Hepatitis E virus (HEV) is an emerging zoonotic pathogen that is currently recognized as one of the major causes of acute human hepatitis worldwide. In Europe, the increasing number of hepatitis E cases is mainly associated with the consumption of animal food products or contact with infected animals. Dogs and cats have been suggested as a zoonotic source of HEV infection. The aim of this study was to assess Orthohepevirus circulation, including HEV‐A, HEV‐B and HEV‐C species, in sympatric urban cats and dogs in southern Spain. Between 2017 and 2020, blood samples were collected from 144 stray cats and 152 dogs, both strays and pets. The presence of antibodies against HEV were tested using a double‐antigen sandwich ELISA and seropositive samples were further analysed by western blot. A RT‐PCR was performed to detect RNA of Orthohepevirus species (HEV‐A, HEV‐B and HEV‐C). A total of 19 (6.4%; 95%CI: 3.6‐9.2) of the 296 animals tested showed anti‐HEV antibodies by ELISA. Seropositivity was significantly higher in dogs (9.9%; 15/152; 95%CI: 5.1‐14.6) than in cats (2.8%; 4/144; 95%CI: 0.1‐5.5). Ten of the 18 ELISA‐positive animals that could be further analysed by western blot, reacted against HEV‐3 and/or HEV‐C1 antigens, which suggest circulation of both genotypes in urban cats and dogs in the study area. However, HEV‐A, HEV‐B and HEV‐C RNA were not detected in any of the tested sera. This is the first study to assess HEV circulation in both stray cats and dogs in Europe. Our results provide evidence of HEV exposure in sympatric urban cat and dog populations in southern Spain. Further studies are needed to determine the role of these species in the epidemiology of HEV.

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A survey study was carried out to identify tick species parasitizing wild lagomorphs in Mediterranean ecosystems in southern Spain and to determine the occurrence of Rickettsia species present in these ticks in this region. A total of 1304 European wild rabbits (Oryctolagus cuniculus) and 58 Iberian hares (Lepus granatensis) were individually examined for the presence of ticks. Ticks were found in 42.9% and 50% of the wild rabbits and hares sampled, respectively. A total of 1122 ticks were collected and five species, including Rhipicephalus pusillus, Hyalomma lusitanicum, Haemaphysalis hispanica, Ixodes ventalloi and Rhipicephalus sanguineus sensu lato (s.l.), were microscopically and molecularly identified at the 16S rRNA gene. This is the first study on Ixodidae parasitizing Iberian hares. The presence of Rickettsia DNA was assessed in 254 tick pools (according to hunting states, lagomorph species, tick species and tick development stage) using PCR assays targeting the rOmpA, rOmpB and gltA. Twenty-seven pools (10.6%) were positive to Rickettsia DNA. Five zoonotic Rickettsia species were identified, being Rickettsia massiliae the most frequent (4.7%), followed by Rickettsia sibirica subsp. mongolitimonae (2.8%), Rickettsia slovaca (2.0%), Rickettsia aeschlimannii (0.8%) and Rickettsia africae (0.4%). The results suggest that wild rabbits and Iberian hares are parasitized by a wide range of tick species and that these lagomorphs may play an important role in the sylvatic cycle of some zoonotic Rickettsia species in Mediterranean ecosystems. Our data represent the first report of R. massiliae, R.

Toxoplasma gondii is an intracellular protozoan parasite of worldwide distribution. This parasite has an indirect life cycle in which domestic cats and other wild felids are definitive hosts, and virtually all warm-blooded species, including human, are intermediate hosts (Dubey, 2010). Up to a third of the human population worldwide is estimated to be chronically infected by this parasite

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A long‐term active epidemiological surveillance programme was conducted to determine seroprevalence to myxoma virus (MYXV), infection prevalence and spatiotemporal patterns and factors associated with MYXV circulation in wild rabbits (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems. A total of 2376 animals were sampled over four study periods: 2009–2012 (P1), 2012–2015 (P2), 2015–2018 (P3) and 2018–2021 (P4). Antibodies against MYXV were detected by a commercial indirect ELISA in 59.9% (1424/2376; 95% CI: 58.0–61.9) of wild rabbits. At least one seropositive animal was detected on 131 (96.3%) of 136 game estates sampled. MYXV infection was confirmed by PCR in 94 of 1063 (8.8%; 95% CI: 7.3–10.7) wild rabbits. Circulation of the novel recombinant MYXV (ha‐MYXV) was not found in wild rabbits analysed during P4. Five statistically significant spatiotemporal clusters of high MYXV seroprevalence were identified using a Bernoulli model: one in P2 and four in P3. A generalized linear mixed model (GLMM) analysis identified sampling season (autumn), age (adult and juvenile), outbreaks of myxomatosis in the month prior to sampling, mean annual temperature, humidity and seropositivity to rabbit haemorrhagic disease virus as factors potentially linked with MYXV seropositivity. GLMM analysis identified outbreaks of myxomatosis in the month prior to sampling, MYXV seropositivity and presence of lesions compatible with myxomatosis as factors associated with MYXV infection. The results indicate high exposure, widespread but non‐homogeneous distribution, and endemic circulation of MYXV in wild rabbit populations in southern Spain during the last decade. Prevalence of antibodies against MYXV showed fluctuations both within the year and over the study periods, revealing variations in the immunity of wild rabbit populations in Mediterranean ecosystems that could increase the risk of MYXV re‐emergence in immunologically naïve populations. The present study highlights the importance of long‐term surveillance to better understand the epidemiology of MYXV in wild lagomorphs.

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