Jerzy Michalik

Borrelia burgdorferi s.l. in Ixodes ricinus (Acari: Ixodidae) Ticks Collected from Vegetation and Small Rodents in Recreational Areas of the City of Poznań

Abstract During 1998–1999, Ixodes ricinus (L.) populations were investigated in three different biotopes (deciduous, mixed, coniferous forest) situated in popular recreational areas in Poznań, Poland. In total, 1,123 questing ticks (1,002 nymphs, 69 males, 52 females) were collected by flagging vegetation. Additionally, in 1998 between May and September small rodents were trapped and inspected for feeding ticks. Altogether, 213 rodents of three species: Apodemus agrarius Pall., A. flavicollis Melchior, Clethrionomys glareolus Schreber were captured. Of 323 engorged ticks, 304 were larvae and 19 nymphs. All ticks collected from vegetation, as well as from rodents were examined for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigenwalt & Brenner s.l. spirochetes by indirect immunofluorescence assay (IFA) using PAB 1B29. The seasonal pattern of activity of questing I. ricinus was always bimodal (May/June and August/September). The most abundant tick population occurred in the deciduous forest. The total infection rate in questing ticks was 16.2%. Differences in mean infection prevalence of host-seeking ticks between three biotopes each year were not significant. On average more larvae parasitized on the genus of Apodemus than on C. glareolus. 17.8% of larvae and 31.6% of nymphs fed on rodents harbored spirochetes. The three rodent species contributed to a different degree in to transmission of the pathogen to subadult stages. ≈27% of larvae infested on A. agrarius, 22% on C. glareolus, and only 4.2% on A. flavicollis contained spirochetes. The results suggest that the prevalence of A. agrarius and C. glareolus in disturbed urban forests used for leisure activities seems to be crucial for the maintenance of B. burgdorferi s.l. in I. ricinus populations.

Wild boars as hosts of human-pathogenic Anaplasma phagocytophilum variants.

To investigate the potential of wild boars to host Anaplasma phagocytophilum, we analyzed bacterial 16S rRNA and ank genes. DNA sequencing identified several A. phagocytophilum variants, including a predominance of strains known to cause human disease. Boars are thus hosts for A. phagocytophilum, notably, strains associated with human granulocytic anaplasmosis.

Borrelia burgdorferi Sensu Stricto in Yellow-Necked Mice and Feeding Ixodes ricinus Ticks in a Forest Habitat of West Central Poland

Abstract Wild rodents and the subadult Ixodes ricinus (L.) ticks infesting them were examined for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner s.l. in a sylvatic habitat in west central Poland during May–September 2002. In total, 818 feeding ticks were recovered from 73 infested yellow-necked mice, Apodemus flavicollis Melchior; in addition, bank voles, Clethrionomys glareolus Schreber, were rarely captured and proved to be weakly parasitized. Only 2.7% of A. flavicollis and 2.2% of 320 engorging larvae were polymerase chain reaction (PCR) positive for the bacterium. All spirochete-PCR-positive samples yielded exclusively B. burgdorferi s.s. This genospecies was also the most prevalent in questing nymphs and accounted for 87.5% of the total number of Borrelia infections in nymphal ticks collected during May and June 2 yr later. The presence of the same genospecies both in naturally engorged larvae and blood-positive animals as well as the high predominance of B. burgdorferi s.s. in questing nymphs strongly differs from most study sites investigated in Europe. This unique pattern of Borrelia-diversity in both rodents and ticks seems to be determined by highly site-specific host vertebrate cenosis, and yellow-necked mice are involved in the maintenance of B. burgdorferi s.s. in the forest habitat. However, the transmission efficiency of this spirochete from the mice to the I. ricinus vector seems to be very low. The research provides additional information on the complexity of B. burgdorferi s.l. ecology in Europe, pointing to the importance of the local host community.

Prevalence of infection with Rickettsia helvetica in feeding ticks and their hosts in western Poland

In Poland, Ixodes ricinus plays an important role as the vector of the TBE virus, Borrelia burgdorferi s.l., Anaplasma phagocytophilum and Babesia spp. Moreover, recent studies have shown that it can also serve as a vector for Rickettsia helvetica, the newly recognised human pathogen of the spotted fever group rickettsiae (SFG), which is widely distributed in questing ticks, with the prevalence ranging from 1.3% to 11.4% [1]. However, reports are still lacking on its occurrence in naturally infected wild animals and ticks feeding on them. So, this study was designed to detect rickettsial organisms in blood samples of various tick hosts that may serve as reservoirs of infection, as well as in I. ricinus parasitising them.

