Molecular Characterization of Anaplasma spp Isolated from Cattle Slaughtered at Central Abattoir Bauchi, Bauchi State, Nigeria

Abstract

Background: This study was conducted to molecularly characterize Anaplasma species isolated from cattle slaughtered at the Bauchi central abattoir. Bovine anaplasmosis is a disease of cattle caused by Anaplasma haemoparasite. It has significant effects on the animal’s health but has been overshadowed by other haemoparasitic diseases like Trypanosomiasis and Babebiosis. The discovery of new species with zoonotic potentials in recent years has given it more significance. Materials and Methods: Blood samples were collected from the severed jugular vein of the animals at the point of slaughter and collected samples were taken to the laboratory for analysis. DNA extraction was carried out using the Quick -DNA™ Miniprep kit using the Quick protocol. Extracted DNA was amplified with specific primers targeting the 16S rRNA (≈430 bp) genes in the Anaplasma parasite.  PCR was performed using BioinGentech Veterinary PCR kits (Concepcion, Chile) and amplified DNA fragments of the 16sRNA gene from the Anaplasma spp. isolates were directly sequenced using the Sanger method. Results: Blood samples positive for Anaplasma specie by microscopy were further examined using molecular method to confirm the isolated parasite by targeting the 16SrRNA gene. 52% were found to be positive for 16srRNA with bands corresponding to 430bp of the Anaplasma primer and the amplified genes were directly sequenced using the Sanger method. Analyzed sequences were compared by BLASTn with those present in the GenBank database and Anaplasma specie obtained from this study clustered into four groups following phylogenetic analysis.  Anaplasma species from Bauchi state clustered with other Anaplasma sequences and based on multiple alignment, the sequence presented nucleotide differences between them suggesting that different strains are circulating in the state based on the analyzed data. The DNA sequencing result of the 16SrRNA also revealed that Anaplasma platys and Anaplasma phagocytophylum are circulating among cattle herds in the state and this is of public health implication because A. phagocytophilum is zoonotic. Conclusion: Five nucleotide sequences were obtained from this study and deposited in Genbank under the following accession numbers: OQ538141 for Anaplasma platys, OQ538142 for Anaplama marginale, OQ538143 for Anaplasma phagocytophilum and OQ538144 to OQ538145 for uncultured Anaplasma species.