• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.3
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• pp.443-450
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• 2020

No established method can be used to select effective antibiotics in antibiotic susceptibility tests for fish bacterial pathogens quickly and accurately. Here, we established the optimal conditions for determining the minimal inhibitory concentration (MIC) of major fish bacterial pathogens (Streptococcus spp., Edwardsiella tarda, Vibrio spp., Aeromonas spp., and Pseudomonas spp.) using the KRAQ1 and CAMPY2 panels. The MIC panel used 18 antibiotics of two types and we conducted experiments to establish the optimal culture medium and temperature for each species. The optimal conditions for incubating Streptococcus spp. were in cation-adjusted Mueller-Hinton broth with TES buffer (CAMHBT) at 28℃, using 5% lysed horse blood (LHB) as recommended by the Clinical Laboratory Standards Institute. For Vibrio spp., the optimal culture conditions were 28℃ in CAMHBT supplemented with 1% NaCl. The optimal conditions for culturing E. tarda, Aeromonas spp., and Pseudomonas spp. were in CAMHBT at 28℃.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.38-44
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• 2006

A simple and easy modification of AST by disk diffusion was tested for the detection of induced clindamycin resistant Staphylococci and their antimicrobial susceptibility at the same time. The incidence of inducible clindamycin resistant staphylococci in blood culture and their MIC characterization at Seoul National University Hospital was analyzed by an AST contained disk approximation test (D-zone test) and Etest, respectively. Of the total 309 staphylococcal isolates, 139 (45%) isolates presented constitutive resistance to ERY and CLI (ERY-R, CLI-R phenotype), and 59 were ERY-I/R and CLI-S phenotypes. Of the 59 isolates, 19 (32%) isolates were inducible resistant to CLI. The incidence was higher in S. aureus (66.7%) than coagulase-negative staphylococci (CNS, 26.0%). Especially, methicillin-resistant staphylococci (MRSA, 100%; MRCNS, 45.5%) presented higher inducibility than methicillin susceptible (MSSA, 50%; MSCNS, 20%). For most of the inducible clindamycin resistant staphylococci (15 of 19 isolates), their ERY MIC were high (>$128_{\mu}g/mL$) and were methicillin resistant. The remaining 4 isolates were methicillin susceptible and their ERY MIC were of intermediate concentrations ($1-4_{\mu}g/mL$). We concluded that suscetibility testing of staphylococci, especially methicillin resistant, should include the D-zone test.

• Journal of Environmental Health Sciences
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• v.47 no.5
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• pp.479-485
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• 2021

Background: Vibrio vulnificus has been frequently detected in seawater, fish, and shellfish mainly in the coastal areas of Chungcheongnam-do Province. Objectives: This study was conducted to investigate the analyzed biochemical properties, genetic characteristics, and distribution of Vibrio vulnificus isolated from environmental sources in coastal areas of Chungcheongnam-do Province from 2019 to 2020. Methods: A total of 1,510 samples were obtained from six different sites in Chungcheongnam-do Province. Isolated strains from the samples were identified by a VITEK 2 system and MALDI-TOF. Antibiotic susceptibility testing for 85 isolates was done by microdilution minimum inhibitory concentration methods, and 11 isolates were analyzed for 16s rRNA sequences in multiple alignments. Results: Among the 1,510 samples taken during the investigation period, 306 strains were isolated and the detection rate of V. vulnificus was 20.3%. One hundred eighty-eight strains (24.6%) from seawater and 118 strains (15.8%) from mud flats were isolated. It was mainly detected in July (17.3%), August (36.5%), and September (28.8%), and the proportion was 82.0%. Based on the CLSI-recommended breakpoints, V. vulnificus isolates were all susceptible to amoxicillin/clavulanic acid, amikacin, gentamicin, ciprofloxacin and trimethoprim/sulfamethoxazole. However, nonsusceptible isolates to ampicillin, ampicillin/sulbactam, cefazolin, cefoxitin, imipenem, tetracycline and chloramphenicol were identified. In the analysis of the nucleotide sequences for 16s rRNA of V. vulnificus isolates, it was confirmed that mutations frequently occurred between nucleotide number 922 and 952, and 98.2% to 100% nucleotide identities between isolates was verified. Conclusions: The results of this study can be used as a basic data for infection control and prevention of Vibrio vulnificus infection by describing the distribution and characteristics of Vibrio vulnificus strains isolated in coastal areas of Chungcheongnam-do Province.

