• Title/Summary/Keyword: Virulence PCR

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Expression of colSR Genes Increased in the rpf Mutants of Xanthomonas oryzae pv. oryzae KACC10859

  • Noh, Young-Hee;Kim, Sun-Young;Han, Jong-Woo;Seo, Young-Su;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.30 no.3
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    • pp.304-309
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    • 2014
  • The rpf genes and $colS_{XOO1207}/colR_{XOO1208}$ were known to require for virulence of Xanthomonas oryzae pv. oryzae (Xoo). In Xoo KACC10331 genome, two more colS/colR genes, $colS_{XOO3534}$ (raxH)/$colR_{XOO3535}$ (raxR) and $colS_{XOO3762}/colR_{XOO3763}$ were annotated. The $colS_{XOO3534}/colR_{XOO3535}$ were known to control AvrXa21 activity and functions of $colS_{XOO3762}/colR_{XOO3763}$ were unknown in Xoo. To characterize the relationship between rpf and colS/colR genes, expression of colS/colR genes in Rpf mutants of Xoo were analyzed with quantitative reverse transcription PCR (qRT-PCR). Expressions of all three colS/colR genes increased in the rpfF mutant in which DSF synthesis is defective. Expression of $colS_{XOO1207}/col-R_{XOO1208}$, $colS_{XOO3534}/colR_{XOO3535}$ and $colS_{XOO3762}/colR_{XOO3763}$ increased 2, 2-7, 3-13 folds respectively. Expression of $colS_{XOO3534}$ and $colS_{XOO3762}$ also increased 2-4 folds in the rpfG mutant in which the signal from DSF is no longer transferred to down-stream. Expression of the other colS/colR genes was not significantly changed in the rpfG mutant compared to the wild type. Since RpfF and RpfG are responsible for DSF synthesis and signal transfer from DSF to down-stream to regulate virulence gene expression, these results suggest that the DSF and DSF-mediated signal regulate negatively three colS/colR genes in Xoo.

Identification and characterization of Shiga toxin-producing Escherichia coli isolated from diarrhea in calves (송아지 설사분변으로부터 Shiga toxin-producing Escherichia coli 의 분리 및 특성규명)

  • Lim, Keum-Gi;Kang, Mun-Il;Kim, Snag-Ki;Nam, Kyung-Woo;Park, Hyun-Joo;Park, Jin- Ryang;Cho, Kyoung-Oh;Lee, Bong-Joo
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.135-142
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    • 2006
  • Shiga toxin (stx) producing Escherichia coli (STEC) causes various clinical signs in animal and human. In this study, 255 fecal samples from calves showing diarrhea were collected from cattle farms in Chonnam province during the period from January 2005 to July 2005. Twenty six STEC (10%) were isolated from 255 fecal samples by PCR. The isolates displayed three different stx combinations (stx1 [69%], stx1 and stx2 [15%], and stx2 [38%]). The isolates were further studied for virulence associated genes and antimicrobial resistance to define the virulence properties. Intimin (eaeA), enterohemolysin (hlyA), and lipopolysaccharide (rfbE) virulence genes were detected in 6 (23%), 7 (26%), and 1 (3.8%) of the isolates, respectively, by PCR. One isolate possessing rfbE gene was typed as E. coli 0157 : H7 by agglutination test with O and H antisera. All 26 isolates showed susceptibility to amikacin (100%) and the majority of isolates showed high susceptibility to gentamicin (88.5%) and chloramphenicol (73.1%). But all isolates were resistant to penicillin. These results may provide the basic knowledge to establish strategies for the treatment and prevention of enteric disease in calves.

