• Title/Summary/Keyword: 16S-rRNA

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Bacteriological Study about the Death of Cultured Doctor Fish, Garra rufa in the Aquarium

  • Lee, Ji-Yoon;Gang, Nam-I;You, Jin-Sol;Ko, Chang-Yong;Lee, Ki-Won;Han, Won-Min;Kim, Eunheui
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.18-24
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    • 2016
  • Since April 2012, doctor fish in the breeding tank and in the quarantine tank in Hanwha Aquaplanet Yeosu Aquarium have been dying, accompanied by diffuse bleeding around the mouth, in the chin, and at the bottom of the abdomen. In this study, the cause of death would be examined through the bacteriological study of doctor fish and the rearing water quality in the aquarium. The water quality and the bacterial counts of the rearing water in the exhibit tank and in the quarantine tank were analyzed once a week, starting from August to November 2014. Water quality was measured based on the following data: temperature was in the range of 24.5~26.8℃, pH at 6.77~7.94, DO at 6.15~8.61 ppm, ammonia at 0~0.93 ppm, nitrite at 0.009~0.075 ppm, and nitrate at 1.1~40.9 ppm. Studies revealed that the differences in these water quality factors were not related to the death of doctor fish. Bacterial counts in the rearing waters of Garra rufa slightly increased to 103~104 CFU/ml, just before the death of the doctor fish. Twelve strains of bacteria were isolated from the dead fish and rearing waters. The isolates were identified as Aeromonas veronii, Citrobacter freundii, Pseudorhodoferax aquiterrae, Shewanella putrefaciens, and Vibrio anguillarum on the basis of 16S rRNA gene sequences. The most dominant species was C. freundii, which showed medium sensitivity to florfenicol and norfloxacin, and was resistant to amoxacillin, doxycycline, oxytetracycline, tetracycline, and trimethoprim. Ten isolates were confirmed to be pathogenic to the doctor fish. Doctor fish infected with C. freundii and S. putrefaciens showed high mortality in the experimental groups. These results indicate that the variation in bacterial numbers in the rearing water was related to the death of doctor fish. C. freundii and S. putrefaciens were directly implicated in causing the death of doctor fish in the aquarium.

Effect of Korean Fermented Food Extracts and Bacteria Isolated from the Extracts for the Control of Rice Seed-borne Fungal Diseases (국산 발효식품 추출물과 발효식품유래 미생물을 활용한 벼 종자전염성 진균병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Lee, Min-Ho;Park, Jong-Ho;Han, Eun-Jung;Choi, Eun-Jung;Bae, Soo-Il;Jee, Hyeong-Jin
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.383-395
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    • 2014
  • When we investigated seed infestation by fungal pathogens from 51 varieties in 9 crops, the contamination rate of rice and sesame seeds was high. Therefore, to control seed-borne diseases, we obtained extracts from commercial products of Kimchi, Gochujang, Doenjang, Ganjang, Makgeolli and Tohajut and their suppressive effects against seed-borne diseases were studied. In addition, bacterial strains were screened to control rice seed-borne diseases in vitro and in vivo. Among forty food extracts, eleven food-extracts suppressed incidence of seedling rots in vitro and five food extracts increased 8-33% of healthy seedling in the greenhouse. Among 218 isolates from 40 fermented foods, 43 isolates showed high antifungal activity against seven fungal pathogens. When we tested 43 isolates for the reduction of rice seed borne disease, 32 isolates were able to reduce the rice seed borne disease. Among 32 isolates, 17 isolates reduced significantly seedling rot and increased healthy seedlings, the other isolates except for Kc4-2 and Mkl 2-2 increased shoot emergence and the percentage of healthy plants. Thirty isolates with high antifungal activity and suppressive effect against rice seedling rots were identified by 16S rRNA sequencing. Twenty one of thirty isolates were identified as Bacillus spp. Three isolates from Makgeolli were identified as Saccharomyces cerevisiae. B. amyloliquifaciens were isolated from six Korean traditional fermented foods except for Ganjang. B. amyloliquifaciens were majority in the effective bacterial population of Gochujang and Jutgal. Relatively diverse Bacillus species including B. subtilis, B. pumilus, and B. amyloliquificiens were isolated from Kimchi. The selected effective microorganisms from Korean fermented foods founded to be effective for controlling seed-borne diseases of rice in vitro and in the greenhouse. We think that Korean fermented foods and their useful microorganisms can be used as biocontrol agents for suppressing rice seed-borne diseases based on above described results.

