Abstract
Spotted fever group (SFG) Rickettsia are zoonotic and emerging pathogens with considerable impact in public and animal health. Greece is an endemic country of diseases caused by SFG Rickettsia. This work aims to evaluate the prevalence of SFG Rickettsia in ticks collected from domestic hosts including sheep, goats and dogs. Several genetic markers for bacterial genes, such as 16S rRNA, ompA, ompB, atpA, gltA, recA, dnaA and dnaK, were amplified and sequenced to accurately classified the rickettsial pathogens in the ticks. Taxonomy and species classification of the Rickettsia was achieved by combining phylogenetic and in silico digestion analysis of the gene sequences obtained. A total of 187 ticks were collected and classified at the species level as Ixodes gibosus, Dermacentor marginatus, Haemaphysalis parva, H. sulcata, H. punctata, Hyalomma scavatum, Rhipicephalus sanguineus, R. bursa and Rhipicephalus sp. The results showed that 7.5% of ticks were infected with at least one SFG Rickettsia including R. massiliae (n = 3), R. slovaca (n = 5), R. raoultii (n = 1) and R. hoogstraalii (n = 5), collected from sheep (n = 4), goats (n = 5) and dogs (n = 3). Molecular analysis revealed the presence of novel genetic variants of R. hoogstraalii (in H. sulcata and H. parva from goat and sheep) and R. raoultii (in D. marginatus from goat). These results proof the presence of SFG Rickettsia in domestic hosts in Greece, and support the need for continued monitoring, surveillance and further analyses of other hosts and study areas.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ammerman NC, Swanson KI, Anderson JM, Schwartz TR, Seaberg EC, Glass GE, Norris DE (2004) Spotted-fever group rickettsia in Dermacentor variabilis, Maryland. Emerg Infect Dis 10:1478–1481. https://doi.org/10.3201/eid1008.030882
Babalis T, Tselentis Y, Roux V, Psaroulaki A, Raoult D (1994) Isolation and identification of a rickettsial strain related to Rickettsia massiliaein Greek ticks. Am J Trop Med Hyg 50:365–372. https://doi.org/10.4269/ajtmh.1994.50.365
Chaligiannis I, Musella V, Rinaldi L, Cringoli G, de la Fuente J, Papa A, Sotiraki S (2016) Species diversity and spatial distribution of ixodid ticks on small ruminants in Greece. Parasitol Res 115:4673–4680. https://doi.org/10.1007/s00436-016-5259-z
Chisu V, Leulmi H, Masala G, Piredda M, Foxi C, Parola P (2017) Detection of Rickettsia hoogstraalii, Rickettsia helvetica, Rickettsia massiliae, Rickettsia slovaca and Rickettsia aeschlimannii in ticks from Sardinia, Italy. Ticks Tick Borne Dis 8:347–352. https://doi.org/10.1016/j.ttbdis.2016.12.007
Chochlakis D, Ioannou I, Sandalakis V, Dimitriou T, Kassinis N, Papadopoulos B, Tselentis Y, Psaroulaki A (2011) Spotted fever group rickettsiae in ticks in cyprus. Microb Ecol 63:314–323. https://doi.org/10.1007/s00248-011-9926-4
Fernández de Mera IG, Zivkovic Z, Bolaños M, Carranza C, Pérez-Arellano JL, Gutiérrez C, de la Fuente J (2009) Rickettsia massiliae in the Canary islands. Emerg Infect Dis 15:1869–1870. https://doi.org/10.3201/eid1511.090521
Fernández de Mera IG, Ruiz-Fons F, de la Fuente G, Mangold AJ, Gortázar C, de la Fuente J (2013) Spotted fever group rickettsiae in questing ticks, central Spain. Emerg Infect Dis 19:1163–1165. https://doi.org/10.3201/eid1907.130005
