key: cord-0000321-oi9j5o0n authors: Skuballa, Jasmin; Oehme, Rainer; Hartelt, Kathrin; Petney, Trevor; Bücher, Thomas; Kimmig, Peter; Taraschewski, Horst title: European Hedgehogs as Hosts for Borrelia spp., Germany date: 2007-06-03 journal: Emerg Infect Dis DOI: 10.3201/eid1306.070224 sha: f5cb9270ebde80bfaf1298576cb7c6fc7076f6b7 doc_id: 321 cord_uid: oi9j5o0n nan . Members of the B. burgdorferi s. l. group are the most common vectorborne pathogens of humans in central Europe (3) . The role of hedgehogs as hosts for these pathogens is, therefore, of considerable epidemiologic interest. Hedgehogs are a common synanthropic species that live in urban, suburban, and rural environments (4) and are known to carry not only the hedgehog tick, Ixodes hexagonus, but also the most common European tick, I. ricinus (2, 5) . Both of these ticks are known vectors of B. burgdorferi s. l. and tickborne encephalitis virus; I. ricinus is the most important vector of both throughout Europe (1, 5) . To date, however, only limited information has been available on the role of the hedgehog as a host or reservoir for B. burgdorferi s. l. in Germany. We report the presence of 3 species of the B. burgdorferi s. l. group in European hedgehogs from Germany. To our knowledge, this is the fi rst report of these species in hedgehogs in this country and the fi rst report of B. spielmanii (A14S) (6) from this host. The investigated hedgehogs came from 2 sources: 9 from the ≈40 in an experimental plot in the city of Karlsruhe, state of Baden-Wuerttemberg, and the remainder from wild hedgehogs that had been brought to hedgehog care centers from various areas of Germany. All hedgehogs had died naturally, and tissue samples were taken from 43 animals (kidneys from 43, heart from 22, bladder from 33). The bodies had been frozen at -17°C before the samples were taken. DNA isolation was done by using the Maxwell 16 Instrument and System (Promega, Madison, WI, USA). Tissue samples were 3×3×3 mm. To detect B. burgdorferi s. l., we used 2 PCR protocols. The fi rst was a nested PCR done according to the method of Rijpkema et al. (7) . The target for the PCR was the spacer region between 5S and 23S rRNA genes of B. burgdorferi s. l. The nested primers generated a product of 226 bp. The amplifi ed products were analyzed by agarose gel electrophoresis. The second protocol, a LightCycler-PCR hybridization assay (Roche Diagnostics, Mannheim, Germany) (8), simultaneously detects and genotypes the 3 genomic groups of B. burgdorferi s. l. This assay was specifi c for B. burgdorferi senso stricto, B. garinii, and B. afzelii (8) but also amplifi ed B. spielmanii and B. valaisiana. The target for the PCR was the OspA gene. The PCR products of both systems were sequenced. For DNA sequencing reaction, the fl uorescence-labeled didesoxynucleotide technology (Applied Biosystems, Darmstadt, Germany) was used. The sequenced fragments were separated, and the data were collected with an ABI PRISM 310 Genetic Analyzer (Applied Biosystems). The obtained sequences were then analyzed and compared by using BLAST (www. ncbi.nlm.nih.gov/BLAST). For 6 hedgehogs, Borrelia spp. could be clearly defi ned by using both gene sequences. Two additional animals had positive results, but sequencing was not possible because of either too little DNA or a mixed infection. B. spielmanii DNA was detected in the kidneys of 2 hedgehogs: 1 from Karlsruhe and 1 from 30 km west of this city in the German federal state of Rhineland-Palatinate. When sequences were compared by using BLAST, 4 BLAST sequences (AM055823, AM055822, DQ133518, AY 995900) showed 100% similarity with B. spielmanii. B. garinii was detected in the heart of 2 animals (from Berlin and Karlsruhe); B. afzelii in 3 animals (in the kidney of 2 from Hamburg and Karlsruhe and in the bladder of 1 from Rhineland-Palatinate). A single animal (from Karlsruhe) had B. afzelii in the kidney and bladder and