Risks of suffering tick-borne diseases in sheep translocated to a tick infested area: A laboratory approach for the investigation of an outbreak

This study was designed to investigate an outbreak of high mortality that occurred in naïve Assaf sheep introduced into a Latxa sheep flock in the Basque Country, a region where piroplasmosis is endemic. To identify the causes of this outbreak, a panel of different methods, including traditional pathological, biopathological and parasitological analyses combined with recently developed molecular methods, was used. These novel molecular methods included a multiplex real-time PCR assay to screen for the presence of the most important tick-borne pathogens (piroplasms and anaplasmas), followed by a second species-specific multiplex real-time PCR assay for the identification of Anaplasma-positive samples. The identification of piroplasm-positive samples was carried out by a multiplexed microsphere-based suspension array using a Luminex® xMAP technology-based procedure. Anaplasmas and/or piroplasms were detected in 7/10 lambs and 11/13 ewes, with Babesia ovis being detected in 12 of the 23 animals, Theileria ovis in 6 and Anaplasma ovis in 4, both as single and mixed infections. Most of the animals infected with B. ovis had a marked decrease in the values of the red blood cell parameters. Ticks collected from the animals were identified as Riphicephalus bursa, recognised vector of B. ovis. Other haemolytic pathologies (clostridial disease, copper poisoning and leptospirosis) were ruled out and, considering all clinical, laboratory and epidemiological data, babesiosis by B. ovis was diagnosed. A detailed description of the clinical outcome, with ca. 60% of mortality, laboratory results and epidemiological findings are provided. The implications of the introduction of naïve animals into a piroplasmosis endemic area are discussed.