Galanakis E, Bitsori M (2011) Rickettsioses in children: a clinical approach. Adv Exp Med Biol 719:144–162. https://doi.org/10.1007/978-1-4614-0204-6_13
Germanakis A, Psaroulaki A, Gikas A, Tselentis Y (2006) Mediterranean spotted fever in crete, Greece: clinical and therapeutic data of 15 consecutive patients. Ann New York Acad Sci 1078:263–269. https://doi.org/10.1196/annals.1374.049
Germanakis A, Chochlakis D, Angelakis E, Tselentis Y, Psaroulaki A (2013) Rickettsia aeschlimannii infection in a man, Greece. Emerg Infect Dis 19:1176–1177. https://doi.org/10.3201/eid1907.130232
Germanakis A, Chochlakis D, Angelakis E, Tselentis Y, Psaroulaki A (2014) Skin lesions and inoculation eschars at the tick bite site in spotted fever group rickettsioses: experience from a patient series in eastern Crete, Greece. Dermatology 228:332–337. https://doi.org/10.1159/000360525
Gortázar C, Reperant LA, Kuiken T, de la Fuente J, Boadella M, Martinez-Lopez B, Ruiz-Fons F, Estrada-Peña A, Drosten C, Medley G, Ostfeld R, Peterson T, VerCauteren KC, Menge C, Artois M, Schultsz C, Delahay R, Serra-Cobo J, Poulin R, Keck F, Aguirre AA, Henttonen H, Dobson AP, Kutz S, Lubroth J, Mysterud A (2014) Crossing the interspecies barrier: opening the door to zoonotic pathogens. PLoS Pathog 10:1–5. https://doi.org/10.1371/journal.ppat.1004129
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 8:33–50. https://doi.org/10.1586/eri.09.118
Kachrimanidou M, Souliou E, Pavlidou V, Antoniadis A, Papa A (2010) First detection of Rickettsia slovaca in Greece. Exp Appl Acarol 50:93–96. https://doi.org/10.1007/s10493-009-9283-x
Katoh K, Standley D (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30:772–780. https://doi.org/10.1093/molbev/mst010
Mediannikov O, Matsumoto K, Samoylenko I, Drancourt M, Roux V, Rydkina E, Davoust B, Tarasevich I, Brouqui P, Fournier PE (2008) Rickettsia raoultii sp. nov., a spotted fever group rickettsia associated with Dermacentor ticks in Europe and Russia. Int J Syst Evol Microbiol 58:1635–1639. https://doi.org/10.1099/ijs.0.64952-0
Merhej V, Raoult D (2011) Rickettsial evolution in the light of comparative genomics. Biol Rev 86:379–405. https://doi.org/10.1111/j.1469-185X.2010.00151.x
Merhej V, Angelakis E, Socolovschi C, Raoult D (2014) Genotyping, evolution and epidemiological findings of Rickettsia species. Infect Genet Evol 25:122–137. https://doi.org/10.1016/j.meegid.2014.03.014
Oteo JA, Portillo A (2012) Tick-borne rickettsioses in Europe. Ticks Tick Borne Dis 3:270–277. https://doi.org/10.1016/j.ttbdis.2012.10.035
Papa A, Dalla V, Petala A, Maltezou HC, Maltezos E (2010) Fatal Mediterranean spotted fever in Greece. Clin Microbiol Infect 16:589–592. https://doi.org/10.1111/j.1469-0691.2009.02910.x
Papa A, Chaligiannis I, Xanthopoulou K, Papaioakim M, Papanastasiou S, Sotiraki S (2011) Ticks parasitizing humans in Greece. Vector Borne Zoonotic Dis 11:539–542. https://doi.org/10.1089/vbz.2010.0036
Papa A, Xanthopoulou K, Kotriotsiou T, Papaioakim M, Sotiraki S, Chaligiannis I, Maltezos E (2016) Rickettsia species in human-parasitizing ticks in Greece. Trans R Soc Trop Med Hyg 110:299–304. https://doi.org/10.1093/trstmh/trw022
Papa A, Tsioka K, Kontana A, Papadopoulos C, Giadinis N (2017) Bacterial pathogens and endosymbionts in ticks. Ticks Tick Borne Dis 8:31–35. https://doi.org/10.1016/j.ttbdis.2016.09.011
