Epidemiological surveillance of Leishmania infantum in wild lagomorphs in Spanish Mediterranean ecosystems.

Wild lagomorphs play a key epidemiological role as reservoirs of Leishmania infantum, causative agent of the largest outbreak of human leishmaniosis in Europe to date. A large-scale survey study was conducted on wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis) populations in Spanish Mediterranean ecosystems to evaluate the exposure of L. infantum and investigate potential risk factors associated with exposure to this zoonotic parasite. Between 2018 and 2021, a total of 631 wild lagomorphs (471 wild rabbits and 160 Iberian hares) were collected in Andalusia (southern Spain) and tested for antibodies against L. infantum using the indirect fluorescent antibody test (IFAT). Spleen samples from 563 of the wild lagomorphs sampled (441 wild rabbits and 122 Iberian hares) were also evaluated by real-time quantitative PCR (qPCR) for detection of Leishmania kDNA. Exposure to L. infantum (positive by IFAT and/or qPCR) was detected in 56.4 % (356/631; 95 %CI: 52.3-60.3) of the lagomorphs analyzed. Anti-Leishmania antibodies were found in 12.8 % (81/631; 95 %CI: 10.2-15.5) of the animals, and L. infantum kDNA was detected in 59.0 % (332/563; 95 %CI: 54.9-63.0) of the spleen samples tested. Phylogenetic analysis revealed high homology (99.9-100 %) between L. infantum sequences obtained and strains previously isolated from humans in Spain. While apparent seroprevalence was significantly higher in Iberian hares (19.4 %; 95 %CI: 13.3-25.5) compared to wild rabbits (10.6 %; 95 %CI: 7.9-13.4), no significant differences in prevalence were found between wild rabbits (61.0 %; 95 %CI: 56.5-65.6) and Iberian hares (51.6 %; 95 %CI: 42.8-60.5). At least one positive animal was found on 64.8 % (70/108) of the hunting grounds sampled, and a high-risk spatial cluster (P < 0.001) was also identified in central Andalusia. The multivariable analysis identified bioclimatic level (meso-Mediterranean climate) and the presence of goats on hunting grounds as risk factors potentially associated with L. infantum exposure in wild lagomorphs. This study shows high, widespread exposure, but heterogeneous distribution of L. infantum in wild lagomorph populations in Mediterranean ecosystems in southern Spain. The results point to the need to promote integrated surveillance programs for the detection of Leishmania spp. in wild lagomorphs in order to establish effective control measures against human leishmaniosis under a One Health approach.

Monitoring the dynamics of consumption of ungulate game by-products in vulture feeding stations in Iberian Mediterranean ecosystems.

Vulture feeding stations (VFS) are strategic sites for the management of ungulate game by-products, with major implications for the conservation of obligate avian scavengers. These feeding points are designed to prevent access by facultative mammalian scavengers and to reduce the risk of trophically transmitted pathogen infection through scavenging behaviour. The aim of this study, using camera trap video recordings, was to evaluate the feeding dynamics around 32 lots of ungulate game by-products deposited in VFS in southern Spain. During the study period (2017-2022), 11 different avian and mammal scavenger species was detected. Also, many feeding events involving all scavenger species (51.1%; 362/709) and mammal scavenger species (23.4%; 166/709) was recorded. A significantly earlier presence of major obligate scavengers (Gyps fulvus and Aegypius monachus) (P = 0.023) and a lower persistence of by-products deposited (P < 0.001) were detected during October-December compared to January-March. Our results confirm that VFS play an important role in vulture conservation in Iberian Mediterranean ecosystems. However, we also point out the potential risk of VFS as hotspots of foodborne pathogens for mammal scavengers, particularly when the hunting offal persists longer. Consequently, we urge the competent authorities to update the regulations affecting VFS management, and hunting estate managers to make every effort to correctly apply those preventing scavenging mammals from gaining access to VFS. Further studies tackling access of non-target species to VFS are also warranted to properly assess the sanitary implications and geographical extent of these problems for mammal species inhabiting areas where VFS are installed.

Monitoring the epidemic of West Nile virus in equids in Spain, 2020-2021.

