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10.1111/j.1469-0691.2008.02277.x

First description of monacensis in in Algeria L. Dib1, I. Bitam2, M. Bensouilah3, P. Parola4 and D. Raoult4

1Institut Ve´te´rinaire, Centre Universitaire El Tarf, El Tarf, 2Unite´ d’Entomologie Me´dicale, Institut Pasteur d’Alge´rie, Alger, 3Faculte´ des Sciences, Universite´ Badji Mokhtar, Annaba, Alge´rie and 4URMITE, UMR CNRS-IRD 6236, WHO Collaborative Centre for Rickettsial Diseases and Other Arthropod-borne Bacterial Diseases, Faculte´ de Me´decine, Marseille, France

629–632-bp fragment [2]. A negative control with distilled INTRODUCTION water instead of DNA template in the PCR mixture and a Tick-borne spotted fever group (SFG) rickettsioses positive control (DNA from Rickettsia montanensis) were included in each test [2]. All obtained sequences were are caused by obligate intracellular Gram-nega- compared to sequences in the GenBank database. tive belonging to the genus Rickettsia. Although some have been known as human pathogens since the beginning of the RESULTS century, tick-borne SFG rickettsioses are also All were identified as Ixodes ricinus (Fig. 1). recognized as important emerging worldwide Using the gltA primers, PCR products of rickett- zoonoses [1]. In Algeria, three species of SFG sial DNA were detected in 35 ticks (59.32%) and rickettsia have been detected in ticks, including from positive controls. All ticks were also shown conorii (the agent of Mediterra- to be positive on the second PCR screening using nean spotted fever) in Rhipicephalus sanguineus, ompA primers. When the sequences of the gltA and two emerging pathogens, i.e. Rickettsia aes- and ompA amplified fragments were obtained, chlimannii in Hyalomma marginatum marginatum, they were shown to correspond to those of and Rickettsia massiliae in Rhipicephalus sanguineus Rickettsia monacenis (100% similarity with Gen- and Rhipicephalus turanicus [2]. We present here Bank accession numbers DQ157778 and the detection and characterization of rickettsiae in DQ100163 for gltA and ompA, respectively). more ticks collected in Algeria using PCR and sequence analysis of amplified products. DISCUSSION

MATERIALS AND METHODS To the best of our knowledge, this is the first report of R. monacensis in Algeria. This rickettsia In total, 59 ticks were collected from cattle in the El Traf region was described when it was isolated from I. ricinus of north-eastern Algeria from December 2005 to March 2006. All ticks were adults and were morphologically identified by ticks collected in 1998 in a city park in Munich, one of us (I.B.) using the usual taxonomic keys. They were kept Germany [3]. R. monacensis has been previously in 70% ethanol at room temperature, until they were tested in Marseille, France. DNA was extracted, and rickettsial DNA was detected by PCR using primers Rp. CS.409p and Rp. CS.1258n, which amplify a 750-bp fragment of the citrate synthase gene (gltA)ofRickettsia [2]. Additionally, all ticks positive for gltA were tested for the ompA gene of Rickettsia using primers Rr. 190.70 and Rr. 190.701, which amplify a

Corresponding author and reprint requests: D. Raoult, Unite´ de Recherche en Maladies Infectieuses et Tropicales Emergen- tes (URMITE), UMR CNRS-IRD 6236, WHO Collaborative Centre for Rickettsial Diseases and Other Arthropod-borne Bacterial Diseases, Faculte´ de Me´decine, 27 Bd Jean Moulin, 13385 Marseille Cedex 5, France E-mail: [email protected] Fig. 1. Ixodes ricinus ticks attached on cattle in the El Traf region of north-eastern Algeria. In the right corner, a male No conflicts of interest declared. on a female.

Ó 2009 The Authors Journal Compilation Ó 2009 European Society of Clinical Microbiology and Infectious Diseases, CMI, 15 (Suppl. 2), 261–262 262 Clinical Microbiology and Infection, Volume 15, Supplement 2, December 2009 reported in the literature under other names, REFERENCES including ‘the Cadiz agent’ in Spain, and Rickett- 1. Parola P, Paddock C, Raoult D. Tick-borne rickettsioses sia IRS3 and IRS4 detected in Slovakia and around the world: emerging diseases challenging old con- Bulgaria [4]. Recently, two human cases of infec- cepts. Clin Microbiol Rev 2005; 18: 719–756. tion due to R. monacensis were documented in 2. Bitam I, Parola P, Matsumoto K et al. First molecular Spain, when investigators succeeded in isolating detection of R. conorii, R. aeschlimannii, and R. massiliae in ticks from Algeria. Ann NY Acad Sci 2006; 1078: 368–372. the agent from the blood of two patients with 3. Simser JA, Palmer AT, Fingerle V, Wilske B, Kurtti TJ, Mediterranean spotted fever-like illnesses [5]. Munderloh UG. Rickettsia monacensis sp. nov., a spotted With our results, R. monacensis joins the list of fever group Rickettsia, from ticks (Ixodes ricinus) collected in autochthonous Rickettsia species confirmed as a European city park. Appl Environ Microbiol 2002; 68: 4559– human pathogens in Algeria. Although a high 4566. 4. Sekeyova Z, Fournier PE, Rehacek J, Raoult D. Character- rate of infection was found here, we cannot ization of a new spotted fever group rickettsia detected in address the prevalence and the distribution of Ixodes ricinus (Acari: Ixodidae) collected in Slovakia. J Med R. monacensis in Algeria. Systematic sampling Entomol 2000; 37: 707–713. should be performed for this purpose. However, 5. Jado I, Oteo JA, Alda´miz M et al. Rickettsia monacensis and human disease, Spain. Emerg Infect Dis 2007; 13: 1405–1407. the high-risk areas for infection will certainly include the cool and humid areas of the country that are associated with the Atlas Mountains. There, I. ricinus ticks usually feed on cattle and sheep, and perhaps on people.

Ó 2009 The Authors Journal Compilation Ó 2009 European Society of Clinical Microbiology and Infectious Diseases, CMI, 15 (Suppl. 2), 261–262