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Background: Paramphistomids are ruminal trematodes that cause important losses in tropical and subtropical regions. However, their presence in Europe has increased significantly in recent decades. In northwestern Spain, this trend has been confirmed in cattle, but data in sheep are scarce and not updated. Moreover, the paramphistomid species affecting sheep in the area have never been molecularly identified. To evaluate the prevalence of paramphistomid infections in sheep from northwestern Spain, 826 faecal samples from 25 sheep farms were collected and analysed via coproscopic techniques. In addition, the rumens of 85 roe deer from the same area were examined to detect adult paramphistomids. The species present were molecularly identified. Multivariate analyses for identifying the risk factors affecting the prevalence and egg shedding of rumen flukes were also performed., Results: Overall, 14% of the animals and 44% of the flocks were positive; the mean egg count was 20.5 eggs per gram of faeces. In contrast, no adult paramphistomids were found in roe deer. Older sheep, those from farms located in the central climatic area, without water throughs available on pastures and using their own manure to fertilize, were considered significantly more susceptible to infection with paramphistomids. With respect to egg shedding, animals from 37 to 72 months of age, farms under semiextensive management, and those coinfected with Fasciola hepatica presented significantly greater egg counts. Molecular identification revealed 100% similarity with Calicophoron daubneyi sequences from other European and Mediterranean countries., Conclusions: The present study confirms the increase in the prevalence of paramphistomid infections in sheep in the area with high dissemination of the parasite, as previously reported in cattle, and represents the first molecular identification of C. daubneyi in sheep from Spain. Our results demonstrate that special attention should be given to adult animals since they are the main carriers and are responsible for environmental contamination. In addition, detecting risk areas and applying effective control management measures such as the installation of watering points on pastures seems essential for limiting infections in livestock, especially in sheep, since they are susceptible to developing clinical paramphistomidosis at any time in their lives. The absence of adult flukes in roe deer suggests that they represent less suitable hosts for this trematode than cattle and sheep, although more robust studies monitoring the situation in sympatric areas with domestic ruminants are needed., Competing Interests: Declarations. Ethics approval and consent to participate: All faecal samples used in this study were collected with the permission of the farm owner. All experimental procedures fully complied with European and Spanish ethics regulations on the protection of animals used for scientific purposes (European Directive 2010/63/EU and Spanish Royal Decree 53/2013) and approved by the ethical committee of the University of Santiago de Compostela. Consent for publication: Not applicable. Competing interests: The authors declare that they have no competing interests., (© 2024. The Author(s).)

Leptospirosis is a bacterial disease of worldwide distribution with relevant implications for animal and human health. Different large wild carnivore species can act as reservoirs of this zoonotic pathogen. This study aimed to evaluate the circulation of Leptospira spp. in free-ranging wolves (Canis lupus) from southern Europe. A total of 281 kidney samples of wolves from Spain and Italy were collected between 2017 and 2023. The presence of Leptospira DNA was analysed by real-time PCR and phylogenetic analyses were carried out using a Bayesian approach. The overall prevalence was 3.2 % (9/281; 95 %CI: 1.1-5.3). Leptospira DNA was detected in nine of the 180 wolves from Spain (5.0 %; 95 %CI: 1.8-8.2), but not in the Italian wolf population (0 %; 0/101). Molecular analyses revealed high homology between the sequences obtained in the present study and isolates of Leptospira interrogans and Leptospira borgpetersenii from different rodent and domestic ungulate species. Our results provide evidence of a low and spatially heterogeneous circulation of this pathogen in wolf populations of southern Europe. The detection of zoonotic Leptospira species in this survey supports the need to consider wolf populations in monitoring programs for leptospirosis with a One Health approach., Competing Interests: Declaration of Competing Interest None of the authors of this study has a financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of the paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Coxiella burnetii, the causative agent of Q fever, is a zoonotic multi-host vector-borne pathogen of major public health importance. Although the European Food Safety Authority has recently made the monitoring of this bacterium in wildlife a priority, the role of wild lagomorphs in the transmission and maintenance of C. burnetii is poorly understood., Aims: The aims of this study were to determine the prevalence and risk factors associated with C. burnetii circulation in European wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis) and to assess the presence of this pathogen in ticks that feed on them in Mediterranean ecosystems in Spain, the country with the highest number of reported cases of Q fever in Europe., Methods: A total of 574 spleen samples were collected from 453 wild rabbits and 121 Iberian hares, and 513 ticks (processed in 120 pools) between the 2017/2018 and 2021/2022 hunting seasons., Results: C. burnetii DNA was detected in 103 (17.9%; 95% CI: 14.8-21.1) of the 574 wild lagomorphs tested. By species, prevalence was 16.3% (74/453; 95% CI: 12.9-19.7) in the European wild rabbit and 24.0% (29/121; 95% CI: 16.4-31.6) in the Iberian hare. At least one positive lagomorph was found on 47.9% of the 96 hunting estates sampled and in every hunting season since 2018/2019. Two risk factors associated with C. burnetii infection were as follows: outbreak of myxomatosis on the hunting estate in the month prior to sampling and high tick abundance observed by gamekeepers on the hunting estate. C. burnetii DNA was also found in 33 of the 120 (27.5%; 95% CI: 19.5-35.5) tick pools tested. The pathogen was detected in 66.7% (4/6), 29.2% (26/89) and 21.4% (3/14) of Haemaphysalis hispanica, Rhipicephalus pusillus and Hyalomma lusitanicum pools respectively., Conclusions: This study provides new epidemiological data on C. burnetii in European wild rabbits and is the first survey on this zoonotic pathogen performed in Iberian hares. Our results indicate widespread endemic circulation of C. burnetii and highlight the importance of both wild lagomorph species as natural reservoirs of this zoonotic bacterium in Mediterranean ecosystems in southern Spain, which may be of public and animal health concern. The high prevalence and wide diversity of positive tick species suggest the possible role of ticks in the epidemiological cycle of C. burnetii, with the potential risk of transmission to sympatric species, including humans., (© 2024 The Author(s). Zoonoses and Public Health published by Wiley‐VCH GmbH.)

Wild lagomorphs can act as reservoirs of several pathogens of public and animal health concern. However, the number of studies assessing the presence of Anaplasma spp. in these species is scarce. The aim of the present study was to molecularly identify Anaplasma spp. in wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis) from Southern Spain and assess their epidemiological role in the maintenance of the bacterium. During 2017-2021, spleen samples of 394 wild rabbits and 145 Iberian hares were collected. Anaplasma DNA was detected using different PCR assays (16S rRNA and groEL) and phylogenetic analyses were carried out by Bayesian approach. The possible influence of lagomorph species, age and sex on the prevalence of Anaplasma spp. was evaluated by a multiple logistic regression model. The 9.4% of the rabbits were positive to Anaplasma bovis, but all the hares were negative. No significant differences were found in Anaplasma spp. prevalence regarding to age or sex. This is the first report of A. bovis in lagomorphs from Europe. The phylogenetic analysis of A. bovis confirms the existence of different clusters suggesting the existence of several lineages. In addition, a high divergence of nucleotide identity was observed within the lineage 4, which could result in the under-detection of some strains when using A. bovis-specific PCR, hindering its detection and characterization. Since this analysis is based on a limited number of nucleotide bases and sequences, more studies are needed for further characterize A. bovis, as well as its relationship with other Anaplasma spp., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)