Abstract
Tortoises of the genus Testudo are the main hosts of Hyalomma aegyptium ticks. This species serves as a vector of several zoonotic pathogens. Therefore, the present study aimed to investigate the presence of four pathogens associated with H. aegyptium ticks obtained from tortoises from Tunisia. Conventional, multiplex and nested PCRs were used for Aanaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii and Babesia spp. screening. The molecular analyses revealed the presence of A. phagocytophilum and Babesia spp. None of the ticks, were infected by E. canis or C. burnetii species. Co-infection was detected in four ticks. As a conclusion, this is the first detection of A. phagocytophilum and Babesia spp. in H. aegyptium ticks collected from tortoises, in Tunisia. Thus, considering these results, the spur-thighed tortoise constitute a potential host of H. aegyptium which plays an important role in the transmission of pathogenic agents affecting both human and animals. In term of public health, a strict control and surveillance should be carried to reduce the circulation of such pathogens between different hosts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akveran GA, Karasartova D, Keskin A et al (2020) Bacterial and protozoan agents found in Hyalomma aegyptium (L., 1758) (Ixodida: Ixodidae) collected from Testudo graeca L., 1758 (Reptilia: Testudines) in Corum Province of Turkey. Ticks Tick Borne Dis. https://doi.org/10.1016/j.ttbdis.2020.101458
Alsaleh A, Pellerin JL, Rodolakis A et al (2011) Detection of Coxiella burnetii, the agent of Q fever, in oviducts and uterine flushing media and in genital tract tissues of the non pregnant goat. Comp Immunol Microbiol Infect Dis 34:355–360. https://doi.org/10.1016/j.cimid.2011.05.002
Altschul SF, Gish W, Miller W et al (1990) Basic local alignment search tool. J Mol Biol 215:403–410. https://doi.org/10.1016/S0022-2836(05)80360-2
Ancelle T (2017) Statistique épidémiologique, 4 eme. MALOINE
Angelakis E, Mediannikov O, Socolovschi C et al (2014) Coxiella burnetii-positive PCR in febrile patients in rural and urban Africa. Int J Infect Dis IJID off Publ Int Soc Infect Dis 28:107–110. https://doi.org/10.1016/j.ijid.2014.05.029
Apanaskevich DA (2003) Towards a diagnostic view of Hyalomma (Hyalomma) aegyptium (Acari, Ixodidae). Parazitologiia 37:47–59
Apanaskevich DA (2004) Host-parasite relationships of the genus Hyalomma Koch, 1844 (Acari, Ixodidae) and their connection with microevolutionary process. Parazitol 38:515–523
Aydin L (2000) Distributions and species of ticks on ruminants in Southern Marmara Region. Türkiye Parazitoloji Derg 24:194–200
Aysul N, Kar S, Yilmazer N et al (2010) Prevalence of Hyalomma aegyptium (Lineaus, 1758) in tortoises (Testudo graeca) in Thrace. Pendik Vet Mikrobiyoloji Derg. 37(1):53–56
Baráková I, Derdáková M, Selyemová D et al (2018) Tick-borne pathogens and their reservoir hosts in northern Italy. Ticks Tick Borne Dis 9:164–170. https://doi.org/10.1016/j.ttbdis.2017.08.012
Barkallah M, Gharbi Y, Ben HA et al (2014) Survey of infectious etiologies of bovine abortion during Mid- to Late Gestation in Dairy Herds. PLoS ONE. https://doi.org/10.1371/journal.pone.0091549
Barlough JE, Madigan JE, DeRock E, Bigornia L (1996) Nested polymerase chain reaction for detection of Ehrlichia equi genomic DNA in horses and ticks (Ixodes pacificus). Vet Parasitol 63:319–329. https://doi.org/10.1016/0304-4017(95)00904-3
