Please cite this article in press as: Belkahia, H., et al., First molecular identification and genetic characterization of Anaplasma ovis in sheep from Tunisia. Small Ruminant Res. (2014), http://dx.doi.org/10.1016/j.smallrumres.2014.07.009 ARTICLE IN PRESS G Model RUMIN-4766; No. of Pages 7 Small Ruminant Research xxx (2014) xxx–xxx Contents lists available at ScienceDirect Small Ruminant Research jou r n al homep age : w w w . elsevier.com/locate/smallrumres First molecular identification and genetic characterization of Anaplasma ovis in sheep from Tunisia Hanène Belkahia a,1 , Mourad Ben Said a,b,1 , Sihem El Hamdi a , Mouna Yahiaoui a , Mohamed Gharbi b , Monia Daaloul-Jedidi a , Moez Mhadhbi b , Mohamed Jedidi b , Mohamed Aziz Darghouth b , Imen Klabi b , Lilia Zribi a , Lilia Messadi a,∗ a Laboratoire de Microbiologie, Ecole nationale de médecine vétérinaire, Université de La Manouba, 2020 Sidi Thabet, Tunisia b Laboratoire de Parasitologie, Ecole nationale de médecine vétérinaire, Université de La Manouba, 2020 Sidi Thabet, Tunisia a r t i c l e i n f o Article history: Received 31 October 2013 Received in revised form 10 July 2014 Accepted 12 July 2014 Available online xxx Keywords: Anaplasma ovis Sheep Molecular detection Loop-mediated isothermal amplification Genetic characterization msp4 gene Tunisia a b s t r a c t The aims of this study consist of first estimating the molecular prevalence of Anaplasma ovis with specific species loop-mediated isothermal amplification (LAMP) method in sheep (n = 204) from Northern and Central Tunisia. This study also characterizes the genotypes of this Anaplasma species by partial msp4 gene sequence. The molecular prevalence of A. ovis used to record 70.1%. Prevalence rate was significantly higher in El Alia locality (governorate of Bizerte, Northern Tunisia), where it indicated 91.1%, than in Sbikha locality (governorate of Kairouan, Central Tunisia), where it indicated 49.5%. In Sbikha, A. ovis prevalence varied significantly according to two factors, i.e., gender and age. In fact, ewes were significantly more infected than ram lambs. Ticks collected on animals allowed recovering 271 ixodid ticks from El Alia. Ticks belonged to 3 species of Rhipicephalus genus: first, R. turanicus (59.78%) which was the dominant species and which was followed by R. sanguineus (36.16%) and R. annulatus (4.06%). In Sbikha, an absence of ticks was noted in all of the examined sheep. The analysis of A. ovis msp4 sequences revealed four different genotypes; two of them were previously described in Italy and two new other genotypes were classified in an independent cluster. The present work is the first published report of A. ovis infection in sheep in Tunisia. The results indicate the presence of A. ovis at a high rate; hence, the need for a survey into the medical and economic impact of the disease, and the identification of vectors of this bacterium in Tunisia must be made. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Anaplasma ovis is a tick-borne obligatory intraery- throcytic bacterium of sheep, goats and wild ruminants ∗ Corresponding author at: Laboratoire de Microbiologie, Ecole nationale de médecine vétérinaire, Université de La Manouba, 2020 Sidi Thabet, Tunisia. Tel.: +216 71 552 200; fax: +216 71 552 441. E-mail address: lilia messadi@yahoo.fr (L. Messadi). 1 These authors have equally contributed to the present work. (Friedhoff, 1997; Yabsley et al., 2005; de la Fuente et al., 2006, 2007). The infection is frequently subclinical but can also cause disease which is more severe among goats than sheep, which is particularly proved in stressed or debilitated animals (Friedhoff, 1997). The acute phase of the disease is characterized by fever, progressive anemia, icterus, weight loss, milk yield decrease, and sometimes death (Splitter et al., 1956; Yasini et al., 2012). In addition, the infection with A. ovis may predispose animals to other infectious or parasitic diseases that aggravate the condition of the animal and can lead to its death (Kocan et al., 2004). http://dx.doi.org/10.1016/j.smallrumres.2014.07.009 0921-4488/© 2014 Elsevier B.V. All rights reserved.