To the Editor: Rickettsia spp. of the spotted fever group cause worldwide emerging human infections known as tick-borne rickettsioses (1). Data on the occurrence and prevalence of Rickettsia in Germany are still limited (2). Six Rickettsia species have been reported to date (2). R. helvetica, R. felis, R. massiliae, and R. monacensis were detected with a relatively low prevalence in Ixodes ricinus ticks collected in southern Germany (2); R. raoultii was identified with high prevalence in the rapidly expanding area where D. reticulatus ticks are found (2). R. raoultii was recently recognized as an agent of tick-borne lymphadenopathy/Dermacentor-borne necrosis and erythema lymphadenopathy (3). Low prevalence of another tick-borne lymphadenopathy agent, R. slovaca, in Dermacentor marginatus ticks collected in southern Germany was recently reported (4)

Dautel, Hans

Graser, Elmara

Hillebrand, Timo

Mantke, Oliver Donoso

Mediannikov, Oleg

Niedrig, Matthias

Roy-Chowdhury, Panchali

Rumer, Leonid

Sheshukova, Olga

Talaska, Thomas

Emerging Infectious Diseases

English

PubMed

Publikationsserver des Robert Koch-Instituts

LETTERS  7.  Tonteri E, Jääskeläinen AE, Tikkakoski T, Voutilainen L, Niemimaa J, Henttonen H, et al. Tick-borne encephalitis virus in wild rodents in winter, Finland, 2008–2009. Emerg Infect Dis. 2011;17:72–75.  8.  Tuomi J, Brummer-Korvenkontio M. Antibodies against viruses of the tick-borne encephalitis group in cattle sera in Finland. Ann Med Exp Biol Fenn. 1965;43:149–54.  9.  Randolph SE, Rogers DJ. Fragile transmis-sion cycles of tick-borne encephalitis vi-rus may be disrupted by predicted climate change. Proc Biol Sci. 2000;267:1741–4. DOI: 10.1098/rspb.2000.120410.  Lukan M, Bullova E, Petko B. Climate warming and tick-borne encephalitis, Slo-vakia. Emerg Infect Dis. 2010;16:524–6. DOI: 10.3201/eid1603.081364Address for correspondence: Anu E. Jääskeläinen, Department of Virology, Haartman Institute, PO Box 21 FI-00014, University of Helsinki, Helsinki 00014, Finland; email: anu.jaaskelainen@helsinki.fi Rickettsia aeschlimannii in Hyalomma marginatum Ticks, GermanyTo the Editor: Rickettsia spp. of the spotted fever group cause world-wide emerging human infections known as tick-borne rickettsioses (1). Data on the occurrence and prevalence of Rickettsia in Germany are still lim-ited (2). Six Rickettsia species have been reported to date (2). R. helvetica, R. felis, R. massiliae, and R. mona-censis were detected with a relatively low prevalence in Ixodes ricinus ticks collected in southern Germany (2); R. raoultii was identifi ed with high prev-alence in the rapidly expanding area where D. reticulatus ticks are found (2). R. raoultii was recently recog-nized as an agent of tick-borne lymph-adenopathy/Dermacentor-borne necrosis and erythema lymphadenop-athy (3). Low prevalence of another tick-borne lymphadenopathy agent, R. slovaca, in Dermacentor marginatus ticks collected in southern Germany was recently reported (4).We report the detection in Germa-ny of the pathogenic SFG species R. aeshlimannii (1), which is phylogenet-ically close to R. raoultii and causes an infection with clinical signs similar to those of Mediterranean spotted fe-ver (1). To determine the prevalence of R. raoultii in the ticks in Berlin/Brandenburg and neighboring re-gions, we collected 294 ticks; 288 had been collected either from vegetation or domestic animals and morphologi-cally identifi ed as adult D. reticulatus ticks. The remaining 6 ticks were de-livered by an ornithologist who had removed them from a bird (belonging to the Acrocephalus scirpaceus spp.) that he had captured in the reeds near Pakendorf and Zerbst, Saxony-Anhalt, in May 2007. These 6 ticks were re-ported as D. reticulatus–like adults but were damaged in the collection process, making an exact morphologic evaluation impossible.DNA was isolated from the com-plete tick body by homogenization in the SpeedMill (Analytik Jena