• Tuberculosis and Respiratory Diseases
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• 제72권6호
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• pp.475-480
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• 2012

Background: Pyrazinamide (PZA) is an effective antitubercular drug that becomes toxic to Mycobacterium tuberculosis when converted to pyrazinoic acid by pyrazinamidase (PZase), encoded by mycobacterial pncA. A strong association was noted between the loss of PZase activity and PZA resistance. The causative organisms in extrapulmonary tuberculosis are rarely cultured and isolated. To detect pncA mutations in specimens from extrapulmonary tuberculosis as confirmative diagnosis of mycobacterial infection and alternative susceptibility test to PZA. Methods: Specimens were collected from clinically proven extrapulmonary tuberculosis. pncA was sequenced and compared with wild-type pncA. Results: pncA from 30 specimens from 23 donors were successfully amplified (56.6% in specimens, 59% in donors). Six mutations in pncA were detected (20.0% in amplified specimens, 26.1% in specimen donors) at nucleotide positions of 169, 248 and 419. The mutation at position 169 results in substitution of aspartic acid for histidine, a possible allelic variation of M. bovis that have intrinsic PZA resistance. The mutation at position 248 changes proline into arginine and that at position 419, arginine into histidine. Conclusion: DNA-based diagnosis using pncA may be simultaneously useful for the early diagnosis of mycobacterial infection and the rapid susceptibility to PZA in extrapulmonary tuberculosis. A potential implication of pncA allelic variation at 169 might be suggested as a rapid diagnostic test for M. bovis infection or Bacille Calmette-Gu$\acute{e}$rin (BCG) reactivation.

• 대한수의학회지
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• 제27권2호
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• pp.227-233
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• 1987

This study was conducted to determine the epidemiological characteristics of Campylobacter enteritis. A total of 187 fecal specimens of Korean native goat were examined for the presence of C. jejuni and C. coli by direct plating. Fifty strains isolated were examined for biochemical and serological properties and susceptibility to 19 chemotherapeutic agents. A total of 29(15.5%) C. jejuni and 21 (11.2%) C. coli were isolated from the fecal specimen of 187 Korean native goats. Of the 50 isolates of C. jejuni and C. coli, 29 isolates of C. jejuni grouped as 7 biotypes (1,2,3,4,6,7 and 8) and biotypes 1(34.5%), 2(17.2%) and 3(20.7%) were encountered most frequently. Twenty-one C. coli strains were differentated into biotype I (61.9% of the isolates) and biotype II (38.1%). Of the 29 C. jejuni strains examined, 24(83.0%) were typable by the Lior serotyping scheme and five isolates were non typable. C. jejuni grouped as 8 serotypes, serotype 4(24.1%) and 26(20.7%) were encountered most frequently. In the case of 21 strains of C. coli grouped as 6 serotypes, the most frequent serotypes were 21(28.6%) and 25(23.8%). Total of 50 strains of isolated were all susceptible to amikacin, clindamycin and tobramycine. Overall 85% of isolates were sensitive to erythromycin, doxycycline, chloramphenicol, flume-quine, kanamycin, gentamicin, nalidixic acid, polymyxin B, colistin, tetracycline and ampicillin, but about 65% of isolates were resistant to cefamandole and ethyl hydrocuprein hydrochloride.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8883-8886
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• 2014

Background: Tetracycline is an antibiotic widely used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing due to increasing bacterial resistance. The aim of this study was to investigate the occurrence of 16S rRNA mutations associated with resistance or reduced susceptibility to tetracycline ofHelicobacter pylori by real-time PCR (RT-PCR) assays from culture. Materials and Methods: Tetracycline susceptibility and minimal inhibition concentration (MIC) was determined by the Epsilometer test (Etest) method. A LightCycler assay developed to detect these mutations was applied to DNA extracted from culture. The 16S rRNA of these isolates was sequenced and resistance-associated mutations were identified. From 104 isolates of H. pylori examined, 11 showed resistance to tetracycline. Results: LightCycler assay was applied to DNA extracted from 11 tetracycline-susceptible and 11 tetracycline resistance H. pylori isolates. In our study the sequencing of the H. pylori wild types in 16 s rRNA gene were AGA 926-928 with MIC (0.016 to $0.5{\mu}g/ml$), while the sequencing and MIC for resistant were GGA and AGC, (0.75 to $1.5{\mu}g/ml$), respectively. Also we found a novel mutation in 2 strains with $84^{\circ}C$ as their melting temperatures and exhibition of an A939C mutation. Conclusions: We conclude that real-time PCR is an excellent method for determination of H. pylori tetracycline resistance related mutations that could be used directly on biopsy specimens.

