• Korean Journal of Clinical Laboratory Science
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• v.47 no.2
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• pp.71-77
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• 2015

Antimicrobial resistant bacteria has recently emerged and been disseminated in livestock environments because of excessive use of antimicrobial agents for the therapeutic and growth promotion purposes to food animals. In particular, there is potential for multidrug-resistant bacteria that can be transmitted from animals to mankind. In this study, we investigated the genotypes of E. coli strains isolated from humans and chickens using multi-locus sequence typing (MLST) and antimicrobial resistance patterns by disk diffusion method along with integron study involving antimicrobial resistance mechanisms. From July 2013 to July 2014, E. coli strains isolated from clinical specimens (n=44) and poultry chickens (n=34). ST131 (n=20) was most common in human-derived E. coli. ST752 (n=7) was most common in chicken-derived E. coli, with four isolates each for ST117, ST189, and ST69. Of the 44 E. coli strains isolated from humans, 25 of had a class 1 integron, as opposed to only 11 of 34 strains in the E. coli isolated from chickens. There were differences in genotypes and antimicrobial resistance patterns between the chicken-derived and the human-derived E. coli.

• Journal of fish pathology
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• v.17 no.3
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• pp.159-169
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• 2004

Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

• Parasites, Hosts and Diseases
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• v.53 no.4
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• pp.395-402
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• 2015

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.

• Research in Plant Disease
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• v.22 no.3
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• pp.158-167
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• 2016

A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.

• The Korean Journal of Mycology
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• v.48 no.3
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• pp.297-312
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• 2020

Wilted soybean plants were collected from soybeans cultivation fields in Korea from 2014 to 2016. Fusarium spp., Colletotrichum spp., Rhizoctonia spp., Macrophomina sp., Phytophthora spp., and Calonectria ilicicola were obtained from the infected samples. Out of these, Fusarium spp. were the dominant species (79.1%). In total, 53 isolates were identified as F. solani species complex, F. oxysporum species complex, F. graminearum species complex, and F. fujikuroi species complex based on mycological characteristics. Sequence typing analysis was conducted using translation elongation factor 1 alpha (TEF) to confirm the identification of isolates. All isolates were identified as F. solani, F. oxysporum, F. commune, F. asiaticum, and F. fujikuroi based on phylogenetic analysis of TEF sequences. Pathogenicity of 44 isolates was tested on three cultivars of soybean using the root dip inoculation method. Out of 5 Fusarium species, only F. asiaticum could not cause the symptoms or be weak. Ten isolates were selected based on pathogenic characters and species identification to investigate the host range and screen soybean cultivars for resistance. Fusarium solani, F. oxysporum, and F. commune were aggressive only to soybean, and F. fujikuroi was aggressive to kidney bean, yellow cowpea, black cowpea, adzuki bean as well as soybean. All 13 Korean soybean cultivars were susceptible to F. commune and F. fujikuroi. Out of 13 cultivars, cv. Janggi, cv. Poongsannamul, and cv. Socheongja were resistant to Fusarium wilt, while cv. Hwanggeumol and Chamol were susceptible to Fusarium wilt.

• Journal of Veterinary Science
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• v.23 no.6
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• pp.12.01-12.10
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• 2022

Background: Staphylococcus aureus and Streptococcus agalactiae are the main cause of clinical mastitis in dairy cattle in Argentina, whereas coagulase-negative staphylococci (CNS) and environmental streptococci are the main cause of subclinical mastitis. Bacteria isolated from infected animals show increasing antimicrobial resistance. Objectives: This study aims to determine the antimicrobial resistance of staphylococci and streptococci isolated from milk with mastitis, and to genotypically characterize the methicillin-resistant (MR) staphylococci. Methods: Isolation was performed on blood agar and identification was based on biochemical reactions. Antimicrobial susceptibility was according to the Clinical and Laboratory Standards Institute guidelines. The antimicrobial resistance genes, SCCmec type and spa type were detected by the polymerase chain reaction method. Results: We isolated a total of 185 staphylococci and 28 streptococci from 148 milk samples. Among the staphylococcal isolates, 154 were identified as CNS and 31 as S. aureus. Among the 154 CNS, 24.6% (n = 38) were resistant to penicillin, 14.9% (n = 23) to erythromycin, 17.5% (n = 27) to clindamycin, 6.5% (n = 10) to cefoxitin and oxacillin. Among the S. aureus isolates, 16.1% (n = 5) were resistant to penicillin, 3.2% (n = 1) to cefoxitin and oxacillin (MRSA). Six MR isolates (5 CNS and 1 MRSA) were positive to the mecA gene, and presented the SCCmec IVa. The MRSA strain presented the sequence type 83 and the spa type 002. Among the 28 streptococcal isolates, 14.3% (n = 4) were resistant to penicillin, 10.7% (n = 3) to erythromycin and 14.3% (n = 4) to clindamycin. Conclusions: The present findings of this study indicate a development of antimicrobial resistance in main bacteria isolated from cows with mastitis in Argentina.

