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Bovine leukemia virus (BLV) is the agent responsible for enzootic bovine leukosis, the most common neoplastic disease in cattle. The horn fly, a major hematophagous pest of cattle, is able to transmit different diseases in cattle. However, its implication in BLV transmission under a natural environment is still discussed. The objectives of this work were to determine the presence of BLV in horn flies (by sequencing) and to evaluate the ability of horn flies to transmit BLV to cattle (through an experimental assay under a natural environment). To demonstrate the presence of BLV in the flies, 40 horn flies were collected from a BLV-positive cow with a sweep net and 10 pools with four horn-fly mouthparts each were prepared. The presence of BLV was determined by nested polymerase chain reaction and sequencing. To demonstrate BLV transmission, other 40 flies were collected from the same BLV-positive cow with a sweep net. Eight homogenates containing five horn-fly mouthparts each were prepared and injected to eight cows of different breeds, and blood samples were collected every 21 days. Then, to evaluate the ability of horn flies to transmit BLV to grazing cattle under natural conditions, both infected and uninfected cattle from the experimental transmission assay were kept together in the same paddock with more than 200 horn flies per animal for 120 days. Blood samples were collected every 20 days and the number of flies was determined. The sequencing results confirmed the presence of the provirus in horn flies. The results also confirmed that BLV transmission is a possible event, at least experimentally. However, the role of horn flies as vectors of BLV under a natural grazing system is still discussed.

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