Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 30, Number 5—May 2024
Research Letter

Crimean-Congo Hemorrhagic Fever Virus in Ticks Collected from Cattle, Corsica, France, 2023

Author affiliations: Unité des Virus Emergents, Aix Marseille Université, Università di Corsica, IRD140, INSERM 207 IRBA, Marseille, France (P. Kiwan, S. Masse, G. Piorkowski, N. Ayhan, M. Gasparine, R.N. Charrel, X. de Lamballerie, A. Falchi); Université de Corse–Institut National de Santé et de la Recherche Médicale, Corte, France (P. Kiwan, S. Masse, G. Piorkowski, N. Ayhan, M. Gasparine, R.N. Charrel, X. de Lamballerie, A. Falchi); Centre National de Référence des Arbovirus, Marseille, France (N. Ayhan, X. de Lamballerie); Université de Montpellier, Montpellier, France (L. Vial)

Cite This Article

Abstract

We report the detection of Crimean-Congo hemorrhagic fever virus (CCHFV) in Corsica, France. We identified CCHFV African genotype I in ticks collected from cattle at 2 different sites in southeastern and central-western Corsica, indicating an established CCHFV circulation. Healthcare professionals and at-risk groups should be alerted to CCHFV circulation in Corsica.

Crimean-Congo hemorrhagic fever (CCHF) is a tickborne disease caused by CCHF virus (CCHFV) (species Orthonairovirus haemorrhagiae, genus Orthonairovirus, family Nairoviridae, order Bunyavirales). Endemic in Africa, the Middle East, Asia, and Eastern Europe, CCHF has expanded to Western Europe (1). Repeated detection of CCHFV in Spain (2) raises questions about its circulation in neighboring countries, such as Portugal, Italy, and France.

In Corsica, a French Mediterranean island, a seroprevalence study of CCHFV conducted in livestock (cattle, goats, and sheep) during 2014–2016 showed an overall seroprevalence of 9.1%, and cattle harbored the highest rates (3). A subsequent surveillance study of 8,051 ticks collected from wild (wild boar, deer, and mouflon sheep) and domestic (cattle, horses, sheep) animals during 2016–2020 failed to detect CCHFV or nairovirus RNA (4).

Figure

Phylogenetic analysis of Crimean-Congo hemorrhagic fever virus in ticks collected from cattle, Corsica, France, 2023. Map at left shows locations of cattle from which ticks were collected at the slaughterhouses of Ponte Leccia in the north and Cuttoli-Cortichiatto in the south during 2022­–2023. Phylogenetic trees show small (A), medium (B), and large (C) RNA segments CCHFV strains. Red font indicates strains detected from Corsica; other sequences are named by GenBank accession number, geographic origin, and sampling year. Evolutionary analyses were conducted in MEGA6 (https://www.megasoftware.net) after best model determination. The optimal tree is shown for each fragment. Trees were constructed using the maximum-likelihood method based on sequences on the small (Tamura-Nei model), medium (general time-reversible model), and large (Tamura-Nei model) segments of the virus. All positions with <95% sequence site coverage were eliminated (i.e., <5% alignment gaps, missing data, or ambiguous bases were allowed at any position [partial deletion option]). Results of bootstrap test (1,000 replicates) are shown next to the branches. Genotypes are indicated by Roman numerals (8) with the equivalent clade nomenclature (9): I, West Africa (Africa 1); II, Central Africa (Africa 2); III, South and West Africa (Africa 3); IV, Middle East/Asia, divided into 2 groups Asia 1 and Asia 2; V (Europe 1), Europe/Turkey (Europe 1); VI, Greece (Europe 2); VII (Europe 3). Scale bars indicate nucleotide substitutions per site. CCHFV, Crimean-Congo hemorrhagic fever virus; L-RNA, large segment of CCHFV RNA.

Figure. Phylogenetic analysis of Crimean-Congo hemorrhagic fever virus in ticks collected from cattle, Corsica, France, 2023. Map at left shows locations of cattle from which ticks were collected at the slaughterhouses...

