• Journal of Animal Science and Technology
• /
• v.64 no.3
• /
• pp.515-530
• /
• 2022

Sequence type (ST) 5 methicillin-resistant Staphylococcus aureus (MRSA) with staphylococcal cassette chromosome mec (SCCmec) type II (ST5-MRSA-II) and ST72-MRSA-IV represent the most significant genotypes for healthcare- (HA) and community-associated (CA) MRSA in Korea, respectively. In addition to the human-type MRSA strains, the prevalence of livestock-associated (LA) MRSA clonal lineages, such as ST541 and ST398 LA-MRSA-V in pigs and ST692 LA-MRSA-V and ST188 LA-MRSA-IV in chickens, has recently been found. In this study, clonotype-specific resistance profiles to cathelicidins derived from humans (LL-37), pigs (PMAP-36), and chickens (CATH-2) were examined using six different ST groups of MRSA strains: ST5 HA-MRSA-II, ST72 CA-MRSA-IV, ST398 LA-MRSA-V, ST541 LA-MRSA-V, ST188 LA-MRSA-IV, and ST692 LA-MRSA-V. Phenotypic characteristics often involved in cathelicidin resistance, such as net surface positive charge, carotenoid production, and hydrogen peroxide susceptibility were also determined in the MRSA strains. Human- and animal-type MRSA strains exhibited clonotype-specific resistance profiles to LL-37, PMAP-36, or CATH-2, indicating the potential role of cathelicidin resistance in the adaptation and colonization of human and animal hosts. The ST5 HA-MRSA isolates showed enhanced resistance to all three cathelicidins and hydrogen peroxide than ST72 CA-MRSA isolates by implementing increased surface positive charge and carotenoid production. In contrast, LA-MRSA strains employed mechanisms independent of surface charge regulation and carotenoid production for cathelicidin resistance. These results suggest that human- and livestock-derived MRSA strains use different strategies to counteract the bactericidal action of cathelicidins during the colonization of their respective host species.

• Food Science of Animal Resources
• /
• v.43 no.5
• /
• pp.792-804
• /
• 2023

Non-aureus staphylococci (NAS), particularly antimicrobial-resistant NAS, have a substantial impact on human and animal health. In the current study, we investigated (1) the species profiles of NAS isolates collected from healthy broilers, farm environments, and farm workers in Korea, (2) the occurrence of antimicrobial-resistant NAS isolates, especially methicillin resistance, and (3) the genetic factors involved in the methicillin and fluoroquinolone resistance. In total, 216 NAS isolates of 16 different species were collected from healthy broilers (n=178), broiler farm environments (n=18), and farm workers (n=20) of 20 different broiler farms. The two most dominant broiler-associated NAS species were Staphylococcus agnetis (23.6%) and Staphylococcus xylosus (22.9%). Six NAS isolates were mecA-positive carrying staphylococcal cassette chromosome mec (SCCmec) II (n=1), SCCmec IV (n=1), SCCmec V (n=2), or nontypeable SCCmec element (n=2). While two mecA-positive Staphylococcus epidermidis isolates from farm workers had SCCmec II and IV, a mecA-positive S. epidermidis isolate from broiler and a Staphylococcus haemolyticus isolate farm environment carried SCCmec V. The occurrence of multidrug resistance was observed in 48.1% (104/216 isolates) of NAS isolates with high resistance rates to β-lactams (>40%) and fusidic acid (59.7%). Fluoroquinolone resistance was confirmed in 59 NAS isolates (27.3%), and diverse mutations in the quinolone resistance determining regions of gyrA, gyrB, parC, and parE were identified. These findings suggest that NAS in broiler farms may have a potential role in the acquisition, amplification, and transmission of antimicrobial resistance.