Pérez-Osorio CE, Zavala-Velázquez JE, Arias JJ, Zavala-Castro JE (2008) Rickettsia felis as emergent global threat for humans. Emerg Infect Dis 14:1019–1023. https://doi.org/10.3201/eid1407.071656
Psaroulaki A, Spyridaki I, Ioannidis A, Babalis T, Gikas A, Tselentis Y (2003) First isolation and identification of Rickettsia conorii from ticks collected in the region of Fokida in Central Greece. J Clin Microbiol 41:3317–3319. https://doi.org/10.1128/JCM.41.7.3317-3319.2003
Psaroulaki A, Ragiadakou D, Kouris G, Papadopoulos B, Chaniotis B, Tselentis Y (2006) Ticks, tick-borne rickettsiae, and Coxiella burnetii in the Greek Island of Cephalonia. Ann N Y Acad Sci 1078:389–399. https://doi.org/10.1196/annals.1374.077
Roux V, Fournier P, Raoult D (1996) Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol 34:2058–2065
Roux V, Rydkina E, Eremeeva M, Raoult D (1997) Citrate synthase gene comparison, a new tool for phylogenetic analysis and its application for the Rickettsiae. Int J Syst Bacteriol 47:252–262. https://doi.org/10.1099/00207713-47-2-252
Ruiz-Fons F, Fernández de Mera IG, Acevedo P, Höfle U, Vicente J, de la Fuente J (2006) Ticks (Acari: ixodidae) parasitizing Iberian red deer (Cervus elaphus hispanicus) and European wild boar (Sus scrofa) from Spain: geographical and temporal distribution. Vet Parasitol 140:133–142. https://doi.org/10.1016/j.vetpar.2006.03.033
Simser JA, Palmer AT, Fingerle V, Wilske B, Kurtti TJ, Munderloh UG (2002) Rickettsia monacensis sp. nov., a spotted fever group Rickettsia, from Ticks (Ixodes ricinus) collected in a European City Park. J Appl Environ Microbiol 68:4559–4566. https://doi.org/10.1128/AEM.68.9.4559-4566.2002
Tamura K (1992) Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+C-content biases. Mol Biol Evol 9:678–687
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Tavaré S (1986) Some probabilistic and statistical problems in the analysis of dna sequences. lectures on mathematics in the life sciences. Am Math Soc 17:57–86
Uchiyama T (2012) Tropism and pathogenicity of rickettsiae. Front Microbiol 3:230. https://doi.org/10.3389/fmicb.2012.00230
Weisburg WG, Dobson ME, Samuel JE, Dasch GA, Mallavia L, Baca O, Mandelco L, Sechrest JE, Weiss E, Woese CR (1989) Phylogenetic diversity of the rickettsiae. J Bacteriol 171:4302–4306
ZeringótaV Maturano R, Luz HR, Oliveira T, Daemon E, Horacio JL, McIntosh D (2017) Molecular detection of Rickettsia rhipicephali and other spotted fever group Rickettsia species in Amblyomma ticks infesting wild birds in the state of Minas Gerais, Brazil. Ticks Tick Borne Dis 8:8189. https://doi.org/10.1016/j.ttbdis.2016.10.001
Acknowledgements
The authors gratefully acknowledge all the veterinarians who contributed in the study by providing the samples. This study was financially supported by the State Scholarship Foundation IKY, NSRF 2007–2013. IGFM is funded from the Own Research Program by University of Castilla-La Mancha.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Moraga-Fernández, A., Chaligiannis, Ι., Cabezas-Cruz, A. et al. Molecular identification of spotted fever group Rickettsia in ticks collected from dogs and small ruminants in Greece. Exp Appl Acarol 78, 421–430 (2019). https://doi.org/10.1007/s10493-019-00392-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10493-019-00392-2