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Background  Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely  Theileria  species,  Anaplasma  species,  Candidatus Mycoplasma haemobos  and  Trypanosoma evansi , and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle.    Methods  Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of  Anaplasma  and  Mycoplasma  species, MPSP genes of  T. orientalis  and  T. sinensis , MSP4 gene of  A. marginale , 16SrRNA gene of  Candidatus Mycoplasma haemobos , and RoTat1.2 VSG gene of  Trypanosoma evansi , in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia.    Results  A total of 44 (72.13%) cattle were infected with more than one blood pathogen.  Theileria  species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83–81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98–100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of  Theileria  species and 16SrRNA gene sequences of  Anaplasma  species revealed  Theileria sinensis  and  Anaplasma platys  respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of  T. sinensis.  The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the  T. evansi and Anaplasma platys + Theileria sinensis  double species co-infected cattle group. Normocytic normochromic anaemia was observed in the  T. sinensis  infected cattle group. Significant ( p  < 0.05) increases in serum liver and kidney parameters, total protein, globulin, total and unconjugated bilirubin and decreased albumin values were observed in the  T. evansi  infected cattle when compared to clinically healthy cattle.    Conclusion  We present the first evidence of  Theileria sinensis -associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of  A. platys , there is an urgent need for appropriate preventive and control measures against these blood pathogens.

bmc veterinary research

Molecular detection of Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos, Trypanosoma evansi and first evidence of Theileria sinensis-associated bovine anaemia in crossbred Kedah-Kelantan x Brahman cattle