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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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Vaccination with CadF, FlaA, FlpA, and trifecta peptides reduces C. jejuni colonization.Panel A: The number of CFU of C. jejuni was determined by serially plating diluted cecal contents on Campy-Cefex plates. The median level of colonization and interquartile range are shown for each group. The number of birds colonized in each group is shown below the graph. NI = no injection. ND = not detected; limit of detection is 103 CFU/gram of cecal contents. Panel B: Sera was isolated from blood collected from the chickens at the time of necropsy and examined via ELISA using microtiter plates coated with a C. jejuni whole-cell lysate. Chicken IgY bound to the coated wells was detected with a rabbit anti-chicken IgY antibody conjugated to HRP. 95% confidence intervals were calculated based on the level of sera reactivity in the non-vaccinated, unchallenged group (solid line) and non-vaccinated, non-challenged groups of chickens (dashed line).
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pone-0114254-g004: Vaccination with CadF, FlaA, FlpA, and trifecta peptides reduces C. jejuni colonization.Panel A: The number of CFU of C. jejuni was determined by serially plating diluted cecal contents on Campy-Cefex plates. The median level of colonization and interquartile range are shown for each group. The number of birds colonized in each group is shown below the graph. NI = no injection. ND = not detected; limit of detection is 103 CFU/gram of cecal contents. Panel B: Sera was isolated from blood collected from the chickens at the time of necropsy and examined via ELISA using microtiter plates coated with a C. jejuni whole-cell lysate. Chicken IgY bound to the coated wells was detected with a rabbit anti-chicken IgY antibody conjugated to HRP. 95% confidence intervals were calculated based on the level of sera reactivity in the non-vaccinated, unchallenged group (solid line) and non-vaccinated, non-challenged groups of chickens (dashed line).

Mentions: The median level of colonization in the C. jejuni-treated, non-vaccinated control group was 5.35×107 CFU/gram (Figure 4, Panel A). The trifecta vaccination resulted in a significant reduction in C. jejuni colonization, as only seven of nine birds were colonized, and the median level of colonization was reduced to 4×104 CFU/gram of cecal content. The FlaA and FlpA vaccinated groups also had a reduction in the number of birds colonized, with eleven of twelve and nine of ten birds colonized, respectively. The median levels of colonization for the FlaA and FlpA vaccinated groups were 2.55×104 and 4.5×104 CFU/gram, respectively. Interestingly, the CadF vaccinated groups did not have any uncolonized birds, although the median level of colonization was reduced to 1.1×106 CFU/gram. The CmeC vaccinated group displayed the greatest range in the level of colonization observed, and only a modest reduction in the median level of colonization. Similar results were generated in a pilot study (Figures S1 and S2).


Reducing Campylobacter jejuni colonization of poultry via vaccination.

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

Vaccination with CadF, FlaA, FlpA, and trifecta peptides reduces C. jejuni colonization.Panel A: The number of CFU of C. jejuni was determined by serially plating diluted cecal contents on Campy-Cefex plates. The median level of colonization and interquartile range are shown for each group. The number of birds colonized in each group is shown below the graph. NI = no injection. ND = not detected; limit of detection is 103 CFU/gram of cecal contents. Panel B: Sera was isolated from blood collected from the chickens at the time of necropsy and examined via ELISA using microtiter plates coated with a C. jejuni whole-cell lysate. Chicken IgY bound to the coated wells was detected with a rabbit anti-chicken IgY antibody conjugated to HRP. 95% confidence intervals were calculated based on the level of sera reactivity in the non-vaccinated, unchallenged group (solid line) and non-vaccinated, non-challenged groups of chickens (dashed line).
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Related In: Results  -  Collection

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Show All Figures
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pone-0114254-g004: Vaccination with CadF, FlaA, FlpA, and trifecta peptides reduces C. jejuni colonization.Panel A: The number of CFU of C. jejuni was determined by serially plating diluted cecal contents on Campy-Cefex plates. The median level of colonization and interquartile range are shown for each group. The number of birds colonized in each group is shown below the graph. NI = no injection. ND = not detected; limit of detection is 103 CFU/gram of cecal contents. Panel B: Sera was isolated from blood collected from the chickens at the time of necropsy and examined via ELISA using microtiter plates coated with a C. jejuni whole-cell lysate. Chicken IgY bound to the coated wells was detected with a rabbit anti-chicken IgY antibody conjugated to HRP. 95% confidence intervals were calculated based on the level of sera reactivity in the non-vaccinated, unchallenged group (solid line) and non-vaccinated, non-challenged groups of chickens (dashed line).
Mentions: The median level of colonization in the C. jejuni-treated, non-vaccinated control group was 5.35×107 CFU/gram (Figure 4, Panel A). The trifecta vaccination resulted in a significant reduction in C. jejuni colonization, as only seven of nine birds were colonized, and the median level of colonization was reduced to 4×104 CFU/gram of cecal content. The FlaA and FlpA vaccinated groups also had a reduction in the number of birds colonized, with eleven of twelve and nine of ten birds colonized, respectively. The median levels of colonization for the FlaA and FlpA vaccinated groups were 2.55×104 and 4.5×104 CFU/gram, respectively. Interestingly, the CadF vaccinated groups did not have any uncolonized birds, although the median level of colonization was reduced to 1.1×106 CFU/gram. The CmeC vaccinated group displayed the greatest range in the level of colonization observed, and only a modest reduction in the median level of colonization. Similar results were generated in a pilot study (Figures S1 and S2).

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