Consecutive artificial membrane feeding across all life stages of Rhipicephalus australis

Ticks are haematophagous ectoparasites of major veterinary and public health significance. Traditionally, tick rearing has relied on live animal hosts for studying tick biology, pathogen transmission and the discovery of drugs and vaccines but increasing concerns regarding animal welfare have driven the development and refinement of artificial tick feeding systems (ATFS). Here, we demonstrate the first successful consecutive artificial feeding of larvae, nymphs and adults of Rhipicephalus australis using an optimised silicone membrane system. Larval attachment and engorgement rates were 61.8 ± 20.4% (range 30.3-95.1%) and 40.1 ± 12.4% (24.0-61.0%), respectively. Nymphs exhibited the highest feeding success, with attachment and engorgement rates of 88.9 ± 8.5% (69.2-98.2%) and 91.8 ± 9.5% (66.7-100.0%), respectively, demonstrating the viability and fitness of nymphs that had moulted from artificially fed larvae. Adult ticks successfully attached (83.7 ± 5.9%; 76.9-87.5%) and engorged (89.1 ± 4.2%; 84.6-92.8%) and yielded viable engorged females. However, oviposition was not observed, and antimicrobial supplementation was required to control microbial contamination. Therefore, this system represents an important initial step towards controlled feeding across all stages of R. australis. Further optimisation, particularly to improve reproductive performance and reduce reliance on antimicrobials, will be required before its broader application to tick-pathogen interaction and vector competence studies.