Experimental and Applied Acarology

, Volume 66, Issue 3, pp 443–452 | Cite as

Molecular detection of Rickettsia, Borrelia, and Babesia species in Ixodes ricinus sampled in northeastern, central, and insular areas of Italy

  • Lyda R. CastroEmail author
  • Simona Gabrielli
  • Albertina Iori
  • Gabriella Cancrini


The aim of the present study was to provide insight into the diversity of tick-borne pathogens circulating in Italy, carried/transmitted by Ixodes ricinus, one of the most abundant tick species in the country. A total of 447 specimens sampled in five areas of northeastern, central and insular Italy were analysed by polymerase chain reaction and sequencing for the presence of rickettsiae, borreliae and babesiae. Several rickettsial species of the spotted fever group of zoonotic concern and other zoonotic pathogens were found, such as Borrelia burgdorferi s.s., Borrelia afzelii, Borrelia garinii, and Babesia venatorum. These findings confirm a wide distribution of tick-borne bacterial and protozoan species in Italy, and highlight the sanitary importance of I. ricinus, often recorded as feeding on humans.


Rickettsia spp. Borrelia spp. Babesia venatorum Italy PCR 



Authors are grateful to Mrs G. Croce and Mr A. Giacomi for the excellent technical assistance and to Prof. D. Piergili Fioretti and P. Calderini for providing some tick samples. This work was financially supported by the Italian Ministry of University and Research (PRIN, Prot. 2004075927_002). We also thank the Erasmus Mundus program of the EulaLinks consortium for the fellowship given to Lyda Castro.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Balmelli T, Piffaretti JC (1995) Association between different clinical manifestations of Lyme disease and different species of Borrelia burgdorferi sensu lato. Res Microbiol 146:329–340CrossRefPubMedGoogle Scholar
  2. Bonnet S, Jouglin M, L’Hostis M, Chauvin A (2007) Babesia sp. EU1 from roe deer and transmission within Ixodes ricinus. Emerg Infect Dis 13:1208–1210CrossRefPubMedCentralPubMedGoogle Scholar
  3. Cancrini G, Calderini P, Gabrielli S, Scaramozzino P, Iori A (2007) Babesiosis and theileriosis: first evidence in domestic and wild animals in Lazio region (Central Italy). Parassitologia 49:80Google Scholar
  4. Casati S, Sager H, Gern L, Piffaretti JC (2006) Presence of potentially pathogenic Babesia sp. for human in Ixodes ricinus in Switzerland. Ann Agric Environ Med 13:65–70PubMedGoogle Scholar
  5. Cassini R, Zanutto S, Frangipane di Regalbono A, Gabrielli S, Calderini P, Moretti A, Tampieri MP, Pietrobelli M (2009) Canine piroplasmosis in Italy: epidemiological aspects in vertebrate and invertebrate hosts. Vet Parasitol 165:30–35CrossRefPubMedGoogle Scholar
  6. Cassini R, Bonoli C, Montarsi F, Tessarin C, Marcer F, Galuppi R (2010) Detection of Babesia EU1 in Ixodes ricinus ticks in northern Italy. Vet Parasitol 171:151–154CrossRefPubMedGoogle Scholar
  7. Cinco M, Barbone F, Grazia Ciufolini M, Mascioli M, Anguero-Rosenfeld M, Stefanel P et al (2004) Seroprevalence of tick-borne infections in forestry rangers from Northeastern Italy. Clin Microbiol Infect 10:1056–1061. doi: 10.1111/j.1469-0691.2004.01026.x CrossRefPubMedGoogle Scholar
  8. Cringoli G, Otranto D, Testini G, Buono V, Di Giulio G, Traversa D, Lia R, Rinaldi L, Veneziano V, Piccini V (2002) Epidemiology of bovine tick-borne diseases in southern Italy. Vet Res 33:421–428CrossRefPubMedGoogle Scholar
  9. Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: more models, new heuristics and parallel computing. Nat Methods 9:772CrossRefPubMedGoogle Scholar
  10. Duh D, Petrovec M, Avsic-Zupanc T (2001) Diversity of Babesia infecting European sheep ticks (Ixodes ricinus). J Clin Microbiol 39:3395–3397CrossRefPubMedCentralPubMedGoogle Scholar
