Epidemiological survey and risk factors associated with Sarcoptes scabiei in wild lagomorphs in Spanish Mediterranean ecosystems

Research in Veterinary Science 177 (2024) 105344 Available online 6 July 2024 0034-5288/© 2024 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/bync/4.0/). Epidemiological survey and risk factors associated with Sarcoptes scabiei in wild lagomorphs in Spanish Mediterranean ecosystems Sabrina Castro-Scholten a , Ignacio García-Bocanegra a , b , * , Natalia Iglesias c , Rosa Casais c , David Cano-Terriza a , b , D´ ebora Jim´ enez-Martín a , Jesús Barbero-Moyano a , Leonor Camacho- Sillero d , Carlos Rouco e , Javier Caballero-G´ omez a , b , f a Departamento de Sanidad Animal, Grupo de Investigaci´ on en Sanidad Animal y Zoonosis (GISAZ), UIC Zoonosis y Enfermedades Emergentes ENZOEM, Universidad de C´ ordoba, 14014 C´ ordoba, Spain b CIBERINFEC, ISCIII – CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III, 28029 Madrid, Spain c Centro de Biotecnología Animal, Servicio Regional de Investigaci´ on y Desarrollo Agroalimentario (SERIDA), 33394 Deva, Spain d Programa de Vigilancia Epidemiol´ ogica de la Fauna Silvestre (PVE), Consejería de Sostenibilidad, Medio Ambiente y Economía Azul, Junta de Andalucía, 29002 M´ alaga, Spain e Departamento Biología Vegetal y Ecología, ´ Area de Ecología, Universidad de Sevilla, 41012 Sevilla, Spain f Grupo de Virología Clínica y Zoonosis, Unidad de Enfermedades Infecciosas, Instituto Maim´ onides de Investigaci´ on Biom´ edica de C´ ordoba (IMIBIC), Hospital Universitario Reina Sofía, Universidad de C´ ordoba, 14004 C´ ordoba, Spain ARTICLE INFO Keywords: S. scabiei Oryctolagus cuniculus Lepus granatensis Zoonoses Surveillance Sarcoptic mange ABSTRACT Although mortality caused by Sarcoptes scabiei has been reported in European wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis), there is a lack of detailed information regarding the exposure of wild lagomorph species to this parasite. Here, we aimed to determine the seroprevalence and potential risk factors associated with S. scabiei exposure in European wild rabbits and Iberian hares in Mediterranean ecosystems of southern Spain. Between 2018/2019 and 2021/2022 hunting seasons, serum samples from 464 wild rabbits and 132 Iberian hares were collected from 100 hunting grounds in Andalusia (southern Spain). Sera were tested using an in-house indirect ELISA to detect specific anti-S. scabiei antibodies based on the immunodominant protein Ssλ20ΔB3. The overall apparent individual seroprevalence was 15.9% (95/596; 95%CI: 13.0–18.9). Antibodies against S. scabiei were detected in 11.6% (54/464; 95%CI: 8.7–14.5) of the European wild rabbits and 31.1% (41/132; 95%CI: 23.2–39.0) of the Iberian hares. Species (Iberian hare), age (adults) and geographical area (western Andalusia) were identified as risk factors potentially associated with S. scabiei exposure using generalized estimating equation analysis. By applying spatial analysis, two significant cluster of high seropositivity were detected in western and central Andalusia, respectively. The seroprevalence values obtained provide evidence of endemic, widespread and heterogeneous exposure to S. scabiei among wild lagomorph populations in Spanish Mediterranean ecosystems. Our findings underscore the importance of implementing integrated surveillance programs for sarcoptic mange in wild lagomorphs as well as in other sympatric species. 