Lyme borreliosis (LB) is the most prevalent tick-borne human infection in Europe, with increasing incidence during the latest decades. Abundant populations of Ixodes ricinus, the main vector of the causative agent, spirochetes from the Borrelia burgdorferi sensu lato (Bbsl) complex, have been observed in urban and suburban areas of Europe, in general, and Slovakia, particularly. Understanding the spread of infectious diseases is crucial for implementing effective control measures. Global changes affect contact rates of humans and animals with Borrelia-infected ticks and increase the risk of contracting LB. The aim of this study was to investigate spatial and temporal variation in prevalence of Bbsl and diversity of its species in questing I. ricinus from three sites representing urban/suburban, natural and agricultural habitat types in Slovakia. Ixodes ricinus nymphs and adults were collected by dragging the vegetation in green areas of Bratislava town (urban/suburban habitat), in the Small Carpathians Mountains (natural habitat) (south-western Slovakia) and in an agricultural habitat at Rozhanovce in eastern Slovakia. Borrelia presence in ticks was detected by PCR and Bbsl species were identified by restriction fragment length polymorphism (RFLP). Borrelia burgdorferi s.l. species in coinfected ticks were identified by reverse line blot. Significant spatial and temporal variability in prevalence of infected ticks was revealed in the explored habitats. The lowest total prevalence was detected in the urban/suburban habitat, whereas higher prevalence was found in the natural and agricultural habitat. Six Bbsl species were detected by RFLP in each habitat type -B. burgdorferi sensu stricto (s.s.), B. afzelii, B. garinii, B. valaisiana, B. lusitaniae and B. spielmanii. Coinfections accounted for 3% of the total infections, whereby B. kurtenbachii was identified by RLB and sequencing in mixed infection with B. burgdorferi s.s, B. garinii and B. valaisiana. This finding represents the first record of B. kurtenbachii in questing I. ricinus in Slovakia and Europe. Variations in the proportion of Bbsl species were found between nymphs and adults, between years and between habitat types. Spatial variations in prevalence patterns and proportion of Bbsl species were also confirmed between locations within a relatively short distance in the urban habitat. Habitat-related and spatial variations in Borrelia prevalence and distribution of Bbsl species are probably associated with the local environmental conditions and vertebrate host spectrum. Due to the presence of Borrelia species pathogenic to humans, all explored sites can be ranked as areas with high epidemiological risk.

Biology Centre Czech Academy of Sciences Institute of Parasitology Branišovská 31 37005 České Budějovice Czech Republic

Institute of Parasitology Slovak Academy of Sciences Hlinkova 3 04001 Košice Slovakia

Institute of Zoology Slovak Academy of Sciences Dúbravská cesta 9 84506 Bratislava Slovakia

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Rizzoli A., Hauffe H.C., Carpi G., Vourc’h G.I., Neteler M., Rosà R. Lyme borreliosis in Europe. Euro Surveill. 2011;16:19906. doi: 10.2807/ese.16.27.19906-en. PubMed DOI

Rudenko N., Golovchenko M., Grubhoffer L., Oliver J.H., Jr. Updates on Borrelia burgdorferi sensu lato complex with respect to public health. Ticks Tick Borne Dis. 2011;2:123–128. doi: 10.1016/j.ttbdis.2011.04.002. PubMed DOI PMC

Steinbrink A., Brugger K., Margos G., Kraiczy P., Klimpel S. The evolving story of Borrelia burgdorferi sensu lato transmission in Europe. Parasitol. Res. 2022;121:781–803. doi: 10.1007/s00436-022-07445-3. PubMed DOI PMC

Wodecka B., Kolomiiets V. Genetic diversity of Borreliaceae species detected in natural populations of Ixodes ricinus ticks in Northern Poland. Life. 2023;13:972. doi: 10.3390/life13040972. PubMed DOI PMC

Wodecka B., Michalik J., Grochowalska R. Red foxes (Vulpes vulpes) are exposed to high diversity of Borrelia burgdorferi sensu lato species infecting fox-derived Ixodes ticks in west-central Poland. Pathogens. 2022;11:696. doi: 10.3390/pathogens11060696. PubMed DOI PMC