  11. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucl Acids Res 32:1792–1797CrossRefPubMedCentralPubMedGoogle Scholar
  12. Favia G, Cancrini G, Carfì A, Grazioli D, Lillini E, Iori A (2001) Molecular identification of Borrelia valaisiana and HGE-like Ehrlichia in Ixodes ricinus ticks sampled in North Eastern Italy. Parassitologia 43:143–146PubMedGoogle Scholar
  13. Gabrielli S, Calderini P, Cassini R, Galuppi R, Tampieri MP, Pietrobelli M, Cancrini G (2014) Human exposure to piroplasms in central and northern Italy. Vet Ital 50(1):41–47PubMedGoogle Scholar
  14. Hengge UR, Tannapfel A, Tyring SK, Erbel R, Arendt G, Ruzicka T (2003) Lyme borreliosis. Lancet Infect Dis 3:489–500. doi: 10.1016/S1473-3099(03)00722-9 CrossRefPubMedGoogle Scholar
  15. Herwaldt BL, Cacció S, Gherlinzoni F, Aspock H, Slemenda SB, Piccaluga P, Martinelli G, Edelhofer R, Hollenstein U, Poletti G, Pampiglione S, Loschenberger K, Tura S, Pieniazek NJ (2003) Molecular characterization of a non-Babesia divergens organism causing zoonotic babesiosis in Europe. Emerg Infect Dis 9:942–948CrossRefPubMedGoogle Scholar
  16. Heyman P, Cochez C, Hofhuis A, van der Giessen J, Sprong H, Porter SR, Losson B, Saegerman C, Donoso-Mantke O, Niedrig M, Papa A (2010) A clear and present danger: tick-borne diseases in Europe. Expert Rev Anti Infect Ther 8:33–50CrossRefPubMedGoogle Scholar
  17. Hilpertshauser H, Deplazes P, Schnyder M, Gern L, Mathis A (2006) Babesia spp. identified by PCR in ticks collected from domestic and wild ruminants in southern Switzerland Appl. Environ Microbiol 72:6503–6507CrossRefGoogle Scholar
  18. Huelsenbeck JP, Ronquist F (2001) MRBAYES: Bayesian inference of phylogeny. Bioinformatics 17:754–755CrossRefPubMedGoogle Scholar
  19. Iori A, Grazioli D, Gabrielli S, Cancrini G (2002) Molecular detection of HGE-like Ehrlichia and four genomic groups of Borrelia burgdorferi s.l. in Ixodes ricinus ticks sampled in Belluno-Feltre areas. Parassitologia 44(Suppl. 1):87Google Scholar
  20. Iori A, Di Giulio A, De Felice S (2005) Zecche d’Italia. In: Crignoli G (ed) Zecche. Rolando, Napoli, pp 1–199Google Scholar
  21. Iori A, Gabrielli S, Calderini P, Moretti A, Pietrobelli M, Tampieri MP, Galuppi R, Cancrini G (2010) Tick reservoirs for piroplasms in central and northern Italy. Vet Parasitol 170:291–296CrossRefPubMedGoogle Scholar
  22. Lindgren E, Jaenson TGT (2006) Lyme borreliosis in Europe: influences of climate and climate change, epidemiology, ecology and adaptation measures. WHO Regional Office for Europe, CopenhagenGoogle Scholar
  23. Manilla G (1998) Fauna d’Italia, Acari Ixodida. Edizioni Calderini, Bologna 280 ppGoogle Scholar
  24. Mannelli A, Cerri D, Buffrini L, Rossi S, Rosati S, Arata T, Innocenti M, Grignolo MC, Bianchi G, Iori A, Tolari F (1999) Low risk of Lyme borreliosis in a protected area on the Tyrrhenian coast, in central Italy. Eur J Epidemiol 15:371–377CrossRefPubMedGoogle Scholar
  25. Marconi RT, Garon CF (1992) Development of polymerase chain reaction primer sets for diagnosis of Lyme disease and for species-specific identification of Lyme disease isolates by 16S rRNA signature nucleotide analysis. Clin Microbiol 30(11):2830–2834Google Scholar
  26. Moretti A, Mangili V, Salvatori R, Maresca C, Scoccia E, Torina A, Moretta I, Gabrielli S, Tampieri MP, Pietrobelli M (2009) Prevalence and diagnosis of Babesia and Theileria infections in horses in Italy: a preliminary study. Vet J. doi: 10.1016/j.tvjl.2009.03.021 PubMedGoogle Scholar
  27. Oteo J, Portillo A (2012) Tick-borne rickettsioses in Europe. Ticks Tick Borne Dis 3:270–277CrossRefGoogle Scholar
  28. Pajoro M, Pistone D, Epis S, Sassera D, Montagna M, Luzzago C, Lanfanchi P, Vigano R, Cerutti MC, Vicari N, Fabbi M, Bandi C (2010) A survey for Bacterial pathogens associated with Ixodes ricinus in north-western Italy. SOIPA XXVI Abstracts. Parassitologia 52(1–2):178Google Scholar
