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Journal Article

PCR-Restriction Fragment Length Polymorphism analysis of fljB gene in Salmonella enterica subspecies enterica serovar Typhimurium isolated from avians.

01 Dec 2010-Iranian journal of microbiology (Tehran University of Medical Sciences)-Vol. 2, Iss: 4, pp 178-184

TL;DR: The results of the present study showed that fljB gene is highly conserved among avians in different geographical regions, suggesting not only the importance of flJB gene in survival of organism in different environmental conditions but also the relation between proteins encoded by flj B gene and serotyping scheme.

AbstractBackground and Objectives: Economic constraint of diseases arising from Salmonella Typhimurium causes the study of this zoonotic organism more important. Most studies on identification and characterization of S. Typhimurium are conducted at DNA level. Flagellin genes (fliC and fljB genes encoding phase-1 and phase-2 flagella, respectively) are useful as a model system for studying genetic differentiation. The objectives of the present study were to identify the polymorphism of fljB among avians in different regions by the PCR-RFLP method. Materials and Methods: Fifty-two S. Typhimurium isolates out of 1,870 intestine samples were identified using culture and serotyping as well as multiplex-PCR (broiler (n = 13), layer (n = 12), duck (n = 5), goose (n = 5), sparrow (n = 8), canary (n = 3), pigeon (n = 5) and casco parrot (n = 1) ). Amplification of fljB gene was performed and amplified products subjected to restriction digestion with Hha I enzyme. Results: Two RFLP patterns generated DNA fragments between approximately 50 to 800 bps. Pattern A was observed in 33 (63.46%) and pattern B in 19 (36.54%) of isolates. Salmonella Typhimurium recovered from 13 broilers (ten with pattern A and 3 with pattern B) and 8 sparrow (three with pattern A and 5 with pattern B) showed both A and B patterns. Twelve layers, 5 pigeons and 3 canaries showed pattern A and 5 ducks, 5 geese and one casco parrot showed pattern B. None of these patterns was allotted for a special region. Conclusion: The results of the present study showed that fljB gene is highly conserved among avians in different geographical regions, suggesting not only the importance of fljB gene in survival of organism in different environmental conditions but also the relation between proteins encoded by fljB gene and serotyping scheme.

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Journal ArticleDOI
TL;DR: The results confirm the potential presence of multidrug-resistant bacteria in canary facilities, suggesting that measures to educate the public about this risk are necessary.
Abstract: The emergence of bacterial resistance to antibiotic use in veterinary practice is considered a source of multidrug-resistant bacterial infections for humans. Although increasing incidence of antimicrobial resistance in small-animal practices has already been noted, limited information is available about the problem in domestic canaries (Serinus canaria domestica). This cross-sectional study describes the prevalence of Gram-negative bacteria among canaries exhibiting clinical disease signs and the antimicrobial sensitivity patterns of the bacterial isolates. During the breeding season, a bacteriological survey was carried out in 50 amateur breeding aviaries from the Messina Province (Sicily, South of Italy) to detect the prevalence of Gram-negative bacteria in sick birds. Fecal samples from breeder canaries were submitted for bacteriological examination. Of 50 breeding aviaries, 43 (86%) were positive for Gram-negative bacteria. Overall, 88 bacterial isolates, representing 12 genera of bacteria, were cultured. The most frequently recovered bacterial species was Escherichia coli (31/88 isolates, 35.2%). Other frequently isolated species were Enterobacter cloacae (9/88 isolates, 10.2%) and Proteus mirabilis (6/88 isolates, 6.8%). Potentially pathogenic species, including Salmonella Typhimurium (n = 5 strains), Enterobacter sakazakii (n = 4 strains), and Pseudomonas aeruginosa (n = 6 strains), were also identified. The 88 isolates displayed significant frequencies of antibiotic resistance. These results confirm the potential presence of multidrug-resistant bacteria in canary facilities, suggesting that measures to educate the public about this risk are necessary.

11 citations


Journal ArticleDOI
TL;DR: The results indicate that MS-H can be used as a sensitive, rapid, and straightforward approach for the typing of Salmonella flagella at the molecular level without antiserum and phase inversion.
Abstract: Forty-three reference strains involving the 24 most common serovars of Salmonella enterica were examined by using a mass spectrometry-based H antigen typing platform (MS-H). The results indicate that MS-H can be used as a sensitive, rapid, and straightforward approach for the typing of Salmonella flagella at the molecular level without antiserum and phase inversion.