Biosolu-tions, Jena, Germany) followed by pu-rifi cation by RapideStripe tick DNA/RNA Extraction Kit (Analytik Jena Biosolutions). Multispacer typing (5) was used for molecular detection and determination of Rickettsia spp. (Fig-ure). DNA sequencing and analysis were performed as described (Figure).In 51.3% of the intact tick isolates, R. raoultii was detected. In each of the 3 damaged isolates, the multispacer type pattern was detected, which was 100% identical to that of R. aeschli-mannii (5) (Figure). Moreover, PCR, which we routinely use as a positive control for molecular identifi cation of D. reticulatus, yielded no product in the damaged isolates (Figure).To determine the species of the damaged ticks, we used 3 tick-specifi c PCRs. One amplifi ed a 16S rRNA frag-ment used for phylogenetic studies of ticks (6). Use of the other 2 PCRs was  Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 2, February 2011 325 Figure. Illustration of multispacer typing. Amplicons 1–4 result from PCRs on DNA obtained from 1 Rickettsia raoultii–infected Dermacentor reticulatus tick isolate; lanes 5–8 result from PCRs on 1 damaged isolate. PCRs amplifying dksA-xerC (lanes 1 and 5), mppA-purC (lanes 2 and 6), and rpmE-tRNA (lanes 3 and 7) intergenic spacers were performed as described (5). PCR amplifying the entire internal transcribed factor 2 (ITS2) locus of D. reticulatus tick (lanes 4 and 8) was involved in each PCR run as a positive control and for validation of D. reticulatus tick identity (the primers will be described elsewhere).The negative result of ITS2 PCR with the damaged isolates (lane 8) indicated that they are not D. reticulatus ticks. Lane M, DNA size marker (100-bp ladder). PCR products were directly sequenced in both directions with respective primers by an ABI PRISM DNA Sequencer (Applied Biosystems, Foster City, CA, USA). DNA Star package (DNA Star, Madison, WI, USA) and the tools offered by the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov) were used for DNA search and analysis.LETTERSbased on the consideration that R. ae-schlimannii is usually found in ticks of the genus Hyalomma, primarily in H. marginatum (1). Therefore, 1 of the PCRs amplifi ed a fragment of the Hya-lomma tick mitochondrial cytochrome oxidase I gene and the other a fragment of the internal transcribed spacer 2 (7).The ITS2 fragment displayed the highest (99%) similarity with the re-spective fragment of H. marginatum, H. dromedarii, H. truncatum, and H. lusitanicum. Cytochrome oxidase sub-unit I fragment was 99% identical to H. marginatum, H. dromedarii, and H. truncatum. The 16S RNA fragment was 98% identical to H. marginatum; its identity to the second closest se-quence belonging to H. lusitanicum was 96%.Earlier, R. aeschlimannii had been detected in sub-Saharan and North Af-rica, southern Europe, and southwest-ern Russia (8). Therefore, the area of Zerbst, the middle of Germany, marks the northernmost point of R. aeschli-mannii detection.Hyalomma spp. ticks are distrib-uted in Africa, the Mediterranean cli-matic zone of southern Europe, and in Asia. The only documented Hyalomma spp. tick in Germany was found on a human in the southern part of the coun-try (Lake Constance area) in May 2006, but the possibility of tick transportation from Spain was not ruled out (9).Acrocephalus scirpaceus birds are migratory birds and live in central Europe from April to October and win-ter in sub-Saharan Africa in the region inhabited by Hyalomma spp. ticks. Therefore, it is reasonable to suggest that the Hyalomma spp. ticks that we examined had been transported by the birds from Africa. The fact that a ran-domly caught bird was infested with R. aeshlimannii –infected ticks is sug-gestive of the intensive stream of new pathogens transported through Europe by migrating birds. The fi rst possible implication of a bird as a vector of in-fected ticks was proposed for Hyalom-ma spp. ticks infected by R. sibirica mongolitimonae (10). Until now, the role