Vaccine efficacy of recombinant BmVDAC on Rhipicephalus microplus fed on Babesia bigemina-infected and uninfected cattle.

Rhipicephalus microplus is the most widely distributed tick worldwide and causes significant economic losses in the livestock industry. It directly affects hosts (especially in large infestations) by feeding on blood and piercing the skin and indirectly affects hosts as a vector of pathogens that cause infectious diseases, such as bovine babesiosis. Current research on the control of ticks is focused on integrated tick control programmes, including vaccination treatment with acaricides and completely blocking pathogen transmission. Our previous studies showed that R. microplus VDAC (BmVDAC) expression is modulated by Babesia bigemina infection. VDAC is a mitochondrial protein with multiple functions in addition to its primary role as a central component of the apoptotic machinery. In this paper, we evaluated BmVDAC as an anti-tick vaccine and its capacity to block the infection of Babesia bigemina in ticks. Our results demonstrate that rBmVDAC is immunogenic and that antibodies specifically recognize the native protein from midguts of R. microplus. Immunization with rBmVDAC afforded an 82% efficacy against R. microplus infestation in the group of vaccinated cattle compared with the control group. In contrast, rBmVDAC showed a lower efficacy of 34% against tick infestation in cattle vaccinated with rBmVDAC, infested with R. microplus and infected with B. bigemina. The main effect on ticks fed in vaccinated and infected cattle was a 34% reduction in egg fertility (DF) compared to ticks fed on the control group. There was no reduction in the B. bigemina parasite levels of ticks fed on rBmVDAC-vaccinated cattle. These results suggest that the rBmVDAC protein could be tested as a vaccine for the control of tick infestation.

Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation.

BACKGROUND: Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane fusion in fertilization processes. The identification and characterization of HAP-2 protein in Babesia would be very significant to understand the biology of the parasite and to develop a transmission-blocking vaccine in the future. RESULTS: To isolate and sequence the hap2 gene DNA from an infected bovine with Babesia bigemina was purified. The hap2 gene was amplified, cloned and sequenced. The sequences of hap2 from four geographically different strains showed high conservation at the amino acid level, including the typical structure with a signal peptide and the HAP2/GSC domain. Antisera anti-HAP2 against the conserved extracellular region of the HAP2 amino acid sequence were obtained from rabbits. The expression of hap2 in the host and vector tissues was analyzed by using semi-quantitative RT-PCR, and the protein was examined by western blot and immunofluorescence. Based on the RT-PCR and WB results, HAP2 is expressed in both, sexual stages induced in vitro, and in infected ticks as well. We did not detect any expression in asexual erythrocytic stages of B. bigemina, relevantly anti-HAP2 specific antibodies were able to block zygotes formation in vitro. CONCLUSION: Babesia bigemina HAP2 is expressed only in tick-infecting stages, and specific antibodies block zygote formation. Further studies regarding the function of HAP2 during tick infection may provide new insights into the molecular mechanisms of sexual reproduction of the parasite.