Characterization of Extended Spectrum Beta-Lactamases (ESBL) Producing Escherichia coli Isolates from Surface Water Adjacent to Pharmaceutical Industries in Bangladesh: Antimicrobial Resistance and Virulence Pattern

  • Taslin Jahan Mou;Nasrin Akter Nupur;Farhana Haque;Md Fokhrul Islam;Md. Shahedur Rahman;Md. Amdadul Huq;Anowar Khasru Parvez
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.268-279
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    • 2023
  • The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes blaTEM, blaCTX-M-13, blaCTX-M-15, and blaSHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

Mutation in clpxoo4158 Reduces Virulence and Resistance to Oxidative Stress in Xanthomonas oryzae pv. oryzae KACC10859

  • Cho, Jung-Hee;Jeong, Kyu-Sik;Han, Jong-Woo;Kim, Woo-Jae;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.89-92
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    • 2011
  • Cyclic AMP receptor-like protein (Clp), is known to be a global transcriptional regulator for the expression of virulence factors in Xanthomonas campestris pv. campestris (Xcc). Sequence analysis showed that Xanthomonas oryzae pv. oryzae (Xoo) contains a gene that is strongly homologous to the Xcc clp. In order to determine the role of the Clp homolog in Xoo, a marker exchange mutant of $clp_{xoo4158}$ was generated. Virulence and virulence factors, such as the production of cellulase, xylanase, and extracellular polysaccharides (EPS) and swarming motility were significantly decreased in the $clp_{xoo4158}$ mutant. Moreover, the mutation caused the strain to be more sensitive to hydrogen peroxide and to over-produce siderophores. Complementation of the mutant restored the mutation-related phenotypes. Expression of $clp_{xoo4158}$, assessed by reverse-transcription realtime PCR and clp promoter activity, was significantly reduced in the rpfB, rpfF, rpfC, and rpfG mutants. These results suggest that the clp homolog, $clp_{xoo4158}$, is involved in the control of virulence and resistance against oxidative stress, and that expression of the gene is controlled by RpfC and RpfG through a diffusible signal factor (DSF) signal in Xanthomonas oryzae pv. oryzae KACC10859.

Association between Beta-lactam Antibiotic Resistance and Virulence Factors in AmpC Producing Clinical Strains of P. aeruginosa

  • Dehbashi, Sanaz;Tahmasebi, Hamed;Arabestani, Mohammad Reza
    • Osong Public Health and Research Perspectives
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    • v.9 no.6
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    • pp.325-333
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    • 2018
  • Objectives: The purpose of this study was to determine the presence of IMP and OXA genes in clinical strains of Pseudomonas aeruginosa (P. aeruginosa) that are carriers of the ampC gene. Methods: In this study, 105 clinical isolates of P. aeruginosa were collected. Antibiotic resistance patterns were determined using the disk diffusion method. The strains carrying AmpC enzymes were characterized by a combination disk method. Multiplex-PCR was used to identify resistance and virulence genes, chi-square test was used to determine the relationship between variables. Results: Among 105 isolates of P. aeruginosa, the highest antibiotic resistance was to cefotaxime and aztreonam, and the least resistance was to colictin and ceftazidime. There were 49 isolates (46.66%) that showed an AmpC phenotype. In addition, the frequencies of the resistance genes were; OXA48 gene 85.2%, OXA199, 139 3.8%, OXA23 3.8%, OXA2 66.6%, OXA10 3.8%, OXA51 85.2% and OXA58 3.8%. The IMP27 gene was detected in 9 isolates (8.57%) and the IMP3.34 was detected in 11 isolates (10.47%). Other genes detected included; lasR (17.1%), lasB (18%) and lasA (26.6%). There was a significant relationship between virulence factors and the OX and IMP genes ($p{\leq}0.05$). Conclusion: The relationship between antibiotic resistance and virulence factors observed in this study could play an important role in outbreaks associated with P. aeruginosa infections.

High Prevalence of Listeria monocytogenes in Smoked Duck: Antibiotic and Heat Resistance, Virulence, and Genetics of the Isolates