• 대한미생물학회지
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• 제13권1호
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• pp.43-48
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• 1978

항균제내성(抗菌劑耐性)인 Salmonella typhi를 공시(供試)하여 chloramphenicol(Cm), tetracycline(Tc), ampicillin(Ap), kanamycin(Km) 및 rifampicin(Rif)의 항균작용(抗菌作用)을 한천평판희석법(寒天平板稀釋法)(평판법(平板法))과 broth 희석법(稀釋法)(broth법(法))으로 비교(比較)하였다. 공시균(供試菌)의 최소발육저지농도(最少發育沮止濃度)(MIC)를 보면 Cm, Tc 및 Rif에 있어서는 평판법(平板法)과 broth법(法) 사이에 유의(留意)할만한 차이(差異)는 없었으나 Km에 있어서는 broth법(法)의 MIC가 평판법(平板法)보다 높은 것이 약(約) 반수(半數)있었고 Ap에 있어서도 소수(少數) 있었다. Cm과 Rif는 대부분(大部分)의 공시균(供試菌)에 대(對)하여 정균적(靜菌的)으로 작용(作用)하였으나 Tc, Ap 및 Km은 살균적(殺菌的)으로 작용(作用)하였고 그 작용(作用)은 균주(菌株)에 따라 차이(差異)가 있었다.

• 한국동물위생학회지
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• 제21권4호
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• pp.451-465
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• 1998

The present study was carried out to investigate the prevalence of brucellosis in Kyungbuk area for the 3 years from 1966 to 1998. Collective milk samples were routinely screened to detect positive farms by using the milk ring test(MRT), and serum agglutination test was performed to detect sero-positive individuals in the MRT positive farms. Attempt were made to isolate the causative organismas from slaughtered sero-positive reactors and some biochemical and polymerase chain reation characters of the isolates were also made to identify the organisms. Seroprevalence to brucellosis in peoples who are close contact with infected dairy herds was also investigated. Brucellosis of dairy cattle was rare before 1997, but has been broken more frequently since early 1998. By the MRT for dairy herds, positive rate was gradually increased every year : 0.6% in 1996, 1.5% in 1997, 3.9% in 1998. Among 262 MRT-positive herds, only 21 herds(8.0%) showed positive brucellosis in serological test. The isolation rates of Brucella sp from tested materials were 51.2% in supramammary glands, 39.5% in milks, and 50.0% in pulmonary Iymphnode, respectively. Isolated strain and biotype were Brucella(B) arbortus biotype 1 in 26 heads, and were B suis biotype 1 in 2 heads. Isolated strain and vaccine strain were very similar in their colony morphology and staining. In drug susceptibility, isolated stains(B abortus) and vaccine strain(B abortus RB-51) were sensitive to ampicillin, gentamycin, kanamycin, neomycin, penicillin, streptomycin, and to tetracycline, but resistant to erythromycin. In the PCR, field strains reacted to BA and IS711 primers, and vaccine strain reacted to BA, IS711, and RB5l primers. In the plate agglutination test of 96 sera of human contacted with animals, serum antibody titer detected 1 : 100 in one person, 1 : 200 in one, and below 1 : 25 in the others.

• 동의생리병리학회지
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• 제19권2호
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• pp.304-314
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• 2005

The major concept of Sasang typology is that the disease susceptibility and drug response as well as physiological characteristics are presumed to be different depending on their Sasang types. Although characterizing fundamental basis of their traits are crucial in this research field, only pathological susceptibility and physical appearances were thoroughly studied. We evaluated their physiological characteristics by tapping psychological, physical and genetic traits of each Sasang types. After determining the Sasang type of one hundred three college students based on the Questionnaire for the Sasang Constitution Classification, the psychological, physical and genetic traits of each type were analyzed with the Myers-Briggs Type Indicator (MBTI), Bioelectrical Impedance Analysis and genetic polymorphism test, respectively. Each of the Sasang types showed significantly different profiles (Generalized estimation equation, coef=11.88, z=2.13, p=0.033), and could be distinctively classified based on their MBTI scores (discriminant analysis Wilks Lambda=0.611, df=8, chi-square=36.7, p<0.001). Subjects with the So-Eum type (Introversion and Judging) and the So-Yang type (Extroversion and Perceiving) showed contrasting psychological features, however they had similar anthropometric characteristics. Subjects with the Tae-Eum type showed bigger Body Mass Index ($R^2$=0.22, df=4, 74, F=5.07, p=0.001) and body shape compared to others. Although there were no significant differences in G-protein beta-3 subunit polymorphism, angiotensin-converting enzyme polymorphism and Methylenetetrahydrofolate reductase polymprhisms among groups with Sasang types, it was shown that the dopamine system could be one for genetic marker for Sasang typology. These results demonstrated distinctive and essential traits of Sasang typology using reproducible psychometric, anthropometric and genetic evaluations. We also found that the Sasang typology was a bio-psychological typology which could show trait-specific guideline for individualized medicine.

