Dataset Information

Molecular Detection and Phylogenetic Analysis of Riemerella anatipestifer in Poultry and Wild Geese in Poland.

ABSTRACT: Riemerella anatipestifer (RA) is one of the most relevant bacterial pathogens of commercial waterfowl from clinical and economic points of view. Our study aimed to evaluate the prevalence of RA infection in different types of commercial poultry in Poland and verify the potential role of wild geese as vectors of this pathogen. We tested a total of 126 poultry flocks, including geese (N = 20), ducks (N = 42), turkeys (N = 64) and 19 wild geese, including greater white-fronted geese (N = 9), greylag geese (N = 5) and Taiga bean geese (N = 5). Tracheal swabs were examined for RA using a PCR targeting a conserved region of the 16S rRNA gene. Selected PCR products were sequenced to perform the phylogenetic analysis. Among the commercial poultry, the highest RA prevalence was found in flocks of ducks (35.7%) and geese (30.0%), whereas the lowest one was found in turkeys (3.2%). Most tested wild geese (94.7%) were RA positive. The phylogenetic analysis showed relatively low genetic diversity of the sequences analyzed, which gathered in two clusters of the phylogenetic tree, and the minimum nucleotide identity was 98.6%. Our results would support the contention that RA isolates from commercial poultry circulate in wild bird populations but are not transmitted back to poultry.

SUBMITTER: Sawicka-Durkalec A

PROVIDER: S-EPMC9965878 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Molecular Detection and Phylogenetic Analysis of <i>Riemerella anatipestifer</i> in Poultry and Wild Geese in Poland.

Sawicka-Durkalec Anna A Tomczyk Grzegorz G Gerilovych Iryna I Kursa Olimpia O

Pathogens (Basel, Switzerland) 20230205 2

<i>Riemerella anatipestifer</i> (RA) is one of the most relevant bacterial pathogens of commercial waterfowl from clinical and economic points of view. Our study aimed to evaluate the prevalence of RA infection in different types of commercial poultry in Poland and verify the potential role of wild geese as vectors of this pathogen. We tested a total of 126 poultry flocks, including geese (N = 20), ducks (N = 42), turkeys (N = 64) and 19 wild geese, including greater white-fronted geese (N = 9), ...[more]

PMID: 36839527

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Project description:Riemerella anatipestifer is an important pathogen in waterfowl, and is generally multidrug resistant. This study assessed the current status of Riemerella anatipestifer antibiotic resistance and antibiotic-resistance genes (ARGs), compared the results of different detection methods, and evaluated a new method of studying the association between antibiotic resistance and ARGs in Riemerella anatipestifer. In this study, 51 strains of Riemerella anatipestifer were isolated from ducks on several farms, their resistance to 28 antibiotics was assessed, and the isolates were subjected to whole-genome sequencing. The number of ARGs carried by Riemerella anatipestifer was predicted, compared, and analyzed, and the consistency between ARGs and antibiotic-resistance phenotypes was assessed. The potential for loss of resistance genes during the sequencing and assembly of genome-wide framework map was assessed, and a new ARG detection method was pilot tested. The 51 strains of Riemerella anatipestifer were multidrug resistant (MDR) and had high level of resistance to aminoglycosides, trimethoprim, lincosamides, polypeptides, and macrolides. Based on the genome-wide framework map of the 51 strains, 3 local databases of ABRicate software and 1 online database of CARD website were used to detect ARGs, and a mean of 4 to 5 ARGs were identified per isolate. Although the detection results differed according to the database used, the general performance was consistent. The online website detected more types of ARGs than the ABRicate software. The association between ARGs and antibiotic-resistance phenotypes was assessed, and the ermF gene was identified as a possible key ARGs regulating macrolide resistance of Riemerella anatipestifer. The method used to investigate and detect Riemerella anatipestifer ARGs was convenient and rapid, and had strong accuracy and pertinence. The ARGs detection method reported here combined the advantages of PCR and genome detection, and could greatly reduce workload and detect ARGs more precisely.

Project description:Duck infectious serositis, also known as Riemerella anatipestifer disease, infects domestic ducks, geese, and turkeys and wild birds. However, the regulatory mechanism of its pathogenicity remains unclear. The PhoPR two-component system (TCS) was first reported in Gram-negative bacteria in our previous research and was demonstrated to be involved in virulence and gene expression. Here, DNA affinity purification sequencing (DAP-seq) was applied to further explore the regulation of PhoPR in relation to pathogenicity in R. anatipestifer. A conserved motif was identified upstream of 583 candidate target genes which were directly regulated by PhoP. To further confirm the genes which are regulated by PhoR and PhoP, single-gene-deletion strains were constructed. The results of transcriptome analysis using next-generation RNA sequencing showed 136 differentially expressed genes (DEGs) between the ΔphoP strain and the wild type (WT) and 183 DEGs between the ΔphoR strain and the WT. The candidate target genes of PhoP were further identified by combining transcriptome analysis and DAP-seq, which revealed that the main direct regulons of PhoP are located on the membrane and PhoP is involved in regulating aerotolerance. Using the in vivo duck model, the pathogenicity of ΔphoP and ΔphoR mutants was found to be significantly lower than that of the WT. Together, our findings provide insight into the direct regulation of PhoP and suggest that phoPR is essential for the pathogenicity of R. anatipestifer. The gene deletion strains are expected to be candidate live vaccine strains of R. anatipestifer which can be used as ideal genetic engineering vector strains for the expression of foreign antigens. IMPORTANCE Riemerella anatipestifer is a significant pathogen with high mortality in the poultry industry that causes acute septicemia and infectious polyserositis in ducks, chickens, geese, and other avian species. Previously, we characterized the two-component system encoded by phoPR and found that R. anatipestifer almost completely lost its pathogenicity for ducklings when phoPR was deleted. However, the mechanism of PhoPR regulation of virulence in R. anatipestifer had not been deeply explored. In this study, we utilized DAP-seq to explore the DNA-binding sites of PhoP as a response regulator in the global genome. Furthermore, phoP and phoR were deleted separately, and transcriptomics analysis of the corresponding gene deletion strains was performed. We identified a series of directly regulated genes of the PhoPR two-component system. The duckling model showed that both PhoP and PhoR are essential virulence-related factors in R. anatipestifer.

Dataset Information

Molecular Detection and Phylogenetic Analysis of Riemerella anatipestifer in Poultry and Wild Geese in Poland.

Publications

Molecular Detection and Phylogenetic Analysis of <i>Riemerella anatipestifer</i> in Poultry and Wild Geese in Poland.

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