1. Introduction Sarcoptes scabiei, the causative agent of sarcoptic mange, is a highly contagious parasite with a worldwide distribution (Gort´ azar et al., 2007). Beside humans, the disease has been reported in 39 different families comprising 148 species of wild and domestic mammals, leading to significant economic, health and conservation issues (Escobar et al., 2022; Menzano et al., 2007; Tarigan and Huntley, 2005). Since the first description of sarcoptic mange in red fox (Vulpes vulpes) (Fürstenberg, 1861), the host range of S. scabiei has exponentially increased, expanding its distribution among wildlife across six continents (Escobar et al., 2022). This expansion, together with the high severity of the disease in some species, has led to recognize sarcoptic mange as a current emerging panzootic in wildlife (Escobar et al., 2022). In Spain, S. scabiei infection has been reported in a wide range of wild mammal species, including ungulates, carnivores, and lagomorphs, with * Corresponding author at: Departamento de Sanidad Animal, Grupo de Investigaci´ on en Sanidad Animal y Zoonosis (GISAZ), UIC Zoonosis y Enfermedades Emergentes ENZOEM, Universidad de C´ ordoba, 14014 C´ ordoba, Spain. E-mail address: [email protected] (I. García-Bocanegra). Contents lists available at ScienceDirect Research in Veterinary Science journal homepage: www.elsevier.com/locate/rvsc https://doi.org/10.1016/j.rvsc.2024.105344 Received 1 April 2024; Received in revised form 10 June 2024; Accepted 30 June 2024 Research in Veterinary Science 177 (2024) 105344 2 certain species demonstrating particular sensitivity to this parasite (Rossi et al., 2019). The first reported epizootic outbreak of sarcoptic mange affecting the Iberian ibex (Capra pyrenaica) occurred in 1987 in the Cazorla Natural Park (Andalusia, southern Spain). This outbreak resulted in a population decline of over 95% affecting an estimated population of nearly 9500 individuals (Le´ on-Vizcaíno et al., 1999). Since then, sarcoptic mange has caused high rates of morbidity and mortality in this wild Caprinae species, especially among populations with no previous exposure to the parasite (Rossi et al., 2019). In Spain, infection by S. scabiei has been documented in several wild ungulate species, including Cantabrian Chamois (Rupicapra pyrenaica parva) (Fern´ andez-Mor´ an et al., 1997), Barbary sheep (Ammotragus lervia) (Gonz´ alez-Candela et al., 2004), roe deer (Capreolus capreolus) (Oleaga et al., 2008a), red deer (Cervus elaphus) (Oleaga et al., 2008b), wild boar (Sus scrofa) (Valldeperes et al., 2021), and European mouflon (Ovis areis musimon) (Le´ on-Vizcaíno et al., 1999). Furthermore, clinical cases and mortality attributed to S. scabiei have also been reported in wild carnivore species, including the Iberian wolf (Canis lupus) (Oleaga et al., 2013), the threatened European wild cat (Felis silvestris silvestris) (N´ ajera et al., 2021), and the Iberian lynx (Lynx pardinus) (Oleaga et al., 2019). Regarding wild lagomorphs, mortality outbreaks by sarcoptic mange were initially observed in European wild rabbit (Oryctolagus cuniculus) populations in 2008 (Mill´ an, 2010), with the first fatal case reported in the Iberian hare (Lepus granatensis) in 2019 (Cardells et al., 2021). Despite the significance of these events, only a single serosurvey on S. scabiei has been conducted worldwide in wild rabbits, dating back 14 years (Mill´ an et al., 2012). Additionally, no studies have assessed the circulation of this pathogen in hare species so far. Given the susceptibility of wild rabbit and Iberian hare to S. scabiei and the paucity of information regarding their exposure to this parasite, the present study aims to determine the seroprevalence, potential risk factors and spatial distribution of S. scabiei in wild lagomorphs in Mediterranean ecosystems of southern Spain. 