Prevalence of infection with Rickettsia helvetica in Ixodes ricinus ticks feeding on non-rickettsiemic rodent hosts in sylvatic habitats of west-central Poland

Ixodes ricinus is the most prevalent and widely distributed tick species in European countries and plays a principal role in transmission of a wide range of microbial pathogens. It is also a main vector and reservoir of Rickettsia spp. of the spotted fever group with the infection level ranging in Poland from 1.3% to 11.4%. Nevertheless, little research has been conducted so far to identify reservoir hosts for these pathogens. A survey was undertaken to investigate the presence of Rickettsia spp. in wild small rodents and detached I. ricinus. Rodents, Apodemus flavicollis mice and Myodes glareolus voles were captured in typically sylvatic habitats of west-central Poland. Blood samples and collected ticks were analyzed by conventional, semi-nested and nested PCRs. Rickettsial species were determined by sequence analysis of obtained fragments of gltA and 16S rRNA genes. A total of 2339 immature I. ricinus (mostly larvae) were collected from 158 animals. Proportion of hosts carrying ticks was 84%, being higher for A. flavicollis than for M. glareolus. Rickettsia helvetica, the only species identified, was detected in 8% of 12 nymphs and in at least 10.7% (MIR) of 804 larvae investigated. Prevalence of infected ticks on both rodent species was comparable (10.8 vs. 9%). None of blood samples tested was positive for Rickettsia spp. The results showed that in sylvatic habitats the level of infestation with larval I. ricinus was higher in A. flavicollis mice in comparison with M. glareolus voles. They show that R. helvetica frequently occurred in ticks feeding on rodents. Positive immature ticks were collected from non-rickettsiemic hosts what might suggest a vertical route of their infection (transovarial and/or transstadial) or a very short-lasting rickettsiemia in rodents. A natural vertebrate reservoir host for R. helvetica remains to be determined.

Differential associations of Borrelia species with European badgers (Meles meles) and raccoon dogs (Nyctereutes procyonoides) in western Poland.

European badgers and raccoon dogs and their associated ticks and lice were assayed for the presence of Lyme borreliosis and relapsing fever-group spirochete DNA in western Poland. Analyses of blood, ear-biopsy and liver samples revealed that 25% of 28 raccoon dogs and 12% of 34 badgers were PCR positive for borreliae. Borrelia garinii was the dominant species in raccoon dogs (62.5%), followed by B. afzelii (25%) and B. valaisiana (12.5%). PCR-positive badgers were infected only with B. afzelii. A total of 351 attached ticks was recovered from 23 (82%) of the raccoon dogs and 13 (38%) of the badgers. Using a nested PCR targeting the ITS2 fragments of Ixodes DNA, four Ixodes species were identified: I. ricinus, I. canisuga, I. hexagonus, and one provisionally named I. cf. kaiseri. Ixodes canisuga and I. ricinus prevailed on both host species. The highest infection prevalence was detected in I. ricinus, followed by I. canisuga and I. cf. kaiseri. Borrelia garinii and B. afzelii accounted for 61.6% and 30.1% of the infections detected in all PCR-positive ticks, respectively. Four other Borrelia species (B. burgdorferi sensu stricto, B. valaisiana, B. lusitaniae and B. miyamotoi) were detected only in I. ricinus from raccoon dogs. Moreover, Borrelia DNA, mostly B. garinii, was detected in 57 (81.4%) of 70 Trichodectes melis lice derived from 12 badgers. The detection of B. afzelii in one-half of PCR-positive biopsies reconfirms previous associations of this species with mammalian hosts, whereas the high prevalence of B. garinii in feeding lice and I. ricinus ticks (including larvae) demonstrates that both carnivores serve as hosts for B. garinii. The lack of B. garinii DNA in the tissues of badgers versus its prevalence in raccoon-dog biopsies, however, incriminates only the latter carnivore as a potential reservoir host.