7 citations


Journal ArticleDOI
TL;DR: The results demonstrate that PCR-RFLP based on these genes showed good typeability but low discriminatory power and suggests the importance of these genes as immunization and diagnostic factors in Salmonella Typhimurium.
Abstract: Restricted fragment length polymorphism (RFLP) was used in analyses on the typing and heterogeneity, typeability and polymorphism of the 16S rRNA, fliC and fimH genes in Salmonella Typhimurium isolates of varied origin. The digestion of PCR products with restriction enzymes EcoRV, ClaI, HaeIII and ScaI (fliC genes), HincII, ClaI, EcoRV and MluI (fimH genes) and EcoRI, SmaI and HaeIII (16S rRNA genes) generated two to four bands of ranging in size from 100 to 1,104 bp. Of all the restriction profiles obtained, only the ClaI profile for fimH could be used to classify Salmonella Typhimurium isolates into different groups. According to this profile, pattern A with uncut fimH was observed in eight isolates (36.36 %) and pattern B with 755- and 253-bp bands was observed in 14 isolates (63.63 %). No pattern was allotted for a special region or source. These results demonstrate that PCR-RFLP based on these genes showed good typeability but low discriminatory power. Moreover, the highly conserved nature of fliC, fimH and 16S rRNA illustrated in our study suggests the importance of these genes as immunization and diagnostic factors in Salmonella Typhimurium. Simultaneously, our results also illustrate the potential of ClaI-based fimH analysis as a marker for the sub-serotype level differentiation of Salmonella Typhimurium isolates.

6 citations


Cites background or result from "PCR-Restriction Fragment Length Pol..."

  • ...Paradoxically, Dilmaghani et al. (2010) found that fljB gene in Salmonella Typhimurium is highly conserved....

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  • ...Our observations corroborate earlier observations of Dilmaghani et al. (2010) who showed that the fljB gene is highly conserved among Salmonella Typhimurium isolates from different geographical regions....

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  • ...No pattern was allotted for a special region or source which is similar to the observation of Dilmaghani et al. (2010) in the fljB gene....

    [...]


Journal Article

2 citations


Cites background from "PCR-Restriction Fragment Length Pol..."

  • ...mutations, deletion and insertions (Dilmaghani et al., 2010)....

    [...]

  • ...Antigenic polymorphism of flagella seems to have been generated by the accumulation of ordinary genetic events in flagellin genes, such as point mutations, deletion and insertions (Dilmaghani et al., 2010)....

    [...]


Journal ArticleDOI
TL;DR: Investigation of the prevalence and molecular epidemiology of S. enterica infections in wild Mallard ducks found that the circulation of virulent Salmonella in wild ducks of Semnan suburbs is high, and more work to assess the correlation of strains from wild life with human and livestock strains is needed.
Abstract: Wild birds serve as major reservoirs for transmission of Salmonella to domestic animals and humans. Given the zoonotic potential of salmonellosis, the main goal of this research was to investigate the prevalence and molecular epidemiology of S. enterica infections in wild Mallard ducks. Faecal samples (n=247) from wild Mallard ducks were tested for the prevalence of Salmonella spp., and genotypes of strains were then differentiated by multiplex PCR. From the 247 faecal samples, 18 (7.29%) were positive for Salmonella spp. Biochemically the most predominant serovars were S. Typhimurium and S. Enteritidis (10 and 6 cases each, respectively), whereas only 2 serovars belonged to S. Infantis. Among the 10 S. Typhimurium serovars, nine strains were positive for rfbJ, fljB, invA, and fliC genes based on multiplex PCR assay and one strain contained only the invA gene. In S. Enteritidis serovars, PCR generated amplification products for spv and sefA genes, and a random sequence in all samples. The two S. Infantis contained the random sequence specific for Salmonella genus. With respect to the circulation of virulent Salmonella in wild ducks of Semnan suburbs, more work to assess the correlation of strains from wild life with human and livestock strains is needed.

1 citations


References
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