• Biomedical Science Letters
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• v.13 no.2
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• pp.141-148
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• 2007

We determined the sequences of the quinolone resistance-determining region (QRDR) of gyrA and gyrB for 21 clinical strains of Enterobacteriaceae resistant to ciprofloxacin, norfloxacin and levofloxacin. The clinical strains were isolated from the specimens of three general hospitals in Busan. In the present study, we found mutations in type II topoisomerase (DNA gyrase) genes for all strains. We confirmed that some genera of Enterobacteriaceae of clinical specimen exhibited decreased sensitivity to fluroquinolone due to changes in Ser-83$\rightarrow$Leu and Asp-87$\rightarrow$Asn types on gyrA and alterations in Glu-465$\rightarrow$Arg and Ser-492$\rightarrow$Asn type on gyrB. All the twenty-one strains had a missense mutation in gyrA (codon 83 and 87). Three of them had an additional mutation in gyrB (codon 465 or 492), but one of them had an additional mutation in gyrB (codon 426, 427, 491, 495 and 496). The strains which had two mutations in type II topoisomerase genes (gyrA and gyrB) were significantly more resistant to fluoroquinolones than those with a single mutation in gyrA (mean MICs of ciprofloxacin: $\geq8\mu$g/ml, mean MICs of levofloxacin: $\geq16\mu$g/ml). Interestingly, the examination of silent nucleotide changes n the gyrA and gyrB genes revealed six different patterns of DNA polymorphism, respectively. Fifteen strains of the twenty-one strains bearing the gyrase A mutation shared the same polymorphism and eleven strains of the twenty-one strains bearing the gyrase B mutation shared the same polymorphism.

• Toxicological Research
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• v.21 no.1
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• pp.39-43
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• 2005

The present study was carried out to investigate the antibacterial activity against Staphylococcus aureus with antibiotic-resistance in vitro and the skin irritation in rabbits with surfactin produced by Bacillus subtilus Complex BC1212. The antibacterial activities of selected antimicrobial agents (surfactin, amoxacillin, colistin, norfloxacin and streptomycin) were evaluated by using the broth microdilution method. As the results, the minimum inhibitory concentration (MIC) of the surfactin was less than 15.6 ㎍/ml. In the skin irritation test, two out of 4 rabbits showed very slight edema at 24 h after the administration of surfactin, and then recovered at 72 h. The change of body weight was normal during the skin irritation test. The primary irritation index in accordance with the Draize evaluation of topical reaction was calculated to be '0.125', which meant not irritating. Based on these results, it could be concluded that the test agent, surfactin, was a non-irritant. We could also think that the surfactin may be useful for the treatment of S. aureus infections such as bovine mastitis.

• Natural Product Sciences
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• v.20 no.1
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• pp.39-43
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• 2014

Papenfussiella kuromo (PK) is a marine plant and an abundant ecological resource for the future; it is found in almost 80% of the terrestrial biosphere. The aim of this study was to investigate the antibacterial activity of PK against methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant pathogen. The minimum inhibitory concentrations (MICs) of PK hexane fraction (PKH) against 7 strains of MRSA ranged from 1.0 to 2.0 mg/mL. In the checkerboard dilution method, a synergistic effect of the PKH and the antibiotics (oxacillin and norfloxacin) was seen. PKH markedly reduced the MIC of each of the 4 antibiotics against MRSA. The time-kill assay showed that the synergistic activity of PKH and an antibiotic reduced the bacterial counts below the lowest detectable limit after 24 h. These findings suggest that PKH has antibacterial activity, and may be important baseline data in future extensive studies of living marine resources as a source of compounds active against MRSA.

• Journal of Microbiology and Biotechnology
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• v.30 no.7
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• pp.982-995
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• 2020

A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drug-sensitive strain of E. coli missing the main multidrug efflux pump AcrB was constructed as a host and the yddA gene was knocked out in wild-type (WT) and drug-sensitive E. coliΔacrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coliΔyddA, E. coliΔacrB and E. coliΔacrBΔyddA strains was compared with minimal inhibitory concentration (MIC) assays and fluorescence tests. MIC assay and fluorescence test results showed that YddA protein was a multidrug efflux pump that exported multiple substrates. Three inhibitors, ortho-vanadate, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and reserpine, were used in fluorescence tests. Ortho-vanadate and reserpine significantly inhibited the efflux and increased accumulation of ethidium bromide and norfloxacin, while CCCP had no significant effect on YddA-regulated efflux. The results indicated that YddA relies on energy released from ATP hydrolysis to transfer the substrates and YddA is an ABC-type multidrug exporter. Functional study of unknown ATP-binding cassette (ABC) superfamily transporters in the model organism E. coli is conducive to discovering new multidrug resistance-reversal targets and providing references for studying other ABC proteins of unknown function.

