In vivo evaluation of a cytochrome P450 gene from poultry red mite, Dermanyssus gallinae, as a vaccine antigen for chicks.

Abstract

BACKGROUND: Dermanyssus gallinae is a prevalent ectoparasite in the poultry farms, inflicting damage on&#xa0;chicken health through blood-sucking.&#xa0;Chemical acaricides commonly used for&#xa0;mite control&#xa0;often show reduced efficacy due to the development of resistance.&#xa0;Therefore, alternative control methods are needed, and vaccination is a promising strategy for controlling D. gallinae. METHODS: The mRNA expression of Deg-CYP-3 in mites at various developmental stages, as well as under fed and starved conditions, was analyzed. Subsequently, recombinant protein rCYP-3 was induced, purified, and employed for immunization. Following immunization, antibodies were analyzed and mite challenge was then conducted. Following a 12&#xa0;h period of blood-feeding on chicks, the mites were collected to evaluate the acaricidal efficacy of the rCYP-3 vaccine. RESULTS: The Deg-CYP-3 gene was expressed across all life stages and maintained stable expression levels under both fed and starved conditions. The recombinant protein rCYP-3 was successfully expressed in Escherichia coli and efficiently secreted into the culture supernatant. Immunization with rCYP-3 induced a specific IgY immune response in chicks, as confirmed by ELISA. Moreover, anti-rCYP-3 serum specifically recognized P450 proteins extracted from D. gallinae, as demonstrated by Western blot analysis. Immunization resulted in an 8.1% reduction in adult mite survival (P&#x2009;>&#x2009;0.05), whereas nymph survival decreased significantly by 22.4% (P&#x2009;<&#x2009;0.01). In addition, oviposition rate, hatching rate, and fecundity were reduced by 2.8%, 2.2%, and 22.0%, respectively, in the immunized group. Overall, vaccine efficacy was calculated to be 30.6% in immunized birds. Furthermore, the expression level of Deg-CYP-3 in mites fed on immunized hosts was significantly lower than that in mites from the unimmunized control group. CONCLUSIONS: Our findings demonstrated that the Deg-CYP-3 gene exhibits high transcriptional activity during both the adult and nymph stages of D. gallinae. Moreover, its expression remains consistent regardless of the feeding status of adult mites. Immunization with rCYP-3 effectively reduced mite survival, reproductive capacity, and gene expression levels, demonstrating its potential as a preventive and control strategy against D. gallinae.