2. Material and methods 2.1. Study design and sampling A cross-sectional epidemiological study was carried out in Andalusia (36◦N-38◦60 ′ N, 1◦75 ′ W-7◦25 ′ W) (southern Spain) between 2018/2019 and 2021/2022 hunting seasons. This region exhibits a Mediterranean climate characterised by hot and dry summers and mild winters. The western region is noted for its higher humid levels and less extreme mean temperatures compared to the central and eastern regions. As a part of the epidemiological surveillance program, the Regional Government of Andalusia has divided the area into 23 hunting areas (Fig. 1) based on biological, environmental, and epidemiological factors, including the presence and abundance of large and small game populations (CAPMA. Consejería de Agricultura Pesca y Medio Ambiente, 2013). Sampling was stratified by hunting area based on the presence and density of wild rabbits in each area (CAPMA. Consejería de Agricultura Fig. 1. Spatial distribution of the seroprevalence of Sarcoptes scabiei in wild lagomorphs in Andalusia (southern Spain). S. Castro-Scholten et al. Research in Veterinary Science 177 (2024) 105344 3 Pesca y Medio Ambiente, 2013). Random selection of sampling hunting grounds was carried out within each hunting area. Hunters contributed between 1 and 33 (mean:12.9) European wild rabbits per hunting ground for sampling (Fig. 1). A total of 464 European wild rabbits were sampled from 36 hunting grounds distributed across 11 hunting areas over the course of four consecutive hunting seasons. Additionally, 132 Iberian hares were opportunistically sampled from 72 hunting grounds across 12 hunting areas within the same study area and period. Blood samples were collected from all animals, extracted from the heart or thoracic cavity, and centrifuged at 400 ×g for 10 min. The resulting serum was stored −80 ◦C until serological analysis was conducted. Whenever possible, information on each animal was recorded, including species, location, date of sampling, age, sex, body condition (based on the kidney fat index) (Flux, 1971) and external and internal lesions (Table 1). Three groups of age were considered according to bodyweight and body length (Morris, 1972): young (<40 days old), subadult (from 40 days to 8 months), or adult (over 8 months). Epidemiological data related to hunting grounds were also gathered through personal interviews with the gamekeepers using a standardized questionnaire. The information obtained included the characteristics of the hunting ground, the presence of disease and control measures, management practices, and presence of other domestic and wild sympatric species (Table 1). In addition, meteorological information for each sampling area [mean and maximum annual temperatures (◦C), humidity (g/m 3 ), and mean annual rainfall (mm)] was collected from the closest official meteorological station (Agencia Estatal de Meteorología (AEMET), 2023). 2.2. Laboratory analysis Sera were tested to detect specific anti-S. scabiei antibodies using an in-house indirect enzyme-linked assay (ELISA) based on the antigen Ssλ20ΔB3 an immunodominant recombinant protein derivates from the polypeptide Ssλ20 (accession number AM286740) (Casais et al., 2007). Ssλ20ΔB3 consists of the 263 amino acid residues from the carboxy terminus of the Ssλ20 polypeptide containing only one complete amino acid repeat present in Ssλ20. The ELISA was performed as previously described for wild rabbits (Mill´ an et al., 2012) with minor modifications. In this study, protein G horseradish peroxidase conjugate, diluted 2000 times in blocking solution, was used as secondary antibody, and TMB as substrate solution to visualize the reaction. The results were expressed as a percentage of the relative optical density (% relative OD 450nm ), calculated using the following formula: OD =[( ODsample–ODnegative control)/( ODpositive control–ODnegative control)]×100. Serum from an experimentally infested rabbit with S. scabei, collected prior to infestation and at week 11 post-challenge (Casais et al., 2014), served as negative and positive controls, respectively, for all plates. The cut-off level for this ELISA test was calculated as the mean of the % relative OD 450nm from 17 scabies-free wild-caught rabbits (Mill´ an et al., 2013) plus three times the standard deviation (Bomstein and Wallgren, 1997; Hollanders et al., 1997). 