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Reducing Campylobacter jejuni colonization of poultry via vaccination.

Neal-McKinney JM, Samuelson DR, Eucker TP, Nissen MS, Crespo R, Konkel ME - PLoS ONE (2014)

Bottom Line: Seven days following challenge, chickens were necropsied and cecal contents were serially diluted and plated to determine the number of C. jejuni per gram of material.The sera from the chickens were also analyzed to determine the concentration and specificity of antibodies reactive against the C. jejuni SECPs.Vaccination of chickens with the CadF, FlaA, and FlpA peptides resulted in a reduction in the number of C. jejuni in the ceca compared to the non-vaccinated C. jejuni-challenged group.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington, United States of America.

ABSTRACT
Campylobacter jejuni is a leading bacterial cause of human gastrointestinal disease worldwide. While C. jejuni is a commensal organism in chickens, case-studies have demonstrated a link between infection with C. jejuni and the consumption of foods that have been cross-contaminated with raw or undercooked poultry. We hypothesized that vaccination of chickens with C. jejuni surface-exposed colonization proteins (SECPs) would reduce the ability of C. jejuni to colonize chickens, thereby reducing the contamination of poultry products at the retail level and potentially providing a safer food product for consumers. To test our hypothesis, we injected chickens with recombinant C. jejuni peptides from CadF, FlaA, FlpA, CmeC, and a CadF-FlaA-FlpA fusion protein. Seven days following challenge, chickens were necropsied and cecal contents were serially diluted and plated to determine the number of C. jejuni per gram of material. The sera from the chickens were also analyzed to determine the concentration and specificity of antibodies reactive against the C. jejuni SECPs. Vaccination of chickens with the CadF, FlaA, and FlpA peptides resulted in a reduction in the number of C. jejuni in the ceca compared to the non-vaccinated C. jejuni-challenged group. The greatest reduction in C. jejuni colonization was observed in chickens injected with the FlaA, FlpA, or CadF-FlaA-FlpA fusion proteins. Vaccination of chickens with different SECPs resulted in the production of C. jejuni-specific IgY antibodies. In summary, we show that the vaccination of poultry with individual C. jejuni SECPs or a combination of SECPs provides protection of chickens from C. jejuni colonization.

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Related in: MedlinePlus

Purified GST- and 6X His-tagged proteins.Purified protein extracts were separated in SDS–12.5% polyacrylamide gels and stained with Coomassie Brilliant Blue R-250. The molecular mass of the protein standards are listed in kDa.
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pone-0114254-g002: Purified GST- and 6X His-tagged proteins.Purified protein extracts were separated in SDS–12.5% polyacrylamide gels and stained with Coomassie Brilliant Blue R-250. The molecular mass of the protein standards are listed in kDa.

Mentions: SDS-PAGE analysis of the purified GST-tagged 90 mers and His-tagged full-length proteins (minus the signal peptide) for CadF, FlaA, FlpA, and CmeC is shown in Figure 2. Also shown in Figure 2 is the purified GST-tagged CadF-FlaA-FlpA trifecta protein. The Mr of the recombinant proteins is similar to the calculated molecular masses (Table S4). The CadF 90 mer peptide contains the FRLS domain, which is required for adherence to fibronectin [55]. The FlaA 90 mer peptide was chosen based on putative surface exposure, as it is located within the D2 and D3 domains of flagellin. The FlpA 90 mer peptide contains a Type III repeat found within fibronectin, as well as the residues WRPHPDFRV, which are required for binding to fibronectin [73]. The CmeC 90 mer peptide is predicted to be exposed to the extracellular environment, based on the hydrophilicity profile. The CadF-FlaA-FlpA trifecta was comprised of 30 residues from each SECP (Table S3).


Reducing Campylobacter jejuni colonization of poultry via vaccination.

Neal-McKinney JM, Samuelson DR, Eucker TP, Nissen MS, Crespo R, Konkel ME - PLoS ONE (2014)

Purified GST- and 6X His-tagged proteins.Purified protein extracts were separated in SDS–12.5% polyacrylamide gels and stained with Coomassie Brilliant Blue R-250. The molecular mass of the protein standards are listed in kDa.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256221&req=5

pone-0114254-g002: Purified GST- and 6X His-tagged proteins.Purified protein extracts were separated in SDS–12.5% polyacrylamide gels and stained with Coomassie Brilliant Blue R-250. The molecular mass of the protein standards are listed in kDa.
Mentions: SDS-PAGE analysis of the purified GST-tagged 90 mers and His-tagged full-length proteins (minus the signal peptide) for CadF, FlaA, FlpA, and CmeC is shown in Figure 2. Also shown in Figure 2 is the purified GST-tagged CadF-FlaA-FlpA trifecta protein. The Mr of the recombinant proteins is similar to the calculated molecular masses (Table S4). The CadF 90 mer peptide contains the FRLS domain, which is required for adherence to fibronectin [55]. The FlaA 90 mer peptide was chosen based on putative surface exposure, as it is located within the D2 and D3 domains of flagellin. The FlpA 90 mer peptide contains a Type III repeat found within fibronectin, as well as the residues WRPHPDFRV, which are required for binding to fibronectin [73]. The CmeC 90 mer peptide is predicted to be exposed to the extracellular environment, based on the hydrophilicity profile. The CadF-FlaA-FlpA trifecta was comprised of 30 residues from each SECP (Table S3).

Bottom Line: Seven days following challenge, chickens were necropsied and cecal contents were serially diluted and plated to determine the number of C. jejuni per gram of material.The sera from the chickens were also analyzed to determine the concentration and specificity of antibodies reactive against the C. jejuni SECPs.Vaccination of chickens with the CadF, FlaA, and FlpA peptides resulted in a reduction in the number of C. jejuni in the ceca compared to the non-vaccinated C. jejuni-challenged group.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington, United States of America.

ABSTRACT
Campylobacter jejuni is a leading bacterial cause of human gastrointestinal disease worldwide. While C. jejuni is a commensal organism in chickens, case-studies have demonstrated a link between infection with C. jejuni and the consumption of foods that have been cross-contaminated with raw or undercooked poultry. We hypothesized that vaccination of chickens with C. jejuni surface-exposed colonization proteins (SECPs) would reduce the ability of C. jejuni to colonize chickens, thereby reducing the contamination of poultry products at the retail level and potentially providing a safer food product for consumers. To test our hypothesis, we injected chickens with recombinant C. jejuni peptides from CadF, FlaA, FlpA, CmeC, and a CadF-FlaA-FlpA fusion protein. Seven days following challenge, chickens were necropsied and cecal contents were serially diluted and plated to determine the number of C. jejuni per gram of material. The sera from the chickens were also analyzed to determine the concentration and specificity of antibodies reactive against the C. jejuni SECPs. Vaccination of chickens with the CadF, FlaA, and FlpA peptides resulted in a reduction in the number of C. jejuni in the ceca compared to the non-vaccinated C. jejuni-challenged group. The greatest reduction in C. jejuni colonization was observed in chickens injected with the FlaA, FlpA, or CadF-FlaA-FlpA fusion proteins. Vaccination of chickens with different SECPs resulted in the production of C. jejuni-specific IgY antibodies. In summary, we show that the vaccination of poultry with individual C. jejuni SECPs or a combination of SECPs provides protection of chickens from C. jejuni colonization.

Show MeSH
Related in: MedlinePlus