Extraction of Saliva, Haemolymph, Salivary Glands, and Midgut from Individual Ticks (Acari: Ixodidae)

Ticks are blood-feeding ectoparasites recognised for their ability to transmit several infectious pathogens to humans and animals. Tick-borne pathogen (TBP) surveillance generally relies on detecting pathogens in homogenates of entire ticks, including those acquired during blood feeding, which may not be transmitted by the tick. To better understand the pathogen transmission mechanism, it is essential to investigate the dissemination and concentration of TBPs in various tick tissues, which is important for elucidating transmission mechanisms in ticks and determining their vector competence. This, in turn, rests on procedures for isolating saliva, haemolymph, salivary glands, and the midgut of individual ticks in order to examine the location of TBPs within these tissues and fluids. In this study, we describe a protocol for tick tissue collection, including a novel procedure for the collection of haemolymph. The results confirm that Rickettsia africae occurs at higher levels in the haemolymph of Amblyomma gemma, affirming the likely role of this tick species as a competent vector for R. africae. In contrast, Hyalomma dromedarii exhibited high rates of R. africae in the midgut, but the bacterium was absent in the haemolymph. The presence of TBPs in the haemolymph is therefore a valuable indicator of vector competence, and rests on the availability of a method that ensures ease of collection.