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Rhipicephalus microplus  is an obligate hematophagous ectoparasite of cattle and an important biological vector of  Anaplasma marginale  in tropical and subtropical regions. The primary determinants for  A. marginale  transmission are infection of the tick gut, followed by infection of salivary glands. Transmission of  A. marginale  to cattle occurs via infected saliva delivered during tick feeding. Interference in colonization of either the tick gut or salivary glands can affect transmission of  A. marginale  to naïve animals. In this study, we used the tick embryonic cell line BME26 to identify genes that are modulated in response to  A. marginale  infection. Suppression-subtractive hybridization libraries (SSH) were constructed, and five up-regulated genes {glutathione S-transferase (GST), cytochrome  c  oxidase sub III (COXIII), dynein (DYN), synaptobrevin (SYN) and phosphatidylinositol-3,4,5-triphosphate 3-phosphatase (PHOS)} were selected as targets for functional  in vivo  genomic analysis. RNA interference (RNAi) was used to determine the effect of tick gene knockdown on  A. marginale  acquisition and transmission. Although RNAi consistently knocked down all individually examined tick genes in infected tick guts and salivary glands, only the group of ticks injected with dsCOXIII failed to transmit  A. marginale  to naïve calves. To our knowledge, this is the first report demonstrating that RNAi of a tick gene is associated with a failure of  A. marginale  transmission.

Knockdown of the Rhipicephalus microplus Cytochrome c Oxidase Subunit III Gene Is Associated with a Failure of Anaplasma marginale Transmission