2.3. Statistical analyses The apparent prevalence of antibodies against S. scabiei was determined from the proportion of positive samples to the total number of examined, using the two-sided exact binomial test, with 95% confidence intervals (95%CI). Coefficients and standard error values generated by an intercept only generalized estimating equation (GEE) binomial logistic regression model, with hunting ground as subject variable, were used to adjust the estimated seroprevalence and 95%CI for clustering at hunting ground level (Dohoo et al., 2003). Associations between seroprevalence with explanatory variables were screened using the Pearson’s Chi-square or Fisher’s exact test, as appropriate. Cut-off points for Table 1 Distribution of the seroprevalence of Sarcoptes scabiei in wild lagomorphs in Andalusia (southern Spain) according to animal and hunting ground variables. Variable Categories No. positives/ Overall* Seroprevalence (%) Data recorded from the sampled animals Species European wild rabbit 54/464 11.6 Iberian hare 41/132 31.1 Age Adult 86/415 20.7 Subadult 6/135 4.4 Young 2/37 5.4 Sex Male 32/278 11.5 Female 61/310 19.7 Kidney fat index 0 35/107 32.7 1 17/148 11.5 2 11/108 10.2 3 13/99 13.1 Bodyweight (kg) 0.4–1.0 6/212 2.8 1.1–1.2 4/129 3.1 1.3–3.1 25/98 25.5 Body length (cm) 19–37 2/144 1.4 38–40 7/153 4.6 41–59 24/115 20.9 Hunting season 2018/2019 13/54 24.1 2019/2020 8/37 21.6 2020/2021 27/432 6.3 2021/2022 47/73 64.4 Hunting ground’s characteristics Geographical area Western 56/206 27.2 Central 27/177 15.3 Eastern 12/213 5.6 Burrow density High 51/413 12.3 Medium 21/61 34.4 Low 2/30 6.7 Predominant age group Adult animals 39/189 20.6 Young animals 9/177 5.1 Similar 24/121 19.8 High abundance of ticks in the hunting ground Yes 20/313 6.4 No 10/140 7.1 High abundance of fleas in the hunting ground Yes 19/279 6.8 No 11/174 6.3 Last restocking (months) No 32/293 10.9 ≤6 8/50 16.0 6–12 18/91 19.8 ≥12 16/70 22.9 Origin of the restocking Same game estate 13/139 9.4 Another game estate 15/49 30.6 Fenced hunting ground Yes 2/36 5.6 No 72/468 15.4 Distance to urban areas (Km) <10 71/459 15.5 10–20 1/28 3.6 Presence of rabbit feeders Yes 47/303 15.5 No 27/201 13.4 Feed supplementation in rabbits Yes 22/215 10.2 No 52/289 18.0 Presence of swamps Yes 7/34 20.6 No 67/470 14.3 Presence of waterholes Yes 32/198 16.2 No 42/306 13.7 Presence of sources Yes 3/142 2.1 No 71/362 19.6 Presence of troughs Yes 57/430 13.3 No 17/74 23.0 Presence of streams Yes 39/258 15.1 No 35/246 14.2 Crops intended for hunting Yes 44/236 18.6 No 30/268 11.2 Detection of clinical cases of other infectious diseases (continued on next page) S. Castro-Scholten et al. Research in Veterinary Science 177 (2024) 105344 4 continuous variables were determined at the 33rd and 66th percentiles, to homogenize the scales of the explanatory variables. All variables with a p <0.05 in the bivariate analysis were selected for further analysis. Collinearity between pairs of variables was then tested by Cramer’s V coefficient, and finally a GEE analysis was carried out to study the effect of the variables selected from the bivariate analysis. The number of positive animals was assumed to follow a binomial distribution, and ‘hunting ground’ was included as the subject variable. Forward selection was used for introduction of variables, starting with the variable with the lowest p-value in the bivariate analysis. The model was re-run until all remaining variables showed statistically significant values (p <0.05). For the choice of the best model, the quasi-likelihood under the independence model criterion (QIC) was considered (Table S1). SPSS 25.0 software (Statistical Package for Social Sciences, Inc., Chicago, IL, USA) was used for all statistical analyses. 