The Optimal Production and Characteristics of an Alginate-degrading Enzyme from Vibrio sp. PKA 1003 (Vibrio sp. PKA 1003의 알긴산 분해 조효소 생산 최적 조건과 조효소의 특성)

  • Kim, Hyun-Jee;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Jeong, Da-Hyun;Jung, Hee-Ye;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.434-440
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    • 2013
  • This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.

Soil Chemical Property and Microbial Community under Organic and Conventional Radish Farming Systems (무 유기재배와 관행재배 토양의 화학성과 미생물 군집 비교)

  • Kang, Ho-Jun;Yang, Sung-Nyun;Song, Kwan-Cheol;Cho, Young-Yuen;Kim, Yu-Kyoung
    • Korean Journal of Organic Agriculture
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    • v.27 no.4
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    • pp.479-499
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    • 2019
  • This study was conducted to investigate the responses of soil properties and microbial communities to different agricultural management and soil types, including organic management in Andisols (Org-A), organic management in Non-andisols (Org-NA), conventional management in Andisols (Con-A) and conventional management in Non-andisols (Con-NA) by using a pyrosequencing approach of 16S rRNA gene amplicon in Radish farms of volcanic ash soil in Jeju island. The results showed that agricultural management systems had a little influence on the soil chemical properties but had significant influence on microbial communities. In addition, soil types had significant influences on both the soil chemical properties and microbial communities. Organic farming increased the microbial density of bacteria and biomass C compared to conventional farming, regardless of soil types. Additionally, Org-NA had the highest dehydrogenase activity among treatments, whereas no difference was found between Org-A, Con-A and Con-NA and had the highest species richness (Chao 1) and diversity (Phyrogenetic diversity). Particularly, Chao 1 and Phyrogenetic diversity were increased in organic plots by 12% and 20%, compared with conventional plots, respectively. Also, regardless of agricultural management and soil types, Proteobacteria was the most abundant bacterial phylum, accounting for 21.9-25.9% of the bacterial 16S rRNAs. The relative abundance of putative copiotroph such as Firmicutes was highest in Org-NA plot by 21.0%, as follows Con-NA (13.1%), Con-A (6.7%) and Org-A (5.1%.), respectively and those of putative oligotrophs such as Acidobacteria and Planctomycetes were higher in Con-A than those in the other plots. Furthermore, LEfSe indicated that organic system enhanced the abundance of Fumicutes, while conventional system increased the abundance of Acidobacteria, especially in Non-andisols. Correlation analysis showed that total organic carbon (TOC) and nutrient levels (e.g. available P and exchangeable K) were significantly correlated to the structure of the microbial community and microbial activity. Overall, our results showed that the continuous organic farming systems without chemical materials, as well as the soil types made by long-term environmental factors might influence on soil properties and increase microbial abundances and diversity.

Prevalence of Anaplasma sp. in Thrushes (Family Turdidae) in Jeju Island, Republic of Korea (제주도와 마라도내 지빠귀과 조류에서 Anaplasma spp. 감염 조사)

