Fatal cases of bovine anaplasmosis in a herd infected with different Anaplasma marginale genotypes in southern Spain

The genus Anaplasma contains various species capable of causing disease in animals and humans. Anaplasma marginale is one of the main tick-borne pathogens of bovines in tropical and subtropical regions; however, these bacteria are now being detected more frequently in other regions of the world including Europe. In July 2017, abortions, mortality and morbidity in Retinta breed of cattle were investigated in southwestern Spain. Based on clinical signs, the provisional clinical diagnosis of bovine anaplasmosis was made. A molecular-phylogenetic approach was used to characterize A. marginale using multiple markers, including 16S rRNA, msp1a, msp4 and msp5 genes. The msp1α sequence was different from the previously described sequences from Spain as well as other countries. The isolates of A. marginale were classified as Genotype C, with two of the five tandem repeats in the amino acid sequences MSP1α being novel. The highest variability was observed in the four sequences of msp5 which was depicted in their clustering into multiple clades on a phylogenetic tree. Comparison of msp5 nucleotide sequences and the corresponding amino acid sequences revealed the co-existence of different strains in the same region. This study highlights the occurrence of clinical bovine anaplasmosis in an endemic region of Spain.The authors gratefully acknowledge all the farm keepers who contributed to this study. AMF and IGFM were funded from the Own Research Program by the University of Castilla-La Mancha.Peer reviewe