Barradas PF, Lima C, Cardoso L et al (2021) Molecular evidence of Hemolivia mauritanica, Ehrlichia spp. and the endosymbiont Candidatus midichloria mitochondrii in Hyalomma aegyptium infesting Testudo graeca tortoises from Doha. Qatar Animals 11:1–8. https://doi.org/10.3390/ani11010030
Bellazreg F, Kaabia N, Hachfi W et al (2009) Acute Q fever in hospitalised patients in Central Tunisia: report of 21 cases. Clin Microbiol Infect off Publ Eur Soc Clin Microbiol Infect Dis 15(Suppl 2):138–139. https://doi.org/10.1111/j.1469-0691.2008.02138.x
Ben Said M, Belkahia H, Messadi L (2018) Anaplasma spp. in North Africa: a review on molecular epidemiology, associated risk factors and genetic characteristics. Ticks Tick Borne Dis 9:543–555
Bitam I, Kernif T, Harrat Z et al (2009) First detection of Rickettsia aeschlimannii in Hyalomma aegyptium from Algeria. Clin Microbiol Infect 15(Suppl 2):253–254. https://doi.org/10.1111/j.1469-0691.2008.02274.x
Brianti E, Dantas-Torres F, Giannetto S et al (2010) Risk for the introduction of exotic ticks and pathogens into Italy through the illegal importation of tortoises, Testudo graeca. Med Vet Entomol 24:336–339. https://doi.org/10.1111/j.1365-2915.2010.00874.x
Bursali A, Tekin S, Orhan M et al (2010) Ixodid ticks (Acari: Ixodidae) infesting humans in Tokat Province of Turkey: species diversity and seasonal activity. J Vector Ecol 35:180–186. https://doi.org/10.1111/j.1948-7134.2010.00075.x
Corales JM, ariell. I, Viloria V V., Venturina VM, Mingala CN, (2014) The prevalence of Ehrlichia canis, Anaplasma platys and Babesia spp. in dogs in Nueva Ecija, Philippines based on multiplex polymerase chain reaction (mPCR) assay. Ann Parasitol 60:267–272
Dumler JS, Choi KS, Garcia-Garcia JC et al (2005) Human granulocytic anaplasmosis and Anaplasma phagocytophilum. Emerg Infect Dis 11:1828–1834
Ereqat S, Nasereddin A, Al-Jawabreh A et al (2016) Molecular detection and identification of spotted fever group Rickettsiae in ticks collected from the West Bank. Palest T Plos Negl Trop Dis 10:e0004348. https://doi.org/10.1371/journal.pntd.0004348
Fares W, Dachraoui K, Najjar C et al (2019) Absence of Crimean-Congo haemorrhagic fever virus in the tick Hyalomma aegyptium parasitizing the spur-thighed tortoise (Testudo graeca) in Tunisia. Parasite. https://doi.org/10.1051/parasite/2019036
Gharbi M, Rjeibi MR, Rouatbi M et al (2015) Infestation of the spur-thighed tortoise (Testudo graeca) by Hyalomma aegyptium in Tunisia. Ticks Tick Borne Dis. https://doi.org/10.1016/j.ttbdis.2015.02.009
Güner ES, Watanabe M, Hashimoto N et al (2004) Borrelia turcica sp. nov., isolated from the hard tick Hyalomma aegyptium in Turkey. Int J Syst Evol Microbiol 54:1649–1652. https://doi.org/10.1099/ijs.0.03050-0
Han R, Yang J, Niu Q et al (2018) Molecular prevalence of spotted fever group Rickettsiae in ticks from Qinghai Province, northwestern China. Infect Genet Evol 57:1–7. https://doi.org/10.1016/j.meegid.2017.10.025
Jin H, Wei F, Liu Q, Qian J (2012) Epidemiology and control of human granulocytic anaplasmosis: a systematic review. Vector-Borne Zoonotic Dis 12:269–274
Jonsson NN, Bock RE, Jorgensen WK (2008) Productivity and health effects of anaplasmosis and babesiosis on Bos indicus cattle and their crosses, and the effects of differing intensity of tick control in Australia. Vet Parasitol 155:1–9
Kar S, Yılmazer N, Midilli K et al (2011) (2011) Presence of the zoonotic Borrelia burgdorferi sl. and Rickettsia spp. in the Ticks from Wild Tortoises and Hedgehogs. J Marmara Univ Inst Health Sci 1(3):166–170
