• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.386-393
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• 2014

Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n=465) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.

• Journal of Veterinary Science
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• v.24 no.6
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• pp.82.1-82.12
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• 2023

Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [bla_TEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

• The Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.35-53
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• 1987

Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.

• Fisheries and Aquatic Sciences
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• v.22 no.10
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• pp.21.1-21.7
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• 2019

Background: Lactococcus garvieae is one of the most important risk factors in the rainbow trout culture. Therefore, the purpose of this study was to identify and detect strains isolated from rainbow trout suspected of having Lactococcus garvieae using biochemical characteristics and PCR and determination of the degree of severity of isolated strains. Methods: In this study, the cause of lactococcosis in selected rainbow trout farms in Kohkilooieh and Boyerahmad province was assayed. Gram-positive and catalase-negative bacterial isolates were first obtained from selected trout fish farms using conventional biochemical tests and PCR assay. The 10-day LD₅₀ method (concentration causing 50% mortality in 10 days) was used to determine the severity of the isolated bacteria. Results: One bacterial isolate was detected from all sampled fish which confirmed as Lactococcus garvieae using a specific PCR assay based on the 16S rDNA gene by producing a single band of 1107 bp. Analysis of the rate of mortality showed that the 10-day LD₅₀ was 4.6 × 10⁵ CFU/fish. The results of this study showed that isolated bacteria had high severity for rainbow trout. The presence of bacteria in internal organs of suspected fish showed a severe systemic infection in challenged fish. Antibiogram assay also indicated that the isolated Lactococcus garvieae were resistant to some mostly used antibiotics in rainbow trout. Conclusions: According to current research, it can be concluded that the condition of lactococcosis in the studied area is not suitable, and despite the presence of disease, there is no proper action to control and prevent the disease. Unfortunately, isolated bacteria from the studied area have a very high severity compared to bacteria isolated from other regions of the country or other countries. Therefore, further investigation is needed to determine the cause of this difference and possibly in the design of the vaccine.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.425-432
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• 2010

Staphylococcus intermedius is a common cause of otitis externa, pyoderma, and wound infections in companion animals. Although S. intermedius infections are rare in humans, it is zoonotic, with several case reports describing fatal human infections. Presently, we sought to isolate S. intermedius strains from various sources at animal hospitals nationwide in Korea, examine their antibiotic susceptibilities, and determine the possibility of horizontal transmission between animals and humans. Pulsed-field gel electrophoresis (pFGE) was used to compare the mecA gene in S. intermedius strains from humans, animals, and the environment in animal hospitals. A total of 119 S. intermedius strains were isolated from 529 samples. Using the disk diffusion method, over 90% of the isolates were found to be susceptible to cephalothin, amoxicillin-clavulanic acid, vancomycin, imipenem, nitroflurantoin, and amikacin, whereas 97.5% and 98.3% of the isolates were resistant to penicillin and ampicillin, respectively. Among the 39 S. intermedius strains harboring mecA, similar PFGE patterns were observed between seven isolates from an animal, two isolates from veterinary staff, and the environment in one animal hospital, and single isolates from an animal and a veterinarian at another hospital. This result suggests the possibility of horizontal transmission of S. intermedius containing mecA between humans, animals, and the environment in animal hospitals and also emphasizes on the importance of S. intermedius with mecA as a possible emerging threat to public health.

• Pediatric Infection and Vaccine
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• v.18 no.2
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• pp.143-153
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• 2011

Purpose : It is known that carriage rates of Staphylococcus aureus (S. aureus) are highest in newborns and that the asymptomatic carriage of methicillin-resistant Staphylococcus aureus (MRSA) is associated with invasive MRSA infection with the colonizing strain. This study was carried out to investigate the carriage rates of MRSA in neonates with neonatal jaundice. Methods : We reviewed the medical records of 545 neonates admitted with neonatal jaundice to neonatal intensive care units between January 2006 and December 2010. Nasal and inguinal swab specimens had been taken from them and cultured for the isolation of S. aureus. Antimicrobial susceptibility tests had been done for such isolates to determine methicillinresistance. Results : Out of 545 neonates, 318 (58.3%) were colonized with S. aureus and 214 (39.3%) were colonized with MRSA. Results of the antibiogram analysis showed that 65.7% of MRSA isolates were likely to be community-associated (CA) MRSA. Conclusion : Based on the MRSA carriage rate of 39.3%, a surveillance program for MRSA colonization is considered necessary in neonates transferred from other clinics or hospitals. Out of MRSA isolates, 65.7% were likely to be CA-MRSA. This suggests that CA-MRSA strains were already present in obstetric clinic environments where the neonates were born. It is thought that MRSA surveillance programs in these environments are also necessary.

