• Journal of Microbiology and Biotechnology
• /
• v.14 no.4
• /
• pp.789-795
• /
• 2004

A promoter-probe shuttle vector pSK1Cat was constructed for the isolation of transcriptional signal sequences from Corynebacterium glutamicum. Besides conferring resistance to kanamycin in Escherichia coli and C. glutamicum, the vector carried a promoterless cat gene to confer resistance to chloramphenicol upon insertion of the appropriate transcriptional signals in the multiple cloning site. By utilizing the vector, a series of transcriptionally active fragments were isolated from the genome of C. glutamicum. The clones, ranging from 200 bp to 1 kb in size, were grouped into 3 classes of strong, medium, and weak, based on the chloramphenicol acetyltransferase (CAT) activity and sensitivity to the chloramphenicol of the clone-carrying C. glutamicum cells. C. glutamicum cells carrying the $P_{19}$ clone, a representative in the strong class, were able to grow on minimal agar plates containing over $40 mg/mell$ chloramphenicol, and showed CAT activity of 10 m㏖/mgㆍmin, performing slightly better than the cells carrying $P_{tac}$ , a strong E. coli promoter. Subcloning analysis of the $P_{19}$ clone identified a 180 bp intergenic fragment ($P_{180}$), which was located upstream of a gene encoding a hypothetical membrane protein. The expression conferred by $P_{180}$ was not affected by either the kinds of carbon sources or changes in temperature. These properties make the $P_{180}$ clone useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation.

• The Plant Pathology Journal
• /
• v.17 no.3
• /
• pp.149-153
• /
• 2001

A total of 2109 isolates of Botrytis cinerea were collected from infected plants fo strawberry, tomato, and cucumber in Korea from 1994 to 1996. Based on in virtotests for mycelial growth on potato-dextrose agar containing fungicides, the esolates were classified into six phenotypic groups : SSR, SRR, RSS, RRS, RSR, and RRR, representing sensitivity (S) or resistance (R) to carbendazim, procymidone, and diethofencarb. In that order the isolation frequencies of the SSR, SRR, RSS, RRS, RSR, and RRR phenotypes were 28.7, 1.1, 28.8, 39.4, 1.0, and 0.9%, respectively. Three isolates from each SSR, SRR, RSS, RRS, and RSR and an isolate of RRR phenotype were selected and evaluated for their fitness-related characteristics such as pathogenic aggressiveness, mycelial growth rate, sporulation, and sclerotial formation. Competitive abilities of the SSR, SRR, RSS, RRS, and RSR phenotypes were also compared by inculating mixtures of conidial suspensions of two phenotypes to cucumber plant, and then determining re-isolation frequencies from lesions. In general, significant differences in fitness-related characteristics, except pathogenic aggressiveness, were found not only between but also within phenotype groups. In the competitiveness tests, carbendazim-sinsitive phenotypes (SSR and SRR) were found to be more competitive than the resistant ones (RSS and RSR), whereas, the procymidone-resistant phenotypes (SRR and RRS) appeared to be more competitive than the sensitive ones (SSR, RSS, and RSR). There was no consistent dominance in competitiveness between the diethofencarb-resistant and sensitive phenotypes. The RSR phenotype was the least competitive among the five phenotypes.

• Korean Journal of Veterinary Service
• /
• v.24 no.1
• /
• pp.51-68
• /
• 2001

The result of studying the epidemiological characteristics of Salmonella strains which have been isolated from the domestic animals in Gyeongbuk province from February 1998 to August 2000 were summarized as follows. The isolation rates of Salmonella strains were 2.0% from cattle feces, 6.3% from cattle lymph node, 9.5% from pig feces, and 25.1% from pig lymph node. In poultry, the isolation rates were 30.3%. The isolates of Salmonella showed positive reaction for MUCAP test, methyl red test, but showed negative reaction for urea test, indole test, Voges Proskauer test. On TSI agar, the isolates showed acid butt, alkaline slant. Also, the isolates were identified as Salmonella strain by API 20E kit. Non H$_2$S Production Salmonella strains isolated from poultry were identified as S gallinarum. As a result of serotyping, B group were the most common in cattle and pig, Dl in chickens. 21 serovars were found. the common serovar from the domestic animals was S typhimurium, S derby, S agona, S schwarzenground, S enteritidis and S gallinarum. The most commonly encountered serovars in cattle were S agona and S typhirimurium in pig, S gallinarum in chicken. As a result of antimicrobial susceptibility test, all Salmonella isolates were susceptible to amikacin, ciprofloxacin, norfloxacin; cefotaxime and polumcin B. The resistance rates to tetracycline and streptomycin was 58% and 56%, respectively. 69.3% of all isolates were resistant to more than one antimicrobial agent. Out of the resistant isolates, the isolates resistant to streptomycin and tetracycline was 36%. There were 24 strains of multiresistant isolates resistant to more than 5 antimicrobial agents. S typhimurium were resistant to all antimicrobial agents, also had a lot of multiresistant strains. Therefore, S typhimurium was considered as a major agent of antimicrobial resistance.