• Microbiology and Biotechnology Letters
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• v.51 no.4
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• pp.517-525
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• 2023

The emergence and spread of multidrug-resistant Pseudomonas aeruginosa (MRPA) have become a serious problem worldwide. The involvement of metallo-β-lactamases (MBLs) in inducing carbapenem resistance is particularly acute. However, unlike other members of the Enterobacteriaceae genus, new clones of P. aeruginosa are constantly emerging and rapidly replacing previously prevalent dominant clones. Therefore, this study aimed to perform antimicrobial resistance gene analysis, integron gene cassette analysis using DNA sequencing, and plasmid transfer analysis by conjugation to investigate the antimicrobial resistance dynamics of 18 P. aeruginosa strains isolated from various medical samples at a general hospital in Busan from September 2017 to September 2019. All 18 strains showed extensively drug-resistant (XDR) phenotype and were resistant to most antibiotics, except colistin (100%) but were susceptible to aztreonam (22.2%) and ceftazidime (16.6%). Approximately 66.7% of the strains had Class 1 integrons showing various antimicrobial resistances. Notably, IMP-6 ST235 (66.7%), VIM-2 ST357 (16.7%), and IMP-1 ST446(16.7%) were identified. The identification of IMP-1-producing ST446, previously unreported in Korea, is noteworthy considering the emergence and prevalence of another MRPA high-risk clone.

• Journal of the Korea Institute of Military Science and Technology
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• v.27 no.4
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• pp.507-515
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• 2024

Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

• Journal of Life Science
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• v.18 no.12
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• pp.1733-1738
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• 2008

SLC6A18, one of the neurotransmitters, was reported the possible relationship to hypertension, and it contained eight blocks of minisatellites. In this study, SLC6A18-MS5 sequence which showed the highest heterozygosity among seven minisatellites was analyzed using the Transfac software, the putative binding sites for the transcription factor Pax4 and HNF4 were discovered as a result. The HNF4 is involved in the diabetes pathway and suggested the relationship to hypertension. Thus, we investigated the putative functional significance of allelic variation in this minisatellites with respect to susceptibility for hypertension. To address this possibility, we analyzed genomic DNA from the blood of 301 hypertension-free controls and 184 cases with hypertension. A statistically significant association was not identified between the allelic distribution of SLC6A18-MS5 and occurrence of hypertension. We then examined the meiotic segregation of SLC6A18-MS5 and it was transmitted following Mendelian inheritance. Therefore, this locus could be useful markers for paternity mapping and DNA fingerprinting. Moreover, we undertook a comprehensive analysis of the genomic sequence to address the evolutionary events of these variable repeats. SLC6A18 minisatellites regions are only conserved in human and primates. This result suggestedthat intronic minisatellites analysis is powerful evolution marker for the non-coding regions in primates and can provide a great insight to the molecular evolution of repeated region in primates.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.286-297
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• 2016

Clostridium perfringens is both a ubiquitous environmental bacterium and a major cause of human gastrointestinal disease, and C. perfringens food poisoning ranks among the most common gastrointestinal diseases in developed countries. 120 isolates of C. perfringens were obtained from food-poisoning outbreaks in 2013~2014, Gyeonggi-do. Using PCR, all 120 isolates were identified as C. perfringens type A. Of the tested isolates, 49 isolates carried the cpe gene, 71 isolates carried the cpb2 gene. The outbreak cases of cpb2 and cpe /cpb2 genes were 7 and 7, whereas the outbreak cases of cpe-gene were 2. The epidemiological relationship between C. perfringens isolates has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The genetics relatedness of the isolates raged from 53.5-100% and 75 district PFGE type were observed. The PFGE results revealed a wide genetic diversity among the 64 cpb2 carrying isolates (except 7 isolates), while 46 cpe-carrying isolates (except 3 isolates) showed a high genetic similarity. The MLST analysis revealed that 14 cpe isolates (cpe-chromosomal isolates) belong to a distinct cluster that is significantly distant from all the other cpb2 isolates (cpe-plasmid carrying and cpe-negative isolates). The isolates carrying a cpb2 appear to be rarely related, and are more variable than chromosomal cpe isolates. The results suggest that the cpe-positive outbreak isolates showed close genetic relation, whereas the cpb2-positive isolates revealed a wide genetic diversity.