We conducted a cross-sectional study in wild boar and extensively managed Iberian pig populations in a hotspot area of Crimean-Congo hemorrhagic fever virus (CCHFV) in Spain. We tested for antibodies ...against CCHFV by using 2 ELISAs in parallel. We assessed the presence of CCHFV RNA by means of reverse transcription quantitative PCR protocol, which detects all genotypes. A total of 113 (21.8%) of 518 suids sampled showed antibodies against CCHFV by ELISA. By species, 106 (39.7%) of 267 wild boars and 7 (2.8%) of 251 Iberian pigs analyzed were seropositive. Of the 231 Iberian pigs and 231 wild boars analyzed, none tested positive for CCHFV RNA. These findings indicate high CCHFV exposure in wild boar populations in endemic areas and confirm the susceptibility of extensively reared pigs to CCHFV, even though they may only play a limited role in the enzootic cycle.
A survey study was carried out to assess the occurrence of selected tick‐borne pathogens (TBP) in wild ungulates in Mediterranean ecosystems in southern Spain. Spleen samples were collected from ...1,132 wild ungulates, including 578 red deer, 269 wild boar, 135 mouflon, 121 fallow deer and 29 roe deer, between 2009 and 2015. Eighty‐nine ticks collected from TBP‐positive animals were also analysed. Samples were tested by PCR and sequenced whenever possible. TBP DNA was detected in 127 of 863 wild ruminants (14.7%; 95% CI: 12.4–17.3) including the following: Anaplasma phagocytophilum (9.2%), Babesia divergens (2.9%), Theileria sp. OT3 (1.7%), Borrelia afzelii (0.7%) and Theileria capreoli (0.2%), but no positive samples were detected in wild boar (0/269). All the strains from mouflon were identified as Theileria sp. OT3, while B. divergens and T. capreoli were mainly found in red deer. Co‐infection with A. phagocytophilum and B. divergens, and A. phagocytophilum and Theileria spp. was detected in red deer and mouflon, respectively. The risk factor analysis showed that the prevalences of A. phagocytophilum and piroplasms were species‐related. Eighty‐nine tick specimens collected from ungulates found to be infected with the selected TBP were identified as Hyalomma lusitanicum (95.5%) and Ixodes ricinus (4.5%). Thirty ticks were positive for Anaplasma/Ehrlichia spp. (33.7%), 25 for Babesia/Theileria (28.1%) and two for B. burgdorferi s.l. (2.3%). Eleven specimens showed co‐infections with Anaplasma/Ehrlichia and Babesia/Theileria (10.1%) or Anaplasma/Ehrlichia and B. burgdorferi s.l. (2.3%). The estimated prevalences obtained in the present study suggest the possible contribution of wild ruminants to the maintenance of some selected TBP in Mediterranean ecosystems in southern Spain, while the role of wild boar in the epidemiology of these pathogens seems to be limited in this region.
Wildlife is an important reservoir of zoonotic pathogens. The objective of the present study was to assess the importance of wild ungulates in the epidemiology of Rickettsia spp. Ticks and spleen ...samples were collected from 262 red deer (Cervus elaphus) and 83 wild boar (Sus scrofa) hunted in southwestern Spain over a 5‐year period. DNA was extracted from tick pools (n = 191) and spleens (n = 345), and two nested PCR assays targeting the rOmpA and rOmpB genes were used to detect Rickettsia DNA. Five tick species were identified (Hyalomma lusitanicum, Dermacentor marginatus, Ixodes ricinus, Rhipicephalus bursa and Haemaphysalis sulcata). Rickettsia DNA was detected in 31 (16.2%) tick pools and two red deer spleen samples (0.8%). Four validated Rickettsia species (R. slovaca, R. monacensis, R. helvetica and R. raoultii), one uncultivated species (Candidatus R. rioja) and two uncharacterized Rickettsia spp. were detected in ticks. R. helvetica and R. slovaca were also detected in spleen samples from red deer. The overall prevalence in ungulate spleen samples was lower than in tick pools suggesting that these ungulates do not play a major role in the transmission of Rickettsia spp. However, their importance as spreaders of positive ticks cannot be ruled out. The results present a challenge for the veterinary and public health communities since most of the Rickettsia spp. detected are pathogenic. Furthermore, the new Rickettsia species could be potential pathogens. For these reasons, identifying Rickettsia species present in ticks and wildlife is of particular interest to clarify their sylvatic cycle and establish appropriate control measures.
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. aeschlimannii, R. slovaca and R africae in ticks collected in wild lagomorphs in Europe, and the first report of not imported R. africae in this continent. Since R. slovaca and R. africae DNA was detected in tick species different to their main vectors, further studies are warranted to unravel the role of wild lagomorphs in the epidemiology of these vector‐borne pathogens.
Background
A cross‐species jump was confirmed in 2018, when a novel recombinant myxoma virus (MYXV) (ha‐MYXV) caused high mortality in Iberian hare (Lepus granatensis) in the Iberian Peninsula.
...Method
The aim of this study was to evaluate the main lesions, tissular distribution and target cells of ha‐MYXV in Iberian hare. Gross postmortem examinations and histological and immunohistochemical studies to detect ha‐MYXV were carried out in 28 animals that were confirmed as ha‐MYXV positive by PCR.