Majerová K., Hönig V., Houda M., Papežík P., Fonville M., Sprong H., Rudenko N., Golovchenko M., Černá Bolfíková B., Hulva P., et al. Hedgehogs, squirrels, and blackbirds as sentinel hosts for active surveillance of Borrelia miyamotoi and Borrelia burgdorferi complex in urban and rural environments. Microorganisms. 2020;8:1908. doi: 10.3390/microorganisms8121908. PubMed DOI PMC

Rudenko N., Golovchenko M. Sexual transmission of Lyme borreliosis? The question that calls for an answer. Trop. Med. Infect. Dis. 2021;6:87. doi: 10.3390/tropicalmed6020087. PubMed DOI PMC

Stanek G., Reiter M. The expanding Lyme Borrelia complex-clinical significance of genomic species? Clin. Microbiol. Infect. 2011;17:487–493. doi: 10.1111/j.1469-0691.2011.03492.x. PubMed DOI

Estrada-Peña A., Ortega C., Sánchez N., Desimone L., Sudre B., Suk J.E., Semenza J.C. Correlation of Borrelia burgdorferi sensu lato prevalence in questing Ixodes ricinus ticks with specific abiotic traits in the western Palearctic. Appl. Environ. Microbiol. 2011;77:3838–3845. doi: 10.1128/AEM.00067-11. PubMed DOI PMC

Strnad M., Honig V., Růžek D., Grubhoffer L., Rego R.O.M. Europe-wide meta-analysis of Borrelia burgdorferi sensu lato prevalence in questing Ixodes ricinus ticks. Appl. Environ. Microbiol. 2017;83:e00609-17. doi: 10.1128/AEM.00609-17. PubMed DOI PMC

Kurtenbach K., De Michelis S., Etti S., Schäfer S.M., Sewell H.S., Brade V., Kraiczy P. Host association of Borrelia burgdorferi sensu lato–the key role of host complement. Trends Microbiol. 2002;10:74–79. doi: 10.1016/S0966-842X(01)02298-3. PubMed DOI

Kurtenbach K., Hanincová K., Tsao J.I., Margos G., Fish D., Ogden N.H. Fundamental processes in the evolutionary ecology of Lyme borreliosis. Nat. Rev. Microbiol. 2006;4:660–669. doi: 10.1038/nrmicro1475. PubMed DOI

O’Keeffe K.R., Oppler Z.J., Brisson D. Evolutionary ecology of Lyme Borrelia. Infect. Genet. Evol. 2020;85:104570. doi: 10.1016/j.meegid.2020.104570. PubMed DOI PMC

Hofmeester T.R., Coipan E.C., van Wieren S.E., Prins H.H.T., Takken W., Sprong H. Few vertebrate species dominate the Borrelia burgdorferi s.l. life cycle. Environ. Res. Lett. 2016;11:043001. doi: 10.1088/1748-9326/11/4/043001. DOI

Heylen D., Lasters R., Adriaensen F., Fonville M., Sprong H., Matthysen E. Ticks and tick-borne diseases in the city: Role of landscape connectivity and green space characteristics in a metropolitan area. Sci. Total Environ. 2019;670:941–949. doi: 10.1016/j.scitotenv.2019.03.235. PubMed DOI

Gern L. Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis: Life in the wilds. Parasite. 2008;15:244–247. doi: 10.1051/parasite/2008153244. PubMed DOI

Humair P.F., Gern L. The wild hidden face of Lyme borreliosis in Europe. Microbes Infect. 2000;2:915–922. doi: 10.1016/S1286-4579(00)00393-2. PubMed DOI

Hanincová K., Schäfer S.M., Etti S., Sewell H.S., Tarageľová V., Žiak D., Labuda M., Kurtenbach K. Association of Borrelia afzelii with rodents in Europe. Parasitology. 2003;126:11–20. doi: 10.1017/S0031182002002548. PubMed DOI

Humair P.F., Gern L. Relationship between Borrelia burgdorferi sensu lato species, red squirrels (Sciurus vulgaris) and Ixodes ricinus in enzootic areas in Switzerland. Acta Trop. 1998;69:213–227. doi: 10.1016/S0001-706X(97)00126-5. PubMed DOI

Hanincová K., Tarageľová V., Koči J., Schäfer S.M., Hails R., Ullmann A.J., Piesman J., Labuda M., Kurtenbach K. Association of Borrelia garinii and B. valaisiana with songbirds in Slovakia. Appl. Environ. Microbiol. 2003;69:2825–2830. doi: 10.1128/AEM.69.5.2825-2830.2003. PubMed DOI PMC