  • Park, Eunyoung;Ha, Jimyeong;Oh, Hyemin;Kim, Sejeong;Choi, Yukyung;Lee, Yewon;Kim, Yujin;Seo, Yeongeun;Kang, Joohyun;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.41 no.2
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    • pp.324-334
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    • 2021
  • This study aimed at determining the genetic and virulence characteristics of the Listeria monocytogenes from smoked ducks. L. monocytogenes was isolated by plating, and the isolated colonies were identified by PCR. All the obtained seven L. monocytogenes isolates possessed the virulence genes (inlA, inlB, plcB, and hlyA) and a 385 bp actA amplicon. The L. monocytogenes isolates (SMFM2018 SD 1-1, SMFM 2018 SD 4-1, SMFM 2018 SD 4-2, SMFM 2018 SD 5-2, SMFM 2018 SD 5-3, SMFM 2018 SD 6-2, and SMFM 2018 SD 7-1) were inoculated in tryptic soy broth (TSB) containing 0.6% yeast extract at 60℃, followed by cell counting on tryptic soy agar (TSA) containing 0.6% yeast extract at 0, 2, 5, 8, and 10 min. We identified five heat resistant isolates compared to the standard strain (L. monocytogenes ATCC13932), among which three exhibited the serotype 1/2b and D-values of 5.41, 6.48, and 6.71, respectively at 60℃. The optical densities of the cultures were regulated to a 0.5 McFarland standard to assess resistance against nine antibiotics after an incubation at 30℃ for 24 h. All isolates were penicillin G resistant, possessing the virulence genes (inlA, inlB, plcB, and hlyA) and the 385-bp actA amplicon, moreover, three isolates showed clindamycin resistance. In conclusion, this study allowed us to characterize L. monocytogenes isolates from smoked ducks, exhibiting clindamycin and penicillin G resistance, along with the 385-bp actA amplicon, representing higher invasion efficiency than the 268-bp actA, and the higher heat resistance serotype 1/2b.

Comparison of O-serogroups, Virulence Factors and Phylogenetic Groups of Uropathogenic Escherichia coli Isolated from Patients with Urinary Tract Infections between 2 Time Periods of 1989 and 2010-2014 at Gangwon Province in Korea

  • Park, Min;Kim, Seong-Mi
    • Biomedical Science Letters
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    • v.28 no.2
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    • pp.127-136
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    • 2022
  • Uropathogenic Escherichia coli (UPEC) is main causative agent of urinary tract infections. They are classified based on various types of O antigen. UPEC strains commonly possess many genes encoding virulece-associated factors. E. coli strains are generally divided into four main phylogenetic groups. The virulence factor (VF) profiles of UPEC are related with their O-serogroups in each strains. A total of 681 strains of UPEC clinical isolates were collected from Korean healthcare facility (1989: 123 strains and 2010-2014: 558 strains). The UPEC clinical isolates were analyzed by polymerase chain reaction (PCR) methods. A total of 14 O-serotypes (O1, O2, O4, O6, O7, O8, O15, O16, O18, O21, O22, O25, O75 and O83), 6 virulence factors (papC, fimG/H, sfaD/E, hly1, cnf1 and usp) and phylogenetic groups were identified. The most prevalent O-serogroups were O6 (11.1%) in 1989 UPEC strains and O25 (21.0%) in 2010-2014 UPEC strains. The identified VFs, phylogenetic groups in 1989 UPEC strains and 2010-2014 UPEC strains were fimG/H and B2 group. In this study, O6 serotype was revealed the close relationships with VFs. Also, the distribution of prevalence O-serogroups of UPEC has been changed from O6 to O25 and virulence of UPEC strains was increased during past twenty-one years.

The Relationship of the Helicobacter pylori Virulence Factor Gene Subtype in Gastric Adenocarcinoma (위선암에서 Helicobacter pylori 독성인자와 유전자 아형의 관련성)

