Identification of a Tick Midgut Protein Involved in Babesia bovis Infection of Female Rhipicephalus microplus Ticks

Rhipicephalus microplus is an important biological vector as it transmits several pathogens, including Babesia bovis, the causative agent of bovine babesiosis. The available strategies for controlling B. bovis are limited, resulting in substantial challenges for both animal health and livestock management. Infection of the tick midgut is the essential first step for the transmission cycle of B. bovis, yet this process remains largely unexamined. To better understand the first step of tick infection, this study employed a proteomic approach to identify a midgut protein that responds to B. bovis infection. We then used RNA interference for gene silencing to determine if the protein is essential for R. microplus infection. The protein we identified, Rm24, is twofold upregulated in the tick midgut during B. bovis infection. We silenced the gene encoding Rm24 and examined the effect of reduced expression on both tick fitness and B. bovis infection. Our results indicated that silencing the Rm24 gene impacted the survivability of adult female ticks, which exhibited a significant reduction in viability as compared to the control and non-injected groups. Importantly, we found that suppressing the gene encoding Rm24 led to a significant decrease in the number of engorged female ticks infected, with only 15% of female ticks testing positive for B. bovis kinetes as compared to over 50% in the control groups. We also detected a significant reduction in vertical transmission of B. bovis to larval progenies. These findings suggest that the Rm24 protein is critical for infection by B. bovis and could serve as a promising target for future transmission-blocking strategies.