Tarageľová V., Koči J., Hanincová K., Kurtenbach K., Derdáková M., Ogden N.H., Literák I., Kocianová E., Labuda M. Blackbirds and song thrushes constitute a key reservoir of Borrelia garinii, the causative agent of borreliosis in central Europe. Appl. Environ. Microbiol. 2008;74:1289–1293. doi: 10.1128/AEM.01060-07. PubMed DOI PMC

Richter D., Matuschka F.R. Perpetuation of the Lyme disease spirochete Borrelia lusitaniae by lizards. Appl. Environ. Microbiol. 2006;72:4627–4632. doi: 10.1128/AEM.00285-06. PubMed DOI PMC

Majláthová V., Majláth I., Víchová B., Derdáková M., Peťko B. Borrelia lusitaniae and green lizards (Lacerta viridis), Karst Region, Slovakia. Emerg. Infect. Dis. 2006;12:1895–1901. doi: 10.3201/eid1212.060784. PubMed DOI PMC

Richter D., Schlee D.B., Allgöwer R., Matuschka F.-R. Relationships of a novel Lyme disease spirochete, Borrelia spielmanii sp. nov., with its hosts in Central Europe. Appl. Environ. Microbiol. 2004;70:6414–6419. doi: 10.1128/AEM.70.11.6414-6419.2004. PubMed DOI PMC

Durand J., Herrmann C., Genné D., Sarr A., Gern L., Voordouw M.J. Multistrain infections with Lyme borreliosis pathogens in the tick vector. Appl. Environ. Microbiol. 2017;83:e02552-16. doi: 10.1128/AEM.02552-16. PubMed DOI PMC

Rudenko N., Grubhoffer L., Golovchenko M. Climate Impact on Lyme Borreliosis and Its Causative Agents. In: Nuttall P., editor. Climate, Ticks and Disease. University of Oxford; Oxford, UK: 2021. pp. 349–354.

Hanincova K., Kurtenbach K., Diuk-Wasser M., Brei B., Fish D. Epidemic spread of Lyme borreliosis, Northeastern United States. Emerg. Infect. Dis. 2006;12:604–611. doi: 10.3201/eid1204.051016. PubMed DOI PMC

Černý J., Lynn G., Hrnková J., Golovchenko M., Rudenko N., Grubhoffer L. Management options for Ixodes ricinus-associated pathogens: A review of prevention strategies. Int. J. Environ. Res. Public Health. 2020;17:1830. doi: 10.3390/ijerph17061830. PubMed DOI PMC

Kahl O., Gray J.S. The biology of Ixodes ricinus with emphasis on its ecology. Ticks Tick Borne Dis. 2023;14:102114. doi: 10.1016/j.ttbdis.2022.102114. PubMed DOI

Rizzoli A., Silaghi C., Obiegala A., Rudolf I., Hubálek Z., Földvári G., Plantard O., Vayssier-Taussat M., Bonnet S., Špitalská E., et al. Ixodes ricinus and its transmitted pathogens in urban and peri-urban areas in Europe: New hazards and relevance for public health. Front. Publ. Health. 2014;2:251. doi: 10.3389/fpubh.2014.00251. PubMed DOI PMC

Gandy S., Kilbride E., Biek R., Millins C., Gilbert L. Experimental evidence for opposing effects of high deer density on tick-borne pathogen prevalence and hazard. Parasit. Vectors. 2021;14:509. doi: 10.1186/s13071-021-05000-0. PubMed DOI PMC

Stanko M., Derdáková M., Špitalská E., Kazimírová M. Ticks and their epidemiological role in Slovakia: From the past till present. Biologia. 2022;77:575–1610. doi: 10.1007/s11756-021-00845-3. PubMed DOI PMC

Kmety E., Rehácek J., Výrosteková V. Investigations of ticks for the presence of Borrelia in Czechoslovakia. Zentralbl. Bakteriol. Mikrobiol. Hyg. A. 1987;263:468–470. doi: 10.1016/S0176-6724(87)80111-6. PubMed DOI

Drgoňová M., Řeháček J. Prevalence of Lyme borrelia in ticks in Bratislava, Slovak Republic. Cent. Eur. J. Public Health. 1995;3:134–137. PubMed

