• Title/Summary/Keyword: API-ZYM

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Isolation and Characterization of Cryptococcus neoformans from Environmental Sources in Busan

  • Oh, Kwang-Seok;Hwang, Soo-Myung
    • Mycobiology
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    • v.33 no.4
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    • pp.188-193
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    • 2005
  • Twenty nine samples of pigeon droppings (n = 12) and soil contaminated with avian excreta (n = 19), collected from different sites in Busan, were examined for isolation and characterization of Cryptococcus neoformans. Of these samples, 5 strains of C. neoformans were recovered from pigeon droppings (5/12 : 41.7%). All isolates were belonged to C. neoformans var. grubii (serotype A). The extracellular enzyme activities of the strains by using the API-ZYM system showed two different enzymatic patterns. The genetic variability among C. neoformans isolates was analyzed by random amplified polymorphic DNA (RAPD) using three 10-mer primers. Two different RAPD patterns, which clearly distinguished the isolates, were identified. Analysis of RAPD patterns provided a good characterization of environmental strains of C. neoformans serotype A as a heterogeneous group and were in good agreement with enzymatic profiles.

Phenotypic Diversity of Vibrio ichthyoenteri Isolated from the Gastrointestinal Tract of Larval Olive Flounder Paralichthys olivaceus

  • Han, Hyun-Ja;Lee, Deok-Chan;Kim, Do-Hyung;Choi, Hye Sung;Jung, Sung Hee;Kim, Jin Woo
    • Fisheries and Aquatic Sciences
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    • v.16 no.2
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    • pp.125-129
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    • 2013
  • Vibrio ichthyoenteri is a facultatively anaerobic gram-negative bacterium with straight or slightly curved rod morphology. The bacterium is an etiological agent of bacterial enteritis of Olive flounder Paralichthys olivaceus. Only a handful of studies, using a limited number of isolates, have investigated the phenotypic and genetic characteristics of V. ichthyoenteri. We isolated 40 V. ichthyoenteri strains, identified based on the 16S rRNA gene sequence, from the diseased flounder larvae and investigated the API 20E and ZYM profiles. The isolates exhibited highly divergent phenotypic characteristics regardless of sampling time point and location, and fish age. Essential enzymes produced by V. ichthyoenteri seemed to be alkaline phosphatase, leucine arylamidase, and N-acetyl-${\beta}$-glucosaminidase. This study reveals a much greater enzymatic and biochemical phenotype diversity than has been evident to date. These results suggest that a given population of V. ichthyoenteri could be heterogeneous in terms of its phenotypic characteristics.

Geographical comparison on different methods for identification of Streptococcus parauberis isolated from cultured olive flounder, Paralichthys olivaceus (양식 넙치에서 분리한 Streptococcus parauberis의 동정방법에 따른 지역적 비교)

  • Cho, Mi-Young;Oh, Yun-Kyeong;Lee, Deok-Chan;Kim, Jae-Hoon;Park, Myoung-Ae
    • Journal of fish pathology
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    • v.20 no.1
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    • pp.49-60
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    • 2007
  • Non-hemolytic Streptococcus parauberis isolated from diseased olive flounder, Paralichthys olivaceus in the South coast of Korea were identified by physiological, biochemical and genetic analysis in order to define the different characteristics geographically. First, twelve strains of S. parauberis were isolated from catalase-negative gram-positive cocci by multiplex PCR assay. Phenotypic identifications were performed with commercially available kit (API 20 Strep and API ZYM system). Analysis of API profiles of the isolates showed that strains were identified as either of Lactococcus lactis, S. constellatus or S. uberis. Moreover, S. parauberis isolated from olive flounder differed from that of turbot (X89967) to the test of not Voges-Proskauer, arginine, hippurate, alkiline phosphatase and pyrroidonyl arylamidase but β-glucuronidase. All S. parauberis isolates were sensitive to florfenicol, ampicillin, ofloxacin and vancomycin but were resistant to oxolinic acid, flumequine, nalidixic acid and sulfisoxazol. However, the 16S rDNA sequences of the isolates showed 99% similarity to S. parauberis KCTC 3651 (AY584477) and a great homogenecity among the flounder isolates.

