Ticks and Tick-borne Diseases 3 (2012) 379–380 Contents lists available at SciVerse ScienceDirect Ticks and Tick-borne Diseases jo u rn al hom epage: www.elsevier.com/locate/ttbdis Short communication Spotted fever group rickettsiae identified in Dermacentor marginatus and Ixodes ricinus ticks in Algeria Tahar Kernif a,b , Dalila Messaoudene a , Soraya Ouahioune a , Philippe Parola b , Didier Raoult b , Idir Bitam a,* a Service d’Ecologie et des Systèmes Vectoriels, Institut Pasteur d’Algérie, Algiers, Algeria b Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198, Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod Borne Bacterial Diseases, Faculté de Médecine, 27 bd Jean Moulin, 13385 Marseille cedex 5, France a r t i c l e i n f o Keywords: Ixodes ricinus ticks Dermacentor marginatus ticks Rickettsia helvetica R. monacensis R. slovaca Algeria a b s t r a c t Our study was carried out using Ixodes ricinus ticks collected from cattle from Tizi-Ouzou and Dermacentor marginatus ticks collected from the vegetation of the Blida region, a tourist site, both regions situated in northern Algeria. The results of real-time quantitative PCR (qPCR) specific for a partial sequence of the citrate synthase gene (gltA) indicate that Rickettsia spp. were present in 11/23 (48%) and 4/9 (44%) of the examined ticks from Tizi-Ouzou and Blida, respectively. The sequences of Rickettsia helvetica and Ri. monacensis were found in I. ricinus ticks using gltA primers. In addition, Ri. slovaca was detected based on the sequences of the gltA and the outer membrane protein (OmpA) genes in D. marginatus ticks. DNA sequencing to identify the species revealed for the first time the presence of Ri. helvetica in I. ricinus ticks and Ri. slovaca in D. marginatus ticks from Algeria and confirmed the presence of Ri. monacensis. © 2012 Elsevier GmbH. All rights reserved. Introduction Spotted fever group (SFG) Rickettsia spp. is important emerg- ing tick-borne human infection agents that are present worldwide (Parola et al., 2005). In Algeria, several Rickettsia spp. that are pathogenic for humans have been detected in various ticks, includ- ing Rickettsia conorii conorii, the causative agent of Mediterranean spotted fever, which is transmitted by Rhipicephalus sanguineus (Mouffok et al., 2009); Ri. aeschlimannii, an emerging pathogen detected in Hyalomma marginatum marginatum; Ri. massiliae, another emerging pathogen transmitted by Rh. sanguineus and Rh. turanicus (Bitam et al., 2006); and Ri. monacensis, which is generally found in Ixodes ricinus (Dib et al., 2009). The aim of the present study was to identify rickettsial species occurring in ticks collected from a specific area of Algeria that had not been previously investigated for tick-borne rickettsiae. Materials and methods In February 2006, we used the flag technique on vegetation close to the road to capture ticks in the mountains of the Blida region (Chréa Mountain) in northern Algeria. In December 2010, ticks were collected from cattle in Bouzguene (Tizi-Ouzou department), * Corresponding author. Tel.: +213 21 673365; fax: +213 21 673365; mobile: +213 771995693. E-mail address: idirbitam@gmail.com (I. Bitam). Algeria. These ticks were identified phenotypically by using cur- rent taxonomic criteria (Estrada-Pe ˜ na et al., 2004), were stored in 70% ethanol, and were sent to the WHO Collaborative Center for Rickettsial Diseases and Other Arthropod-borne Bacterial Diseases in Marseille, France. DNA from these specimens and negative con- trols (uninfected lice maintained in colonies in our laboratory) was extracted using a QIAamp Tissue Kit (QIAGEN, Hilden, Germany), according to the manufacturer’s instructions. Rickettsial DNA was detected using a Rickettsia genus-specific real-time PCR (qPCR) with a 25-bp probe targeting the partial sequence of the citrate syn- thase gene (gltA) using a 7900 HT Fast instrument (Varagnol et al., 2009). The positive control was DNA from Ri. montanensis (qPCR). Ticks analyzed by qPCR were considered to be positive when the cycle threshold was less than or equal to 30. All ticks identified as positive by qPCR were confirmed by 2 different standard PCRs and sequencing for fragments of the gltA and OmpA genes. DNA sequencing reactions were performed for all samples amplified by standard PCR, as previously described (Sarih et al., 2008). Data were collected with an ABI Prism 3130xl Genetic Analyzer capil- lary sequencer (ABI PRISM, PE Applied Biosystems, USA). Sequences were edited and assembled using Chromas Pro 1.34 (Technelysium Pty. Ltd., Tewantin, Australia). BLAST searches were performed to identify the obtained sequences. Results The ticks collected from the Blida region were divided into 2 groups: The first group contained I. ricinus and Haemaphysalis spp. 1877-959X/$ – see front matter © 2012 Elsevier GmbH. All rights reserved. http://dx.doi.org/10.1016/j.ttbdis.2012.10.012