  • Oh, Mi-Rae;Moon, Kyoung-Ha;Kim, So-Yeon;Kim, Yun-Gi;Choi, Chang-Yong;Kang, Chang-Wan;Kim, Hwa-Jung;Lee, Kyoung-Kap;Yun, Young-Min
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.206-211
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    • 2014
  • Anaplasmosis is a rickettsial zoonosis mediated by blood-sucking arthropods, such as ticks, flies, and mosquitos. Migratory birds are common hosts of ticks that are mediators of anaplasmosis, in particular, the tick infection rate in thrushes (family Turdidae) has been known to be high. The main purpose of this study is to survey the occurrence and prevalence of Anaplasma spp. from the migratory thrushes in Jeju island. We collected blood samples from 6 thrushes rescued at the Jeju Wildlife Rescue Center and from 34 wild-caught thrushes on Mara island which is a satellite island of Jeju. As a result, the nested PCR confirmed that seven out of 40 individuals (17.5%) were infected by Anaplasma spp. and all of them were identified as A. phagocytophilum based on sequences obtained from partial 16S rRNA. All the infected birds were on their northward migration in spring, our results suggest that the Turdidae family, which is a common and abundant migrant group passing through Jeju island, may act a role as active reservoir and disperser of A. phagocytophilum causing potential influx of the zoonotic pathogens from its wintering grounds in lower latitude to the mainland Korea as well as Jeju.

Identification of Novel Bacillus subtilis IDCC 9204 Producing a High-Level Fibrinolytic Enzyme and Properties of NK-IL9204 (고농도 혈전용해효소를 생산하는 신규 Bacillus subtilis IDCC 9204의 분리 및 NK-IL9204의 효소학적 특성)

  • Lee, Seung-Hun;An, Gwangmin;Kim, Heu-Hang;Kang, Jae-Hoon;Kang, Dae-Jung
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.600-606
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    • 2012
  • A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

Production of a Fermented Korean Pear Puree using a New Strain Leuconostoc mesenteroides KACC 91495P Isolated from Kimchi (김치에서 신규 Leuconostoc mesenteroides KACC 91495P 균주의 분리 및 이를 이용한 배 발효물의 제조)

  • In, Man-Jin;Kim, Hye-Min;Jin, Hea-Jin;Kim, Dong-Chung;Oh, Nam-Soon;Chae, Hee-Jeong
    • Journal of Applied Biological Chemistry
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    • v.53 no.1
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    • pp.51-55
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    • 2010
  • A lactic acid bacterial strain showing fast growth and high acid production in Korean pear puree was isolated from Kimchi. This strain was analyzed by API 50 CHL kit and 16S rRNA sequencing analysis and identified as Leuconostoc (Ln.) mesenteroides KACC 91495P. Korean pear puree was fermented using Ln. mesenteroides KACC 91495P strain at $30^{\circ}C$ for 18 h. The changes of pH, titratable acidity and viable cell number during fermentation were investigated. The pH and titratable acidity were reached to pH 3.86 and 1.09% after 18 h fermentation, respectively. The viable cell population of Ln. mesenteroides KACC 91495P was rapidly increased to $2.0{\times}10^9\;CFU/g$ during the 9 h of cultivation. The contents of lactic acid, acetic acid and malic acid were determined to be 0.213, 0.259, and 0.217% after 18 h fermentation, respectively. The content of polyphenolic compounds, known as antioxidants, in pear puree were enhanced by Ln. mesenteroides KACC 91495P cultivation. The level of total polyphenolic compounds was increased to around 140% of initial concentration. When the fermented pear puree was kept at $4^{\circ}C$, pH, titratable acidity and number of viable cells population were nearly maintained for 13 days.

Biocontrol Potential of Streptomyces griseus H7602 Against Root Rot Disease (Phytophthora capsici) in Pepper

