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Background  Buffalo flies ( Haematobia irritans exigua ) (BF) and closely related horn flies ( Haematobia irritans irritans ) (HF) are invasive haematophagous parasites with significant economic and welfare impacts on cattle production.  Wolbachia  are intracellular bacteria found widely in insects and currently of much interest for use in novel strategies for the area wide control of insect pests and insect-vectored diseases. In this paper, we report the transinfection of BF towards the development of area-wide controls.    Methods  Three stages of BF; embryos, pupae and adult female flies, were injected with different  Wolbachia  strains ( w AlbB,  w Mel and  w MelPop). The success of transinfection and infection dynamics was compared by real-time PCR and FISH and fitness effects were assessed in transinfected flies.    Results  BF eggs were not easily injected because of their tough outer chorion and embryos were frequently damaged with less than 1% hatch rate of microinjected eggs. No  Wolbachia  infection was recorded in flies successfully reared from injected eggs. Adult and pupal injection resulted in higher survival rates and somatic and germinal tissue infections, with transmission to the succeeding generations on some occasions. Investigations of infection dynamics in flies from injected pupae confirmed that  Wolbachia  were actively multiplying in somatic tissues. Ovarian infections were confirmed with  w Mel and  w MelPop in a number of instances, though not with  w AlbB. Measurement of fitness traits indicated reduced longevity, decreased and delayed adult emergence, and reduced fecundity in  Wolbachia -infected flies compared to mock-injected flies. Effects varied with the  Wolbachia  strain injected with most marked changes seen in the  w MelPop-injected flies and least severe effects seen with  w AlbB.    Conclusions  Adult and pupal injection were the most suitable methods for transinfecting BF and all three strains of  Wolbachia  successfully replicated in somatic tissues. The  Wolbachia- induced fitness effects seen in transinfected BF suggest potential for use of the  w Mel or  w MelPop strains in  Wolbachia -based biocontrol programmes for BF.

Transinfection of buffalo flies (Haematobia irritans exigua) with Wolbachia and effect on host biology