Molecular characterization of Anaplasma marginale based on the msp1a and msp1b genes.

Anaplasma marginale is an intracellular rickettsial bacterium causing anaplasmosis in ruminants. A. marginale is transmitted biologically by ticks and mechanically by blood-sucking vectors. Anaplasmosis occurs in tropical and subtropical areas of the world. This disease causes huge economic losses due to decreasing meat yield and milk production. The aims of this study were to determine the genetic diversity and antigenicity of A. marginale based on the msp1a and msp1b genes in cattle in Thailand. The A. marginale msp1a and msp1b genes were amplified by the polymerase chain reaction (PCR). There have been four copies of MSP1a tandem repeats among A. marginale Thailand strain, and thirteen different MSP1a tandem repeats were found including repeats B, 25, 27, M, 3, S, C, H, β, 80, 4, TH1 and TH2. Notably, this study showed two copies of the novel conserved tandem sequences namely Thailand Type 1 (TH1) and Type 2 (TH2). The phylogenetic analysis revealed that A. marginale msp1a and msp1b genes were genetically diverse and showed 9 and 5 clades with similarity ranging from 98 to 100% and 79.5 to 100%, respectively, when compared within the isolates of this study. The results of diversity analysis showed 18 and 16 haplotypes of the msp1a and msp1b genes, respectively. The entropy analyses of msp1a and msp1b nucleic acid sequences showed 39 and 900 high entropy peaks with values ranging from 0.35 to 0.85 and from 0.41 to 1.48, respectively, while those of MSP1a and MSP1b amino acid sequences exhibited 75 and 72 high entropy peaks with values ranging from 0.35 to 1.06 and from 0.41 to 1.55, respectively. In addition, B-cell and T-cell epitopes have also been investigated in this study. Hence, our results could be employed to improve the insight input of molecular phylogenetics, genetic diversity and antigenicity of A. marginale Thailand strain.