• Food Science of Animal Resources
• /
• v.42 no.2
• /
• pp.225-239
• /
• 2022

As commensal colonizers in livestock, there has been little attention on staphylococci, especially non-aureus staphylococci (NAS), contaminating meat production chain. To assess prevalence of staphylococci in retail pork and slaughterhouse carcass samples in Korea, we collected 578 samples from Korean slaughterhouses (n=311) and retail markets (n=267) for isolation of staphylococci and determined antimicrobial resistance phenotypes in all the isolates. The presence of and prevalence of fusB-family genes (fusB, fusC, fusD, and fusF) and mutations in fusA genes were examined in fusidic acid resistant isolates. A total of 47 staphylococcal isolates of 4 different species (Staphylococcus aureus, n=4; S. hyicus, n=1; S. epidermidis, n=10; Mammaliicoccus sciuri, n=32) were isolated. Fusidic acid resistance were confirmed in 9/10 S. epidermidis and all of the 32 M. sciuri (previously S. sciuri) isolates. Acquired fusidic acid resistance genes were detected in all the resistant strains; fusB and fusC in S. epidermidis and fusB/C in M. sciuri. Multi-locus sequence type analysis revealed that ST63 (n=10, 31%) and ST30 (n=8, 25%) genotypes were most prevalent among fusidic acid resistant M. sciuri isolates. In conclusion, the high prevalence of fusB-family genes in S. epidermidis and M. sciuri strains isolated from pork indicated that NAS might act as a reservoir for fusidic acid resistance gene transmissions in pork production chains.

• Pediatric Infection and Vaccine
• /
• v.26 no.3
• /
• pp.148-160
• /
• 2019

Purpose: This study aimed to investigate the molecular epidemiology of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak at a newborn nursery and neonatal intensive care unit (NICU). Methods: During the outbreak, from August to September 2017, MRSA isolates collected from neonates and medical staff underwent genotyping and screened for virulence factors. Antibiotic susceptibilities were tested. Results: During the study period, 41 neonates were admitted at the nursery (n=27) and NICU (n=14). Of these, 7 had MRSA infections (skin infection [n=6] and sepsis [n=1]) and 4 were colonized with MRSA. Associated medical staff (n=32) were screened; three were nasal MRSA carriers. Staphylococcal chromosomal cassette mec (SCCmec) type II, sequence type (ST) 89, spa type t375 was found to be the skin infection outbreak causing strain, with multi-drug resistance including low-level mupirocin resistance. SCCmec type IVa, ST 72, and a novel spa type designated t17879, was the cause of MRSA sepsis. Many different types of MRSA were colonized on the neonates; however, SCCmec type IVa, ST 72, spa type t664 was colonized in both neonates and a NICU nurse. All MRSA isolates from colonized infants were positive for the Panton-Valentine leukocidin (PVL) toxin gene. Conclusions: The strain causing an outbreak of skin infections had multi-drug resistance. Also, MRSA colonized in the neonates were found to carry the PVL toxin gene. Because different strains are present during an outbreak, molecular epidemiologic studies are important to identify the outbreak strain and colonized strains which aid in effective control and prevention of future MRSA outbreaks.

• Pediatric Infection and Vaccine
• /
• v.25 no.2
• /
• pp.61-71
• /
• 2018

Purpose: Research on the clinical role of Staphylococcus aureus as a pathogen in acute gastroenteritis (AGE) in children has been scarce. This study aimed to clarify the prevalence and clinical correlation of S. aureus detection in children with AGE. Methods: Fecal samples were collected from children with symptoms of AGE who visited a secondary hospital between January 2012 and December 2015. The samples were sent to the Seoul Metropolitan Government Research Institute of Public Health and Environment to test for pathogenic organisms. Clinical patterns were analyzed through medical record review. Results: Among the 663 participants, the bacteria detection rate was 26.2% (n=174), the virus detection rate was 29.7% (n=197), and the non-detection rate was 43.1% (n=286). S. aureus was tested positive from 102 cases and was confirmed as a single pathogen in 53 cases. It was the third most common pathogen. The prevalence by age was highest (45.3%) in 0-2 year-olds. Most cases occurred in summer. Symptoms included diarrhea (71.7%), vomiting (67.9%), fever (49.1%), and abdominal pain (37.7%). Only vomiting showed a significant difference between the S. aureus group and the non-detection group (67.9% vs. 43.0%; P=0.001). Among enterotoxins, the higher incidence of vomiting was associated with classical staphylococcal enterotoxins (SEA, SEB, SEC, SED, and SEE) and SEH (P=0.027). Conclusions: S. aureus was the bacteria commonly isolated from children with AGE. Our study identified cases of staphylococcal AGE in children based on fecal samples and confirmed the characteristic symptoms, affected age groups, seasonal distribution, and correlation with enterotoxins.