  29. Parola P, Raoult D (2001) Ticks and tickborne bacterial diseases in humans: an emerging infectious threat. Clin Infect Dis 32:897–928CrossRefPubMedGoogle Scholar
  30. Parola P, Paddock CD, Raoult D (2005) Tick-borne rickettsioses around the world: emerging diseases challenging old concepts. Clin Microbiol Rev 18:719–756CrossRefPubMedCentralPubMedGoogle Scholar
  31. Parola P, Paddock CD, Socolovschi C, Labruna MB, Mediannikov O, Kernif T et al (2013) Update on tick-borne rickettsioses around the world: a geographic approach. Clin Microbiol Rev 26:657–702. doi: 10.1128/CMR.00032-13 CrossRefPubMedCentralPubMedGoogle Scholar
  32. Pietrobelli M, Cancrini G, Moretti A, Tampieri MP (2007) Animal babesiosis: an emerging zoonosis also in Italy? Parassitologia 49:33–38PubMedGoogle Scholar
  33. Pistone D, Pajoro M, Fabbi M, Varari N, Marone P, Genchi C, Novati S, Sassera D, Epis S, Bandi C (2010) Lyme Borreliosis, Po River Vallley, Italy. Emerg Infect Dis 16(8):1289–1291CrossRefPubMedCentralPubMedGoogle Scholar
  34. Raoult D (2004) A new tick-borne rickettsiosis in the USA. Clin Infect Dis 38:812–813CrossRefPubMedGoogle Scholar
  35. Raoult D, Fournier PE, Fenollar F et al (2001) Rickettsia africae, a tick-borne pathogen in travelers to sub-Saharan Africa. N Engl J Med 344:1504–1510CrossRefPubMedGoogle Scholar
  36. Rauter C, Hartung T (2005) Prevalence of Borrelia burdorferi Sensu lato genospecies in Ixodes ricinus ticks in Europe: a metaanalysis. Appl Envirn Microbiol 71(11):7203–7216CrossRefGoogle Scholar
  37. Ravagnan S, Montarsi F, Porcellato R, Ariani P, Cassini R, Granato A, Capelli G (2010) Borrelia burgdorferi genospecies in Ixodes ricinus ticks form Lyme borreliosis endemic area of Italy. SOIPA XXVI Abstracts. Parassitologia 52(1–2):180Google Scholar
  38. Regnery RL, Spruill CL, Plikaytis BD (1991) Genotypic identification of rickettsiae and estimation of interspecies sequence divergence for portions of two rickettsial genes. J Bacteriol 173:1576–1589PubMedCentralPubMedGoogle Scholar
  39. Ronquist F, Huelsenbeck J (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574CrossRefPubMedGoogle Scholar
  40. Skotarczak B, Wodecka B, Hermanowska-Szpakowicz T (2002) Sensitivity of PCR method for detection of DNA of Borrelia burgdorferi sensu lato in different isolates. Prz Epidemiol 56:73–79Google Scholar
  41. Sparagano OAE, Allsopp MTEP, Mank RA, Rijpkema SGT, Figueroa JV, Jongejan F (1999) Molecular detection of pathogen DNA in ticks (Acari: Ixodidae): a review. Exp Appl Acarol 23:929–960CrossRefPubMedGoogle Scholar
  42. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefPubMedCentralPubMedGoogle Scholar
  43. Torina A, Caracappa S (2007) Babesiosis in Italy: an overview. Parassitologia 49:23–28PubMedGoogle Scholar
  44. Van Dam HK, Vos K, Widjojokusumo A, De Jongh BM, Spanjaard L, Ramselaar ACP, Kramer MD, Dankert J (1993) Different genospecies of Borrelia burgdorferi are associated with distinct clinical manifestations of Lyme borreliosis. Clin Infect Dis 17:708–717CrossRefPubMedGoogle Scholar
  45. Veronesi F, Rapicetta S, Gabrielli S, Iori A, Diaferia M, Piergili Fioretti D (2006) Preliminary data on the distriburion of Rickettsia rickettsia and Rickettsia conorii in Umbria region. Parassitologia 48:328Google Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Lyda R. Castro
    • 1
    Email author
  • Simona Gabrielli
    • 2
  • Albertina Iori
    • 2
  • Gabriella Cancrini
    • 2
  1. 1.Grupo de Investigación Evolución, Sistemática y Ecología Molecular, Lab 2, IntropicUniversidad del MagdalenaSanta MartaColombia
  2. 2.Dip. di Sanità Pubblica e Malattie InfettiveUniversità “Sapienza”RomeItaly

Personalised recommendations