Since 2022, we have continued CCHFV surveillance by collecting ticks from cattle at 2 slaughterhouses >2 times/month. Cattle originate from a broad geographic area, and the national ear-tag identification system enables tracing of each animal’s origin and farm owner (Figure). We identified ticks by using taxonomic keys, then pooled ticks by species, sex, development stage, study site, and animal host, as previously reported (4). We spiked each pool, consisting of 1–6 ticks, with a predefined amount of MS2 bacteriophage for monitoring nucleic acid extraction, reverse transcription PCR (RT-PCR), and nucleic acid amplification (5). We used MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit (Thermo Fisher Scientific, https://www.thermofisher.com) to purify nucleic acids. We tested each sample by using 2 real-time RT-PCRs, 1 targeting the large (L) RNA segment (2) and 1 targeting the small (S) RNA segment (6). We used the SuperScript IV One-Step RT-PCR System Kit (ThermoFisher) to design 28 CCHFV-specific pairs of primers to amplify the S, medium (M), and L segments (Appendix). We sequenced PCR products by using S5 Ion Torrent technology (ThermoFisher). We determined the best model by using the maximum-likelihood method and performed phylogenetic analyses by using MEGA6 software (7) (Figure).

During June 2022–July 2023, we collected 5,165 ticks from 465 cattle and grouped ticks into 1,491 pools. Tick species consisted of 2,390 (46.27%) Rhipicephalus bursa, 1,103 (21.35%) Hyalomma marginatum, 750 (14.52%) Boophilus annulatus, 507 (9.81%) Hyalomma scupense, 238 (4.60%) Haemaphysalis punctata, 127 (2.45%) Ixodes ricinus, 48 (0.92%) Rhipicephalus sanguineus, and 2 (0.03%) Dermacentor marginatus. A total of 24 (1.70%) pools collected from 5 cattle from southern Corsica tested positive by the L-RNA assay (Table). Nineteen of the 24 tick pools were collected from 1 animal (no. 4039) (Table). Partial sequences for S (1,340 bp), M (4,894 bp), and L (11,275 bp) segments were obtained from animal nos. 2478 (pool 417) and 4039 (pool 1207) (Table). The effective detection of CCHFV genome is strongly supported by the formal exclusion of contamination because no CCHFV strain or genome had been previously processed in the laboratory, the PCR systems used can distinguish genomic RNA from the positive control (6), and the CCHFV sequences obtained were original and unambiguous.

The obtained S and M segment sequences constituted a monophyletic group belonging to genotype I (Africa 1), whereas the L segment sequence grouped with genotype III strains (Africa 3) (Figure). The sequences of all 3 segments of the CCHFV from Corsica are closely related to 2 sequences from Senegal corresponding to strains identified in the 1970s and likely represent strains reassorted in Senegal. Whether those strains are typical of strains from Senegal or have been circulating in other parts of Africa requires additional investigations.

Our results suggest that CCHFV strains circulating in Corsica and Spain have distinct origins. In Spain, genotype III is the most widespread and is most often detected in H. lusitanicum ticks (2), a species not yet identified in Corsica. Trans-Saharan migratory birds carrying H. marginatum ticks are the most likely source of CCHFV strains entering Corsica (6). Examination of the main bird migration routes suggests that 2 different migratory corridors link Spain and Corsica to Africa; mainly, but not exclusively, West Africa for Spain and Central Africa for Corsica (6). Those migration routes also could explain the different origin of CCHFV strains circulating in Corsica and in Spain.

Our results provide evidence for established CCHFV circulation in Corsica because detection occurred at 2 distinct sites in the southeastern and central western parts of the island. In addition, our results provide evidence for infection in cattle because multiple CCHFV-positive ticks were found on the same animal. CCHFV detection in feeding ticks is indicative of virus circulation within the cattle population but does not elucidate the role of ticks in virus transmission. Our results must be interpreted by considering previous serologic evidence of CCHFV circulation in cattle in Corsica (3), the presence of competent vectors locally (4), and recent reports of CCHFV detection in southern mainland France (10). The threat of possible continuous expansion and circulation of the virus over Western Europe should not be disregarded. Healthcare professionals and other groups at risk for infection, including hunters and farmers, should be informed about CCHFV circulation in Corsica.

Ms. Kiwan is a PhD student at the Unité des Virus Emergents, University of Corsica Pascal Paoli and Aix-Marseille University, France. Her primary research interests focus on tickborne viruses via a One Health approach.

Top

Acknowledgments

We thank the staff of the slaughterhouses for their help in collecting ticks from cattle. We thank Gregory Mollé, Laurence Thirion, Pierre Combe, and Cecile Baronti for the production and validation of diagnostic reagents.

