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Background   Anaplasma marginale  is an obligate intracellular bacterium and the main cause of bovine anaplasmosis in tropical and subtropical regions. In Egypt, data regarding the prevalence of  A. marginale  in ruminant hosts and of the circulating genotypes is lacking. This study therefore aimed to (i) investigate the presence, epidemiology and genotypes of  A. marginale  in cattle and buffaloes in Egypt, (ii) to evaluate suitable diagnostic tools and (iii) to identify co-infections of  A. marginale  with other selected tick-borne pathogens.    Methods  Blood samples were collected from 394 animals (309 cattle and 85 buffaloes) from three different areas in Egypt. For the detection of  A. marginale  infection, several tests were compared for their sensitivity and specificity: blood smear analysis, enzyme-linked immunosorbent assay (ELISA), PCR, real-time PCR and reverse line blot (RLB) assay. Co-infections with  A. marginale,  piroplasms and other  Anaplasmataceae  were surveyed by RLB while  A. marginale  genotypes were identified by amplifying and sequencing the partial  msp1α  gene.    Results   Anaplasma marginale  DNA was amplified by qPCR in 68.3% of cattle and 29.4% of buffaloes. RLB showed infection with  A. marginale  in 50.2% of cattle and 42.5% of buffaloes. Blood smear analysis detected this agent in 16.2% of cattle and 2.4% of buffaloes. ELISA showed specific antibodies against  A. marginale  in 54.9% of cattle.  Anaplasma marginale  was associated, in cattle and buffaloes, with several tick-borne pathogens ( Theileria annulata ,  Babesia bovis ,  Babesia bigemina ,  Babesia occultans  and  Anaplasma platys ). A significant difference of  A. marginale  infection level was noticed in cattle, where animals between 3–5-years-old had a higher prevalence (79.2%) compared to those older than 5 years (36.4%) and younger than 3 years (59.7%) and one year (64.5%), respectively ( P  = 0.002281). Microsatellite analysis identified 15 different genotypes.    Conclusions  The epidemiological findings revealed high prevalence of  A. marginale  in cattle and buffaloes in all the investigated areas. The circulation of diverse genotypes was observed, most of these  A. marginale  genotypes being specific for Egypt. The qPCR assay was confirmed to be the most sensitive tool for detection of  A. marginale  in cattle and buffaloes even in the carrier state, highlighting the importance of using suitable diagnostic tests.

Epidemiology and genotyping of Anaplasma marginale and co-infection with piroplasms and other Anaplasmataceae in cattle and buffaloes from Egypt