Sequences were aligned, edited and analysed selleckchem at the URL http://asparagin.cenargen.embrapa.br/phph/ using MEGA 4.0 software. The

identity of each sequence was confirmed by comparison with other sequences available at GenBank using BLAST software. Blood smears and PCV could be evaluated for just 12 blood samples (nine M. gouazoubira and three B. dichotomus) since the remaining nine presented haemolysis during storage in the fridge. Seven out of the 12 blood smears (58.3%) presented erythrocytes infected with protozoa in the form of small trophozoites (<2 μm). The positives blood smears were the free-living M. gouazoubira. However, the infected animals presented low parasitemia, which varied in the range 0.0125–0.200%. The mean PCV value for M. gouazoubira was 30.6% (interval 17–50%), whilst the mean value for B. dichotomus was 27%. According

to the nPCR assays, 15 (71.4%) of the 21 blood samples (13 from M. gouazoubira and two from B. dichotomus) were infected with hemoprotozoa. BLAST analysis of the amplicon sequences showed that the protozoan DNA extracted from one B. dichotomus and nine M. gouazoubira samples presented high similarity with T. cervi DNA (AY735135.1), namely, MGI12 (accession number HM466922) (99%), MGI2 (accession number HM466923), MGI5 (accession number HM466928), MGI6 (accession number HM466929), MGE1 (accession number HM466930), MGZBH1 (accession number HM466926) and BDZBH3 (accession number HM466927) (98%), MGI3 (accession number HM466923) (97%), MGI8 (accession number HM466925) (96%) and MGI11 (accession number BMN 673 ic50 HM466920) (91%). Amplicon sequences from a further three M. gouazoubira samples, namely, MGI1, MGI13 and MGI9 (accession number HM466921), exhibited 97 to 98% similarity with Theileria sp. (FJ668374.1), whilst that from M. gouazoubira sample MGE2 (accession number HM466918) presented 99% similarity with B. bigemina (EF458206.1). The amplicon

sequence from B. dichotomus Cell press sample BDZBH4 (accession number HM466919) exhibited 96% similarity with Babesia bovis (EF458215.1). Nested PCR assays of the pools of tick salivary glands showed negative although the control was positive. Although the nested PCR primers had been designed based on Babesia sequences, the sequencing from the amplified products showed that all these sequences share some homology, and by the blast search it was shown, undoubtedly, that these sequences came from different organisms. Actually, these results were serendipity founds, as we were searching for Babesia species. After the products had been identified as Theileira, the sequences between Babesia and Theileiria were aligned, showing that they present homology to the primers region. There was general concordance between the results of the nPCR assays and those of blood smears, in that the nPCR-positive samples MGI5, MGI8, MGI11, MGI12, MGE1 and MGE2 were also positive for the presence of hemoprotozoa in the blood smears. In the case of the adult female M.