B. garinii in the heart. Preliminary results have also shown that ticks collected from hedgehogs from the Karlsruhe site were infected with B. afzelii (an I. hexagonus nymph and an I. ricinus female) and with B. spielmanii (an I. ricinus female, a nymph, and a larva) (Skuballa et al., unpub. data). These results show, that hedgehogs harbor at least 3 of the 5 recognized Borrelia genospecies found in Germany, all of which are known (B. afzelii, B. garinii) or are strongly suspected (B. spielmanii) of being pathogens for humans (9, 10) . To our knowledge, ours is the fi rst report of B. spielmanii from hedgehogs, a Borrelia sp. that is usually associated with rodents, especially with garden and hazel dormice (10) . That Borrelia spp. infections commonly occur in European hedgehogs is likely. However, questions remain about the role of these pathogens in regulating the populations of European hedgehogs and about the status of these common synanthropic mammals as a reservoir host of B. burgdorferi s. l. in periurban and rural environments. Survival strategy of tick-borne encephalitis virus: cellular basis and environmental determinants Transmission cycles of Borrelia burgdorferi sensu lato involving Ixodes ricinus and/or I. hexagonus ticks and the European hedgehog, Erinaceus europaeus, in suburban and urban areas in Switzerland Epidemiology and diagnosis of Lyme borreliosis Acquisition of Borrelia burgdorferi by Ixodes ricinus ticks fed on the European hedgehog, Erinaceus europaeus Phenotypic and genetic characterization of a novel Borrelia burgdorferi sensu lato isolate from a patient with Lyme borreliosis Simultaneous detection and genotyping of three genomic groups of Borrelia burgdorferi sensu lato in Dutch Ixodes ricinus ticks by characterization of the amplifi ed intergenic spacer region between 5S and 23S rRNA genes Distribution of clinically relevant Borrelia genospecies in ticks assessed by a novel, single-run, real-time PCR Borrelia burgdorferi infection prevalences in questing Ixodes ricinus ticks (Acari: Ixodidae) in urban and suburban Bonn, western Germany Delineation of Borrelia burgdorferi sensu lato species by multilocus sequence analysis and confi rmation of the delineation of Borrelia spielmanii sp. nov We thank Miriam Pfäffl e, Heiko Fischer, Agnes Giniewski, and the staff at the hedgehog rehabilitation centers, especially Sigrun Goroncy and Elisabeth Swoboda, for their help.We acknowledge fi nancial support from the Konrad-Krieger Foundation and a foundation of the Landesbank Baden-Wuerttemberg, Germany. email: jasmin.skuballa@bio.uka.de To the Editor: Tumor necrosis factor-α (TNF-α) antagonists are immunosuppressants that have shown effi cacy in treating infl ammatory disorders. However, a recent meta-analysis of controlled trials has shown evidence of increased risk for serious infections in patients with rheumatoid arthritis treated with TNF-α antagonists (1).Adalimumab is a human monoclonal antibody to TNF-α approved by the US Food and Drug Administration (FDA) for treatment of rheumatoid arthritis. The Spanish registry of adverse events of biologic therapies in rheumatic diseases reported that 1,080 patients were treated with adalimumab from 2003 through 2006 and no cases of cryptococcosis were recorded (2) . No cases of cryptococcosis have been detected in 10,050 treated patients in the US postmarketing database for adalimumab (3) . We report invasive cryptococcosis in a patient receiving adalimumab. This case underscores the relationship between TNF antagonists and emergence of severe and diffi cult-to-treat opportunistic infections.A 69-year-old woman with rheumatoid arthritis diagnosed in 2002 was referred to our hospital for severe acute infl ammation of the second fi nger of the left hand. She had been treated with oral corticosteroids (prednisone, 7.5 mg/day) and several disease-modifying antirheumatic drugs, including chloroquine, methotrexate,