Iberian wild leporidae as hosts of zoonotic enteroparasites in Mediterranean ecosystems of Southern Spain

Authors: REGO, LAURA - National Center For Microbiology, The Institute Of Health Carlos Iii; CASTRO-SCHOLTEN, SABRINA - University Of Cordoba; CANO, CARMEN - National Center For Microbiology, The Institute Of Health Carlos Iii; JIMENEZ-MARTIN, DEBORA - University Of Cordoba; KOSTER, PAMELA - National Center For Microbiology, The Institute Of Health Carlos Iii; CABALLERO-GOMEZ, JAVIER - University Of Cordoba; BAILO, BEGONA - National Center For Microbiology, The Institute Of Health Carlos Iii; DASHTI, ALEJANDRO - National Center For Microbiology, The Institute Of Health Carlos Iii; HERNANADEZ-CASTRO, CAROLINA - National Center For Microbiology, The Institute Of Health Carlos Iii; CANO-TERRIZA, DAVID - University Of Cordoba; VIOQUE, FATIMA - National Center For Microbiology, The Institute Of Health Carlos Iii; Maloney, Jenny; Santin-Duran, Monica

Submitted to: Zoonoses and Public Health
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/5/2023
Publication Date: 12/19/2023
Citation: Rego, L., Castro-Scholten, S., Cano, C., Jimenez-Martin, D., Koster, P.C., Caballero-Gomez, J., Bailo, B., Dashti, A., Hernanadez-Castro, C., Cano-Terriza, D., Vioque, F., Maloney, J.G., Santin, M. 2023. Iberian wild leporidae as hosts of zoonotic enteroparasites in Mediterranean ecosystems of Southern Spain. Zoonoses and Public Health. 70(3):223-237. https://doi.org/10.1111/zph.13018.
DOI: https://doi.org/10.1111/zph.13018

Interpretive Summary: Wildlife such as wild rabbits and hares can act as carriers of parasitic diseases which can also infect humans. However, studies on the epidemiology and public health significance of intestinal eukaryotic parasites in wild rabbits and hares are needed to understand the potential for transmission between wildlife and humans. To address this research need, we examined fecal samples from European wild rabbits (n = 438) and Iberian hares (n = 111) collected in southern Spain from 2012–2021. These samples were tested for the presence of intestinal parasites of veterinary and public health importance including Blastocystis sp., Cryptosporidium spp., Giardia duodenalis, Encephalitozoon intestinalis, and Enterocytozoon bieneusi. Testing was performed using PCR followed by Sanger or next generation amplicon sequencing to explore the zoonotic potential of positive samples. Giardia duodenalis was the most prevalent species found (27.8%, 153/550), followed by Cryptosporidium spp. (1.3%, 7/550), Blastocystis sp. (1.1%, 6/550), and E. intestinalis (0.2%, 1/550). All samples tested negative for E. bieneusi. Sequence analyses revealed the presence of G. duodenalis sub-assemblage BIV (n = 1), Cryptosporidium cuniculus (n = 6), and Cryptosporidium andersoni (n = 1). The six Blastocystis-positive samples were identified as ST2 (n = 3) and ST1+ST2 (n = 3). Our data suggest that wild rabbits and hares are primarily infected by host-adapted species of microeukaryotic pathogens. However, Blastocystis sp. subtypes ST1 and ST2, G. duodenalis sub-assemblage BIV, and E. intestinalis are all potentially zoonotic. Thus, wild rabbits and hares could be sources of parasites with public health relevance. This study expands current knowledge on the epidemiology of microeukaryotic pathogens that will be useful to other scientists, veterinarians, and public health agencies in understanding zoonotic potential and public health importance of these parasites.

Technical Abstract: Wild lagomorphs including rabbits and hares can act as natural carriers or reservoirs of bacterial and parasitic zoonotic diseases. However, little is known on the epidemiology and potential public health significance of intestinal eukaryotes in wild leporids. We examined faecal samples from European wild rabbits (Oryctolagus cuniculus, n = 438) and Iberian hares (Lepus granatensis, n = 111) collected in the Autonomous Region of Andalusia in southern Spain during 2012–2021. We searched for the presence of DNA from the main intestinal protist and microsporidial pathogens of veterinary and public health concern using molecular methods (PCR followed by Sanger and next generation sequencing). Giardia duodenalis was the most prevalent species found (27.8%, 153/550; 95% CI: 24.1–31.8), followed by Cryptosporidium spp. (1.3%, 7/550; 95% CI: 0.5–2.6), Blastocystis sp. (1.1%, 6/550; 95% CI: 0.4–2.4), and Encephalitozoon intestinalis (0.2%, 1/550; 95% CI: 0.0–10.1). All samples tested negative for Enterocytozoon bieneusi. Sequence analyses revealed the presence of sub-assemblage BIV (n = 1) within G. duodenalis, and C. cuniculus (n = 6) and C. andersoni (n = 1) within Cryptosporidium. The presence of ruminant-adapted C. andersoni is indicative of a potential cross-species transmission event, although a spurious infection (mechanical carriage) cannot be ruled out. Samples assigned to C. cuniculus belonged to the gp60 subtype families Va (n = 3) and Vb (n =2). The six Blastocystis-positive samples were identified as ST2 (n = 3) and ST1+ST2 (n = 3). Our molecular results suggest that wild rabbits and hares were primarily infected by leporid-adapted species of eukaryotic pathogens. However, the occasional findings of zoonotic G. duodenalis sub-assemblage BIV, Blastocystis sp. ST1 and ST2, and E. intestinalis could be of public health relevance.