ERIC-PCR genotyping of field isolates of<i>Salmonella enterica</i>subsp.<i>enterica</i>serovar Gallinarum biovars Gallinarum and Pullorum

Souza, Andrei Itajahy Secundo de; Neto, Oliveiro Caetano de Freitas; Batista, Diego Felipe Alves; Estupinan, Anny Lucia del Pilar Celis; Almeida, A. Bugalho de; Barrow, Paul; Berchieri, Ângelo

doi:10.1080/03079457.2015.1086975

Cited by 11 publications

(6 citation statements)

References 20 publications

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“…Our results suggest that these bvGA strains are probably circulating in the field for a long time, since the 1850s for 287/91 and since the 1950s for BR_RS12. These results are in agreement with previous reports that demonstrated more than one pattern of S. Gallinarum bvGA spreading in Brazil by ERIC-PCR and PFGE (Secundo de Souza et al, 2015;Celis-Estupiñan et al, 2017). Noteworthy, the results presented here are based only in two Brazilian strains, what might not be representative of all genetic diversity of bvGA field strains in the country.…”

Section: Discussionsupporting

confidence: 92%

“…Moreover they clearly clustered outside the SG9 bvGA sub-clade, so they did not originated from a recent event of SG9R vaccine reversion. Another studies also found genetic differences among the field strains circulating in Brazil and the vaccine strain (Secundo de Souza et al, 2015;Celis-Estupiñan et al, 2017).…”

Section: Discussionmentioning

confidence: 89%

“…Despite PNSA, there was a new and high increase of cases between 2005 and 2016 and more than one hundred outbreaks of avian typhoid salmonelosis in layers and broilers flocks (most of them related to FT) were reported in different poultry-producing states of the country (OIE, 2016). Two main possibilities have been raised to explain the high incidence of FT in recent years: (i) virulence reversion of the live vaccine SG9R, as previously described (Van Immerseel et al, 2013); and (ii) failure in the farm biosecurity programs (Secundo de Souza et al, 2015;Celis-Estupiñan et al, 2017). Furthermore, vertical transmission of S. Gallinarum strains in the hatchery, increasing the presence in breeder flocks, could not be discarded as a possible reason to spread this bacterial pathogen in layers and broilers (Gast, 2008;Kwon et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

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Molecular and phylogenetic analyses of Salmonella Gallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms

Carli

Gräf

Kipper

et al. 2017

Veterinary Microbiology

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Fowl typhoid (FT) and pullorum disease (PD) are two important poultry infections caused by Salmonella enterica subsp. enterica serotype Gallinarum (S. Gallinarum). S. Gallinarum strains are adapted to birds and classified into biovars Gallinarum (bvGA) and Pullorum (bvPU) as they are the causative agent of FT and PD, respectively. In Brazil, FT/PD outbreaks have been reported along the last 50 years, but there was a recent increase of FT field reports with the suspicion it could be due to virulence reversion of the attenuated live vaccine SG9R. In this study, we applied molecular biology assays and phylogenetic methods to detect and investigate S. Gallinarum isolates from commercial poultry flocks in order to understand the evolutionary history and origin of the recent FT outbreaks in Brazil. S. Gallinarum isolates were obtained from thirteen different poultry flocks with clinical signs of FT/PD from 2013 to 2015. These isolates were serotyped, tested with three specific PCR (for the detection of bvGA, bvPU and live vaccine strain SG9R) and submitted to sequencing of a variable genome region (ISR analysis). The complete genome of one bvGA strain (BR_RS12) was also compared to other S. Gallinarum complete genomes (including other two Brazilian ones: bvGA 287/91 and bvPU FCVA198). PCR detected all thirteen isolates as S. Gallinarum (eight bvGA and five bvPU), none positive for SG9R strain. ISR analysis revealed that all eight bvGA isolates showed exactly the same nucleotide sequences with 100% similarity to reference strains, while two patterns were observed for bvPU. Genome phylogeny demonstrated distinct clades for bvGA and bvPU, with the bvGA clade showing a clear subdivision including three genomes: SG9R vaccine, the respective SG9 parent strain and one SG9R revertant field isolate (MB4523). The evolutionary rate of the total S. Gallinarum genome was calculated at 6.15×10 substitutions/site/year, with 2.8 observed substitutions per year per genome (1 SNP per 4292 bases). Phylodynamics analysis estimated that at least two introductions of S. Gallinarum bvGA happened in Brazil, the first in 1885 and the second in 1950. The Brazilian bvGA genomes 287/91 and BR_RS12 analyzed here were related to the early and the late introductions, respectively. In conclusion, these results indicate the occurrence of S. Gallinarum strains associated with FT outbreaks that have been circulating for more than 50 years in Brazil and are not originated from virulence reversion of the SG9R vaccine.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

