[Show abstract][Hide abstract] ABSTRACT: Background. Recently isolated Vibrio parahaemolyticus strains have displayed multiple antibiotic resistance. Alternatives to conventional antibiotics are needed, especially for the multiple-antibiotic-resistant V. parahaemolyticus pandemic strain.Methods. A bacteriophage, designated pVp-1, showed effective infectivity for multiple-antibiotic-resistant V. parahaemolyticus and V. vulnificus, including V. parahaemolyticus pandemic strains. The therapeutic potential of the phage was studied in a mouse model of experimental infection using a multiple-antibiotic-resistant V. parahaemolyticus pandemic strain. We monitored the survivability and hispathological changes, quantified the bacterial and phage titers during phage therapy, and observed the immune response induced by phage induction.Results. Phage-treated mice displayed protection from a V. parahaemolyticus infection and survived lethal oral and intraperitoneal bacterial challenges.Conclusions. This is the first report, to the best of our knowledge, of phage therapy in a mouse model against a multiple-antibiotic-resistant V. parahaemolyticus pandemic strain infection.
The Journal of Infectious Diseases 02/2014; · 5.85 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Thelohanellus kitauei (Myxobolidae) infects cyprinid fish. The evolution of species derived from common ancestors results in the sharing of biological features. To reveal the origin of T. kitauei biological features, the correlation between phylogeny and biological features of Myxobolidae was investigated by Bayesian inference tree and Bayesian tip association significance testing. The results demonstrated that host specificity and infection site tropism were correlated with the phylogeny of Myxobolidae, and that the biological features of T. kitauei originated from the ancient Myxobolidae as exhibited by the non specific infection site tropism and the ability to infect cyprinids.
Molecular Phylogenetics and Evolution 01/2014; · 4.07 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Philasterides dicentrarchi is a causative agent of scuticociliatosis in olive flounder Paralichthys olivaceus, aquaculture in Korea. In this study, a cDNA encoding a cathepsin L-like cysteine protease (PdCtL) of P. dicentrarchi (synonym Miamiensis avidus) was identified. To express the PdCtL recombinant protein in a heterologous system, 10 codons were redesigned to conform to the standard eukaryotic genetic code using polymerase chain reaction (PCR)-based site-directed mutagenesis. The recombinant P. dicentrarchi procathepsin L (proPdCtL) was expressed at high levels in E. coli Rosetta (DE3) pLysS with a pPET21a vector, and successfully refolded, purified, and activated into a functional and enzymatically active form. The optimal pH for protease activity was 5. Similar to other cysteine proteases, enzyme activity was inhibited by E64 and leupeptin. Immunogenicity of recombinant PdCtL was assessed by enzyme-linked immunosorbent assay, western blot, and specific anti-recombinant PdCtL antibodies were detected. Our results suggest that the biochemical characteristics of the recombinant ciliate proPdCtL protein are similar to those of the cathepsin L-like cysteine protease, that the PCR-based site-direct mutated ciliate gene was successfully expressed in a biochemically active form, and that the recombinant PdCtL acted as a specific epitope in olive flounder.
Parasitology International 12/2013; · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We describe coagulase-negative staphylococci (CoNS) isolates collected from ducklings exhibiting tremor in South Korea over the period of 2010 to 2011. Screening of antimicrobial susceptibility and analysis of SCCmec elements of CoNS were also investigated.
Staphylococcus cohnii was the most frequent staphylococcus (9 isolates) and S. sciuri (4 isolates), S. lentus (3 isolate), S. simulans (1 isolate) and S. epidermidis (1 isolate) were also detected. Among the 15 antimicrobials tested in this study, resistance against oxacillin (15 isolates, 83.3%) was most frequently observed, but only one isolate (SNUDS-1) possessed mecA. This isolate was shown to possess SCCmec type III; the type 3 ccr complex and the class A mec complex.
