1.39 kGy. This study demonstrated that gamma irradiation could be used to improve the safety of dried seafood products.

• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.448-456
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• 2012

Thirty-seven carbofuran-degrading bacteria were isolated from agricultural soils, and their genetic and phenotypic characteristics were investigated. The isolates were able to utilize carbofuran as a sole source of carbon and energy. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Rhodococcus, Sphingomonas, and Sphingobium, including new types of carbofuran-degrading bacteria, Bosea and Microbacterium. Among the 37 isolates, 15 different chromosomal DNA patterns were obtained by polymerase chain reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences. Five of the 15 representative isolates were able to degrade carbofuran phenol, fenoxycarb, and carbaryl, in addition to carbofuran. Ten of the 15 representative isolates had 1 to 8 plasmids. Among the 10 plasmid-containing isolates, plasmid-cured strains were obtained from 5 strains. The cured strains could not degrade carbofuran and other pesticides anymore, suggesting that the carbofuran degradative genes were on the plasmid DNAs in these strains. When analyzed with PCR amplification and dot-blot hybridization using the primers targeting for the previously reported carbofuran hydrolase gene (mcd), all of the isolates did not show any positive signals, suggesting that their carbofuran hydrolase genes had no significant sequence homology with the mcd gene.

• Journal of Crop Science and Biotechnology
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• 제10권3호
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• pp.167-174
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• 2007

Spongy Alphonso mangoes were found to be infected with Staphylococcus bacteria. A Gram positive Staphylococcus strain was isolated from spongy pulp and identified from CABI Bioscience, UK, by partial 16S rDNA sequence analysis and by morphological and biochemical characterization through IMTECH, Chandigarh, India. Although identification by both of these methods indicated the organism belonged to same genus, different species names were given. Changes in total phenolics, reducing, and non-reducing sugars, respiration rate, total carotenoids, peroxidase(POX), and catalase activities were monitored during ripening of these fruits. The climacteric rise in spongy fruits was marked by an increase in respiration rate and a decrease in sugar content. Total phenolics content increased in spongy fruits as compared to ripe non-spongy fruits. Development of corky white tissue in spongy fruits was associated with about a 2.5-fold reduction in total carotenoids and a concomitant increase in lipoxygenase-mediated, $\beta$-carotene co-oxidation. A marked decrease in soluble protein content and about a 1.5-fold increase in POX activity was observed. Maximum POX activity was confined to 50-70%$(NH_4)_2SO_4$ fraction. The intense dark bands visible after POX specific substrate staining of the Native gel indicated a high expression of isoenzymes of POX in spongy fruits. Similarly, changes in levels of catalase activity were also observed in spongy fruits. The results suggest that infection of Alphonso mangoes with Staphylococcus bacteria affects the normal ripening processes of the fruit interfering with the carbohydrate and carotenoid metabolism. Also, the studies indicate the expression of POX and catalase enzymes as a plant defense response to microbial invasion.

• 한국미생물·생명공학회지
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• 제32권4호
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• pp.312-316
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• 2004

토양 우점능이 강한 생물학적 방제제 제조를 위해 경북지역 토양에서 길항균주를 분리하고 이들 중 Fusarium oxysporum, Phytophthora capsici 에 강력한 길항능을 보이는 Pseudomonas sp. 4059 를 선발, 동정하였다. Pseudomonas sp. 4059 의 시들음 병균 Fusarium oxysporum, 고추 역병균 Phytophthora capsici 에 대한 길항기작은 내열성 저분자의 항생물질과 철이온을 특이적으로 흡착하는 siderophore의 생산에 의한 것이다. Pseudomonas sp. 4059 는 항진균성 항생물질 Phenazine 생산 유전자를 소유하며 Salkowski test에 양성인 옥신류 생산도 한다는 것을 확인하였다. Pseudomonas sp. 4059 는 bioochemical tests, API test, MicroLogTM system을 통해 Pseudomonas fluorescens (biotype A)으로 98% 상동성을 보였으므로 이를 Pseudomonas fluorescens (biotype A) 4059 로 명명하였다. 선발된 길항균 Pseudomonas fluorescens (biotype A) 4059는 고추를 기주식물로 하였을 때 고추역병균인 Phytophthora capsici가 원인이 되는 고추역병을 in vivo 상에서도 충분히 억제할 수 있는 생물방제능을 나타내었다.

• 한국축산식품학회지
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• 제32권5호
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• pp.635-643
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• 2012

As an appraisal for the application of a new starter culture, more than 200 lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains showing excellent growth and acid production ability in 10% skim milk media were selected and identified as Lactobacillus casei based on the results of their API carbohydrate fermentation patterns, as well as 16S rDNA sequence analysis. To assess the effect of L. casei strains on irritable bowel disease (IBD), the inhibitory effect of the selected strains against the nitric oxide (NO) production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells was measured. Among the tested L. casei strains, L. casei MCL was observed to have the greatest NO inhibitory activity. Additionally, L. casei MCL was found to inhibit mRNA expression of pro-inflammatory cytokines (interleukin-$1{\beta}$, IL-6, TNF-${\alpha}$), as well as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) involved in pathophysiologic processes such as inflammation. The mRNA expression of anti-inflammatory cytokines, including IL-10 and transforming growth factor-$1{\beta}$ (TGF-${\beta}$) of L. casei MCL, was confirmed using quantitative real-time PCR. In conclusion, L. casei MCL showed decreases in the expression of pro-inflammatory cytokines and up-regulated expression of the anti-inflammatory cytokine.

