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Uropathogenic Escherichia coli are the major causative agent of urinary tract infection – they may simultaneously express a number of virulence factors to cause disease. The aim of this study was to investigate the relation between virulence factors content of fifteen UPEC isolates and their pathogenic potential. The isolates belonged to the five serotypes O78:K80, O114:K90, O142:K86, O164 and O157. Nine of the virulence factors have been explored, ibeA, pap, sfa/foc, cnf1, hly, fyuA, pil, ompT and traT. Virulence factors profiling of the isolates revealed a different content ranging from 22% to 100% of the virulence genes explored. The pathogenic capacity of all fifteen isolates when tested on Vero cells showed that the cytotoxicity for all tested strains on Vero cells was approximately equal and enhanced after growth in syncase broth, leading mainly to cell lysis. The toxic effects reduced slightly after heat treatment of the toxin, and greatly after formalin detoxification, but not all the deleterious effect was abolished. Endotoxin also has cytotoxic effects on Vero cells, but longer time is needed for cytolysis which is greatly diminished with formalin treatment. In conclusion, our study revealed that pathogenic strains of UPEC can exert their pathogenic effect on live cells or system with limited virulence factors gene content.
A total of 71 Aeromonas strains were isolated in the south of Jiangsu Province China in order to analyze the difference of Aeromonas spp. distribution between diseased fish and water environment. The sequence of 16S rDNA and gyrB demonstrated that the 71 Aeromonas isolates could be divided into 4 species, including A. veronii (55), A. hydrophila (11), A. salmonicida (3) and A. media (2). A. veronii was the most common species isolated from fish and water environment. All Aeromonas isolates were screened for three putative virulence genes, aer, hly and alt. hly was the most common gene among three virulence genes.
In this study, a total of 54 Vibrio alginolyticus strains were analyzed. The isolates were recovered from different compartments of the Ruditapes decussatus hatchery in the National Institute of Marine Sciences and Technologies, Monastir, Tunisia. All isolates were biochemically identified (API 20E and API ZYM strips), characterized by amplification of the Hsp-40 gene polymerase chain reaction (PCR) and analyzed by enterobacterial repetitive intergenic consensus (ERIC)-based genotyping to evaluate genetic relationship between the isolated strains. We also looked for the presence of ten V. cholera virulence genes (toxRS, toxR, toxT, toxS, tcpP, tcpA, ace, vpi, zot and ctxA) in the genomes of Vibrio isolates. The antibiotics susceptibility, exoenzymes production and in vitro cytotoxic activitiy against HeLa cell line were also carried out for all tested bacteria. Most of V. alginolyticus isolates showed significant antimicrobial resistance rates to at least ten antibacterial agents. For most isolates, the minimum inhibitory concentration (MIC) data showed that tetracyclin and streptomycin were the most effective antibiotics. Construction of the phylogenetic dendogram showed that studied isolates were in general genetically heterogeneous; however some Vibrio strains were present in different structures of the R. decussatus hatchery. The V. cholerae virulence genes investigation showed a wild distribution of toxS (49/54), toxR (45/54) and toxT (22/54) genes among V. alginolyticus strains isolated from the R. decussatus rearing system. Cytotoxic effects of several Vibrio extracellular products (28/54) were also observed on HeLa cells.
The aim of this study was to evaluate and compare two genotyping methods for the examination of genetic diversity in C. jejuni and C. coli strains isolated from poultry. Moreover, the molecular identification of putative virulence factors was additionally included to establish a correlation between the isolates. Both typing methods, RAPD-PCR and ERIC-PCR, were found to have a high discriminatory power, D=0.95 and 0.94, respectively. The isolates were clustered to 28 groups by RAPD-PCR and to 29 by ERIC- PCR. Sometimes the ERIC or RAPD assays were able to further discriminate the isolates, which had the same virulence profile generated by the second technique. These PCR-based differentiation methods demonstrated the potential for initial rapid epidemiology studies, and when used in conjunction with distribution of virulence markers provide a high resolution strategy for accurately defining subtypes of Campylobacter.
The presence of the flaA, cadF, cdtB and iam genes of Campylobacter spp. was determined with the PCR method. The materials to investigate were 56 C. jejuni and 23 C. coli strains isolated from clinical samples (children and domestic animals). It was found that all of the Campylobacter spp. isolates from children with diarrhoea and domestic animals had cadF gene, responsible for adherence. The flaA gene was present in all Campylobacter spp. isolates derived from children and cats. Occurrence of flaA gene was confirmed in 100% of C. jejuni strains obtained from dogs. The high prevalence of the cdtB gene associated with toxin production was observed in this study (100%-Campylobacter spp. isolates obtained from dogs and cats, 97.9%-Campylobacter spp. isolates from children). The isolates showed a wide variation for the presence of iam gene. The lowest prevalence (23.5%) was detected in Campylobacter spp. obtained from dogs. The highest rates of iam detection (91.6%) were revealed in C. coli isolates from children.