Kar S, Rodriguez SE, Akyildiz G et al (2020) Crimean-Congo hemorrhagic fever virus in tortoises and Hyalomma aegyptium ticks in East Thrace, Turkey: potential of a cryptic transmission cycle. Parasit Vectors 13:201. https://doi.org/10.1186/s13071-020-04074-6
Kawahara M, Rikihisa Y, Lin Q et al (2006) Novel genetic variants of Anaplasma phagocytophilum, Anaplasma bovis, Anaplasma centrale, and a novel Ehrlichia sp. in wild deer and ticks on two major islands in Japan. Appl Environ Microbiol 72:1102–1109. https://doi.org/10.1128/AEM.72.2.1102-1109.2006
Köseoğlu AE, Can H, Güvendi M et al (2021) Molecular investigation of bacterial and protozoal pathogens in ticks collected from different hosts in Turkey. Parasit Vectors 14:270. https://doi.org/10.1186/s13071-021-04779-2
Kumar S, Stecher G, Li M et al (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35:1547–1549. https://doi.org/10.1093/molbev/msy096
Liu Z, Ma M, Wang Z et al (2012) Molecular survey and genetic identification of Anaplasma species in goats from central and southern China. Appl Environ Microbiol 78:464–470. https://doi.org/10.1128/AEM.06848-11
Manoj RRS, Mendoza-Roldan JA, Latrofa MS et al (2021) Molecular detection of zoonotic blood pathogens in ticks from illegally imported turtles in Italy. Acta Trop 222:106038. https://doi.org/10.1016/J.ACTATROPICA.2021.106038
Najjar C, Kaabi B, Younsi H et al (2020) Ticks parasitizing the spur-thighed tortoise (Testudo graeca) population of Tunisia. J Wildl Dis 56:815–822. https://doi.org/10.7589/2019-09-219
Nieto NC, Foley JE, Bettaso J, Lane RS (2009) Reptile infection with Anaplasma phagocytophilum, the causative agent of granulocytic Anaplasmosis. J Parasitol 95:1165–1170. https://doi.org/10.1645/GE-1983.1
Norte AC, Harris DJ, Silveira D et al (2021) Diversity of microorganisms in Hyalomma aegyptium collected from spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Transbound Emerg Dis. https://doi.org/10.1111/TBED.14188
Nowak M, Cieniuch S, Stańczak J, Siuda K (2010) Detection of Anaplasma phagocytophilum in Amblyomma flavomaculatum ticks (Acari: Ixodidae) collected from lizard Varanus exanthematicus imported to Poland. Exp Appl Acarol 51:363–371. https://doi.org/10.1007/s10493-009-9332-5
Orkun Ö, Emir H (2020) Identification of tick-borne pathogens in ticks collected from wild animals in Turkey. Parasitol Res 119:3083–3091. https://doi.org/10.1007/S00436-020-06812-2
Paperna I, Kremer-Mecabell T, Finkelman S (2002) Hepatozoon kisrae n. sp. infecting the lizard Agama stellio is transmitted by the tick Hyalomma cf. aegyptium.. Parasite 9:17–27. https://doi.org/10.1051/parasite/200209117
Paștiu AI, Matei IA, Mihalca AD et al (2012) (2012) Zoonotic pathogens associated with Hyalomma aegyptium in endangered tortoises: evidence for host-switching behaviour in ticks? Parasites Vectors 51(5):1–6. https://doi.org/10.1186/1756-3305-5-301
Roest HJ, Bossers A, Rebel JMJ (2013) Q fever diagnosis and control in domestic ruminants. Dev Biol (basel) 135:183–189. https://doi.org/10.1159/000188081
Rybářová M, Honsová M, Papoušek I, Široký P (2017) Variability of species of Babesia Starcovici, 1893 in three sympatric ticks (Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna) at the edge of Pannonia in the Czech Republic and Slovakia. Folia Parasitol (praha) 64:28. https://doi.org/10.14411/fp.2017.028
Schwartz D (1993) Méthodes statistiques à l’usage des médecins et des biologistes, 4th edn. Médecine Sciences Publications, Paris