This work was supported in part by the VHFMODRAD project (no. 823666, call H2020-JTI-IMI2-2015-08-single-stage) of the H2020 program by the Innovative Health Initiative (IHI) agency of the European Commission; by the European Commission European Virus Archive Global project (EVA GLOBAL, grant agreement no. 871029) of the Horizon 2020 Research and Innovation Programme; and by Centre de Coopération Internationale en Recherche Agronomique pour le Développement (provision of service no. E2F03F6F). The reagent material was provided by the European Virus Archive-Marseille (EVAM) under the label technological platforms of Aix-Marseille University.

Top

References

  1. Lorenzo Juanes  HM, Carbonell  C, Sendra  BF, López-Bernus  A, Bahamonde  A, Orfao  A, et al. Crimean-Congo hemorrhagic fever, Spain, 2013–2021. Emerg Infect Dis. 2023;29:2529. DOIPubMedGoogle Scholar
  2. Sánchez-Seco  MP, Sierra  MJ, Estrada-Peña  A, Valcárcel  F, Molina  R, de Arellano  ER, et al.; Group for CCHFv Research. Widespread detection of multiple strains of Crimean-Congo hemorrhagic fever virus in ticks, Spain. Emerg Infect Dis. 2021;28:394402. DOIPubMedGoogle Scholar
  3. Grech-Angelini  S, Lancelot  R, Ferraris  O, Peyrefitte  CN, Vachiery  N, Pédarrieu  A, et al. Crimean-Congo hemorrhagic fever virus antibodies among livestock on Corsica, France, 2014–2016. Emerg Infect Dis. 2020;26:10414. DOIPubMedGoogle Scholar
  4. Cicculli  V, Maitre  A, Ayhan  N, Mondoloni  S, Paoli  JC, Vial  L, et al. Lack of evidence for Crimean-Congo hemorrhagic fever virus in ticks collected from animals, Corsica, France. Emerg Infect Dis. 2022;28:10358. DOIPubMedGoogle Scholar
  5. Ninove  L, Nougairede  A, Gazin  C, Thirion  L, Delogu  I, Zandotti  C, et al. RNA and DNA bacteriophages as molecular diagnosis controls in clinical virology: a comprehensive study of more than 45,000 routine PCR tests. PLoS One. 2011;6:e16142. DOIPubMedGoogle Scholar
  6. Estrada-Peña  A, D’Amico  G, Fernández-Ruiz  N. Modelling the potential spread of Hyalomma marginatum ticks in Europe by migratory birds. Int J Parasitol. 2021;51:111. DOIPubMedGoogle Scholar
  7. Tamura  K, Stecher  G, Peterson  D, Filipski  A, Kumar  S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 2013;30:27259. DOIPubMedGoogle Scholar
  8. Chamberlain  J, Cook  N, Lloyd  G, Mioulet  V, Tolley  H, Hewson  R. Co-evolutionary patterns of variation in small and large RNA segments of Crimean-Congo hemorrhagic fever virus. J Gen Virol. 2005;86:333741. DOIPubMedGoogle Scholar
  9. Carroll  SA, Bird  BH, Rollin  PE, Nichol  ST. Ancient common ancestry of Crimean-Congo hemorrhagic fever virus. Mol Phylogenet Evol. 2010;55:110310. DOIPubMedGoogle Scholar
  10. Anses – National Agency for Food, Environmental and Occupational Health Safety. Crimean-Congo hemorrhagic fever: first detection of the virus in cattle farms in southern France [in French] [cited 2023 Dec 18]. https://www.anses.fr/fr/content/fievre-hemorragique-crimee-congo-detection-virus-elevages-bovins

Top

Figure
Table

Top

Cite This Article

DOI: 10.3201/eid3005.231742

Original Publication Date: April 11, 2024

Table of Contents – Volume 30, Number 5—May 2024

EID Search Options
presentation_01 Advanced Article Search – Search articles by author and/or keyword.
presentation_01 Articles by Country Search – Search articles by the topic country.
presentation_01 Article Type Search – Search articles by article type and issue.

Top

Comments

Please use the form below to submit correspondence to the authors or contact them at the following address:

Alessandra Falchi, Unité des Virus Emergents, IRD 190-Inserm 1207, University of Corsica Pascal Paoli and Aix-Marseille University, Laboratoire de Virologie, Faculté des Sciences, Campus Grimaldi, Corte 20250, France

Send To

10000 character(s) remaining.

Top

Page created: March 28, 2024
Page updated: April 24, 2024
Page reviewed: April 24, 2024
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
file_external