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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.
Abstract: Background 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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Citations
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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.

16 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 ArticleDOI
TL;DR: It is demonstrated that using HhaI restriction endonuclease could differentiate Salmonella enteritidis andSalmonella colindale but there is similarity between pattern of SalmoneLLA typhimurium and Salmoneella infantis.
Abstract: The genus of Salmonella is very polymorphic and comprised of a number of genetically closely related serotypes. It is one of the emerging pathogen in food-borne disease which is often found in contaminated chicken eggs. Salmonella enterica is considered one of the major pathogens in public health worldwide. A total of 31 Salmonella isolates identified by specific antisera, which included Salmonella enteritidis (51.6%), Salmonella typhimurium (25.8%), Salmonella infantis (19.4%) and Salmonella colindale (3.2%). DNA was extracted using phenol- chloroform- isoamylalchol method. All the isolates showed fliC gene (1500bp) by using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. PCR- RFLP results showed 3 patterns between all isolates. Our research gained in this study demonstrated that using HhaI restriction endonuclease could differentiate Salmonella enteritidis and Salmonella colindale but there is similarity between pattern of Salmonella typhimurium and Salmonella infantis .

2 citations


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

  • ...Dilmaghani et al (2010) identified the polymorphism of fljB gene among avian in different regions by PCR-RFLP method....

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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)....

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References
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TL;DR: This book is very definitely a must for anyone who calls him or herself a microbiologist, whether clinical, medical, or technological in persuasion.
Abstract: The publication of the third edition of "Cowan and Steel," almost 20 years after the last, is like the return of a very dear, old friend. The beauty is that despite everything that has happened, the friend is at heart just the same-the mission "to help those who have isolated a bacterium (from a "medical" source) and want to identify it". The book begins with five general chapters on classification and nomenclature; culture media (with Frau Hesse still in her rightful place); isolation techniques; characterisation; and identification. I found the relative dismissal of the antibiogram as an aid to identification unsubtle, and would have liked to have more on the perils of over-reliance on kits. I believe, however, that the warning to the medical microbiologists of the future that the "materials and methods (that) are simply to be taken from the refrigerator or shelf as kits" may be a fool's paradise is timely. The essence of the book for the DIY enthusiast is to be found in chapters 6 and 7 in which all the major medical bacteria are considered in the familiar series of first, second, and third stage tables. Of course they are longer than before; there are, for example, now 26 staphylococcal species compared with eight last time. It seemed to me that a potential inconsistency which the authors challenge us to find concerns motility in Enterococcus faecium. On page 26 it is implied that many strains of E faecium are motile. Table 6.3b has Efaecium non-motile (up to 15% possibly motile according to the key to the tables), but E casseliflavus and E gallinarum motile. On page 66 they refer to E faecium "with its five varieties". What are the facts please? The authors continue a tradition of not being afraid to be different. They have not really given up much in relation to Kiebsiella (the little essay on the genus is great fun) and hang on to Acinetobacter iwoffii. Tables 7.5 and 7.6 are masterpieces of active speciation, being not quite up to date. However, they could have been a little more daring with the old Bacteroides genus, and Mobiluncus dosen't fare well. Overall, however, most of the organisms that turn up in a medical laboratory find a home here, even if only among such taxonomic evasions as "A group of difficult organisms". The Appendices are far from incidental and contain a great deal of valuable information on media and stains, characterisation tests and test organisms (all useful ammunition in current European discussions on standardisation), and on information processing. There are three sections on the bacteriological law in relation to taxonomy, and finally a useful glossary. There are almost 50 pages of references. This book is very definitely a must for anyone who calls him or herself a microbiologist, whether clinical, medical, or technological in persuasion. IAN PHILLIPS Quality Management fo: Safety, Environmental Hes No. 141. IPCS. (Pp 112; Sw Health Organization. 1 92-4-157141-1

917 citations

Journal ArticleDOI
TL;DR: It is hoped that an appreciation of the differences that exist in the way these host-adapted serotypes of Salmonella interact with the host will lead to a greater understanding of the complex host–parasite relationship that characterizes salmonella infections.
Abstract: Salmonella constitutes a genus of zoonotic bacteria of worldwide economic and health importance. The current view of salmonella taxonomy assigns the members of this genus to two species: S. enterica and S. bongori. S. enterica itself is divided into six subspecies, enterica , salamae , arizonae , diarizonae , indica , and houtenae , also known as subspecies I, II, IIIa, IIIb, IV, and VI, respectively. Members of Salmonella enterica subspecies enterica are mainly associated with warm-blooded vertebrates and are usually transmitted by ingestion of food or water contaminated by infected faeces. The pathogenicity of most of the distinct serotypes remains undefined, and even within the most common serotypes, many questions remain to be answered regarding the interactions between the organism and the infected host. Salmonellosis manifests itself in three major forms: enteritis, septicaemia, and abortion, each of which may be present singly or in combination, depending on both the serotype and the host involved. Although currently over 2300 serovars of Salmonella are recognized, only about 50 serotypes are isolated in any significant numbers as human or animal pathogens and they all belong to subspecies enterica . Of these, most cause acute gastroenteritis characterized by a short incubation period and a severe systemic disease in man or animals, characterized by septicaemia, fever and/or abortion, and such serotypes are often associated with one or few host species. It is the intention of this review to present a summary of current knowledge of these host-adapted serotypes of S. enterica . The taxonomic relationships between the serotypes will be discussed together with a comparison of the pathology and pathogenesis of the disease that they cause in their natural host(s). Since much of our knowledge on salmonellosis is based on the results of work on Typhimurium, this serotype will often be used as the baseline in discussion. It is hoped that an appreciation of the differences that exist in the way these serotypes interact with the host will lead to a greater understanding of the complex host–parasite relationship that characterizes salmonella infections.

459 citations

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