  • Shin Jong Min;Han Sang Young;Keum Dong Joo;Kim Kwang Jin;Jee Sam Ryong;Hong Gi Bong;Lee Jong Hun;Choi Seok Ryeol;Shin Woo Won
    • Journal of Gastric Cancer
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    • v.2 no.1
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    • pp.12-19
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    • 2002
  • Purpose: The H. pylori cagA gene, vacA gene and iceA gene are considered to be important virurence factors that have been implicated in the development of gastric adenocarcinoma. It was reported that the presence of IS605 elements may be responsible for rearrangements and lead to partial or total deletions of the cag pathogenicity island (PAI) and the virulence of cag PAI may be changed. However, different results regarding the association between these virulence factors and clinical disease have been reported from different geographic regions. This study evaluated the relationship between H. pylori virulence factors such as cagA, vacA, iceA, IS605 and gastric adenocarcinoma. Materials and Methods: H. pylori isolates were obtained from 54 infected patients (24 cases of gastric adenocarcinoma, 30 cases of control). H. pylori isolates were identified by PCR with ureC gene and 16S rRNA. PCR was performed to examine cagA, vacA, iceA and IS605 genotypes. Results: Significant difference was found in the negative rates of cagA between gastric adenocarcinoma group and control ($62.5\%\;vs.\;33.3\%$ P=0.033). No significant difference was found in the prevalence of iceA, vacA between gastric adenocar cinoma and control. The genotype of cagA+ vacA s1-m1 iceA1 was predominant in H. pylori isolates irrespective of the clinical outcome. IS605 in PAI was not found in gastric adenocarcinoma gruop and control. The positive rates of IS605 in genome were $33.3\%$ in gastric adenocarcinoma group and $36.7\%$ in control (P>0.05). In gastric carcinoma, the positive rate of $cagA^{+}/IS605$ was lower than in control ($12.5\%\;vs\;40.0\%$, P=0.025) and the positive rate of cagA-/IS605 was higher than in control ($54.2\%\;vs\;23.3\%$, P=0.02). Conclusion: H. pylori virulence factors had not related significantly with gastric adenocarcinoma. Further study is needed to examine the specificity of H. pylori strains.

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Virulence genes and antimicrobial resistance of pathogenic Escherichia coli isolated from diarrheic weaned piglets in Korea

  • Do, Kyung-Hyo;Byun, Jae-Won;Lee, Wan-Kyu
    • Journal of Animal Science and Technology
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    • v.62 no.4
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    • pp.543-552
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    • 2020
  • For efficient prevention and treatment of enteric colibacillosis, understanding about latest virulence factors and antimicrobial resistance of Escherichia coli is essentially needed. The aim of this study was to survey antimicrobial resistance and determine the prevalence of fimbriae and enterotoxin genes among 118 pathogenic E. coli isolates obtained from Korean pigs with diarrhea between 2016 and 2017. The genes for the toxins and adhesins were amplified by polymerase chain reaction (PCR). The susceptibility of the E. coli isolates to antimicrobials were tested using the standard Kirby-Bauer disk diffusion method. The most prevalent fimbrial antigen was F18 (40.7%), followed by F4 (16.9%), and the most prevalent combinations of toxin genes were Stx2e (21.2%), STb:EAST-1 (19.5%), and STa:STb (16.9%), respectively. Among the pathotypes, enterotoxigenic E. coli (ETEC) was the most predominant (67.8%), followed by Shiga-toxin producing E. coli (STEC, 23.7%). We confirmed high resistance rates to chloramphenicol (88.1%), tetracycline (86.4%), streptomycin (86.4%), and ampicillin (86.4%). And the majorities of isolates (90.7%) showed multi-drug resistance which means having resistance to 3 or more subclasses of antimicrobials. Results of this study can be a source of valuable data for investigating the epidemiology of and control measures for enteric colibacillosis in Korean piggeries.

Characterization of Diarrheagenic Escherichia coli Isolated from Fresh Beef, Pork, and Chicken Meat in Korean Markets

  • Cho, Yong Sun;Koo, Min Seon;Jang, Hye Jin
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.121-128
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    • 2020
  • Diarrhea is a major public health concern associated with pathogenic Escherichia coli infections. Food-borne pathogenic E. coli can lead to large diarrheal outbreaks and hence, there is a need to estimate the frequency of pathogenic E. coli load in the various types of meat available in markets. In the present study, we classified and characterized diarrheagenic E. coli isolates collected from 399 raw meat samples from retail sources in Korea. Shiga toxin-producing E. coli (STEC) were detected in 11 (9.7%) samples, including nine strains (8.0%) in beef and two strains (1.8%) in chicken. The frequency of the detected virulence markers were as follows: astA, 28.3%; escV,18.6%; eaeA,17.7%; ent, 7.0%; EHEC-hly, 4.4%; stx1, 3.5%; and stx2, 3.5%. We did not observe any typical EPEC, EIEC, or ETEC virulence determinants in any of the samples. The STEC serotype O26 was detected in one sample, but no other serogroups (O91, O103, O128, O157, O145, O111, and O121) were found. Further research is needed to better understand the virulence mechanism of STEC serotypes, their ecology, and prevalence in animals, food, and the environment. These results will help improve risk assessment and predict the sources of food poisoning outbreaks.