• Journal of Veterinary Clinics
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• v.20 no.2
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• pp.177-184
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• 2003

The present study was conducted to investigate isolation and antimicrobial drug susceptibility of clinical mastitic milking total 610 (897 quaters) dairy cattles from 36 dairy farms in Gyeongnam south area (Cosung, Masan) during the period from March 1999 to August 2002. The results obtained were summerized as follows . 1. Incidence of bacterial infection in four quaters was showed that right anterior quarter was 178(19.8%), right posterior quarter was 292(32.6%), left anterior quarter was 148 (16.2%), and left posterior quarter was 279 (31.1%), respectively. Isolation rate of posterior two quarters was higher 2 times than anterior two quarters. 2. Incidence of double infections of 897 clinical mastitic milk were showed that single infection was 549 (61.2%), double infection was 167(18.6%), triple infection was 9(1%) and no isolation was 172(19.2%). 3. Isolation of infected etiologic bacteria was showed that Streptococci spp., was 267(31%), Staphylococci spp., was 267(41%), S aureus, was 48(5.6%), G(-) bacillus was 126(5.6%), and Corynebactrium spy. was 52(6%), respectively, from total 861 samples. 4. The results of antimicrobial drug susceptibility of all isolates were showed that Streptococci spp., Staphylococci spp., S aureus, (G)(-) bacillus, and Corynebactrium spp. were susceptible to cefuroxime, cefoperazone, amoxicillin, cefazolin, ampicilin, penicillin, gentamicin, erythromycin, cloxacilin, ciprofloxacin, sulfamethoxasole/trimetoprim, teteracyclin, and norfloxacin (> 70%), but some of them were resistant to neomycin, streptomycin colistin, and cephalothin(> 60). 5. The results of drug susceptibility obtained from each farms had different susceptibility, even though, etiological microorganisms were same in each farms.

• Korean Journal of Veterinary Service
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• v.29 no.2
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• pp.111-122
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• 2006

A direct, accurate and sensitive chromatographic analytical method for quantitative determination of four fluoroquinolones (norfloxacin, cirprofloxacin, danofloxacin and enrofloxacin) in chicken, pork and beef edible muscle is proposed in the present study. The developed method was successfully applied to the determination of enrofloxacin, as the main component of commercially available veterinary drugs. The samples were homogenized and the antimicrobials were added, then they were extracted twice with dichloromethane. Fluoroquinolone antibiotics were separated on an agilent $250x4mm,\;C_{18},\;5{\mu}m$, analytical column, at $25^{\circ}C$. The mobile phase consisted of a mixture of DW : acetonitrile : triethylamine(80:19:1%, v/v, pH 3.0) leading to retention times less than 14 min. at a flow rate 0.5 ml/min. These fluoroquinolones were detected by liquid chromatography with fluorescence at 290 nm excitation and 465 nm emission. The limits of quantification in each edible muscle (chicken, pork, and beef) were 0.32-6.54 ng/g. Using 0.5 g of each sample, average recovery rates at fortification levels of 0.05, 0.1 and 0.2 ${\mu}g/ml$ ranged 70.14-71.71% for NFX, 71.87-73.89% for CFX, 82.16-92.35% for DFX, and 90.13-98.12 for EFX This is a simple and economic method to quantify the presence of NFX, CFX, EFX and DFX in edible muscle of animal origin.

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.281-286
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• 2009

Fluoroquinolones in muscle and egg were separated by liquid extraction and determined. The analysis was carried out using following conditions; C18 column ($150{\times}4.6mm$, $5{\mu}m$), mobile phase composed of D.W. (containing 0.4% triethylamine and phosphoric acid) : methanol : acetonitrile (780:100:120, v/v/v), quarternary pump at a flow rate of 0.9ml/min and $20{\mu}l$ of injection volume, fluorescence detector with EX 278nm/Em 456nm. The calibration range of seven fluoroquinolones showed linearity ($r^2{\geq}0.999$) at concentration range of $0.025{\sim}0.8{\mu}g/ml$. The recoveries in fortified muscle and egg represented more than 81.3%. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin, saraloxacin and orbifloxacin were 3.1, 2.5, 3.6, 1.7, 0.9, 2.5 and $2.1{\mu}g/kg$, respectively. We also monitored fluoroquinolones residue in the sample (chicken muscle 182, cattle muscle 140, pig muscle 139, egg 212) using EEC-plate (E. coli ATCC 11303) screening and HPLC confirmation methods. The screening test results, fluoroquinolones, antibacterial substances were all negative.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.305-313
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• 2008