Gern L., Hu C.M., Kocianova E., Vyrostekova V., Rehacek J. Genetic diversity of Borrelia burgdorferi sensu lato isolates obtained from Ixodes ricinus ticks collected in Slovakia. Eur. J. Epidemiol. 1999;15:665–669. doi: 10.1023/A:1007660430664. PubMed DOI

Štepánová-Tresová G., Peťko B., Štefančíková A., Nadzamová D. Occurrence of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii in the Ixodes ricinus ticks from Eastern Slovakia. Eur. J. Epidemiol. 2000;16:105–109. doi: 10.1023/A:1007606623892. PubMed DOI

Lenčáková D., Hizo-Teufel C., Petko B., Schulte-Spechtel U., Stanko M., Wilske B., Fingerle V. Prevalence of Borrelia burgdorferi s.l. OspA types in Ixodes ricinus ticks from selected localities in Slovakia and Poland. Int. J. Med. Microbiol. 2006;296((Suppl. S40)):108–118. doi: 10.1016/j.ijmm.2005.12.012. PubMed DOI

Pangrácová L., Derdáková M., Pekárik L., Hviščová I., Víchová B., Stanko M., Hlavatá H., Peťko B. Ixodes ricinus abundance and its infection with the tick-borne pathogens in urban and suburban areas of Eastern Slovakia. Parasit. Vectors. 2013;6:238. doi: 10.1186/1756-3305-6-238. PubMed DOI PMC

Rusňáková Tarageľová V., Mahríková L., Selyemová D., Václav R., Derdáková M. Natural foci of Borrelia lusitaniae in a mountain region of Central Europe. Ticks Tick Borne Dis. 2016;7:350–356. doi: 10.1016/j.ttbdis.2015.12.006. PubMed DOI

Chvostáč M., Špitalská E., Václav R., Vaculová T., Minichová L., Derdáková M. Seasonal patterns in the prevalence and diversity of tick-borne Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia spp. in an urban temperate forest in south western Slovakia. Int. J. Environ. Res. Public Health. 2018;15:994. doi: 10.3390/ijerph15050994. PubMed DOI PMC

Vaculová T., Derdáková M., Špitalská E., Václav R., Chvostáč M., Rusňáková Tarageľová V. Simultaneous occurrence of Borrelia miyamotoi, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia helvetica in Ixodes ricinus ticks in urban foci in Bratislava, Slovakia. Acta Parasitol. 2019;64:19–30. doi: 10.2478/s11686-018-00004-w. PubMed DOI

Mtierová Z., Derdáková M., Chvostáč M., Didyk Y.M., Mangová B., Rusňáková Tarageľová V., Selyemová D., Šujanová A., Václav R. Local population structure and seasonal variability of Borrelia garinii genotypes in Ixodes ricinus ticks, Slovakia. Int. J. Environ. Res. Public Health. 2020;17:3607. doi: 10.3390/ijerph17103607. PubMed DOI PMC

Subramanian G., Sekeyova Z., Raoult D., Mediannikov O. Multiple tick-associated bacteria in Ixodes ricinus from Slovakia. Ticks Tick Borne Dis. 2012;3:406–410. doi: 10.1016/j.ttbdis.2012.10.001. PubMed DOI

Hamšíková Z., Coipan C., Mahríková L., Minichová L., Sprong H., Kazimírová M. Borrelia miyamotoi and co-infection with Borrelia afzelii in Ixodes ricinus ticks and rodents from Slovakia. Microb. Ecol. 2017;73:1000–1008. doi: 10.1007/s00248-016-0918-2. PubMed DOI

Rosà R., Andreo V., Tagliapietra V., Baráková I., Arnoldi D., Hauffe H.C., Manica M., Rosso F., Blaňarová L., Bona M., et al. Effect of climate and land use on the spatio-temporal variability of tick-borne bacteria in Europe. Int. J. Environ. Res. Public Health. 2018;15:732. doi: 10.3390/ijerph15040732. PubMed DOI PMC

Svitálková Z., Haruštiaková D., Mahríková L., Berthová L., Slovák M., Kocianová E., Kazimírová M. Anaplasma phagocytophilum prevalence in ticks and rodents in an urban and natural habitat in South-Western Slovakia. Parasit. Vectors. 2015;8:276. doi: 10.1186/s13071-015-0880-8. PubMed DOI PMC