Based on these results, isolate SNUDS-1 was shown to possess SCCmec type III; the type 3 ccr complex and the class A mec complex. Although the SCCmec type III is not predominant in human, MR-CoNS (Methicillin resistance Coagulase-negative staphylococci) in food animals should be monitored to prevent the dissemination of antimicrobial resistance genes and resistant pathogens to the community.
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to investigate sequence-based genotyping methods to distinguish 27 Riemerella anatipestifer isolates from ducklings in South Korea. The 16S rRNA sequences of the 27 R. anatipestifer isolates showed 99�100% similarities to each other and to reference sequences from Genbank (AY871822.2, AY871834.2, CP002562.1, EU715016.1, EU016548.1, EU715000.1, EU715008.1 and EU715011.1). In addition, the ompA gene sequences of 25 of the 27 R. anatipestifer isolates were 100% identical to each other, and these sequences were also 100% identical to reference sequences (CP002562.1, GQ415419.1, DQ059079, FJ765034.1, AY606207.1, AF104937.1, and FJ765033.1). Alternatively, four housekeeping genes (mdh, gdh, pgi, and rpoB) and three virulence-associated genes (prtC, hagA, and sspA) were used for a multi-locus sequence typing (MLST) and a single-locus sequence typing (SLST) among R. anatipestifer isolates. Compared to 16S rRNA and the ompA gene, seven genes showed higher genetic divergence patterns, and the isolates were separated into three distinct groups in phylogenetic trees.
Housekeeping genes, MLST, Phylogenetic analysis, Riemerella anatipestifer, SLST, Virulence-associated genes.
The New Microbiologica: official journal of the Italian Society for Medical Virology (SIVIM) 10/2013; 36(4):395-404. · 1.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Shigellosis is a global health problem, and Shigella flexneri is the major cause of this disease. In this study, we isolated a virulent Siphoviridae bacteriophage (phage), pSf-1, that infects S. flexneri. This phage was isolated from the Han River in Korea and was found to infect S. flexneri, S. boydii, and S. sonnei. One-step growth analysis revealed that this phage has a short latent period (10 min) and a large burst size (86.86 PFU/cell), indicating that pSf-1 has good host infectivity and effective lytic activity. The double-stranded DNA genome of pSf-1 is composed of 51,821 bp with a G + C content of 44.02%. The genome encodes 94 putative ORFs, 71 putative promoters, and 60 transcriptional terminator regions. Genome sequence analysis of pSf-1 and comparative analysis with the homologous Shigella phage Shfl1 revealed that there is a high degree of similarity between pSf-1 and Shfl1 in 54 of the 94 ORFs of pSf-1. The results of this investigation indicate that pSf-1 is a novel Shigella phage and that this phage might have potential uses against shigellosis.
Research in Microbiology 09/2013; · 2.89 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A lytic bacteriophage (phage), designated SAH-1, was isolated from sewage effluent near a dairy cow farm in Gwacheon, South Korea to search for biocontrol agents against Staphylococcus aureus infections. The SAH-1 was morphologically classified as Myoviridae and possessed an approximate 144kb double-stranded genomic DNA. The phage showed broad host ranges within S. aureus strains including methicillin-resistant strains, and its latent period and burst size were approximately 20min and 100PFU/cell, respectively. Moreover, morphologic and genomic analysis of SAH-1 revealed that the phage was closely related to other Myoviridae phages infecting Staphylococcus species. The bacteriolytic activity of phage SAH-1 at a multiplicity of infection (MOI) 1 and 100 indicated its efficiency for reducing bacterial growth. Based on these results, phage SAH-1 could be considered a potential therapeutic or prophylactic candidate against S. aureus infections.
Research in Veterinary Science 06/2013; · 1.77 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Thelohanellus kitauei was isolated from the koi Cyprinus carpio haematopterus, and the 18S rRNA gene of T. kitauei was amplified by optimised nested-PCR. The PCR product was sequenced and compared with other 18S rRNA genes of Thelohanellus species to investigate the relationships between their host specificities and infection sites. Based on the 18S rRNA sequences, T. kitauei is most closely related to T. hovorkai (which can infect the intestine). Phylogenetic analysis revealed that T. kitauei was clustered with other Thelohanellus spp. infecting Cyprininae. The present study suggests that the infection site and the host specificity (subfamily level) are reflected in the genetic relationships among Thelohanellus species.