of migrating birds in distribu-tion of tick-borne pathogens has been poorly understood (9). The changing climate and environment in central Europe may facilitate the establish-ment of pathogen-carrying tick spe-cies transported by birds. These new pathogens can be directly transmitted from infected birds to the species of the local fauna.AcknowledgmentWe thank Yuliya Dobrydneva for crit-ical reading of the manuscript.Leonid Rumer, Elmara Graser, Timo Hillebrand, Thomas Talaska, Hans Dautel, Oleg Mediannikov, Panchali Roy-Chowdhury, Olga Sheshukova, Oliver Donoso Mantke, and Matthias NiedrigAuthor affi liations: Robert Koch Institut, Berlin, Germany (L. Rumer, P. Roy-Chow-dhury, O. Sheshukova, O. Donoso Mantke, M. Niedrig); AJInnuscreen GmbH, Berlin (E. Graser, T. Hillebrand); Practice for Mi-crobiology and Epidemiology of Infectious Diseases, Lindow, Germany (T. Talaska); IS Insect Services GmbH, Berlin (H. Dautel); and Université de la Méditerranée, Mar-seille, France (O. Mediannikov)DOI: 10.3201/eid1702.100308References  1.  Parola P, Paddock C, Raoult D. Tick-borne rickettsioses around the world: emerging diseases challenging old concepts. Clin Microbiol Rev. 2005;18:719–56. DOI: 10.1128/CMR.18.4.719-756.2005  2.  Dobler G, Wölfel R. Typhus and oth-er rickettsioses. Dtsch Arztebl Int. 2009;106:348–54.  3.  Parola P, Rovery C, Rolain JM, Brouqui P, Davoust B, Raoult D. Rickettsia slovaca and R. raoultii in tick-borne rickettsioses. Emerg Infect Dis. 2009;15:1105–8. DOI: 10.3201/eid1507.081449  4.  Pluta S, Tewald F, Hartelt K, Oehme R, Kimmig P, Mackenstedt U. Rickettsia slo-vaca in Dermacentor marginatus ticks, Germany. Emerg Infect Dis. 2009;15:2077–8. DOI: 10.3201/eid1512.090843  5.  Fournier PE, Raoult D. Identifi cation of rickettsial isolates at the species level us-ing multi-spacer typing. BMC Microbiol. 2007;7:72. DOI: 10.1186/1471-2180-7-72   6.  Caporale DA, Rich SM, Spielman A, Tel-ford SR 3rd, Kocher TD. Discriminating between Ixodes ticks by means of mito-chondrial DNA sequences. Mol Phylo-genet Evol. 1995;4:361–5.   7.  Rees DJ, Dioli M, Kirkendall LR. Mole-cules and morphology: evidence for cryp-tic hybridization in African Hyalomma (Acari: Ixodidae). Mol Phylogenet Evol. 2003;27:131–42.   8.  Shpynov S, Rudakov N, Tohkov Y, Ma-tushchenko A, Tarasevich I, Raoult D, et al. Clin Microbiol Infect. 2009;15(Sup-pl 2):S315–6. Detection of Rickettsia aeschlimannii in Hyalomma margin-atum ticks in western Russia. DOI: 10.1111/j.1469-0691.2008.02256.x  9.  Kampen H, Poltz W, Hartelt K, Wölfel R, Faulde M. Detection of a questing Hyalomma marginatum marginatum adult female (Acari, Ixodidae) in southern Ger-many. Exp Appl Acarol. 2007;43:227–31. DOI: 10.1007/s10493-007-9113-y10.  Raoult D, Roux V. Rickettsioses as para-digms of new or emerging infectious dis-eases. Clin Microbiol Rev. 1997;10:694–719.Address for correspondence: Thomas Talaska, Facharzt für Mikrobiologie und Infektionsepidemiologie, Georg-Schacht Str 23, 15295 Brieskow-Finkenheerd, Germany; email: thomas.talaska@t-online.deDogs as Reservoirs for Leishmania braziliensisTo the Editor: I have read the re-view by Sousa and Pearson (1), which provides a fascinating historical ac-count of the Great Drought and the smallpox epidemic of the 1870s and their association with the emergence of cutaneous leishmaniasis in Ceará, Brazil. In their review, the authors went back to the 19th century, remem-bering the hard years experienced by those who faced the Great Drought, 326 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 2, February 2011

Rickettsia aeschlimannii in Hyalomma marginatum Ticks, Germany

Leonid Rumer

Elmara Graser

Timo Hillebrand

Thomas Talaska

Hans Dautel

Oleg Mediannikov

Panchali Roy-Chowdhury

Olga Sheshukova

Oliver Donoso Mantke

Matthias Niedrig

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Rickettsia aeschlimannii in Hyalomma marginatum Ticks, Germany

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