• 한국동물위생학회지
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• 제39권2호
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• pp.131-136
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• 2016

Clostridium novyi (C. novyi) is a gram positive, non-capsulated, motile, obligatory anaerobe that produces endospores. Both C. novyi type A and B produce a bacteriophage encoded lethal alpha toxin which belongs to a family of large clostridial cytotoxins. These large clostridial cytotoxins of C. novyi bind to the uncharacterized receptors on host vascular endothelial cells, which leads to the loss of integrity of the vascular endothelium with subsequent edema, refractory hypotension, organ failure, and sudden death. A total of 13 sudden death cases were submitted to Chonbuk National University-Veterinary Diagnostic Center between June and October, 2015. The samples, mainly liver, were collected in sterile vials after necropsy and processed within 12~24 hours for diagnosis, isolation and identification of C. novyi. All of the 4 gram positive samples showed amplification by PCR. Out of 4 positive samples, 3 were detected to be C. novyi type B and 1 was detected as C. novyi type A. Based on the 16S rDNA sequence analysis, 1 case (150564) showed 99% similarity with C. novyi type A while other 3 cases (150388, 150557 and 150775) presented 99% similarity with C. novyi type B. Based on the results, C. novyi was found to be prevalent in Korean pig farms and causes sudden death to finishing pigs or sows during summer season. Thus, C. novyi should be considered for differential diagnosis on sudden death cases during the summer season.

• Mycobiology
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• 제36권1호
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• pp.13-18
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• 2008

Eight distinct bacteria were isolated form diseased mycelia of the edible mushroom, Pleurotus eryngii. 16S rDNA sequence analysis showed that the isolates belonged to a variety of bacterial genera including Bacillus (LBS5), Enterobacter (LBS1), Sphingomonas (LBS8 and LBS10), Staphylococcus (LBS3, LBS4 and LBS9) and Moraxella (LBS6). Among them, 4 bacterial isolates including LBS1, LBS4, LBS5, and LBS9 evidenced growth inhibitory activity on the mushroom mycelia. The inhibitory activity on the growth of the mushroom fruiting bodies was evaluated by the treatment of the bacterial culture broth or the heat-treated cell-free supernatant of the broth. The treatment of the culture broths or the cell-free supernatants of LBS4 or LBS9 completely inhibited the formation of the fruiting body, thereby suggesting that the inhibitory agent is a heat-stable compound. In the case of LBS5, only the bacterial cell-containing culture broth was capable of inhibiting the formation of the fruiting body, whereas the cell-free supernatant did not, which suggests that an inhibitory agent generated by LBS5 is a protein or a heat-labile chemical compound, potentially a fungal cell wall-degrading enzyme. The culture broth of LBS1 was not inhibitory. However, its cell-free supernatant was capable of inhibiting the formation of fruiting bodies. This indicates that LBS1 may produce an inhibitory heat-stable chemical compound which is readily degraded by its own secreted enzyme.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.437-443
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• 2013

The diversity of culturable non-actinobacterial (NA) bacteria associated with four species of South China Sea gorgonians was investigated using culture-dependent methods followed by analysis of the bacterial 16S rDNA sequence. A total of 76 bacterial isolates were recovered and identified, which belonged to 21 species of 7 genera, and Bacillus was the most diverse genus. Fifty-one percent of the 76 isolates displayed antibacterial activities, and most of them belonged to the Bacillus genus. From the culture broth of gorgonian-associated Bacillus methylotrophicus SCSGAB0092 isolated from gorgonian Melitodes squamata, 11 antimicrobial lipopeptides including seven surfactins and four iturins were obtained. These results imply that Bacillus strains associated with gorgonians play roles in coral defense mechanisms through producing antimicrobial substances. This study, for the first time, compares the diversity of culturable NA bacterial communities among four species of South China Sea gorgonians and investigates the secondary metabolites of gorgonian-associated B. methylotrophicus SCSGAB0092.

• Preventive Nutrition and Food Science
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• 제18권4호
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• pp.273-279
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• 2013

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.256-263
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• 2012

A strain of bacterium producing antifungal antibiotic was isolated and identification of the strain was attempted. We could identify the bacterium as being a Bacillus sp., based on morphological observation, physiological characteristics, and 16S rDNA sequence analysis, thus leading us to designate the strain as Bacillus sp. AH-E-1. The strain showed potent antibiotic activity against phytopathogenic and human pathogenic fungi by inducing mycelial distortion and swelling and inhibiting spore germination. The antibiotic metabolite produced by the strain demonstrated excellent thermal and pH (2-11) stability, but was labile to autoclaving. From these results, we could find a broader antifungal activity of Bacillus genus. Isolation and characterization of the active agent produced by the strain are under progress.