• Pediatric Infection and Vaccine
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• v.26 no.1
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• pp.42-50
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• 2019

Purpose: We investigated the clinical features and epidemiology of staphylococcal scalded skin syndrome (SSSS) from year 2006 to 2015 in Changwon city, Korea. Methods: We reviewed medical records of 69 patients diagnosed with SSSS from year 2006 to 2015. Antibiotic susceptibility testing was performed by agar dilution method. Methicillinresistant Staphylococcus aureus (MRSA) was phenotypically identified by oxacillin susceptibility testing and genotypically confirmed by the existence of the mecA gene. Results: The median age of patients was 2.0 years (range 0.2-6 years). Three (4.3%), 53 (76.8%), and 13 (18.9%) patients showed the generalized type, the intermediate type, and the abortive type, respectively. Patients occurred throughout the year, but most patients occurred between July and October. MRSA was isolated from 54 of the 60 patients regardless of the clinical types. All patients recovered without any complications. Conclusions: There was a constant occurrence of SSSS patients caused by MRSA in Changwon area during 2006 and 2015. It is needed to constantly monitor the occurrence of patients with SSSS.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.334-340
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• 2010

It is well known that smoking as well as drinking is a factor of stomatopathy, however there are few investigations about comparison of oral flora between smokers and non-smokers. In this study, we isolated the oral flora of 30 smokers and 30 non-smokers and cultured them on blood agar plates. The isolated pathogenic microorganisms were tested for antibiotic susceptibility and resistance using the Kirby-Bauer antibiotic testing method. Each colony was stained using the Gram staining method and was identified by an automatic identifier, known as the VITEK system. We isolated 41 colonies from smokers' oral cavity, and they were sorted as 63% of Gram-positive cocci, 29% of Gram-negative cocci, 3% of Gram-positive bacilli, and 5% of Gram-negative bacilli by gram staining, whereas 38 colonies were isolated from non-smoters' oral cavity, and their proportions were 55% of Gram-positive cocci, 26% of Gram-negative cocci, 3% of Gram-positive bacilli, and 16% of Gram-negative bacilli. The VITEK system revealed specific distribution of bacteria species that Streptococcus mutans (6/41), Gemella morillorum (6/41), Streptococcus oralis (2/41), Streptococcus pneumoniae (1/41), Staphylococcus aureus (3/41), Streptococcus anginosus (1/41), Streptococcus intermedius (1/41), Streptococcus uberis (1/41), and Streptococcus sanguinis (1/41) in smokers oral cavity whereas Streptococcus sanguinis (8/38), Staphylococcus aureus (1/38), Staphylococcus auricularis (1/38), Streptococcus uberis (1/38), Streptococcus intermedius (1/38), Streptococcus mutans (1/38), and Streptococcus oralis (1/38) in those of non-smokers'. Three cases of Staphylococcus aureus from smokers produced Beta-lactamase and were identified methicillin-resistance Staphylococcus aureus (MRSA). However one case of Staphylococcus aureus from non-smoker did not produce Beta-lactamase and was sensitive to methicillin. In conclusion, the distribution of oral flora was different between smokers' and non-smokers' oral cavity, especially Gemella morillorum and MRSA were predominantly found in smoker's oral cavity. These results are useful in the treatment and prevention of patients with stomatopathy caused by smoking.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.26-33
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• 2006

Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.4
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• pp.382-388
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• 2019