• Microbiology and Biotechnology Letters
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• v.50 no.1
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• pp.89-94
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• 2022

Triclosan (TCS) is an antimicrobial agent used in various human personal care products against both gram-positive and gram-negative bacteria. The purpose of this study was to evaluate the presence of TCS-resistant bacteria in sewage water in Peshawar, Khyber Pakhtunkhwa (KPK), Pakistan, for the first time. TCS-supplemented Luria Bertani (LB) agar was used to isolate TCS-tolerant bacteria. A total of 17 TCS-resistant isolates were randomly selected from a large pool of bacteria that showed growth on TCS-supplemented LB agar. Based on gram staining and physiochemical characteristics, the isolated strains were identified as Salmonella typhi (n = 6), Escherichia coli (n = 4), Citrobacter freundii (n = 4), Proteus mirabilis (n = 1), Enterobacter cloacae (n = 1), and Pseudomonas aeruginosa (n = 1). The Triclosan mean minimum inhibitory concentrations (MICs) for the isolates of Salmonella typhi, Escherichia coli, Citrobacter freundii, Proteus mirabilis, Enterobacter cloacae, and Pseudomonas aeruginosa were 23.66 ㎍ ml^-1, 18.75 ㎍ ml^-1, 42 ㎍ ml^-1, 32 ㎍ ml^-1, 64 ㎍ ml^-1, and 128 ㎍ ml^-1, respectively. The antibiogram revealed that all isolates were resistant to penicillin G (100%) and linezolid (100%), followed by ampicillin (94%), tetracycline (76%), tazobactam (76%), sulbactam/cefoperazone (64%), polymyxin PB (58%), amikacin (29.41%), aztreonam (29.41%), imipenem (5%), and gentamicin (5%). This is the first known study regarding the isolation of TCS-tolerant bacteria from sewage water in Peshawar, KPK, Pakistan. It was concluded that all the TCS-resistant isolates were multidrug resistant (MDR) gram-negative rod-shaped bacteria, mostly belonging to the Enterobacteriaceae family.

• Pediatric Infection and Vaccine
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• v.31 no.1
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• pp.46-54
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• 2024

Purpose: This study aimed to identify the pathogens of bloodstream infection in children with underlying hemato-oncologic diseases, analyze susceptibility patterns, compare temporal trends with those of previous studies, and assess empirical antimicrobial therapy. Methods: Retrospective review study of children bacteremia in hemato-oncologic diseases was conducted at Seoul National University Bundang Hospital from January 2013 to July 2023. Results: Overall, 98 episodes of bacteremia were observed in 74 patients. Among pathogens isolated, 57.1% (n=56) were Gram-positive bacteria, 38.8% (n=38) were Gram-negative bacteria, and 4.1% (n=4) were Candida spp. The most common Gram-positive bacteria were coagulase-negative staphylococci (n=21, 21.4%) and Staphylococcus aureus, (n=14, 14.3%) whereas the most common Gram-negative bacteria were Klebsiella pneumoniae (n=16, 16.3%) and Escherichia coli (n=10, 10.2%). The susceptibility of Gram-positive bacteria to penicillin, oxacillin, and vancomycin was 11.5%, 32.7%, and 94.2%, respectively and the susceptibility of Gram-negative bacteria to cefotaxime, piperacillin/tazobactam, imipenem, gentamicin, and amikacin was 68.6%, 80%, 97.1%, 82.9%, and 91.4%, respectively. Methicillin-resistant S. aureus was detected in 1 strain and among Gram-negative strains, extended spectrum β-lactamase accounted for 28.9% (12/38). When analyzing the antibiotic susceptibility and empirical antibiotics, the mismatch rate was 25.5% (n=25). The mortality rate of children within 30 days of bacteremia was 7.1% (n=7). Conclusions: Empirical antibiotic therapy for bacteremia in children with hemato-oncologic diseases should be based on the local antibiogram in each institution and continuous monitoring is necessary.

• Clinical and Experimental Pediatrics
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• v.46 no.1
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• pp.24-32
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• 2003

Purpose : Nosocomial infection with Staphylococcus aureus, especially methicillin resistant S. aureus, has become a serious concern in the neonatal intensive care unit. The aim of this study is to investigate the virulence factors, and the relationship between the antibiotic resistance and the associated genes of Staphylococcus aureus isolated from nasal cavity of neonates. Methods : Fifty one isolates of S. aureus were obtained from nasal swab taken in 28 neonates in the NICU and nursery of Pusan National University Hospital between February and May, 2001. They were tested in regard to antibiotic susceptibility, coagulase test and typing, plasmid DNA profile, as well as reactivity to enterotoxin A-E(sea, seb, sec, sed, see) genes and toxic shock syndrome toxin-1(tst) gene by polymerase chain reaction(PCR). Associated genes such as mecA, mecR1, mecI, and femA were also determined by PCR. The origin of MRSA strains was assessed using DNA fingerprinting by arbitrarily-primed polymerase chain reaction(AP-PCR). Results : Twenty three(45.1%) and six(11.8%) isolates were resistant to oxacillin and vancomycin respectively. Multidrug resistance to three or more of the antibiotics tested was observed in 51.0% of the isolates. Forty two isolates were coagulase positive and twenty two isolates had mecA gene. Sixteen isolates had both mecA and femA genes and had type I-III plasmids. 64.7% of isolates carried sec gene, and 80.4% carried tst gene. DNA fingerprinting by AP-PCR for 12 MRSA strains showed 10 distinct patterns, suggesting different origins. Conclusion : We confirmed that the prevalence of nasal carriage of S. aureus and the incidence of antimicrobial-resistant S. aureus, especially vancomycin resistance, is very high in neonates who were admitted in NICU and nursery. It is possible that these pathogens are responsible for serious nosocomial infections in neonates. The need for improved surveillance and continuous control of pathogens is emphasized.

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.386-395
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• 2007

A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.