• Title/Summary/Keyword: bacterial identification

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Characterization of typical Aeromonas salmonicida isolated from Sea-Chum Salmon (Oncorhynchus keta) (해수에 순치된 첨연어(Oncorhynchus keta)에서 분리된 정형 에로모나스 살모니시다(Aeromonas salmonicida)에 대한 특성 분석)

  • Jongwon Lim;Sungjae Ko;Youngjun Park;Do-il Ahn;Suhee Hong
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.263-275
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    • 2023
  • Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ70), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

The Microbiological Assessment and Identification of Food Utensils and Food Service Facilities in School (학교 급식설비 및 집기류의 미생물학적 위해요소 분석)

  • Hong, Seung-Hee
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.189-194
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    • 2014
  • This study was conducted to evaluate microorganism contamination of food utensils and service facilities in school and to prevent hazards by food poisoning occurrence. As a result, the highest number of microorganism growth plate ($12.3{\pm}2.6$) was detected in total bacteria test plate, and also observed $10.3{\pm}3.9$ growth plates in Staphylococcus aureus test plate and $9.5{\pm}3.9$ growth plates in E. coli and coliform bacteria test plate. But we could detect to the lowest number of growth plates ($1.5{\pm}1.0$) in Vibrio test plate. We also assessed that floors were appeared to the highest microorganism contamination rate in food utensils and service facilities. Therefore, $4.5{\pm}0.6$ growth plates was detected in pre-operation floor and $4.3{\pm}1.0$ growth plates in floor. And high level of microorganism contamination also observed in tables as $3.3{\pm}1.0$ growth plates in cooking table and $3.0{\pm}0.0$ growth plates in dining table. The level of microorganism contamination of food utensils such as kitchen knife, cutting board, and food tray were lower than that in food service facilities. We analysed microorganism contamination according to purpose of use in kitchen knifes and cutting boards. The microorganism contamination rate in fish kitchen knife ($2.0{\pm}0.8$) and fish cutting board ($1.3{\pm}1.5$) were slightly higher than that of others purpose of use. As a result of microorganism identification, various strains of microorganism were contaminated in food service facilities and some strains could detected more than two times. Especially, Staphylococcus aureus was repeatedly identified in cooking table, trench, and kitchen knife. Bacillus cereus was identified in kitchen knife, and then Pseudomonas fluorescens and Pseudomonas aeruginosa were also detected in food utensils and service facilities as known to food spoilage microorganisms. Klebsiella pneumoniae was detected four times repeat, which widely distribute natural environment as normal bacterial flora but sometimes cause acute pneumonia. These results suggest that food utensils and service facilities are contaminated with not only major food poisoning microorganisms such as Staphylococcus aureus, but also food spoilage microorganisms. Taken together, strict personal hygiene control and efficient food service facilities management will be needed to enhance food safety in school feeding and to improve student health.

A novel cold-active lipase from Psychrobacter sp. ArcL13: gene identification, expression in E. coli, refolding, and characterization (새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구)

  • Koo, Bon-Hun;Moon, Byung-Hern;Shin, Jong-Suh;Yim, Joung-Han
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.192-201
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    • 2016
  • Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.

Microbiological Identification and Distribution of Metal Components in Suspended Particulate Matter during Yellow Sand Phenomena at TaeAn Region in 2003 (2003년 태안지역에서 황사 부유분진의 미생물학적 동정과 금속 성분 및 농도)

  • Bae, Kang Woo;Kim, Jong Ho;Kim, Youn Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.2
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    • pp.167-173
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    • 2005
  • Background : Airborne particles during Yellow Sand phenomena are known to be associated with the respiratory disease. The purpose of this study was to evaluate the concentration and metal component properties of Yellow Sand particles and compare with airborne microbial concentration and species in non Yellow Sand and Yellow Sand phenomena. Methods : Samplings were carried out in 2002 in Seosan, during non Yellow Sand and Yellow Sand phenomena. Samples were taken using the 8-stage Cascade impactor and metallic elements were analyzed by XRF. Those were culture on the media for bacterial and fungal culture and celline for virus. Results : The concentration of total suspended particulate matter were respectively $80.2{\mu}g/m^3$, $40.3{\mu}g/m^3$ in non Yellow Sand and Yellow Sand phenomena. The concentration of metallic elements such as Ca, Fe, Cu and Zn in Yellow Sand phenomena were higher than its in non Yellow Sand. Two bacteria, Bacillus species and Staphylococcus were grown in two periods. In both periods, several fungal spores(Mucor species, Cladosporum, Alternaria, Aspergillus, Penicillium, and Alternaria species) were identified. The differences of bacteria and fungus species not observed in Yellow Sand and non Yellow Sand. Any viruses were not isolated in between both periods. Conclusions : The concentration of total suspended particulate matter and some metallic elements in Yellow Sand phenomena were higher than its in non Yellow Sand. The difference of bacteria and fungus species was not observed in non Yellow Sand and Yellow Sand phenomena.