Commensal, intestinal E.coli microflora plays a role in maintenance of intestinal balance of the host, is responsible for defending against pathogenic E.coli. This study encompasses the analysis of BOX-PCR fingerprinting patterns, phylogenetic grouping and virulence genes prevalence among commensal E.coli isolates derived from healthy pigs. Altogether, 274 unique E.coli isolates were identified, 110 from weaned piglets (Piglets I and Piglets II) and 164 from adult sows (Sows I and Sows II). BOX-PCR analysis distinguished isolates from pigs in different age and indicated that during maturation the changes in E.coli microflora occurred. Phylogenetic grouping revealed significant differences between distribution of four phylogenetic groups among isolates derived from piglets and sows. In phylogenetic structure of isolates from the piglets group B1 prevailed significantly, while among isolates derived from the sows the majority of them were classified into phylogenetic group A. The identification of 17 virulence factors in E.coli isolates derived from healthy pigs was performed. Three of 13 intestinal (escV, ehxA, estII) and four extra-intestinal virulence genes (VGs) (hlyA, fimH, papA, sfaS) were detected in the porcine isolates. The percentage of VGs positive isolates among piglets is higher than among sows, moreover, the VGs occurring in E.coli isolates from piglets revealed greater diversity than that detected among isolates from sows.
The genetic features of each isolate were determined by enterobacterial repetitive intergenic consensus (ERIC) primer sequences used in PCR and by searching for six virulence genes (alg D, las B, tox A, plc H,plc N, exo S). 49 (79%) of the isolates were distributed in three ERIC PCR subgroups and showed 62% of similarity. The remaining 13 strains generated unique patterns. The first subgroup was primarily composed of isolates from faeces, these strains indicated over 70% relationship with the next subgroup, and primarily contained strains isolated from wounds and bronchial washings and the last subgroup contained strains isolated from wounds and urine. The unique strains were isolated mainly from urine. Statistical analysis indicated that variations in distribution of virulence genes in P. aeruginosa isolates with respect to strain origin and genomic subgroups were not significant. In the group of 49 strains, 100% gave a positive reaction to alg D, las B and plc H genes, 91.8% to tox A and plc N genes and 83.7% to exo S gene. Among the strains that generated unique (ERIC-PCR) patterns, 69.2% gave a positive reaction to alg D gene, 84.6% to las B gene, 76.9% to tox A, plc N and plc H genes, and 46.15% to exo S gene.
The 150 Y. enterocolitica strains isolated from humans and from pigs belonged to biotypes 4 (68.7%), 1A (18.7%) and 2 (4%), or were biochemically untypeable (8.6%). Biotype 4 was comprised of Y. enterocolitica strains representing serotype 0:3, within biotype 1A the strains either belonged to serotypes 0:5 and 0:6 or were untypeable, and biotype 2 was represented by the strains of serotype 0:9. The strains which were biochemically untypeable belonged to serotypes 0:5, 0:6 and 0:3. Among the strains tested there also were those of an unidentified biotype and serotype. Nearly all the strains of biotype 1A represented genotype ystB+myfA+, and few belonged to genotype ystB+. The presence of the ystB gene in the strains of biotype 1A and only occasional occurrence of the gene in the other biotypes makes ystB a distinguishing marker of biotype 1A. The strains of genotype ystA+ail+myfA+yadA+ predominated in biotype 4 (serotype 0:3). The strains of biotype 2 (serotype 0:9) represented genotype ystA+ail+myfA+, and the plasmid yadA gene was detected in some of them. Within the group of biochemically untypeable strains ystB- and myfA-specific PCR products were mainly obtained. The genotypes determined for the tested biotypes and serotypes of Y. enterocolitica, based upon the selected genes of virulence, can be applied as distinguishing markers and indicators of the potential virulence of Y. enterocolitica strains, excluding bioserotyping.
The association between Escherichia coli virulence factors and chronic intestinal disorders is mostly unknown. The presented study compared the distribution of virulence genes and phylogroups among E. coli isolated from chronic intestinal disorders such as Crohn’s disease and irritable bowel syndrome (IBS) with strains isolated from patients with acute diarrhea as a control group. The presence of 159 virulence genes corresponding to known E. coli pathotypes was determined among 78 E. coli archive strains isolated from IBS, acute diarrhea and Crohn’s disease using CGH microarray. E. coli isolated from IBS demonstrated a mosaic of virulence genes specific to enteropathogenic, enterotoxigenic, enterohemorrhagic E. coli strains and Shigella species. In contrast, virulence factors and phylogroups distribution among E. coli isolated from children with acute diarrhea was similar to extraintestinal E. coli strains that probably acquired some virulence genes. The acquisition of virulence genes might have an impact on diarrheagenic potential of these strains. On the other hand, E. coli isolated from children with Crohn’s disease seem to be similar to adherent-invasive E. coli strains (AIEC), as it lack most known virulence genes. The presented study showed that these analyzed groups of E. coli strains differed from each other with the respect to the distribution of virulence genes. The differences in gene content support the idea that the participation of E. coli in chronic intestinal diseases is mostly related to virulence potential of these strains.
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