The largest epidemic of West Nile virus (WNV) reported ever in Spain in both humans and equines occurred in 2020, affecting 77 humans and 139 equine herds. Here, we aimed to monitor the outbreaks detected in equid herds in Andalusia (southern Spain), the Spanish region where 89.9% of the outbreaks were reported, and to evaluate the virus circulation and risk factor associated with WNV exposure in the affected herds. The first WNV case was detected in mid-July 2020, the number of outbreaks peaked in mid-August and the last one was confirmed on 26th October 2020. WNV lineage 1 was detected in 12 clinically affected horses using real time RT-PCR. Molecular analysis evidenced high nucleotide identity with WNV sequences obtained from humans, birds and mosquitoes from Spain and Italy between 2020 and 2022. Between five and eight months after the WNV epidemic, a total of 724 equids (including 485 unvaccinated and 239 vaccinated animals) from 113 of the 125 affected herds in Andalusia were sampled. IgM and IgG antibodies against WNV were detected in 1.6% (8/485; 95%IC: 0.0-2.5) and 61.9% (300/485; 95%IC: 58.3-65.5) of the unvaccinated individuals, respectively. The seropositivity in vaccinated horses was 86.6% (207/239). The main risk factors associated with WNV exposure in unvaccinated equids were the breed (crossbreed), the location of animals in spring-summer (outside), and the presence of natural water ponds close to the surveyed herds. The high individual seroprevalence obtained in the affected herds indicates that WNV circulation was more widespread than the reported by passive surveillance during the WNV epidemic in 2020. The re-emergence of WNV in 2020 in southern Spain evidenced the needed to improve integrated surveillance systems, minimizing the impact of future cases in equids and humans in high-risk areas.

High exposure of West Nile virus in equid and wild bird populations in Spain following the epidemic outbreak in 2020.

A cross-sectional study was conducted to assess the circulation and risk factors associated with West Nile virus (WNV) exposure in equine and wild bird populations following the largest epidemic outbreak ever reported in Spain. A total of 305 equids and 171 wild birds were sampled between November 2020 and June 2021. IgG antibodies against flaviviruses were detected by blocking enzyme-linked immunosorbent assay (bELISA) in 44.9% (109/243) and 87.1% (54/62) of unvaccinated and vaccinated equids, respectively. The individual seroprevalence in unvaccinated individuals (calculated on animals seropositive by both bELISA and virus microneutralization test [VNT]) was 38.3% (95%CI: 33.1-43.4). No IgM antibodies were detected in animals tested (0/243; 0.0%; 95%CI: 0.0-1.5) by capture-ELISA. The main risk factors associated with WNV exposure in equids were age (adult and geriatric), breed (crossbred) and the absence of a disinsection programme on the facilities. In wild birds, IgG antibodies against flaviviruses were found in 32.7% (56/171; 95%CI: 26.8-38.6) using bELISA, giving an individual WNV seroprevalence of 19.3% (95%CI: 14.3-24.3) after VNT. Seropositivity was found in 37.8% of the 37 species analysed. Species group (raptors), age (>1-year old) and size (large) were the main risk factors related to WNV seropositivity in wild birds. Our results indicate high exposure and widespread distribution of WNV in equid and wild bird populations in Spain after the epidemic outbreak in 2020. The present study highlights the need to continue and improve active surveillance programmes for the detection of WNV in Spain, particularly in those areas at greatest risk of virus circulation.

Outbreak of Yersinia pseudotuberculosis in capybaras (Hydrochoerus hydrochaeris) kept in captivity.

On January 2020, an outbreak of acute mortality was detected in the capybara (Hydrochoerus hydrochaeris) population kept in the Cordoba Zoo Conservation Center (CZCC) in Spain. Morbidity and mortality rates were 71%, and the fatality rate was 100%. Macroscopic examination revealed multifocal nodular lesions with fibrinonecrotic exudate in lungs and mesenteric lymph nodes. Microscopic lesions were characterized as severe acute fibrinopurulent bronchopneumonia and generalized vascular changes. Cultures and biochemical analyses yielded Yersinia pseudotuberculosis (YPT). This species was confirmed by both PCR and MALDI-TOF. This report describes the first outbreak of fatal YPT infection in zoo animals in Spain and confirms that capybaras are highly susceptible to this pathogen, which could be of animal health, conservation, and public health concern. Future studies are warranted to determine potential sources of YTP infection in zoos to prevent future cases in susceptible species, including humans.