A comparison of the hydrolase activities of excretory-secretory products and somatic extracts from fish parasitic nematodes, Anisakis simplex sensu stricto and Anisakis pegreffii larvae (어류 기생성 선충 Anisakis simplex sensu stricto와 Anisakis pegreffii 유충의 excretory-secretory products 및 somatic extracts의 가수분해효소 활성 비교)

  • Jeon, Chan-Hyeok;Wi, Seong;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.25-33
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    • 2014
  • Hydrolase activities of excretory-secretory products (ESP) and somatic extracts (SE) from Anisakis simplex sensu stricto (s.s.) and Anisakis pegreffii larvae were investigated by using API ZYM kit. In esterase group, acid phosphatase showed high activity from both of A. simplex (s.s.) and A. pegreffii. Esterase (C4) showed activity only from SE and A. simplex (s.s.) showed higher activity than A. pegreffii. Alkaline phosphatase, acid phosphatase and naphthol-AS-BI-phosphohydrolase showed higher activity in 3rd stage larvae than in 4th stage larvae of both species. In aminopeptidase group, only leucine arylamidase showed remarkable activity in SE of both anisakid species, and A. simplex (s.s.) SE showed higher activity than A. pegreffii SE. In glycosidase group, N-acetyl-${\beta}$-glucosaminidase, ${\alpha}$-mannosidase, ${\alpha}$-fucosidase showed higher activity in A. simplex (s.s.) than A. pegreffii, and 4th larvae showed higher activity than 3rd larvae. These differences in hydrolase activity of anisakid nematodes larvae are thought to be due to different metabolism such as growth, moulting, digestion and feeding.

Serotype and Enzymatic Profile of Crypfococcus neoformans Isolates from Clinical and Environmental Sources in Korea (한국의 임상과 자연환경에서 분리된 Cryptococcus neoformans의 혈청형과 효소생성능)

  • Hwang, Soo-Myung;Oh, Kwang-Seok;Lee, Kyung-Won
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.257-264
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    • 2006
  • Fifty eight Cryptococcus neoformans strains isolated from clinical and environmental sources in Korea were examined for their serotypes and extracellular enzyme activities. Among the 51 strains isolated from clinical sources, 48 strains were serotype A (94.1%), 2 strains were serotype B (3.92%), and 1 strain was serotype D (1.96%). All seven environmental strains isolated from pigeon excreta were identified as serotype A. All isolates of C. neoformtans were positive for the production of extracellular proteinase and phospholipase. In the API-ZYM system, all fifty eight isolates produced alkaine phosphatase, esterase C4, esterase lipase: C8, leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrase, $\alpha$-glucosidase and $\beta$-glucosidase. Thirty nine isolates (67.2%) of C. neoformans produced N-acetyl-$\beta$-glucosidase. Two isolates, serotype B, and B only one serotype A produced $\beta$-glucuronidase. Analysis of enzymatic profiles to 21 enzymes revealed four biotypic patterns among the 58 strains. The enzymatic patterns of C. neoformans isolated from clinical and environmental sources represented a significant relationship with the serotypes.

Histopathological reaction of red marbled rockfish, Sebastiscus tertius infected by Aeromonas salmonicida (Aeromonas salmonicida에 감염된 붉은쏨뱅이, Sebastiscus tertius의 조직병리학적 반응)

  • Kim, Seok-Ryel;Jin, Young Guk;Park, Jung Jun
    • Journal of fish pathology
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    • v.32 no.2
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    • pp.81-89
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    • 2019
  • As an epidemiological survey, mortality of marbled rockfish, Sebastiscus tertius observed from a fish farm in Gyeongnam province of South Korea. The major macroscopic sign of the diseased fish was severe multifocal dermal ulceration. Histological observation revealed inflammation, necrosis and colonization of bacteria in various tissues (gill, liver, spleen and kidney). Bacteria was isolated from spleen and kidney in moribund and mortality fish. Seven bacterial isolates from the diseased fish were identified as Aeromonas salmonicida subsp. salmonicida using API 20E and 20NE, API 50CH API ZYM system. Under light microscopy, infected marbled rockfish showed the lifting of the lamella epidermal layer, edematous changes and hypertrophy of epithelial cell in the gill filament. The atrophy of the mucosal fold, erythema in the intestine, and the necrosis of hematopoietic tissue and renal tubule cells with karyolysis were observed in the kidney. In this study was demonstrated the histological reaction of red marbled rockfish infected by Aeromonas salmonicida. Furthermore, this is the first account of extensive dermatitis in Sebastiscus tertius due to atypical A. salmonicida infection, which has high potential in aquaculture among native fish species.

Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-1 Gene (Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정)

  • Yoon, Sang-Hong;Kim, Dong-Gwan;Lee, Young-Han;Shin, Soon-Young;Kwon, Soon-Woo;Lee, Chang-Muk;Kang, Han-Chul;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.119-125
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    • 2011
  • The Egr-1 gene is known to be a transcription factor for activating the expression of many tumor-repressing genes. In this study, three strains activating the promoter of the Egr-1 gene were selected, through the use of Egr-1 luciferase reporter assay and western blotting, from amongst approximately 3,800 oligotrophic bacteria isolated from the cultivated soils of various regions within Korea. These strains were identified as Pseudomonas koreensis on the basis of phylogenetic tree analysis of their 16S ribosomal DNA sequences and biochemical characteristics analyses using a variety of commercial kits (API 20NE, ID 32GN, API ZYM kits). In addition, we discovered that these strains produced anti-bacterial activity against Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes.