[Show abstract][Hide abstract] ABSTRACT: Vibrio parahaemolyticus is the leading cause of food-borne diseases, and several pathogenic strains cause global gastroenteritis outbreaks. Here, we report a draft genome sequence of V. parahaemolyticus SNUVpS-1, which was isolated from seafood in a fishery market in the Republic of Korea and contained TL, toxR, and toxRS(old) genes. The current draft genome sequence will contribute to the effort to monitor the spread of V. parahaemolyticus seafood isolates and clinical isolates.
[Show abstract][Hide abstract] ABSTRACT: We present the genome of a clinical isolate, Aeromonas hydrophila SNUFPC-A8, from a moribund cherry salmon. The completed draft genome of this strain shows high sequence homology to the reference strain A. hydrophila ATCC 7966 (NC008570.1) and known plasmids pAsa2 and pAAk1 from other Aeromonas species (NC004925.1 and NC019014.1).
[Show abstract][Hide abstract] ABSTRACT: Cyprinid herpes virus 3 (CyHV-3) diseases have been reported around the world and are associated with high mortalities of koi (Cyprinus carpio). Although little work has been conducted on the molecular analysis of this virus, glycoprotein genes identified in the present study seem to be valuable targets for genetic comparison of this virus. Three envelope glycoprotein genes (ORF25, 65 and 116) of the CyHV-3 isolates from the USA, Israel, Japan and Korea were compared, and interestingly, sequence insertions or deletions were observed in these target regions. In addition, polymorphisms were presented in microsatellite zones from two glycoprotein genes (ORF65 and 116). In phylogenetic tree analysis, the Korean isolate was remarkably distinguished from USA, Israel, Japan isolates. These findings may be suitable for many applications including isolates differentiation and phylogeny studies.
[Show abstract][Hide abstract] ABSTRACT: We present the draft genome sequence of Aeromonas salmonicida subsp. achromogenes strain AS03, an atypical A. salmonicida strain that causes erythrodermatitis in crucian carp (Carassius carassius). This is the first genome sequence report of A. salmonicida subsp. achromogenes, one of the four subspecies of atypical A. salmonicida.
[Show abstract][Hide abstract] ABSTRACT: To date, a number of bacteriophages (phages) infecting Acinetobacter species have been reported and characterized. However, Acinetobacter phages which infect A. soli have not been investigated yet. Here, we report the complete genome sequence of Acinetobacter phage phiAC-1, which belongs to the Myoviridae, infecting Acinetobacter soli strain KZ-1.
Journal of Virology 12/2012; 86(23):13131-2. · 5.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Among the abundant bacteriophages that belong to the order Caudovirales in the ocean, the genome sequences of marine siphoviruses are poorly investigated in comparison to those of myo- or podoviruses. Here we report the complete genome sequence of Vibrio phage pVP-1, which belongs to the family Siphoviridae and infects Vibrio parahaemolyticus ATCC 33844.
Journal of Virology 06/2012; 86(12):7013-4. · 5.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To date, a number of Myoviridae bacteriophages that infect Aeromonadaceae have been identified and characterized. However, the genome sequences of Aeromonas phages that not belong to the Myoviridae have not been investigated yet. Herein, we report the complete genome sequence of Aeromonas phage phiAS7, which belongs to the Podoviridae and infects Aeromonas salmonicida subsp. salmonicida.