• Korean Journal of Microbiology
• /
• v.41 no.1
• /
• pp.38-46
• /
• 2005

To determine the actual hygienic status of domestic chicken meats sold in public markets (conventional markets and department stores), microbial contamination levels (Total cells, Coliforms and Staphylococcal cells) and zoonotic pathogens (Salmonella species, Campylobacter species, Listeria species, and Staphylococcus aureus) isolation tests were conducted. Chicken meats and eggs tested were collected from the conventional markets (Si-Jang) and department-stores located in Seoul and Kyung-gi regions in 1996. In total cells and coliforms contamination tests, chicken meats sold in department stores were much lesser contamination status than those of Si-Jang, but staphylococcal cells level was much more higher than that of conventional markets. Salmonella isolation frequency was investigated as $68.8\%$, but Campylobacter jejuni and Listeria monocytogenes isolation frequency were appeared both $64.0\%\;and\;63.3\%$. In case of eggs sold in public markets, one of S. gallinarum strain $(0.7\%)$ was isolated only on the egg-shell part among the four-hundred and fourty-six. In comparison with foreign imported chicken meats, there were no big differences in microbial contamination status. On the other hand, both Salmonella and L. monocytogenes were isolated only in the chicken wings from Korea and China, but not from U.S.A. This data suggest that more hygienic control system in order to produce the safe and hygienic chicken meats and eggs is need in our country as soon as possible.

• Current Research on Agriculture and Life Sciences
• /
• v.2
• /
• pp.115-121
• /
• 1984

Some investigations on chronic mastitis in dairy cattle in Taegu-Kyungpook Provinces from the beginning of October, 1984 till the end of August, 1985 were conducted with the particular regard to the causative agents and their drug susceptibility. Milk samples from 83 isolated cases of chronic mastitis cattle were investigated bacteriologically and the causative organisms recovered were examined for their antibiotic susceptibility by using disc diffusion susceptibility technique against the major antibiotics of current veterinary use. Major causative agents involved in chronic mastitis in Taegu-Kyungpook Provinces were in order of prevalence Staphylococcus spp. (48.2 %), Escherichia coli (18.1 %), Candida spp. (10.8 %) and Corynebacterium spp. (8.4 %), Streptococcus agalactiae (3.6 %), Bacillus cereus (3.6 %) and Pseudomonas aeruginosa (2.4 %) were found to be one of the minor agents. The majority of staphylococcal isolates and E. coli were highly resistant to the most antibiotics tested. The percentages of staphylococcal cultures resistant to penicillin, methicillin, lincomycin, novobiocin, ampicillin and tetracycline were 87.2 %, 78.7 %, 68.1 %, 61.7% and 57.4 %, respectively, while the majority of them were susceptible to gentamicin(78.7 %), cephalothin(76.6 %) and chloramphenicol (74.5%). E. coli isolates were found to be highly resistant to streptomycin, cephalothin, tetracycline and ampicillin while the majority of them were susceptible to colistin (83.3 %), gentamicin (77.8 %) and chloramphenicol (66.7 %). Corynebacterium spp. were susceptible to ampicillin, chloramphenicol, erythromycin, gentamicin, oleandomycin and tetracycline although they showed resistance to novobiocin and penicillin. Two cultures of Pseudomonas aeruginosa recovered from mastitis milk were highly resistant to the antibiotics employed in the present study.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.3
• /
• pp.312-320
• /
• 2000

Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

• Korean Journal of Clinical Laboratory Science
• /
• v.48 no.3
• /
• pp.202-209
• /
• 2016

This study is aimed to determine the correlation between the toxin gene types and antibiotic resistance from MRSA (methicillin-resistant Staphylococcus aureus). Fifty-two strains of MRSA, between January 2014, and December 2014, were isolated from clinical specimens obtained from 2,664 cases in the intensive care unit of a hospital in Suncheon, Jeonnam, Korea. Genes encoding mecA, enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET), and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Toxin genes (seg and sei) were present in 40 strains (76.9%), followed by tst in 34 strains (65.4%). Other genes (eta, etb, sea, sed, see, seh, sej, and pvl) were not detected. Forty strains (76.9%) of MRSA had 2 or more toxin genes simultaneously; 5 coexistent toxin-genes (seb, sec, seg, sei, tst) were the most common in 28 strains (53.8%), and 6 strains (11.5%) had seg and sei genes. The coexistence of genes were 72.5~100%, showing a high correlation among genes (seb, sec, seg, sei and tst). As strains (seb, sec, tst) that had particular toxin genes (seb, sec, seg, sei, tst) in multiple showed 100% resistance to ciprofloxacin, clindamycin, erythromycin, we were able to find that seb, sec, and tst genes have a close relationship to the aforementioned antibiotics. It showed a higher resistance to ciprofloxacin, clindamycin, erythromycin, and tetracycline compared with strains that had toxin genes independent from multiple toxin genes.

• Korean Journal of Food Science and Technology
• /
• v.20 no.6
• /
• pp.856-861
• /
• 1988

A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.