• Korean Journal of Ecology and Environment
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• v.46 no.3
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• pp.360-366
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• 2013

This study was it conducted on effluent or river water close to discharge locations from a treatment plant, which were analyzed for the presence, phenotype and antibiotic resistance rates of Vancomycin Resistant Enterococci (VRE). The test results for isolation and identification of Enterococcus spp. showed that they all were VRE positive, with a total of 32 strains detected. Multiplex PCR was conducted for 32 strains, each of which were identified as E. faecium, and the results indicated that they were all confirmed as VRE, corresponding to the Van A phenotype. The results of E. faecium concentrations measured at various locations indicated that they were, on average, higher at the location of sewage treatment plants. The frequency of positive tests as well as the number of colonies was higher downstream of treatment plants. The minimum inhibitory concentration was inspected for 26 strains of discharged water samples from sewage water treatment plants, and 6 strains of river samples. Out of 19 antibiotics, 14 and 5 species showed resistance and sensitivity, respectively.

• Journal of Veterinary Clinics
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• v.32 no.2
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• pp.191-195
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• 2015

A 7-year-old castrated male Korean Shorthair cat was referred with lethargy and anorexia. Laboratory examination revealed moderate degenerative changes of peripheral neutrophils on blood smear examination and decreased levels of free and total thyroxine ($T_4$) as well as bacterial growth on blood culture. Molecular analyses of the 16S ribosomal RNA gene and heat shock protein 60 gene confirmed the bacterium as Enterobacter cloacae. A minimal inhibitory concentration test showed multidrug resistance of the bacterium against 16 antibiotics. Polymerase chain reaction (PCR) and subsequent sequencing specifically for $bla_{TEM}$, $bla_{SHV}$, $bla_{CTX-M}$, and plasmid-mediated ampC genes revealed positive results to $bla_{TEM-1}$, $bla_{CTX-M-15}$, and plasmid-mediated $bla_{ACT-1}$ genes, indicating that the isolated bacterium contains plasmids containing genes encoding extended-spectrum beta-lactamase and plasmid-mediated ampC beta-lactamase. After 1 month of treatment with antibiotics and levothyroxine, the cat's condition improved; both the thyroid function test and the blood culture showed no abnormalities. This is the first report of community-acquired multidrug-resistant E. cloacae-induced euthyroid sick syndrome in a cat. By the prompt diagnostic procedures and properly selected antibiotic therapy, the cat was recovered from the multidrug-resistant bacterium-induced septicaemia.

• Journal of Korean Medicine Rehabilitation
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• v.25 no.2
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• pp.1-13
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• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.305-310
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• 2008

Staphylococcus aureus is one of the most significant pathogens and a causative agents of nosocomial infections. The emergence of methicillin resistant S. aureus (MRSA), in particular, has become a major clinical and epidemiological problems worldwide. In this study, we analyzed the toxin genes and investigated molecular epidemiological characteristics of S. aureus isolated from stools of diarrheal patients at the hospitals in Incheon. Of the 609 strains from 2,281 specimens, 173 strains retained enterotoxin; 68 isolates (39.30%), 100 isolates (57.80%) were classified to A and C type, respectively. In the antibiotic susceptibility, all of enterotoxin positive isolates were resistant to oxacillin. Eighty eight strains (50.86%) of 173 MRSA isolate possessed tsst gene, but eta and eth genes were not detected at all. In the detection of MRSA associated genes by PCR method, mecA genes were detected in 167 strains (96.53%). From the result of PFGE analysis, we classified tsst-positive MRSA to 10 types and 24 subtypes. Type A, H and F were the major strains comprised of 57.95% (51 strains), 10.22% (9 strains) and 9.09% (8 strains) respectively.

