• Title/Summary/Keyword: Bacterial Diffusion

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Prevalence of Methicillin-resistant Staphylococci Isolates from Horses and Horse-related Personnel in Korea (말 및 말관련 종사자의 methicillin 내성 포도상구균의 유병율 조사)

  • Lee, Sang-Kyu;Han, Jae-Ik;Kim, Ill-Hwa;Na, Ki-Jeong;Kang, Hyun-Gu
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.194-198
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    • 2014
  • Methicillin-resistant staphylococci (MRS) are emerging as important pathogens in humans and animals worldwide. The aim of this study was to investigate the prevalence of MRS in the racehorse population and in horse-related personnel in Korea. A total of 195 horses and 18 humans (eight veterinarians, three veterinary hospital staff, and seven horse-handlers) from racehorse farms in Korea were included in the study. The samples were collected from nasal cavities using bacterial transport medium and were cultivated on tryptic soy agar with 5% sheep blood for 3 days at $37^{\circ}C$ to confirm the presence of Staphylococcus spp. Presumptive Staphylococcus spp. isolates were identified by 16S ribosomal RNA gene analysis. The coagulase test and oxacillin susceptibility tests were performed using the tube dilution and disk diffusion methods, respectively. The presence of the mecA gene was determined using a polymerase chain reaction assay. Of the 195 horses, 29 (15.6%) yielded 29 MRS isolates. Twelve (66.7%) of the 18 horse-related personnel yielded 12 MRS isolates. All of the MRS isolates from horses or horse-related personnel were identified as methicillin-resistant coagulase-negative staphylococci (MRCNS). The result of this study suggest that the prevalence of MRS increased with the duration of antibiotic use (p = 0.002). This study also provides evidence for the zoonotic transmission of MRCNS between horses and humans, although further investigations are needed.

Specific Detection of Serratia marcescens Based on a PCR Assay and Antimicrobial Susceptibility of S. marcescens Isolated from Boar Semen (Serratia marcescens 검출을 위한 PCR 기법 개발 및 돼지정액 유래균주에 대한 항생제 감수성 양상)

  • Jung, Ji-A;Kim, Aeran;Seo, Byoung Joo;Jung, Suk Chan;Kim, In Cheul;Chung, Ki Hwa;Jung, Byeong Yeal
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1133-1139
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    • 2013
  • During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

Characterizations of Class 1 Integrons in Proteus mirabilis Isolated from Chickens at Chungcheong Province (충청지역의 닭으로부터 분리된 Proteus mirabilis 균주에 존재하는 Class 1 Integron의 유전형 분석)

  • Sung, Ji Youn;Byeon, Yong Gwan
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.65-70
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    • 2015
  • Antimicrobial agents or additives have commonly been used in domestic animals for the prevention and treatment of bacterial diseases. Unfortunately, this has resulted in the overgrowth of bacteria that is resistant to antimicrobial agents used by humans, and these might get disseminated to humans via the food. In this study, we investigated the prevalence of integrons, and characterized gene cassette arrays in Proteus mirabilis isolates obtained from chickens in Chungcheong province of Korea. Additionally, the correlation between gene cassette arrays and antimicrobial resistance rate was studied. A total of 26 Proteus mirabilis isolates were recovered from chickens in Chungcheong province in Korea. Antimicrobial susceptibility was determined by disk diffusion method. PCR and DNA sequencing were performed to characterize the gene cassette arrays. In addition, we employed repetitive extragenic palindromic sequence-based PCR (REP-PCR) method for clonality analysis of P. mirabilis strains. Of the 26 P. mirabilis isolates tested, 14 (53.8%) isolates carried class 1 integrons, while class 2 and class 3 integrons were not detected in our study. The class 1 integrons harbored genes encoding resistance to aminoglycosides (aacCA5, aadA2, aadA5 and aadA7), trimethoprim (dfrA17, and dfrA32), lincosamides (linF) and erythromycin (ereA). In particular, the presence of class 1 integron had a significant correlatation to a high resistance rate of aminoglycoside and trimethoprim. We confirmed that class 1 integrons are widely disseminated in P. mirabilis isolates from chickens, contributing to the resistance to diverse antimicrobial agents in Korea. To prevent further spreading of antimicrobial resistant genes among P. mirabilis isolates, constant monitoring and clinical policing will become necessary.