Prevalence and habitat preference of quill mites (Acari, Syringophilidae) parasitizing forest passerine birds in Poland

We surveyed the prevalence of quill mites in wild passerines captured in Wielkopolski National Park, Poland. A total of 438 birds of 12 species were mist-netted during three consecutive spring seasons from 2006–2008. Eight bird species were infested with syringophilid mites: Erithacus rubecula (L.) (index of prevalence, IP = 16.2%), Turdrus merula L. (IP = 22.4%), T. philomelos Brehm (IP = 10.5%), Fringilla coelebs L. (IP = 36.2%), Coccothraustes coccothraustes (L.) (IP = 7.4%), Sturnus vulgaris L. (IP = 53.6%), Sylvia atricapilla (L.) (IP = 6.1%) and Troglodytes troglodytes (L.) (IP = 7.1%). In four cases, multiple infestation was observed, where one bird species was parasitized by 2–3 species of quill mites occupying separate niches in the plumage of the host.

Correction to: Bartonella and Rickettsia Infections in Haematophagous Spinturnix myoti Mites (Acari: Mesostigmata) and their Bat Host, Myotis myotis (Yangochiroptera: Vespertilionidae), from Poland

Hematophagous Spinturnix myoti mites and their host, the greater mouse-eared bat (Myotis myotis), were tested for the presence of Bartonella spp., Rickettsia spp., and Anaplasma phagocytophilum. In total, Bartonella spp. DNA was amplified in 28% of 134 mite pools and in 25% of 59 bats tested by PCR targeting a fragment of citrate synthase gltA gen. Adult mites were at least threefold more frequently infected compared to immature stages. The overall infection prevalence among mite pools from cave-dwelling bats was higher than for those collected from attic shelters. Three distinct genotypes were detected. The most prevalent genotype in mites and bats matched closely with Candidatus Bartonella hemsundetiensis identified in bats from Finland and was relatively distant from bat-borne Bartonella strains described in the UK and France. Importantly, most sequences were close to those reported in forest workers from Poland. The presence of identical genotype among S. myoti samples and M. myotis bats suggests that bartonellae can be shared between mites and their bat hosts. In this case, wing mites could serve as vectors, whereas their hosts as reservoirs. One blood sample was positive by PCR for the msp2 gene of A. phagocytophilum. Two mite pools yielded Rickettsia spp. DNA. Both sequences were distinct from any known species but can be classified as spotted fever group Rickettsia spp. Our findings expanded our knowledge on the role of spinturnicid mites in the ecology and epidemiology of bacterial infections associated with vespertilionid bats, especially regarding the genus Bartonella.

Human behaviors elevating the risk of exposure to Ixodes ricinus larvae and nymphs in two types of lowland coniferous forests in west-central Poland

Ixodes ricinus L. is the commonest tick encountered by humans in Central Europe and the most important vector of tick-borne diseases (TBD) in Europe. Foresters represent a group at elevated risk of exposure to I. ricinus throughout Europe. Here, we employed an experimental design similar to one used previously to determine what behavioral factors heighten exposure to questing Ixodes pacificus nymphs in a hardwood forest in the far-western United States. Five activities were evaluated systematically in a coniferous forest and a mixed broadleaved forest in west-central Poland: (i) walking in low-cut running shoes; (ii) sitting atop logs; (iii) sitting on leaf litter; (iv) sitting against tree trunks; and (v) gathering firewood. In total, 540 I. ricinus ticks were collected (198 larvae, 327 nymphs, 15 adults) in spring and late summer. Many more ticks were collected in spring than in late summer, and in the mixed broadleaved forest versus the coniferous forest. The riskiest behavior for acquiring nymphs in both spring and late summer was gathering firewood, though sitting atop logs was nearly as perilous in late summer. In contrast, the riskiest behaviors for encountering larvae in spring and late summer were sitting against trunks and sitting on leaf litter, respectively. Pairwise comparisons revealed only two significant associations among the variables explored for the nymphs: season and human gender, and season and type of forest. For larval ticks, the most significant associations were gender and sitting behaviors and for type of forest and sitting on leaf litter or against tree trunks. The most commonly infested body region was the lower legs.