• Journal of the Computational Structural Engineering Institute of Korea
• /
• v.33 no.5
• /
• pp.311-318
• /
• 2020

This study introduces a technique for improving the elastomeric-isolator performance using modular devices. The modular devices are shear resistance block, polymer spring, displacement acceptance guide, and anti-falling block. They are installed on the elastomeric isolator as a supplementary device. Each modularized device improves the isolator performance by performing step-by-step actions according to the seismic intensity and displacement. The PRB isolation device works in four stages, depending on the seismic magnitude, to satisfy the target performance. It is designed to accommodate design displacement in the first stage and large magnitude of earthquakes in the second and third stages. This design prevents superstructures from falling in the fourth stage due to large-magnitude earthquakes by increasing the capacity limit of the elastomeric isolator. In this study, the PRB isolation device is analyzed using finite element analysis to verify that the PRB isolation device works as intended and it can withstand loads corresponding to large-magnitude earthquakes. The performance of the PRB isolation device is validated by the analysis, which is further corroborated by actual experiments.

• Plant Biotechnology Reports
• /
• v.5 no.4
• /
• pp.331-344
• /
• 2011

The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.

• Korean Journal of Veterinary Service
• /
• v.39 no.1
• /
• pp.51-57
• /
• 2016

Fifty and forty two Salmonella enterica subspecies enterica serovar 52 Typhimurium (ST) strains were isolated from chicken and pigs, respectively, collected from markets throughout Korea from 2008 to 2011. The isolates were investigated for the presence of antimicrobial resistance and multi-drug patterns. All 50 ST isolates from chicken and 42 ST isolates from pigs were resistant to at least one of 13 antibiotics used in this study, 92.0% of ST isolates from chicken and 88.1% of ST isolates from pigs were resistant to three or more antimicrobials. As many as 3 isolates of ST isolates from chicken were resistant to 11 of 13 antimicrobials tested in this study. Only one isolate of ST isolates from pigs was resistant to 10 of 13 antimicrobials. The ACSSuT resistance phenotype was observed in 34% of the 50 isolates and 23.8% of the 42 isolates. Especially, 1 isolate from pigs had the ACSSuTAu. The high rate of antimicrobial-resist and multi-drug resistant (MDR) ST isolation may give rise to crucial public health problems. Therefore, control of antimicrobial use, and continuous monitoring of antimicrobial resistance and MDR patterns among Salmonella isolates in chicken and pig farms is necessary to ensure public health.

• Korean Journal of Veterinary Service
• /
• v.32 no.4
• /
• pp.325-334
• /
• 2009

Salmonella spp. are the important pathogens both economically and clinically in animals as well as human. Some of them have highly zoonotic potentials even though they are asymptomatic in animals. Therefore, the prevalence of Salmonella spp. in animals is highly concerned for human health. The present study was carried out to investigate the prevalence, antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area. The overall isolation rate of Salmonella spp. from cloaca and cecum specimens was 7.3 % (37/510). Thirty seven isolates of Salmonella spp. were identified to 5 serotypes; S. Enteritidis, S. Newport, S. Typhimurium, S. Gallinarum, and S. Derby with prevalence of 46.0%, 40.5%, 8.1%, 2.7%, and 2.7%, respectively. Resistance to nalidixic acid was found in 97.3% of Salmonella spp. isolated, followed by streptomycin (16.2%), tetracycline (16.2%), ampicillin (5.4%). Only 6 isolates (16.2%) showed resistance to more than two antimicrobials. In PFGE analysis of chicken and human isolates with Xba I, S. Enteritidis isolates from chicken showed very high similarity over 82.8% and also the similarity was very high in the comparison with human isolates. However, the higher similarity (100%) was observed among chicken isolates of S. Typhimurium. These results suggest the close genetic relatedness of Salmonella spp. isolated from chickens with human.