• Annals of Laboratory Medicine
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• 제38권6호
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• pp.545-554
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• 2018

Background: The increasing morbidity and mortality rates associated with Acinetobacter baumannii are due to the emergence of drug resistance and the limited treatment options. We compared characteristics of colistin-resistant Acinetobacter baumannii (CR-AB) clinical isolates recovered from patients with and without prior colistin treatment. We assessed whether prior colistin treatment affects the resistance mechanism of CR-AB isolates, mortality rates, and clinical characteristics. Additionally, a proper method for identifying CR-AB was determined. Methods: We collected 36 non-duplicate CR-AB clinical isolates resistant to colistin. Antimicrobial susceptibility testing, Sanger sequencing analysis, molecular typing, lipid A structure analysis, and in vitro synergy testing were performed. Eleven colistin-susceptible AB isolates were used as controls. Results: Despite no differences in clinical characteristics between patients with and without prior colistin treatment, resistance-causing genetic mutations were more frequent in isolates from colistin-treated patients. Distinct mutations were overlooked via the Sanger sequencing method, perhaps because of a masking effect by the colistin-susceptible AB subpopulation of CR-AB isolates lacking genetic mutations. However, modified lipid A analysis revealed colistin resistance peaks, despite the population heterogeneity, and peak levels were significantly different between the groups. Conclusions: Although prior colistin use did not induce clinical or susceptibility differences, we demonstrated that identification of CR-AB by sequencing is insufficient. We propose that population heterogeneity has a masking effect, especially in colistin non-treated patients; therefore, accurate testing methods reflecting physiological alterations of the bacteria, such as phosphoethanolamine-modified lipid A identification by matrix-assisted laser desorption ionization-time of flight, should be employed.

• Infection and chemotherapy
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• 제50권4호
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• pp.357-361
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• 2018

While carbapenems are the drug of choice to treat extended-spectrum-${\beta}$-lactamase (ESBL)-producing strains, some alternative carbapenem-sparing regimens are suggested for antibiotic stewardship. We experienced a case of ciprofloxacin treatment failure for acute pyelonephritis caused by an apparently susceptible Escherichia coli. A 71-year-old woman presented the emergency department with fever for 7 days and bilateral flank pain for 2 days. The laboratory results and abdominopelvic computed tomography finding were compatible with acute pyelonephritis. During 3-day ciprofloxacin therapy, the patient remained febrile with persistent bacteremia. After the change in antibiotics to ertapenem, the patient's clinical course started to improve. ESBL-producing E. coli isolates were identified in all three consecutive blood samples. Pulsed-field gel electrophoresis (PFGE) patterns, serotypes, and sequence types showed the three isolates were derived from the identical strain. The isolates produced CTX-M-14 type ESBL belonging to the ST69 clonal group. Despite in vitro susceptibility, the failure was attributed to a gyrA point mutation encoding Ser83Leu within quinolone resistance-determining regions. This case suggests that ciprofloxacin should be used cautiously in the treatment of serious infections caused by ciprofloxacin-susceptible, ESBL-producing E. coli, even in acute pyelonephritis because in-vitro susceptibility tests could fail to detect certain genetic mutations.

• Journal of Veterinary Science
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• 제21권4호
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• pp.57.1-57.11
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• 2020

Background: Streptococcus dysgalactiae subspecies equisimilis (SDSE) acts as an etiological agent for lameness, neurological signs, and high mortality in pigs. Despite its importance in pig industries and zoonotic potential, little is known about the effects of this pathogen. Objectives: This study aimed to determine the molecular characteristics and antimicrobial resistance of SDSE strains isolated from diseased pigs. Methods: A total 11 SDSE isolates were obtained from diseased pigs. Bacterial identification, PCR for virulence genes, emm typing, and antimicrobial resistance genes, multilocus sequence typing, and antimicrobial susceptibility test were performed. Results: Nine isolates were from piglets, and 8 showed lameness, sudden death, or neurological signs. The isolates were PCR-positive for sla (100%), sagA (100%), and scpA (45.5%), and only 1 isolate amplified the emm gene (stL2764). Eight different sequence types were detected, categorized into 2 clonal complexes and 4 singletons. All the isolates in this study were included in a small cluster, which also contained other strains derived from humans and horses. The minimum inhibitory concentrations for the tested beta-lactams were low, while those for macrolides, tetracyclines, and fluoroquinolones were relatively high. PCR analysis of the macrolide and tetracycline resistance genes demonstrated that the isolates carried erm(B) (18.2%, n = 2), mef(A/E) (9.1%, n = 1), tet(M) (18.2%, n = 2), and tet(O) (90.2%, n = 10). Two isolates presented a mutation in parC, which is associated with fluoroquinolone resistance. Conclusion: This study provided insight into swine-derived SDSE, as it is related to veterinary medicine, and elucidated its zoonotic potential, in the context of molecular epidemiology and antimicrobial resistance in public health.

• Journal of Veterinary Science
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• 제24권6호
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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.