Antimicrobial Effect of Extract of Glycyrrhiza uralensis on Methicillin-Resistant Staphylococcus aureus (감초 추출물이 항생제 내성균주의 항균활성에 미치는 영향)

  • Lee, Ji-Won;Ji, Young-Ju;Yu, Mi-Hee;Im, Hyo-Gwon;HwangBo, Mi-Hyang;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.456-464
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    • 2005
  • Antimicrobial drug-resistance is natural response to antimicrobial stress based on selection, which weakens chemotherapy effect. Introduction of large numbers of chemotherapeutic agents to clinical practice has generated strains of microorganisms that survive and multiply in vivo with high-drug concentrations. Methicillin-resistant Staphylococcus aureus (MRSA), bacteria found in normal daily life, can be easily ingested through milk vegetables, and meats, etc. MRSA emerged in many port of the world, increasing complex clinical problems. Therefore, new agents are needed to treat MRSA. Glycyrrhiza uralensis was extracted using 80% MeOH to investigate its antimicrobial activity against MRSA stains KCCM 11812, 40510, and 40512 through bacterial measurement, disc diffusion, and O.D. methods, MIC values, MRSA gene expression investigation, and scanning electron microscope observation. Results revealed MecA, Mecl, MecRI, and FemA were the most highly manifested MRSA genes. Methanolic extract of G. uralensis significantly inhibited MRSA and thus could be used in development of antibacteria.

Comparison of Virulence Factors, Phylogenetic Groups and Ciprofloxacin Susceptibility of Escherichia coli Isolated from Healthy Students and Patients with Urinary Tract Infections in Korea

  • Park, Min;Park, Soon-Deok;Kim, Sa-Hyun;Woo, Hyun-Jun;Lee, Gyu-Sang;Kim, Hyun-Woo;Yang, Ji-Young;Cho, Eun-Hee;Uh, Young;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.146-151
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    • 2012
  • Urinary tract infection (UTI) is one of the most common bacterial infections and is predominantly caused by uropathogenic Escherichia coli (UPEC). UPEC strains generally possess several genes encoding virulent factors, which are mostly adhesins, toxins, bacteriocin and siderophores. E. coli is composed of four main phylogenetic group (A, B1, B2, D) and virulent extra-intestinal strains mainly belong to groups B2 and D. Prescription of ciprofloxacin, a kind of fluoroquinolone group antibiotics, is increasing now a days, but resistance to this drug is also increasing. A total of 188 strains of E. coli were collected. Thirteen strains were collected from healthy students in 2011 and 175 strains from patients with urinary tract infection in 2010. Virulence factor genes (papC, fimG/H, sfaD/E, hlyA, cnf1, and usp) were amplified by polymerase chain reaction (PCR) methods for phylogenetic group (A, B1, B2, D) detection. Ciprofloxacin susceptibility test was performed by disk diffusion method. The identified virulence factors (VFs), phylogenetic groups and ciprofloxacin resistance in 13 E. coli strains isolated from healthy students were papC (15.4%), fimG/H (76.9%), sfaD/E (30.8%), hlyA (23.1%), cnf1 (23.1%), usp (7.7%), phylogenetic group A (23%), B1 (8%), B2 (46%), D (23%) and ciprofloxacin resistance (7.7%), while those of in 175 E. coli strains isolated from patients with UTI were papC (41.1%), fimG/H (92.5%), sfaD/E (30.3%), hlyA (10.3%), cnf1 (30.3%), usp (27.4%), phylogenetic group A (9.1%), B1 (5.1%), B2 (60.6%), D (25.1%) and ciprofloxacin resistance (29.7%). In this study, 10 out of 13 E. coli strains (76.9%) from healthy students were found to possess more than one virulence factor associated with adhesion. In addition, one E. coli strain isolated from healthy students who had never been infected with UPEC showed ciprofloxacin resistance. According to these results between the virulence factors and phylogenetic groups it was closely associated, and UPEC strains isolated from patients showed high level of ciprofloxacin resistance.

Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Isolation and characterization of Bacillus subtilis NO12 from button mushroom substrates (양송이 배지로부터 분리된 Bacillus subtilis NO12의 특성)

  • Kim, Hye Soo;Park, Hyun Young;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.249-253
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    • 2017
  • Twelve strains of bacteria with cellulase and xylanase activities were isolated from spent mushroom substrates collected from button mushroom cultivation farm, Buye, Chungcheongnam-do in Korea. Among them, one strain, designated NO12, with higher cellulase and xylanase activities was selected by agar diffusion method. The strain NO12 was identified to be a Bacillus sp. by biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA gene sequence analysis showed that strain NO12 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99.2%. Based on its physiological properties, biochemical characteristics, and phylogenetic distinctiveness, strain NO12 was classified within the genus Bacillus, for which the name Bacillus subtilis NO12 was proposed. The cellulase and xylanase activities of B. subtilis NO12 were slightly increased according to bacterial population from exponential phase to stationary phase in the growth curve for B. subtilis NO12. The xylanase activity continuously increased from the beginning of the exponential phase and exhibited maximum activity in the middle of the exponential phase.

Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.

Isolation of Anaerobic Bacteria from Clinical Specimens in Veterinary Medical Teaching Hospital (대학 동물병원 임상 검체로부터 분리된 혐기성 세균과 항생제 감수성 양상)

  • Park, Se-won;Chung, Jin-young;Hwang, Cheol-yong;Youn, Hwa-young;Han, Hong-ryul
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.15-19
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    • 2004
  • The emergence of bacterial resistance to antibiotics during therapy is a matter of great problem in clinical medicine. This may be because many veterinarians have used inappropriate antibiotics without bacteriological culture. Therefore, the purpose of this study is to determine isolation of anaerobic bacteria as pathogens from veterinary clinical specimens as well as susceptibility pattern for choosing antibiotics. Various anaerobic bacteria were isolated from clinical specimens of dogs, cats, rabbits at Veterinary Medical Teaching Hospital of Seoul National University from May 2001 to October 2002. The total number of isolated anaerobic bacteria was 13 isolates; Bacteroides spp. (3 isolates), Fusobacterium spp. (2 isolates), Peptostreptococcus spp. (2 isolates), Porphyromonas gingivalis (2 isolates), Prevotella spp. (3 isolates), and Propionibacterium acnes (1 isolate). For evaluating the antibiotic susceptibility patterns of the isolates disk diffusion method was used. All isolates were susceptable to all tested antibiotics except only one Fusobacterium varium was resistant to norfloxacin.

Clonal Dissemination of Multidrug Resistant Acinetobacter baumannii Isolates Harboring blaOXA-23 at One University Hospital in Daejeon, Korea (대전지역 소재 대학병원에 blaOXA-23 유전자를 가지고 있는 다제내성 Acinetobacter baumannii의 확산)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.94-101
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    • 2016
  • Acinetobacter species isolates are important opportunistic pathogens and commonly implicated in nosocomial infections. The therapeutic options for treatment of the bacterial infections are limited because the bacteria isolates are usually multidrug resistant (MDR). In the current study, we investigated various carbapenemase genes in 68 Acinetobacter species isolates. Antimicrobial susceptibilities were tested using the disk diffusion method. Screening of carbapenemase genes was performed via multiplex PCR. In addition, PCR and DNA sequencing were used to identify the carbapenemase genes. Repetitive extragenic palindromic-PCR (REP-PCR) was also performed to assess the clonality of isolates. In our study, A. baumannii isolates were highly resistant to all agents tested while all non-A. baumannii isolates were susceptible to all agents tested, with the exception of aztreonam and cefotaxime. All 51 A. baumannii isolates contained the $bla_{OXA-51}$ gene and 37 (72.5%) isolates also harbored the $bla_{OXA-23}$ gene. In addition, 39 MDR A. baumannii isolates were identified in our study and 37 isolates contained the $bla_{OXA-23}$ gene. The 37 MDR strains harboring $bla_{OXA-23}$ showed type I (n=22) or type II (n=15) banding patterns on their REP-PCR profiles. Our results suggest clonal relation and horizontal spreading of MDR A. baumannii isolates containing the $bla_{OXA-23}$ gene at the hospital located in Daejeon. Continuous investigation of antimicrobial resistant determinants and monitoring emergence and dissemination of MDR isolates is required to prevent and control infection and colonization of MDR A. baumannii isolates.