ORIGINAL ARTICLE High Prevalence of Anaplasma spp. in Small Ruminants in Morocco H. Ait Lbacha 1 , S. Alali 1 , Z. Zouagui 1 , L. El Mamoun 1 , A. Rhalem 1 , E.Petit 2 , N. Haddad 2 , C. Gandoin 2 , H-J. Boulouis 2 and R. Maillard 2,3 1 Institut Agronomique et V et erinaire Hassan II, Rabat, Morocco 2 Anses, ENVA, INRA, BIPAR, Maisons-Alfort, France 3 INP-ENVT, Toulouse, France Keywords: Anaplasma; co-infection; 16S rRNA; msp2; ruminants; Morocco Correspondence: R. Maillard. unit e de pathologie des ruminants,  Ecole Nationale V et erinaire de Toulouse, 23 chemin des capelles, F-31076 Toulouse, France. Tel.: +33 5 61 1 9 23 16; Fax: +33 5 61 19 38 34; E-mail: r.maillard@envt.fr This work was carried out at: ‘Institut Agronomique et V et erinaire Hassan II’, ‘  Ecole Nationale V et erinaire d’Alfort-BIPAR’ (ENVA) and ‘  Ecole Nationale V et erinaire de Toulouse’ (INP-ENVT). Received for publication November 13, 2014 doi:10.1111/tbed.12366 Summary The prevalence of infection by Anaplasma spp. (including Anaplasma phagocyto- philum) was determined using blood smear microscopy and PCR through screen- ing of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Indi- vidual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were col- lected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocy- tophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic mea- sures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemo- parasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases. Introduction Anaplasmosis refers to the disease caused by bacteria of the genus Anaplasma. Anaplasma spp. are gram-negative bacte- ria of the family Anaplasmataceae, the Rickettsiales order and the alpha-Proteobacteria class. They have a worldwide distribution (Kocan et al., 2004; Rar and Golovljova, 2011). Small ruminants host several Anaplasma species, including Anaplasma ovis, Anaplasma marginale and the zoonotic agent Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) causing human granulocytic anaplasmosis (HGA) (Dumler et al., 2001; Torina et al., 2010, 2012; Alessandra and Santo, 2012; Chi et al., 2013). Although less frequently reported in humans, A. ovis also may be a zoo- notic agent (Psaroulaki et al., 2008; Chochlakis et al., 2009b, 2010). Anaplasma phagocytophilum has been known for a long time in veterinary medicine: A. phagocytophilum infection © 2015 Blackwell Verlag GmbH • Transboundary and Emerging Diseases. 1 Transboundary and Emerging Diseases