Journal of Virology 03/2012; 86(5):2894-5. · 5.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The principal objective of this study was to investigate the incidence, risk assessment, antibiotic resistance, and genotyping of Vibrio parahaemolyticus in Korean seafood. The incidence of V. parahaemolyticus in seafood obtained from several fish markets in Korea was investigated from May to December of 2009, except between July and September. Two selective mediums (TCBS [thiosulfate, citrate, bile salts, and sucrose] agar and CHROMagar™ Vibrio) were used, and the V. parahaemolyticus strains were identified via polymerase chain reaction (PCR) amplification (Vp. flaE, tl, and toxR). 16S rRNA gene sequencing and their virulence were analyzed via the detection of tdh, trh, ORF8, toxRS/old, and toxRS/new genes. We collected 24 strains of V. parahaemolyticus: 19 seafood isolates, three environmental isolates, and two clinical (human) isolates. Among these strains, two tdh+ strains, two ORF8+ strains, 16 toxRS/old+ strains, and one toxRS/new+ strain were isolated. Twenty-two commercial antibiotics were used to assess the antibiotic susceptibility of isolates, and all the strains evidenced resistance to more than four antibiotics. The strains harboring antibiotic-resistant genes such as TetA (25%) and strB (4.16%) were detected via PCR. Repetitive extragenic palindromic sequence (REP)-PCR analysis revealed differences in the V. parahaemolyticus strains from other species and intraspecific strains.
Foodborne Pathogens and Disease 01/2012; 9(3):224-31. · 2.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Antimicrobial resistance patterns in a collection of 33 motile Aeromonas species were described in this study. Quinolone has been frequently employed for treatment of Aeromonas-related diseases, and prolonged use of antimicrobial compounds has led to development of resistant strains. In a sample of diseased fish and environmental water, we evaluated nalidixic acid (n = 19) and ciprofloxacin (n = 4) resistance via minimum inhibitory concentration (MIC) assays and the genetic basis was also investigated. Among the isolated Aeromonas spp., 17 strains encoded for chromosomal mutations of quinolone resistance-determining regions (QRDRs) in gyrA, 11 strains encoded for mutations of QRDRs in parC, 1 strain harbored plasmid-mediated quinolone resistance (PMQR) qnrS1-like gene and 4 strains harbored the PMQR qnrS2 gene. In particular, the new variant (qnrS1-like) differed from qnrS1 by 6 amino acid substitutions at positions 5 (Asn(5)→Arg(5)), 120 (Ser(120)→Thr(120)), 148 (Asn(148)→His(148)), 206 (Leu(206)→Glu(206)), 207 (Ile(207)→ Leu(207)), and 216 (Tyr(216)→Phe(216)), and the gene was designated qnrS5. These resistant strains may function as reservoirs of quinolone resistance.
Research in Microbiology 01/2012; 163(1):73-9. · 2.89 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this study, we report one lytic Myoviridae bacteriophage (phage) infecting Aeromonas salmonicida subsp. salmonicida. The phage (named as phiAS5) was isolated from environmental river waters in Korea, and showed broad infectivity to other bacterial species in the family Aeromonadaceae as well as antibiotic-resistant A. salmonicida subsp. salmonicida strains. The biological properties and complete genome of phiAS5 were simultaneously investigated. The complete genome of phiAS5 composed of linear double-stranded DNA of 225,268 bp with G+C content of 43.0%, and encoded 343 putative ORFs, 69 putative promoters, 33 transcriptional terminator regions and 24 tRNA-encoding genes. A high degree of similarity to other T4-like Aeromonas phage was found in most ORFs of phiAS5. Therefore, the genome of phiAS5 was further compared with T4 phage and the closest relative, Aeromonas phage Aeh1, and the result demonstrated that it could be classified as a new member of the T4-like group. The bacteriolytic activity of phiAS5 against A. salmonicida subsp. salmonicida was evaluated at different doses of multiplicity of infection using one each of virulent strain that possesses the ascV gene and multi-drug resistant strain, and the results proved to be efficient for the reduction of bacterial growth. Based on these results, phiAS5 may have the potential for reducing the impacts of virulent or antibiotic-resistant A. salmonicida subsp. salmonicida in aquaculture and may also advance our understanding of the biodiversity of T4-like Aeromonas phages.