• Korean Journal of Veterinary Research
• /
• v.39 no.2
• /
• pp.343-352
• /
• 1999

Vancomycin resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989 the Center for Disease Control, United States, has reported a rapid increase in the incidence of enterococcal bacteremia and endocarditis infection by VRE. It was suggested that the use of avoparcin was associated with the appearance of VRE in animal husbandry. To date, several detection methods have been used based on conventional methods of culture and gene detection. However, these methods have some limitations such as time-consuming, laborious and additional differential needs. Therefore, In this study a multiplex PCR method was established to detect and differentiate resistance types of enterococci which specifically amplify the four van genes encoding vancomycin resistance elements. Using the method, we investigated the incidence rates and types of VRE from farms using or not using avoparcin. A total of 1091 animal fecal samples were collected from 70 pig and 32 poultry farms. A total of 425 of enterococci were isolated from samples. Of the 425 isolates, 11 of the them showed a pattern of high-level vancomycin resistance (MIC : $64{\sim}256{\mu}g/ml$) which was associated with the presence of the vanA or vanB gene. Fifty-seven isolates showed a pattern of low-level vancomycin resistance (MIC : $3{\sim}8{\mu}g/ml$) associated with the vanC-1 or vanC-2 gene. Interestingly, all isolates with high-level vancomycin resistance were from farms that have never used avoparcin. Moreover, the high-level VRE isolation rate in Korea (2.58%) was much lower than that of other countries (50% in England, 7% in Belgium) where avoparcin have been used. In conclusion, the multiplex PCR method established in this study could be applied for detection of VRE.

• Korean Journal of Veterinary Service
• /
• v.40 no.2
• /
• pp.101-105
• /
• 2017

The purpose of this study was to investigate prevalence and antimicrobial resistance patterns of Campylobacter spp. from duck feces in northern area of the Gyeongnam province, Korea. Samples of 121 duck feces were taken from April to December 2014 for this survey. Samples were examined by bacteria isolation and reverse transcriptase-polymerase chain reaction assay for detection of Campylobacter spp. Campylobacter were isolated in 37 samples (30.6%). Among these samples, C. jejuni and C. coli were isolated in 35 samples and 2 samples, respectively. Minimum inhibitory concentration (MIC) test is performed to investigate antimicrobial resistance patterns of Campylobacter spp. C. jejuni were resistant to ciprofloxacin (85.7%), nalidixic acid(82.9%), tetracycline (77.1%), gentamicin (57.1%), azithromycin (40.0%), clindamycin (34.3%), erythromycin (22.9%), and florfenicol (8.6%). These data support a database of pollution and antimicrobial resistance of Campylobacter spp. from duck feces and provide a basic information of reducing the secondary damage of antibiotic misuse.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.56 no.5
• /
• pp.626-633
• /
• 2023

Vibrio spp. are aquatic bacteria that are ubiquitous in warm estuarine and marine environments. Especially, V. vulnificus and V. cholerae are currently known to cause potentially fatal infections in humans. This study investigated the distribution and antibiotic resistance of V. vulnificus and V. cholerae isolated from coastal areas of Gyeongsangnam-do in 2022. A total of 252 samples of water, shellfish and coastal sediment were collected from 7 locations along the coast, and 124 samples of fishery products were collected from markets. Among the 252 samples, forty-four V. vulnificus (11.7%) and fourteen V. cholerae non-O1/non-O139 (3.7%), none of which carried the ctx gene, were isolated. Out of the 124 samples, 6 (4.8%) tested positive for V. vulnificus and V. cholerae was not detected. The isolation rates of V. vulnificus and V. cholerae showed a significant correlation with environmental factors such as seawater temperature and salinity. In an antibiotic resistance test, V. vulnificus was susceptible to amikacin, gentamicin, imipenem trimethoprim/sulfamethoxazole, and ciprofloxacin, but resistant to cefoxitin (100.0%), followed by tetracycline (9.1%). Multidrug resistance was also observed. Continuous monitoring of Vibrio pathogens with water temperature and salinity is expected to help reduce the outbreaks, and rational use of antibiotic agents is needed to prevent the accession of antibiotic-resistant microorganisms in aquatic ecosystems.