Etiologic Survey on the Acute Diarrhea of Puppies in Seoul (서울지역에서 문제되고 있는 강아지의 급성설사증에 관한 병인학적 연구)

  • Han Hong-Ryul;Park Hee-Myung;Lee Jin-Hee;Oh Tae-Ho;Jeong Soon-Wuk;Youn Sin-Keun;Park Cheol-Man
    • Journal of Veterinary Clinics
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    • v.10 no.2
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    • pp.171-184
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    • 1993
  • In Seoul area, there are so many kennel clubs, veterinary hospital, pet establishment and breeding confers that various problems have occurred. They are crowed pet houses, poor sanitation, stress to puppies, sudden environmental changes to puppies and unvaccination against parvovirus, canine distemper virus, parainfluenza virus, infectious hepatitis caused by Adenovirus type I and Leptospira. Several studies were made to survey the infectious agents involved in acute diarrhea of poppies in Seoul are, such as history taking, physical examination, complete blood count and serum chemistry, histopathological finding, bacterial isolation and identification, and hemagglutination test in feces and hemagglutination inhibition test in serum against parvovirus, respectively. The results obtained are summarized as followed. 1. The percent of PCV (30.5$\pm$5.6) and concentration of Total protein(5.0$\pm$0.8) resulted from statistical analysis are significantly lower than normal values (p<0.05), respectively. In addition, fibrinogen (505$\pm$326) was significantly higher an normal value (p<0.001), Band neutrophil (22.9$\pm$12.7) showed signifiant difference (P<0.01). decreased monocyte (3.2$\pm$2.1) and eosinophil (0.7$\pm$0.8) values appeared statistically significant (p<0.001), lymphocyte (16.7$\pm$9), as well. 2. The concentrations of calcium .(8.0$\pm$2.8), glucose (40.1$\pm$31.4), and albumin (2.0$\pm$0.39) were lower than normal values (p<0.01, p<0.01, p<0.001), respectively. Also Inorganic phosphate (7.1$\pm$2.4), pH (p<7.9f:0.2), and Blood Urea Nitrogen (40.41=37.1) were significantly higher than normal values (p<0.001, p(0.001, p<0.05), respectively. 3. Simple and mixed infections occupied 18% and 82% in the distribution of causes in puppies with acute diarrhea, respectively. 4. As puppies got older, incidence of acute diarrhea caused by Staphylococcus aureus was decreased to 13% and infection of canine distemper virus was increased to 53%, but E coli and canine parvovirus always showed high frequency of outbreak in the body weight ranged from 35g to 7.8Kg. 5. As showed in table 5, infections of E coli and Canine Parvovirus showed high outbreak regardless of the age which is classified into three stages, 35~50 day, 60 day and 75~day to 1 year, canine distemper virus appeared increased, but in case of Staphylococcus aureus, visa versa. 6. In comparison wi methods for the laboratory diagnosis diagnosis parvovirus, Hemagglutination test showed positive reaction in 25% and mean serum antibody titer measured by Hemagglutination inhibition test showed 2779 (n=20). In addition, positive reaction was 90% (18/20). 7. In histopathological studies, enteritic and pneumonic lesions were indicated in 53.7% and 39.5% of samples, respectively. 8. Etiologic diagnosis based on the history taking, clinical signs and histopathological findings in puppies with acute diarrhea and vomiting indicated that 50% of puppies were infected by canine parvovirus and distemper virus. 9. In the parasitological examination made by simple flotation with saturated zinc sulfate tour parasites found were Isospora canis, Toxocara canis, Ancylostoma spp and Toxocara spp. Isospora canis and Toxocara canis were more frequently found among those parasites of diarrhoeic causes in puppies ranged from 35 days to 1 year. Their infestation rates were 15% and 13% respectively.

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Effective Ways of Performing Surveillance Surface Cultures in Extremely Low Birth Weight Infants

  • Lee, Ju-Young;Kim, Ee-Kyung;Lee, Jin-A;Choi, Chang-Won;Kim, Han-Suk;Kim, Beyong-Il;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.18 no.2
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    • pp.240-247
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    • 2011
  • Purpose: The rationale for skin surface cultures is that bacterial colonization precedes infection and, as a result, that identification of a potential pathogen is predictive of later systemic infection in preterm infants. We aimed to analyze results of surveillance surface cultures in extremely low birth weight (ELBW) infants and seek for effective ways of performing surveillance surface cultures. Methods: We analyzed the surveillance surface cultures of 113 ELBW infants over a 4-year period. Surveillance cultures were obtained routinely from five sites: axilla, external ear canal, nasopharynx, throat (or tracheal aspirate if intubated) and anus. Each surface culture obtained during the 13 days, prior to the date of the blood culture, was compared with the blood culture obtained when sepsis was suspected. The sensitivity, specificity and positive predictive value (PPV) of the surveillance cultures were calculated among 1894 blood-surface culture pairs by surface sites, recovered organisms and interval between surveillance samples and blood cultures. Results: The overall sensitivity, specificity and PPV of surface cultures were 45.9, 22.4 and 6.8%, respectively. The PPV was highest for the throat/tracheal cultures (11.0%) and lowest for the anal cultures (2.3%). As the time of culturing progressed toward the day of blood culturing, the sensitivity and specificity of the surface cultures significantly increased. Only axillary and throat/tracheal cultures were useful in predicting the microorganisms causing sepsis. Conclusion: Surface cultures could help to predict sepsis pathogens and infection surveillance in preterm infants could be continued with a reduced number of cultured sites focusing on the axilla and throat/trachea.