Kazimírová M., Hamšíková Z., Kocianová E., Marini G., Mojšová M., Mahríková L., Berthová L., Slovák M., Rosá R. Relative density of host-seeking ticks in different habitat types of south-western Slovakia. Exp. Appl. Acarol. 2016;69:205–224. doi: 10.1007/s10493-016-0025-6. PubMed DOI

Špitalská E., Stanko M., Mošanský L., Kraljik J., Miklisová D., Mahríková L., Bona M., Kazimírová M. Sesonal analysis of Rickettsia species in ticks in an agricultural site of Slovakia. Exp. Appl. Acarol. 2016;68:315–324. doi: 10.1007/s10493-015-9941-0. PubMed DOI

Stanko M., Mošanský L., Fričová J., Čisláková L., Schniererová E. Long-Time Study of Natural Focus and Its Structure in Eastern Slovakia. In: Buczek A., Blaszak C., editors. Stawonogi Roznorodnosc Form i Oddzialywan. Koliber; Lublin, Poland: 2005. pp. 113–119.

Siuda K. Monografie Parazytologiczne. Polskie Towarzystwo Parazytologiczne; Warszawa, Poland: 1993. Kleszcze Polski (Acari: Ixodida). Część II. Systematyka i Rozmieszczenie; pp. 1–375.

Filippova N.A. Fauna of the USSR: Arachnida, Subfamily Ixodinae 4(4) Nauka Publisher; Leningrad, Russia: 1977.

Mydlová L., Svitálková Z., Slovák M., Kazimírová M. Variability in Prevalence and Distribution of Borrelia burgdorferi s.l. In: Kocianová E., Kazimírová M., Lúčanová V., editors. Ixodes ricinus Ticks across Bratislava Forest Park and Adjacent Sites (SW Slovakia). In Zborník z Konferencie III. Labudove dni. Virologický ústav SAV; Bratislava, Slovakia: 2013. pp. 26–29.

Derdáková M., Beati L., Peťko B., Stanko M., Fish D. Genetic variability within Borrelia burgdorferi sensu lato genospecies established by PCR-single-strand conformation polymorphism analysis of the RrfA-RrlB intergenic spacer in Ixodes ricinus ticks from the Czech Republic. Appl. Environ. Microbiol. 2003;69:509–516. doi: 10.1128/AEM.69.1.509-516.2003. PubMed DOI PMC

Rudenko N., Golovchenko M., Grubhoffer L., Oliver J.H., Jr. Borrelia carolinensis sp. nov., a new (14th) member of the Borrelia burgdorferi sensu lato complex from the southeastern region of the United States. J. Clin. Microbiol. 2009;47:134–141. doi: 10.1128/JCM.01183-08. PubMed DOI PMC

Bunikis J., Garpmo U., Tsao J., Berglund J., Fish D., Barbour A.G. Sequence typing reveals extensive strain diversity of the Lyme borreliosis agents Borrelia burgdorferi in North America and Borrelia afzelii in Europe. Microbiology. 2004;150:1741–1755. doi: 10.1099/mic.0.26944-0. PubMed DOI

Clark K., Hendricks A., Burge D. Molecular identification and analysis of Borrelia burgdorferi sensu lato in lizards in the southeastern United States. Appl. Environ. Microbiol. 2005;71:2616–2625. doi: 10.1128/AEM.71.5.2616-2625.2005. PubMed DOI PMC

Alekseev A.N., Dubinina H.V., Van De Pol I., Schouls L.M. Identification of Ehrlichia spp. and Borrelia burgdorferi in Ixodes ticks in the Baltic regions of Russia. J. Clin. Microbiol. 2001;39:2237–2242. doi: 10.1128/JCM.39.6.2237-2242.2001. PubMed DOI PMC

Blazejak K., Raulf M.K., Janecek E., Jordan D., Fingerle V., Strube C. Shifts in Borrelia burgdorferi (s.l.) geno-species infections in Ixodes ricinus over a 10-year surveillance period in the city of Hanover (Germany) and Borrelia miyamotoi-specific Reverse Line Blot detection. Parasit. Vectors. 2018;11:304. doi: 10.1186/s13071-018-2882-9. PubMed DOI PMC

Gern L., Douet V., López Z., Rais O., Cadenas F.M. Diversity of Borrelia genospecies in Ixodes ricinus ticks in a Lyme borreliosis endemic area in Switzerland identified by using new probes for reverse line blotting. Ticks Tick Borne Dis. 2010;1:23–29. doi: 10.1016/j.ttbdis.2009.11.001. PubMed DOI

