Israeli spotted fever Rickettsia in Sicilian Rhipicephalus sanguineus ticks. (Letters).

To the Editor: Mediterranean spotted fever (MSF) is endemic in Italy, where it is a reportable disease. From 1992 to 1998, the Italian Ministry of Health was notified of approximately 8,500 cases of human rickettsioses presumed to be MSF. MSF occurs more commonly in some central (Lazio) and southern (Sardinia, Sicily, and Calabria) regions (1,2); in 1998, an average of 8.8 cases occurred for every 100,000 persons in Sicily, compared with the national average of 1.6 cases per 100,000 persons. Rickettsia conorii has been thought to be the only pathogenic Rickettsia of the spotted fever group in Sicily (3,4) or the western Mediterranean area. Recently, three different spotted fever group rickettsiae, including R. helvetica, were detected in Ixodes ricinus ticks from central and northern Italy. This finding suggests that bacteria other than R. conorii are involved in rickettsial diseases in Italy (5).

To investigate whether unusual tick-transmitted rickettsiae are also present in Sicily, we used molecular-sequence-based identification techniques to study two strains isolated from the hemolymph of Rhipicephalus sanguineus ticks collected in 1990 in western Sicily. These isolates had been previously identified by serologic tests as belonging to the spotted fever group rickettsiae. We obtained bacterial DNA and performed polymerase chain reaction (PCR) for ompA gene and restriction analysis under conditions previously described by Roux et al. (6). Our observation of a peculiar PstI profile allowed a presumptive identification of one of the two tick isolates as belonging to the Israeli spotted fever rickettsiae, while the other showed a restriction profile corresponding to that of R. conorii strain Seven. To confirm the identification of the Israeli spotted fever Rickettsia isolate, we sequenced the PCR-amplified fragment of ompA gene (MWG-Biotech AG, Ebersberg, Germany) and aligned sequence data with homologous sequences of reference strains of the spotted fever group rickettsiae retrieved from the GenBank database. Sequence analysis showed 100% similarity with the homologous sequence of Israeli spotted fever Rickettsia reference strain ISTT CDC1 (GenBank accession no. U43797). The Israeli spotted fever Rickettsia belongs to the R. conorii complex (7,8) and was first isolated in 1974 from ticks and humans. Initially, Israeli spotted fever rickettsiae distribution appeared to be restricted to Israel (9), but more recently the organism has also been isolated from patients with MSF in Portugal (10). Our finding of Israeli spotted fever Rickettsia infection in a R. sanguineus tick, the main vector for MSF in Sicily, also suggests that the geographic distribution of Israeli spotted fever might be wider than previously thought, including not only Israel and the Iberian Peninsula but also Italy.

Molecular analysis of spotted fever group Rickettsia isolates from Sicilian MSF patients is under way to verify this hypothesis. Because initial signs and symptoms of Israeli spotted fever are particularly uncharacteristic, awareness of the presence of Israeli spotted fever Rickettsia in our geographic area may hasten provision of the appropriate treatment. The Sicilian ompA gene sequence described in this study has been deposited in the GenBank database (accession no. AY 197565).

References

(1.) Scaffidi V. Current endemic expansion of boutonneuse fever in Italy. Minerva Med 1981 ;72:2063-70.

(2.) Tringali G, Intonazzo V, Perna AM, Mansueto S, Vitale G, Walker DH. Epidemiology of Boutonneuse fever in western Sicily. Distribution and prevalence of spotted fever group rickettsial infection in dog ticks (Rhipicephalus sanguineus). Am J Epidemiol 1986; 123:721-7.

(3.) Mansueto S, Vitale G, Lavagnino A, Di Rosa S, Merulla R. Rickettsiae of the spotted fever group in dog fleas (Ctenocephalides spp.) in western Sicily. Ann Trop Med Parasitol 1989;83:325.

(4.) Vitale G, Di Stefano R, Damiani G, Mansueto S. Characterization of Sicilian strains of spotted fever group rickettsiae by using monoclonal antibodies. J Clin Microbiol 1989;27:1081-5.

(5.) Beninati T, Lo N, Noda H, Esposito F, Rizzoli A, Favia G, et al. First detection of spotted fever group Rickettsiae in Ixodes ricinus from Italy. Emerg Infect Dis 2002;8:983-6.

(6.) Roux V, Fournier PE, Raoult D. Differentiation of spotted fever group Rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR-amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol 1996;34:2058-65.

(7.) Regnery RL, Spruill CL, Plikaytis BD. Genotypic identification of Rickettsiae and estimation of interspecies sequence divergence for portions of two rickettsial genes. J Bacteriol 1991; 173:1576-89.

(8.) Fournier PE, Roux V, Raoult D. Phylogenetic analysis of spotted fever group Rickettsiae by study of the outer surface protein rOmpA, Int J Syst Bacteriol 1998;48:839-49.

(9.) Roux V, Raoult D. Phylogenetic analysis and taxonomic relationships among the genus Rickettsia. In: Raoult D, Brouqui P, editors. Rickettsiae and rickettsial diseases at the turn of the third millennium. Paris: Elsevier; 1999. p. 52-66.

(10.) Bacellar F, Beati L, Franca A, Pocas J, Regnery R, Filipe A. Israeli spotted fever rickettsia (Rickettsia conorii complex) associated with human disease. Emerg Infect Dis 1999;5:835-6.

Address for correspondence: Giovanni M. Giammanco, Department of Hygiene and Microbiology, Universita di Palermo, via del Vespro 133, 1-90127, Palermo, Italy; fax: [phone]; email: [e-mail]

Giovanni M. Giammanco, *

Serafino Mansueto, *

Pietro Ammatuna, *

and Giustina Vitale *

* Universita di Palermo, Palermo, Italy