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Molecular and phylogenetic analyses of Salmonella Gallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms

Carli

Gräf

Kipper

et al. 2017

Veterinary Microbiology

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“…The enterobacterial repetitive intergenic consensus (ERIC) is a repetitive sequence that is highly conserved and located in the intergenic zones; it has a variable distribution along the bacterial chromosome, separated by different lengths of intragenic sequences, which allows these primers to offer different band profiles [ 8 , 9 ]. The REP-PCR technique has also been widely applied in Salmonella studies [ 8 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of antimicrobial resistance and enterobacterial repetitive intergenic consensus-PCR as a molecular typing tool for Salmonella spp. isolated from poultry and humans

2020

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Background and Aim: Salmonella spp. are one of the most important food-borne pathogens in the world, emerging as a major public health concern. Moreover, multidrug-resistant (MDR) strains have been isolated from salmonellosis outbreaks, which compromise its treatment success. This study was conducted to characterize the phenotypic and genotypic antibiotic resistance profile of Salmonella strains isolated from broilers and humans from the regions of Tolima and Santander (Colombia). Materials and Methods: Salmonella spp. strains (n=49) were confirmed through molecular detection by amplification of the invA gene. Phenotypic antibiotic resistance was determined by the automated method and the agar diffusion method, and the presence of resistance genes was evaluated by PCR. Genotypic characterization was conducted using the enterobacterial repetitive intergenic consensus (ERIC)-PCR method, from which a dendrogram was generated and the possible phylogenetic relationships were established. Results: Salmonella isolates were classified as MDR strains exhibiting resistance to four antibiotic classes, penicillins, aminoglycosides, sulfonamides, and cephalosporins, and the human strains were resistant to gentamicin. At the genotypic level, the isolates contained the genes blaCMY2, blaCTX-M, blaPSE-1, blaTEM, aadA1, srtB, dfrA1, sul2, and floR. The genotyping results obtained by ERIC-PCR allowed the grouping of strains according to the source of isolation. Conclusion: The Salmonella spp. strains exhibited resistance to multiple antibiotics, as well as multiple genes associated with them, and the ERIC-PCR method was a technique that was helpful in generating clusters with biological significance.

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“…Molecular typing is a useful method for distinguishing among different bacterial isolates that can be used to trace the origins of pathogenic bacteria ( Clark et al, 2012 ). For instance, the enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) analysis is useful to highlight relationships among strains of Salmonella isolated from different sources ( Swanenburg et al, 1998 ; de Souza et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Detection and Characterization of Salmonella Serotypes in the Production Chain of Two Pig Farms in Buenos Aires Province, Argentina

et al. 2018

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The aim of the present study was to determine the prevalence of Salmonella in the pork production chain and to characterize Salmonella isolates. From 764 samples, 35 (4.6%) were positive for Salmonella spp., as determined by biochemical tests and the presence of the invA gene. From these, 2.6, 2.0, 8.8, and 8.0% corresponded to samples collected from farms, slaughterhouses, boning rooms and retail markets, respectively. Salmonella strains were classified into five serotypes and distributed as follows: S. Typhimurium in the pork production chain, S. Kentucky in farms and slaughterhouses, S. Brandenburg in slaughterhouses, S. Livingstone in farms and S. Agona in boning rooms and retail markets. Interestingly, the antimicrobial susceptibility testing indicated that all 35 Salmonella spp.-positive isolates were resistant to at least one antimicrobial agent, and 30 were multidrug-resistant (MDR) and resistant to different classes of antibiotics. The enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) analysis showed clonal relatedness among strains isolated from farms, boning rooms and retail markets. The presence of antibiotic-resistant Salmonella in food poses a potential health hazard to consumers.

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ERIC-PCR genotyping of field isolates ofSalmonella entericasubsp.entericaserovar Gallinarum biovars Gallinarum and Pullorum

Cited by 11 publications

References 20 publications

Molecular and phylogenetic analyses of Salmonella Gallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms

Molecular and phylogenetic analyses of Salmonella Gallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms

Molecular characterization of antimicrobial resistance and enterobacterial repetitive intergenic consensus-PCR as a molecular typing tool for Salmonella spp. isolated from poultry and humans

Detection and Characterization of Salmonella Serotypes in the Production Chain of Two Pig Farms in Buenos Aires Province, Argentina

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