Springer A., Jordan D., Glass A., Kahl O., Fingerle V., Girl P., Chitimia-Dobler L., Strube C. Borrelia infections in ageing ticks: Relationship with morphometric age ratio in field-collected Ixodes ricinus nymphs. Microorganisms. 2022;10:166. doi: 10.3390/microorganisms10010166. PubMed DOI PMC

Fukunaga M., Okada K., Nakao M., Konishi T., Sato Y. Phylogenetic analysis of Borrelia species based on flagellin gene sequences and its application for molecular typing of Lyme disease borreliae. Int. J. Syst. Bacteriol. 1996;46:898–905. doi: 10.1099/00207713-46-4-898. PubMed DOI

Postic D., Assous M.V., Grimont P.A., Baranton G. 1994. Diversity of Borrelia burgdorferi sensu lato evidenced by restriction fragment length polymorphism of rrf (5S)-rrl (23S) intergenic spacer amplicons. Int. J. Syst. Bacteriol. 1994;44:743–752. doi: 10.1099/00207713-44-4-743. PubMed DOI

Wang G., van Dam A.P., Schwartz I., Dankert J. 1999. Molecular typing of Borrelia burgdorferi sensu lato: Taxonomic, epidemiological, and clinical implications. Clin. Microbiol. Rev. 1999;2:633–653. doi: 10.1128/CMR.12.4.633. PubMed DOI PMC

Hammer O., Harper D.A.T., Ryan P.D. PAST: Paleontological Statistics Software Package for Education and Data Analysis. Palaeontol. Electron. 2001;4:1–9.

Jouda F., Perret J.L., Gern L. Density of questing Ixodes ricinus nymphs and adults infected by Borrelia burgdorferi sensu lato in Switzerland: Spatio-temporal pattern at a regional scale. Vector Borne Zoonotic Dis. 2004;4:23–32. doi: 10.1089/153036604773082960. PubMed DOI

Reye A.L., Hübschen J.M., Sausy A., Muller C.P. Prevalence and seasonality of tick-borne pathogens in questing Ixodes ricinus ticks from Luxembourg. Appl. Environ. Microbiol. 2010;76:2923–2931. doi: 10.1128/AEM.03061-09. PubMed DOI PMC

Venclíková K., Betášová L., Sikutová S., Jedličková P., Hubálek Z., Rudolf I. Human pathogenic borreliae in Ixodes ricinus ticks in natural and urban ecosystem (Czech Republic) Acta Parasitol. 2014;59:717–720. doi: 10.2478/s11686-014-0296-1. PubMed DOI

Hönig V., Svec P., Halas P., Vavruskova Z., Tykalova H., Kilian P., Vetiskova V., Dornakova V., Sterbova J., Simonova Z., et al. Ticks and tick-borne pathogens in South Bohemia (Czech Republic)—Spatial variability in Ixodes ricinus abundance, Borrelia burgdorferi and tick-borne encephalitis virus prevalence. Ticks Tick Borne Dis. 2015;6:559–567. doi: 10.1016/j.ttbdis.2015.04.010. PubMed DOI

Honig V., Carolan H.E., Vavruskova Z., Massire C., Mosel M.R., Crowder C.D., Rounds M.A., Ecker D.J., Ruzek D., Grubhoffer L., et al. Broad-range survey of vector-borne pathogens and tick host identification of Ixodes ricinus from Southern Czech Republic. FEMS Microbiol. Ecol. 2017;93:fix129. doi: 10.1093/femsec/fix129. PubMed DOI PMC

Raileanu C., Moutailler S., Pavel I., Porea D., Mihalca A.D., Savuta G., Vayssier-Taussat M. Borrelia diversity and co-infection with other tick borne pathogens in ticks. Front. Cell. Infect. Microbiol. 2017;7:36. doi: 10.3389/fcimb.2017.00036. PubMed DOI PMC

Kubiak K., Dziekońska-Rynko J., Szymańska H., Kubiak D., Dmitryjuk M., Dzika E. Questing Ixodes ricinus ticks (Acari, Ixodidae) as a vector of Borrelia burgdorferi sensu lato and Borrelia miyamotoi in an urban area of north-eastern Poland. Exp. Appl. Acarol. 2019;78:113–126. doi: 10.1007/s10493-019-00379-z. PubMed DOI