• Journal of Veterinary Clinics
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• v.19 no.3
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• pp.303-311
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• 2002

Isolation and identification of causative microorganisms and susceptibility testing are important in selecting appropriate antimicrobial agent. The purpose of this study was to determine the antimicrobial susceptibility patterns and identification of bacteria for the selection of a therapeutic antibiotic agent for treatment. Specimens were cultured aerobically from dog patients brought to the veterinary medical teaching hospital of Seoul National University between July 1999 and September 2000. A total of 157 isolates were from skin(63), urine(45), ear canal(31) and conjunctiva(18). The result is that the most common organisms isolated from dog patients were S. intermedius was the most common isolates from the skin, ear canal, and conjunctiva. E. coli was the most common isolated from urine. Most of gram-positive isolates were resistant to ampicillin(80.6%), erythromycin(68.8%), penicillin(86.2%), tetracycline(89.2%). Otherwise most of gam-negative isolates were resistant to ampicillin(73.4%), trimethoprim-sulfa(53.3%). E. coli was resistant to ciprofloxacin(61.5%), piperacillin (69.2%). Antimicrobial susceptibility pattern by the sampling site was not remarkably different except 5. aureus isolated from urine.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.200-208
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• 2011

Most problematic antibiotic resistance mechanism for MLS (macrolide-lincosamide-streptogramn B) antibiotics encountered in clinical practice is mono- or dimethylation of specific adenine residue at 2058 (E. coli coordinate) of 23S rRNA which is performed by Erm (erythromycin ribosome resistance) protein through which bacterial ribosomes reduce the affinity to the antibiotics and become resistant to them. ErmSF is one of the four gene products produced by Streptomyces fradiae to be resistant to its own antibiotic, tylosin. Unlike other Erm proteins, ErmSF harbors idiosyncratic long N-terminal end region (NTER) 25% of which is comprised of arginine well known to interact with RNA. Furthermore, NTER was found to be important because when it was truncated, most of the enzyme activity was lost. Based on these facts, capability of NTER peptide to inhibit the enzymatic activity of ErmSF was sought. For this, expression system for two different proteins to be expressed in one cell was developed. In this system, two plasmids, pET23b and pACYC184 have unique replication origins to be compatible with each other in a cell. And expression system harboring promoter, ribosome binding site and transcription termination signal is identical but disparate amount of protein could be expressed according to the copy number of each vector, 15 for pACYC and 40 for pET23b. Expression of NTER peptide in pET23b together with ErmSF in pACYC 184 in E. coli successfully gave more amounts of NTER than ErmSF but no inhibitory effects were observed suggesting that there should be dynamicity in interaction between ErmSF and rRNA rather than simple and fixed binding to each other in methylation of 23S rRNA by ErmSF.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1340-1349
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• 2017

Objective: This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease. Methods: Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical electrical conductivity of milk (ECM), and lactoferrin content in milk. Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation). Results: The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus haemolyticus, and Escherichia coli. It was observed that positive samples in the microbiological examination showed total bacterial count between $9.10{\times}10^3$ to $6.94{\times}10^6$ colony forming units/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis. Conclusion: Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp., and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.3
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• pp.85-93
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• 2017

The urinary tract infection (UTI) is one of the most important infections in hospital. The overuse and misuse of antimicrobial agents and the resulting emergence of resistant microorganisms have made choices regarding antimicrobial therapy more difficult. This study examined the changes in the antibiotic susceptibility to the causative organisms of urinary tract infections to provide useful information on the choice of adequate drugs in the treatment of urinary tract infections. The medical records of 2,707 patients with more than $10^5/ml$ microorganism in urine culture between January 2010 and December 2015 were reviewed retrospectively. The most common pathogenic organism was E. coli (28.1%). In the case of E.coli, there were no differences in frequency from 2010 to 2015 in men, but since 2014, the frequency decreased gradually since 2014 in women. For E. coli, the resistance rates to antibiotics were 72.2% in ampicillin, 44.9% in trimethoprim/sulfamethoxazole (TMP/SMX), and 41.3% in ciprofloxacin, but the 2nd, 3rd, and 4th cephalosporin (5%) had low antibiotic resistance rates. The pathogens of urinary tract infection are becoming diverse and their frequencies are also changing over time. These results suggest that the recommended drugs for UTI should be selected more carefully for in-patients and out-patients.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.79-86
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• 2012

ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.