Isolation and Identification of Oceanisphaera sp. JJM57 from Marine Red Algae Laurencia sp. (Ceramiales: Rhodomelaceae) (해양 홍조류 Laurencia sp. (Ceramiales: Rhodomelaceae)에서 분리한 Oceanisphaera sp. JJM57의 분리 및 동정)

  • Kim, Man-Chul;Dharaneedharan, S.;Moon, Young-Gun;Kim, Dong-Hwi;Son, Hong-Joo;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.58-63
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    • 2013
  • A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

지류 및 섬유질 문화재의 미생물에 관한 연구 (紙類 및 纖維質 文化財의 微生物에 關한 硏究) ­경남지방(慶南地方)을 중심(中心)으로-

  • Min, Gyeong-Hui;An, Hui-Gyun
    • Korean Journal of Heritage: History & Science
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    • v.14
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    • pp.225-250
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    • 1981
  • The investigation of air fungal population in the storages to keep papers and textiles that are designated as important folk life materials or treasures was carried out from Dec. 17 to. 23, 1980. Isolation media was used for malt extract agar with chloramphnicol to prevent bacterial contamination. Isolation and identification of air fungi from the four preserved rooms were Cladosporium cladosporioides, Alternaria chlamydospora, Aspergillus fumigatus, A. versicolor, Eurotium chevalieri, Penicillium charlesii var. rapidum, P. oxalicum. P. viridicatum, Trichoderma viride, Acremomium sp., Mucor sp. and Yeast. It was found that nine species in eight genera was isolated. Among them, underscribed species in Korea was two species ; Eurotium chevalieri and Penicillium visidicatum. The fungal population of four storages was showed to be dominant species such as Cladosporium cladosporioides and the order was Acremonium, Penicillium, Aspergillus, Trichoderma, Alternaria and Eurotium. Eurotium chevalieri was ascomycetous fungi including distinctive ascospores in cleistothecia, the filamentous fungi was directly isolated from the papers and cellulose materials showing to be fourteen species in eight genera. The most species of the fungi isolated was also Cladosporium cladosporioides and the other fungi were found as Acremonium, Penicillium, Aspergillus and Trichoderma. It was confirmed that underscribed fungi were two species ; Mucor racemosus and Penicillium spinulosvm. The effect of four antifungal agents, benzoic acid, sorbic acid, dehydroacetic acid and thymol was also examined on eight species of Aspergillus, Penicillium, Cladosporium. and Tricoderma. this results were shown that more than 0.5% concentration of thymol inhibited the grow of all fungalspecies and other three chemicals appeared various inhibition zones of fungal growth depending in their different concentrations.

Incidence of hearing loss and importance of risk factors in the neonatal intensive care unit (신생아 중환자실에서 난청의 발생빈도 및 위험요소의 중요성)

  • Kong, Seung Hyun;Kang, Jang Hee;Hwang, Kwang Su;Kim, Joong Pyo;Lee, Hyeon Jung;Choi, Hyeon;Mok, Ji Sun;Kim, Jung Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.8
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    • pp.845-850
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    • 2006
  • Purpose : To assess the incidence of neonatal hearing loss in a neonatal intensive care unit and the relative importance of risk factors for hearing imparement in a neonatal intensive care unit which the Joint Committee on Infant Hearing(JCIH) had recommended. Methods : One thousand, two hundred and one newborns admitted to the Good Moonhwa Intensive Care Unit from May 2003 to December 2005 were assesed using the automated auditory brainstem response(AABR). The screening was performed on those aged more than 36 weeks and weighing more than 2,200 g. We divided the infants into two groups, 'pass' and 'refer'. The 'refer' group were retested one month later, and if classified as 'refer' during the retest, were referred to a hearing impairment clinic. Results : From the 1,201 neonates, 1,187(98.8 percent) passed the test and 14(1.2 percent) failed. 293(24.4 percent) of the 1,201 neonates had a risk factor for hearing impairment; 282(96.2 percent) passed the test and 11(3.8 percent) failed. The group with risk factors were shown to have a higher incidence of hearing loss(P<0.001). The neonates in the refer group were shown to have a higher incidence of ototoxic drugs(P<0.001), low birth weight(<1,500 g)(P<0.001) and craniofacial anomalies(P=0.007). On the other hand, there were no statistical differences between the pass and refer groups in congenital infection, hyperbilirubinemia, bacterial meningitis, low Apgar scores, prolonged mechanical ventilation and syndromes known to include hearing loss. Conclusion : In order to identify hearing-impaired infants within an appropriate period, neonatal hearing screening tests and identification of the risk factors for neonatal hearing loss are important.

Developmental Genetic Analysis of Avian Primordial Germ Cells and the Application to Poultry Biotechnology

  • Kagami, H.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.135-142
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    • 2001
  • A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

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