  • Nguyen, Xuan-Hoa;Naing, Kyaw-Wai;Lee, Young-Seong;Tindwa, Hamisi;Lee, Geon-Hyoung;Jeong, Byoung-Kon;Ro, Hee-Myeong;Kim, Sang-Jun;Jung, Woo-Jin;Kim, Kil-Yong
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.282-289
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    • 2012
  • The root rot of pepper (Capsicum annuum L.) caused by Phytophthora capsici is one of the most important diseases affecting this crop worldwide. This work presents the evaluation of the capacity of Streptomyces griseus H7602 to protect pepper plants against Phytophthora capsici and establishes its role as a biocontrol agent. In this study, we isolated an actinomycete strain H7602 from rhizosphere soil, identified it as Streptomyces griseus by 16S rRNA analysis and demonstrated its antifungal activity against various plant pathogens including P. capsici. H7602 produced lytic emzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase and protease. In addition, crude extract from H7602 also exhibited destructive activity toward P. capsici hyphae. In the pot trial, results showed the protective effect of H7602 against pepper from P. capsici. Application of H7602 culture suspension reduced 47.35% of root mortality and enhanced growth of pepper plants for 56.37% in fresh root and 17.56% g in fresh shoot as compared to control, resulting in greater protection to pepper plants against P. capsici infestation. Additionally, the enzymatic activities, chitinase and ${\beta}$-1,3-glucanase, were higher in rhizosphere soil and roots of pepper plants treated with H7602 than other treated plants. Therefore, our results indicated a clear potential of S. griseus H7602 to be used for biocontrol of root rot disease caused by P. capsici in pepper.

Changes of Biological and Chemical Properties during Composting of Livestock Manure with Isolated Native Microbe (토착미생물별 가축분 퇴비화 과정중 생물화학적 특성 변화)

  • Han, Hyo-Shim;Lee, Kyung-Dong
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1126-1135
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    • 2012
  • In order to produce high-quality fermenting composts, bacteria strains with high activities of extracellular enzymes (cellulase, chitinase, amylase, protease and lipase) were isolated from the soils in 6 provinces of Korea, and characterized by 16S rRNA gene sequence analysis and properties. The selected 7 stains inoculated to livestock manure for 2' fermenting time, and experimental treatment divided into 3 groups, B1, B2 and B3, according to microbial activity and enzyme type. Our results showed that microbe applications (B1, B2 and B3) can increase (p<0.05) both rhizomes (17-38%) and enzyme activities (50-81%) in compost after fermenting time, respectively, compared to non-microbe treatment (control). The microbe application also decreased significantly (p<0.05) the $NH_3$ and $H_2S$ gas contents 13.4 and 27.3% compared with control, and the Propionic acid and Butyric acid gas contents 14.5 and 19.6%, respectively, as compared to the control. The microbial degradation rate (%) of pesticides and heavy metals increased significantly (p<0.05) after fermenting time, respectively, as compared to the control. Especially, microbe applications were more effective in total rhizomes yields and bioactivities than non-microbe treatment. Thus the results of this study could help in development of potential bioinoculants and composting techniques that maybe suitable for crop production, and protectable for earth environment under various conditions.

Utilization of Leuconostoc mesenteroides 310-12 Strain in the Fermentation of a Traditional Korean Rice-based Beverage (Leuconostoc mesenteroides 310-12 균주를 이용한 전통 쌀 발효 음료의 제조)

  • Kim, Dong-Chung;Choi, Jin-Woong;In, Man-Jin
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.21-25
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    • 2011
  • A lactic acid bacterial strain showing high acid production in saccharified-rice suspension was isolated from Kimchi. This strain was analyzed by API 50 CHL kit and 16S rRNA sequencing analysis and identified as Leuconostoc mesenteroides 310-12. Saccharified-rice suspension was fermented using L. mesenteroides 310-12 strain at $30^{\circ}C$ for 15 h. The changes of pH, titratable acidity and viable cell number during fermentation were determined. The pH and titratable acidity were reached to pH 3.57 and 0.40% after 15 h fermentation, respectively. The viable cell population of L. mesenteroides 310-12 was rapidly increased to $8.9{\times}10^8$ CFU/g during the 15 h of cultivation. The contents of lactic acid and acetic acid were determined to be 0.077% and 0.065% after 15 h fermentation, respectively. The rice-based fermented beverage was manufactured by blending L. mesenteroides 310-12 fermented broth and some food additives. When this beverage was stored at $4^{\circ}C$, the viable cells population was decreased to $1.0{\times}10^7$ CFU/g and pH was nearly maintained for 25 days.