The present study was conducted to investigate the epidemiological characteristics such as isolation rate and antimicrobial drug susceptibility of etiological agents. The specimens (stool or intestine) were collected from 319 calves with clinical diarrhea from 195 farms in Gyeongnam south area (Gosung, Tongyung, Hadong) from June 2005 to August 2006. The isolation rate of Salmonella spp was higher in summer (8.4%) than in winter (4.8%) and the average was 7.2% (23/319 head). Some of Salmonella spp isolated were resistant to penicillin, oxytetracycline, tetracycline, and cephalexin (>90%), but some of them were susceptible to norfloxacin, ciprofloxacin, sulfamethoxazole/trimethoprim and amikacin(>30%). There was no statistical difference in the isolation rate of Eimeria spp between summer(48.9%) and winter(42.3%). For the evaluation of infection level of Eimeria spp oocyst per gram of feces (OPG) was examined, and severe, moderate and light infection level were 11.9%, 12.5% and 22.3%, respectively. In the isolation rates of Eimeria spp the calves under 19 days was lowerthan those over 60 days, but there was not different among herd size.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.4
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• pp.322-328
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• 2009

Three-hundred and sixteen Escherichia coli strains from seawater, and a variety of farmed fishes, including oliver flounder (Paralichthys olivaceus), black rock fish (Sebastes schlegeli), red sea bream (Pagrus major) and sea bass (Lateolabrax japonicus) between May to October in 2004, were tested by agar dilution method to determine their susceptibility patterns to 17 antimicrobial agents. Overall, 92.1% of Escherichia coli isolates from samples showed antimicrobial resistance to at least one antimicrobial agent and the multiple resistance was seen in 173 isolates (54.7%). The resistance of E. coli isolates to tetracycline (74.1%) was highest, followed by cephalothin (69.9%), doxycycline (66.5%), streptomycin (47.2%), ampicillin (46.2%), cefazolin (31.6%), enrofloxacin (31.0%). norfloxacin (28.2%). The most frequent resistance pattern was TE-D-CF-CIP-ENO-NOR-AM-S-C-SXT-AmC-CZ (14.7%), followed by CF (6.2%), TE (5.1%), TE-CF (4.5%) in 177 isolates from fishes and TE-D-CF (7.2%) followed by TE-D-CF-S (5.8%), CF and TE-D-S (3.6%) in 139 isolates from seawater.

• Journal of Microbiology
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• v.43 no.2
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• pp.133-143
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• 2005

Shigellosis is a global human health problem. Four species of Shigella i.e. S. dysenteriae, S. flexneri, S. boydii and S. sonnei are able to cause the disease. These species are subdivided into serotypes on the basis of O-specific polysaccharide of the LPS. Shigella dysenteriae type 1 produces severe disease and may be associated with life-threatening complications. The symptoms of shigellosis include diarrhoea and/or dysentery with frequent mucoid bloody stools, abdominal cramps and tenesmus. Shigella spp. cause dysentery by invading the colonic mucosa. Shigella bacteria multiply within colonic epithelial cells, cause cell death and spread laterally to infect and kill adjacent epithelial cells, causing mucosal ulceration, inflammation and bleeding. Transmission usually occurs via contaminated food and water or through person-to-person contact. Laboratory diagnosis is made by culturing the stool samples using selective/differential agar media. Shigella spp. are highly fragile organism and considerable care must be exercised in collecting faecal specimens, transporting them to the laboratories and in using appropriate media for isolation. Antimicrobial agents are the mainstay of therapy of all cases of shigellosis. Due to the global emergence of drug resistance, the choice of antimicrobial agents for treating shigellosis is limited. Although single dose of norfloxacin and ciprofloxacin has been shown to be effective, they are currently less effective against S. dysenteriae type 1 infection. Newer quinolones, cephalosporin derivatives, and azithromycin are the drug of choice. However, fluoroquinolone-resistant S. dysenteriae type 1 infection have been reported. Currently, no vaccines against Shigella infection exist. Both live and subunit parenteral vaccine candidates are under development. Because immunity to Shigella is serotype-specific, the priority is to develop vaccine against S. dysenteriae type 1 and S. flexneri type 2a. Shigella species are important pathogens responsible for diarrhoeal diseases and dysentery occurring all over the world. The morbidity and mortality due to shigellosis are especially high among children in developing countries. A recent review of literature (KotIoff et al.,1999) concluded that, of the estimated 165 million cases of Shigella diarrhoea that occur annually, $99\%$ occur in developing countries, and in developing countries $69\%$ of episodes occur in children under five years of age. Moreover, of the ca.1.1 million deaths attributed to Shigella infections in developing countries, $60\%$ of deaths occur in the under-five age group. Travellers from developed to developing regions and soldiers serving under field conditions are also at an increased risk to develop shigellosis.