Results
The main macroscopic lesions were bilateral blepharoconjunctivitis, epistaxis, intense congestion and oedema in several organs and some internal haemorrhages. Visible myxomas were not found. Histopathological examination revealed hyperplastic epidermis with predominant hyperkeratosis and myxoid matrix in the dermis. ha‐MYXV‐positive keratinocytes showed hydropic degeneration and cytoplasmic inclusion bodies. Alveolar oedema, interstitial pneumonia, dramatic lymphoid depletion in the spleen and necrosis in the liver and testis were observed. ha‐MYXV was mainly detected in epithelial and myxoma cells in the skin, and also in macrophages, lymphocytes, fibroblasts and endothelial cells in several organs, as well as in hepatocytes and Leydig cells.
Limitations
A non‐homogeneous number of samples were included in all the animals. Future experimental studies with controlled variables are necessary.
Conclusion
These findings correspond to an unusual form of myxomatosis, characterised by an acute or hyperacute presentation.
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 concerns 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 Cryptosporidium cuniculus (n = 6) and Cryptosporidium 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 Encephalitozoon intestinalis could be of public health relevance.
A cross‐sectional study was carried out to assess the circulation of bluetongue virus (BTV) and Schmallenberg virus (SBV) within the wild and domestic ungulate host community in Doñana National Park ...(southwestern Spain). A total of 440 animals, including 138 cattle, 102 red deer (Cervus elaphus), 101 fallow deer (Dama dama) and 99 wild boar (Sus scrofa), were sampled in 2015 during the seasonal peak of Culicoides spp. (summer–autumn). Serum and spleen samples were analysed to detect exposure (using commercial blocking ELISAs) and infection (by RT‐PCR), respectively, to BTV and SBV. Cattle were not tested by BTV‐bELISA because all were previously vaccinated against BTV serotypes 1 and 4. High BTV seroprevalences were found in red deer (97.0%) and fallow deer (64.7%). Antibodies against SBV were detected in 37.0% of cattle, 16.8% of red deer, 23.5% of fallow deer and 2.0% of wild boar. Thirty‐eight of the 203 deer (18.7%; 17 red deer and 21 fallow deer) were co‐exposed to both viral agents. BTV‐4 RNA was confirmed in four red deer and two fallow deer. SBV RNA was found in two fallow deer. Co‐infections were not detected. Our results indicate high exposure, widespread distribution and active circulation of BTV and SBV in the ruminant community in the study area. We provide additional evidence for the potential role of wild cervids as reservoirs of these Culicoides‐borne viruses in two different epidemiological scenarios: with vaccination (BTV) and without vaccination (SBV) of sympatric livestock. This study highlights the importance of wildlife surveillance, particularly of cervid species, for the proper execution of control programmes of Culicoides‐borne diseases in extensively reared livestock.
The possibility of Toxoplasma gondii transmitted from game meat to humans is of public health concern. Here we determined seroprevalence and risk factors associated with T. gondii in large game ...ungulates that cohabit in Sierras de Cazorla, Segura and Las Villas Natural Park (SCSV‐NP) (Southern Spain), a natural park with high human–animal interaction. Antibodies against T. gondii in 328 wild ungulates were assayed by the modified agglutination test (MAT ≥ 1:25). Antibodies were found in 39 (11.9%, 95% CI: 8.4–15.4) wild ungulates, with seroprevalence levels of 20.8% in wild boars (Sus scrofa) (5/24), 19.0% in fallow deer (Dama dama) (12/63), 13.9% in Iberian ibex (Capra pyrenaica hispanica) (14/101), 7.9% in red deer (Cervus elaphus) (6/76), and 3.1% in mouflons (Ovis aries musimon) (2/64). Significantly higher seroprevalence was observed in fallow deer and wild boars compared to mouflons. Animals living close to urban areas (<2 km) had 4.6‐times higher risk compared to those living at >5 km of urban areas. The results indicate high circulation of T. gondii in wild ungulates in SCSV‐NP, which is of animal and public health concern. The increased seroprevalence of T. gondii detected in wildlife ungulates living close to urban areas may increase human infection in those areas if meat from infected animals is consumed raw or undercooked.
Toxoplasmosis, caused by the obligate intracellular protozoan Toxoplasma gondii, is a worldwide parasitic zoonosis. A cross‐sectional study was carried out to determine the exposure to T. gondii in ...equids in Europe. Serum samples from 1399 equids (1085 horses, 238 donkeys, and 76 mules/hinnies) bred in four European countries (Italy, Spain, United Kingdom UK, and Ireland) were collected during the period of 2013–2021. The overall seroprevalence of T. gondii was 18.9% (95% Confidence Interval CI: 16.9–21.0) by using the modified agglutination test (MAT) at a cut‐off of 1:25. Seropositivity by country was 27.1% in Italy, 16.6% in Spain, 12.0% in UK and 7.0% in Ireland. Anti‐T. gondii antibodies were detected in 12.8% of the horses, 43.7% of the donkeys, and in 28.9% of the mules/hinnies. A Generalized Estimating Equation (GEE) analysis was carried out to study the associations between seropositivity and explanatory variables related to individuals, herds, and management measures on these herds, selected based on the bivariate analysis. The risk for being seropositive for T. gondii was 5.3 and 2.7 times higher in donkeys and mules/hinnies than in horses, respectively. In addition, significantly higher seropositivity was observed in horses from herds that used disinfectants less than once a week (13.9%; p = 0.038, odds ratio OR = 1.6; 95% CI: 1.03–2.62) compared with those from herds that performed weekly disinfection of the facilities (9.4%). This is the first large‐scale seroepidemiological study on T. gondii comprising horses, donkeys, and mules/hinnies in Europe and the first report of T. gondii exposure in horses from Ireland and UK. We found a widespread distribution of T. gondii among equid populations in different European countries. The seroprevalence found in these species, especially in donkeys and mules/hinnies, highlights the potential risk of human infection through the consumption of their raw/undercooked milk or meat.
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.
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