Zubriková D., Wittmann M., Hönig V., Švec P., Víchová B., Essbauer S., Dobler G., Grubhoffer L., Pfister K. Prevalence of tick-borne encephalitis virus and Borrelia burgdorferi sensu lato in Ixodes ricinus ticks in Lower Bavaria and Upper Palatinate, Germany. Ticks Tick Borne Dis. 2020;11:101375. doi: 10.1016/j.ttbdis.2020.101375. PubMed DOI

Hartemink N., van Vliet A.J.H., Gort G., Gassner F., Jacobs F., Fonville M., Takken W., Sprong H. Seasonal patterns and spatial variation of Borrelia burgdorferi (sensu lato) infections in Ixodes ricinus in the Netherlands. Parasit. Vectors. 2021;14:121. doi: 10.1186/s13071-021-04607-7. PubMed DOI PMC

Răileanu C., Silaghi C., Fingerle V., Margos G., Thiel C., Pfister K., Overzier E. Borrelia burgdorferi sensu lato in questing and engorged ticks from different habitat types in southern Germany. Microorganisms. 2021;9:1266. doi: 10.3390/microorganisms9061266. PubMed DOI PMC

Knoll S., Springer A., Hauck D., Schunack B., Pachnicke S., Fingerle V., Strube C. Distribution of Borrelia burgdorferi s.l. and Borrelia miyamotoi in Ixodes tick populations in Northern Germany, co-infections with Rickettsiales and assessment of potential influencing factors. Med. Vet. Entomol. 2021;35:595–606. doi: 10.1111/mve.12537. PubMed DOI

Król N., Obiegala A., Imholt C., Arz C., Schmidt E., Jeske K., Ulrich R.G., Rentería-Solís Z., Jacob J., Pfeffer M. Diversity of Borrelia burgdorferi sensu lato in ticks and small mammals from different habitats. Parasit. Vectors. 2022;15:195. doi: 10.1186/s13071-022-05326-3. PubMed DOI PMC

Rauter C., Hartung T. Prevalence of Borrelia burgdorferi sensu lato genospecies in Ixodes ricinus ticks in Europe: A metaanalysis. Appl. Environ. Microbiol. 2005;71:7203–7216. doi: 10.1128/AEM.71.11.7203-7216.2005. PubMed DOI PMC

Trávniček M., Guryčová D., Halánová M., Kožuch O., Nadzamová D., Miško J. Presence of antibodies to some zoonoses in mouflons (Ovis musimon Pall.) and fallow deer (Dama dama L.) in eastern Slovakia. Vet. Med. 1999;44:215–219.

Štefančíková A., Derdáková M., Lenčáková D., Ivanová R., Stanko M., Čisláková L., Peťko B. Serological and molecular detection of Borrelia burgdorferi sensu lato and Anaplasmataceae in rodents. Folia Microbiol. 2008;53:493–499. doi: 10.1007/s12223-008-0077-z. PubMed DOI

Nadzamová D., Petko B., Stefancíková A., Cisláková L., Stĕpánová G., Seba M. Potential foci of Lyme borreliosis in towns. Cent. Eur. J. Public Health. 2000;8:176–178. PubMed

Peťko B., Štefančíková A., Nadzamová D., Stanko M., Siuda K., Karbowiak G., Winiarczyjk S. 2001. Epizootiological Aspects of Lyme Borreliosis in the City Agglomerations of the Carpathian Regions of Slovakia and Poland and Their Peripheral Parts. In: Buczek A., Błaszak C., editors. Stawonogi Pasożyty i Nosiciele. Wyd. KGM; Lublin, Poland: 2001. pp. 157–164.