Silaghi C, Woll D, Hamel D et al (2012) Babesia spp and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents—Analyzing the host-pathogen-vector interface in a metropolitan area. Parasites Vectors 5:191. https://doi.org/10.1186/1756-3305-5-191
Silaghi C, Nieder M, Sauter-Louis C et al (2018) Epidemiology, genetic variants and clinical course of natural infections with Anaplasma phagocytophilum in a dairy cattle herd. Parasit Vectors. https://doi.org/10.1186/s13071-017-2570-1
Široký P, Mikulíček P, Jandzík D et al (2009) Co-distribution pattern of a haemogregarine Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) and its vector Hyalomma aegyptium (Metastigmata: Ixodidae). J Parasitol 95:728–733. https://doi.org/10.1645/GE-1842.1
Široký P, Bělohlávek T, Papoušek I et al (2014) Hidden threat of tortoise ticks: high prevalence of Crimean-Congo haemorrhagic fever virus in ticks Hyalomma aegyptium in the Middle East. Parasit Vectors 7:101. https://doi.org/10.1186/1756-3305-7-101
Siroký P, Erhart J, Petrželková KJ, Kamler M (2011) Life cycle of tortoise tick Hyalomma aegyptium under laboratory conditions. Exp Appl Acarol 54:277–284. https://doi.org/10.1007/s10493-011-9442-8
Smrdel KS, Serdt M, Duh D et al (2010) Anaplasma phagocytophilum in ticks in Slovenia. Parasit Vectors 3:102. https://doi.org/10.1186/1756-3305-3-102
Stańczak J, Gabre RM, Kruminis-Łozowska W et al (2004) Ixodes ricinus as a vector of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti in urban and suburban forests. Ann Agric Environ Med 11:109–114
Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 10:512–526. https://doi.org/10.1093/OXFORDJOURNALS.MOLBEV.A040023
Toma L, Di Luca M, Mancini F et al (2017) Molecular characterization of Babesia and Theileria species in ticks collected in the outskirt of Monte Romano, Lazio Region, Central Italy. Ann Ist Super Sanita 53:30–34. https://doi.org/10.4415/ANN_17_01_07
Wasfi F, Dowall S, Ghabbari T et al (2016) Sero-epidemiological survey of Crimean-Congo hemorrhagic fever virus in Tunisia. Parasite. https://doi.org/10.1051/parasite/2016010
Acknowledgements
We are grateful to Mr Sassi Limam and Mr Mohamed Jedidi for their support.
Funding
This work was co-funded by the laboratory of «Laboratoire d'épidémiologie d'infections enzootiques des herbivores en Tunisie: application à la lutte» (Ministère de l'Enseignement Supérieur et de la Recherche Scientifique, Tunisia) [LR16AGR01] and the Deutsche Forschungsgemeinschaft project “Molecular epidemiology network for promotion and support of delivery of live vaccines against Theileria parva and Theileria annulata infection in eastern and northern Africa” (SE862/2–1).
Author information
Authors and Affiliations
Contributions
MRR designed the work, performed the laboratory work and wrote the manuscript. SA contributed to the laboratory work and the writing of the manuscript. MM. contributed to the collection of samples and the molecular analysis. MR revised the paper, approved the version to be published. MG supervised all the work and approved the final version of the article.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
No ethical approval was required for this study.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Rjeibi, M.R., Amairia, S., Mhadhbi, M. et al. Detection and molecular identification of Anaplasma phagocytophilum and Babesia spp. infections in Hyalomma aegyptium ticks in Tunisia. Arch Microbiol 204, 385 (2022). https://doi.org/10.1007/s00203-022-02995-7
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-02995-7