Streptococcosis caused by Streptococcus parauberis is a very important disease in farmed olive flounder Paralichthys olivaceus. For most fish pathogens, including S. parauberis, there are no analytical criteria to distinguish antibioticsusceptible strains from antibiotic-resistant strains. In this study, epidemiological cut-off ($CO_{WT}$) values were generated to classify 75 strains of S. parauberis isolated from 1999 to 2018 as wild type (WT) and non-wild type (NWT) using disc diffusion data and normalized resistance interpretation (NRI) analysis. The susceptibility of the isolates to 16 antibiotics was evaluated using CLSI guideline M42-A. The wild-type cut-off values for amoxicillin, erythromycin, oxytetracycline, and florfenicol for S. parauberis were ${\geq}35$, 31, 28, and 27 mm, respectively. The NWT ratios of S. parauberis strains to treatment with GEN, FFC, ENR, SXT, EFT, VAN, and CHL were 17% or less, indicating that these antibiotics may be used to treat streptococcosis caused by S. parauberis. For recent S. parauberis isolates, the NWT ratios for AMX, ERY, OTC and FFC are much higher than for strains isolated from 1999-2007. The $CO_{WT}$ data from this study will assist aquatic animal disease professionals in prescribing appropriate antibiotics for the treatment of streptococcosis caused by S. parauberis, which will help reduce the misuse and abuse of antibiotics in the aquaculture sector.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.104-110
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• 2004

To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

• Journal of Veterinary Clinics
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• v.32 no.3
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• pp.268-271
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• 2015

About 2-year old northern goshawk (Accipiter gentilis) which is designated as natural monument (#323-1) in Republic of Korea was rescued by a local farmer and presented with a 2-weeks history of pain, swelling, stiffness and limping. On physical examination, plantar pododermatitis and digit IV weakness were observed. Radiographic findings also showed bone lyses with soft tissue swelling in the foot. A definite diagnosis was made as stage III bumble foot after multidisciplinary approach of the patient. Bacterial culture was performed, and concurrently antibiotic susceptibility testing is determined using wound site exudates specimen. Bacterial isolates were identified as Staphylococcus aureus, known normal skin flora. Treatment was initiated with surgical incision and necrotized tissues removal. Lavage-drainage and ball bandage were applied with topical mupirocin ointment application. Doughnut shaped pad was attached on bottom of the ball bandage to reduce weight bearing. After three weeks of intensive care, the wound site completely healed but digit IV weakness remained permanently. The goshawk returned to nature after eight weeks after treatment.

• Journal of Microbiology
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• v.44 no.4
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• pp.423-431
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• 2006

Among 53 Acinetobacter baumannii isolates collected in 2004, nine imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Korea. Nine carbapenemase-producing isolates were further investigated in order to determine the mechanisms underlying resistance. These isolates were then analyzed via antibiotic susceptibility testing, microbiological tests of carbapenemase activity, pI determination, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. One outbreak involved seven cases of infection by A. baumannii producing OXA-23 ${\beta}-lactamase$, and was found to have been caused by a single ERIC-PCR clone. During the study period, the other outbreak involved two cases of infection by A. baumannii producing IMP-1 ${\beta}-lactamase$. The two clones, one from each of the outbreaks, were characterized via a modified cloverleaf synergy test and an EDTA-disk synergy test. The isoelectric focusing of the crude bacterial extracts detected nitrocefin-positive bands with pI values of 6.65 (OXA-23) and 9.0 (IMP-1). The PCR amplification and characterization of the amplicons via direct sequencing showed that the clonal isolates harbored $bla_{IMP-1}$ or $bla_{oxA-23}$ determinants. The two clones were characterized by a multidrug resistance phenotype that remained unaltered throughout the outbreak. This resistance encompassed penicillins, extended-spectrum cephalosporins, carbapenems, monobactams, and aminoglycosides. These results appear to show that the imipenem resistance observed among nine Korean A. baumannii isolates could be attributed to the spread of an IMP-lor OXA-23-producing clone. Our microbiological test of carbapenemase activity is a simple method for the screening of clinical isolates producing class D carbapenemase and/or class B $metallo-{\beta}-lactamase$, in order both to determine their clinical impact and to prevent further spread.