Phenotypic Characteristics of Pseudomonas syringae pv. actinidiae Strains from Different Geographic Origins (지리적 기원이 다른 Pseudomonas syringae pv. actinidiae 균주들의 표현형적 특성)

  • Choi, Eun Jin;Lee, Young Sun;Kim, Gyoung Hee;Koh, Young Jin;Jung, Jae Sung
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.245-248
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    • 2014
  • Pseudomonas syringae pv. actinidiae is the causal agent of bacterial canker in kiwifruit (genus Actinidia). Multilocus sequence analysis of seven housekeeping and 11 type III effector genes differentiated the virulent P. syringae pv. actinidiae isolates worldwide into three groups designated as Psa1-Psa3. In this work, a total of 12 P. syringae pv. Actinidiae strains, including three Psa1, three Psa2, three Psa3 strains isolated from Korea and three Psa3 strains from Italy, were compared based on their phenotypic properties. Strains with different geographic origins had unique growth patterns as demonstrated by growth rate at several temperatures; all tested strains exhibited maximum growth at temperatures below $22^{\circ}C$, while the growth of Psa3 strains was completely inhibited above $30^{\circ}C$. Psa3 strains isolated from Korea had longer lag phases than the Psa3 strains from Italy. The Psa2 strains were different from Psa1 and Psa3 strains in the API 20NE test, in which the Psa2 strains could not utilize potassium gluconate, capric acid and trisodium citrate. Psa3 strains isolated from Korea could hydrolyze esculin. The API ZYM test showed that ${\beta}$-glucosidase activity was detected only from Psa3 strains. The strains belonging to the three Psa groups differed with regard to their susceptibility to ampicillin, novobiocin, and oleandomycin.

Studies on the Ornithobacterium rhinotracheale infection in the chickens (닭에서 Ornithobacterium rhinotracheale 감염증에 대한 연구)

  • Kwon, Yong-Kuk;Jeon, Woo-Jin;Kim, Jae-Hong
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.121-127
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    • 2003
  • Ornithobacterium rhinotracheale (OR) is a bacterium responsible for a respiratory disease in turkeys and chickens, and has been identified as one of the emerging respiratory bacterial pathogens. Ten cases of four hundred cases submitted to National Veterinary Research & Quarantine Service for diagnosis in 2001 and in 2002 were diagnosed as OR infection. The major clinical signs of chickens infected with OR were respiratory symptoms including sneezing, sniveling, wet eyes, and swelling of the sinus infraorbitalis at 3 to 4 weeks of age. At necropsy, gross lesions were commonly found to foamish, white, and yoghurt-like exudates in the peritonium and abdominal air sacs. Microscopically, epithelial metaplasia and proliferation of air sacs were prominant with accompaning inflammatory reactions characterized by heterophils, fibrins, and bacterial colonization. Ten field isolates were obtained from air sacs and peritonium of these affected chickens, and were identified as OR, resulted from by gram-staining, catalase, oxidase, API NE and API ZYM Kit. In additon, using a previously reported primer targeted to 16S rRNA of ORT, 784bp fragment was successfully amplified from templates extracted from the isolates and a reference strain. This report describes an occurrence of Ornithobacterium rhinotracheale infection in chickens in Korea.

Probiotic Properties of Lactobacillus plantarum NK181 Isolated from Jeotgal, a Korean Fermented Food

  • Lee, Na-Kyoung;Kim, Hyoun-Wook;Chang, Hyo-Ihl;Yun, Cheol-Won;Kim, Seung-Wook;Kang, Chang-Won;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.227-231
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    • 2006
  • Strain NK181 was isolated for probiotic use from jeotkal and based on results of API 50 CHL kit and 16S rDNA sequencing was tentatively named Lactobacillus plantarum NK181. L. plantarum NK181 was highly resistant to artificial gastric juice (pH 2.5) and bile acid and demonstrated strong adherence to Caco-2 cells. In test using API ZYM kit, eight enzymes were produced. Supernatant of L. plantarum NK181 exhibited about 30% 1,1-diphenyl-2-picyryl hedrazyl (DPPH) radical-scavenging activity and reduced cholesterol by 70%. These results demonstrate potential use of L. plantarum NK181 as health-promoting probiotic.