Derdakova M., Halanova M., Stanko M., Stefancikova A., Cislakova L., Pet’ko B. Molecular evidence for Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato in Ixodes ricinus ticks from eastern Slovakia. Ann. Agric. Environ. Med. 2003;10:269–271. PubMed

Smetanová K., Schwarzová K., Kocianová E. Detection of Anaplasma phagocytophilum, Coxiella burnetti, Rickettsia spp., and Borrelia burgdorferi s.l. in ticks and wild living animals in western and middle Slovakia. Ann. N. Y. Acad. Sci. 2006;1078:312–315. doi: 10.1196/annals.1374.058. PubMed DOI

Smetanová K., Burri C., Pérez D., Gern L., Kocianová E. Detection and identification of Borrelia burgdorferi sensu lato genospecies in ticks from three different regions in Slovakia. Wien. Klin. Wochenschr. 2007;119:534–537. doi: 10.1007/s00508-007-0851-y. PubMed DOI

Danielová V., Daniel M., Schwarzová L., Materna J., Rudenko N., Golovchenko M., Holubová J., Grubhoffer L., Kilián P. Integration of a tick-borne encephalitis virus and Borrelia burgdorferi sensu lato into mountain ecosystems, following a shift in the altitudinal limit of distribution of their vector, Ixodes ricinus (Krkonose mountains, Czech Republic) Vector Borne Zoonotic Dis. 2010;10:223–230. doi: 10.1089/vbz.2009.0020. PubMed DOI

Picken M.M., Cheng Y., Strle F., Picken R.N. Patient isolates of Borrelia burgdorferi sensu lato with genomic and phenotypic similarities of strain 25015. J. Infect. Dis. 1996;174:1112–1115. doi: 10.1093/infdis/174.5.1112. PubMed DOI

Strle F., Picken R.N., Cheng Y., Cimperman J., Maraspin V., Lotric-Furlan S., Ruzic-Sabljic E., Picken M.M. Clinical findings for patients with Lyme borreliosis caused by Borrelia burgdorferi sensu lato with genomic and phenotypic similarities of strain 25015. Clin. Infect. Dis. 1997;25:273–280. PubMed

Hansford K.M., Wheeler B.W., Tschirren B., Medlock J.M. Questing Ixodes ricinus ticks and Borrelia spp. in urban green space across Europe: A review. Zoonoses Public Health. 2022;69:153–166. doi: 10.1111/zph.12913. PubMed DOI PMC

Hornok S., Meli M.L., Gönczi E., Halász E., Takács N., Farkas R., Hofmann-Lehmann R. Occurrence of ticks and prevalence of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. in three types of urban biotopes: Forests, parks and cemeteries. Ticks Tick Borne Dis. 2014;5:785–789. doi: 10.1016/j.ttbdis.2014.05.010. PubMed DOI

Richtrová E., Míchalová P., Lukavská A., Navrátil J., Kybicová K. Borrelia burgdorferi sensu lato infection in Ixodes ricinus ticks in urban green areas in Prague. Ticks Tick Borne Dis. 2022;13:102053. doi: 10.1016/j.ttbdis.2022.102053. PubMed DOI

Glass A., Springer A., Raulf M.K., Fingerle V., Strube C. 15-year Borrelia prevalence and species distribution monitoring in Ixodes ricinus/inopinatus populations in the city of Hanover, Germany. Ticks Tick Borne Dis. 2023;14:102074. doi: 10.1016/j.ttbdis.2022.102074. PubMed DOI

Rollins R.E., Yeyin Z., Wyczanska M., Alig N., Hepner S., Fingerle V., Margos G., Becker N.S. Spatial variability in prevalence and genospecies distributions of Borrelia burgdorferi sensu lato from ixodid ticks collected in southern Germany. Ticks Tick Borne Dis. 2021;12:101589. doi: 10.1016/j.ttbdis.2020.101589. PubMed DOI

Kowalec M., Szewczyk T., Welc-Faleçiak R., Siński E., Karbowiak G., Bajer A. Ticks and the city—Are there any differences between city parks and natural forests in terms of tick abundance and prevalence of spirochaetes? Parasit. Vectors. 2017;10:573. doi: 10.1186/s13071-017-2391-2. PubMed DOI PMC

Gray J.S., Kahl O., Janetzki C., Stein J. Studies on the ecology of Lyme disease in a deer forest in County Galway, Ireland. J. Med. Entomol. 1992;29:915–920. doi: 10.1093/jmedent/29.6.915. PubMed DOI

Zeman P., Daniel M. Mosaic pattern of Borrelia infection in a continuous population of the tick Ixodes ricinus (Acari: Ixodidae) Exp. Appl. Acarol. 1999;23:327–335. doi: 10.1023/A:1006167001554. PubMed DOI

Prevalence of Borrelia and Rickettsia in Ixodes ricinus from Chosen Urban and Protected Areas in Poland and the Czech Republic