2.4. Spatial cluster analysis The Bernoulli spatial scan statistic was applied to detect the presence of aggregations of hunting grounds with significantly higher S. scabiei seropositivity (Kulldorff et al., 2006), using SaTScanTM v10.1.2 software. The number of Monte Carlo simulations was set to 1000 for the cluster scan statistic. SaTScan was used to estimate relative risk (RR), representing the relative frequency of seropositive individuals compared to baseline, for each cluster. Clusters were considered significant at p < 0.05. 3. Results Antibodies against S. scabiei were found in 95 out of the 596 (15.9%; 95%CI: 13.0–18.9) wild lagomorphs tested. The apparent individual seroprevalence was 11.6% (54/464; 95%CI: 8.7–14.5) in European wild rabbit, and 31.1% (41/132; 95%CI: 23.2–39.0) in Iberian hare. After adjustment for clustering, the estimated individual seroprevalences were 11.8% (95%CI: 5.3–24.3) and 29.1% (95%CI: 21.6–38.0) in European wild rabbit and Iberian hare, respectively. None of the analysed animals showed lesions compatible with sarcoptic mange. At least one seropositive animal was found in 38.0% (38/100) and 71.4% (10/14) of the sampled hunting grounds and hunting areas, respectively. The Bernoulli model identified two statistically significant cluster in the study area. One was in western Andalusia with a radius of 16.2 km and included 7 hunting grounds (RR) =9.2; p <0.001). The second cluster was located in central Andalusia with a radius of 33.3 km and included 10 hunting grounds (RR: 3.1; p =0.022) (Fig. 1). Seroprevalence varied among hunting season, ranging from 6.3% (27/432; 95%CI: 4.0–8.5) in the 2020–2021 hunting season to 64.4% (47/63; 95%CI: 53.4–75.4) during the 2021–2022 season (Table 1). The explanatory variables obtained from the epidemiological questionnaire and the results of the bivariate analysis are summarized in Table 1 and Table S2. The final GEE model identified three variables Table 1 (continued) Variable Categories No. positives/ Overall* Seroprevalence (%) Outbreaks of myxomatosis in the last year Yes 74/470 15.7 No 0/34 0.0 Outbreaks of RHD** in the last year Yes 66/382 17.3 No 8/122 6.6 Outbreaks of myxomatosis in the last month Yes 60/324 18.5 No 14/180 7.8 Outbreaks of RHD** in the last month Yes 23/93 24.7 No 51/411 12.4 Presence of other sympatric species in the hunting ground Presence of wild boar (Sus scrofa) Yes 56/334 16.8 No 38/244 15.6 Presence of red deer (Cervus elaphus) Yes 14/45 31.1 No 80/533 15.0 Presence of European wildcat (Felis silvestris silvestris) Yes 28/155 18.1 No 46/349 13.2 Presence of badger (Meles meles) Yes 71/436 16.3 No 3/68 4.4 Presence of Iberian lynx (Lynx pardinus) Yes 45/86 52.3 No 29/418 6.9 Presence of mongoose (Herpestes ichneumon) Yes 67/372 18.0 No 1/125 0.8 Presence of genet (Genetta genetta) Yes 70/441 15.9 No 4/63 6.3 Presence of polecat (Mustela putorius) Yes 45/246 18.3 No 29/258 11.2 Presence of domestic cat (Felis silvestris catus) Yes 66/456 14.5 No 8/48 16.7 Presence of dog (Canis familiaris) Yes 62/389 15.9 No 12/115 10.4 Presence of cattle (Bos taurus) Yes 2/47 4.3 No 27/389 6.9 Presence of goat (Capra aegagrus hircus) Yes 17/171 9.9 No 12/265 4.5 Presence of sheep (Ovis aries) Yes 15/223 6.7 No 14/213 6.6 Presence of farmed rabbit (Oryctolagus cuniculus) Yes 32/54 59.3 No 42/450 9.3 Presence of domestic pig (Sus scrofa domesticus) Yes 3/38 7.9 No 26/398 6.5 Climate characteristics of the hunting ground Mean temperature (◦C) 12.3–16.9 23/220 10.5 17.0–17.5 15/116 12.9 17.6–18.5 5/159 3.1 Max temperature (◦C) 18.9–23.0 13/186 7.0 23.1–24.0 15/147 10.2 24.1–27.7 15/154 9.7 Mean annual rainfall (mm) 273.3–586.7 12/201 6.0 586.8–600.0 69/202 34.2 600.1–1135.0 6/143 4.2 Humidity (g/m 3 ) 56–64 9/79 11.4 65–66 13/120 10.8 67–100 4/64 6.3 * Missing values omitted. ** RHD: rabbit hemorrhagic disease. Table 2 Generalized estimating equations analysis of risk factors associated with exposure to Sarcoptes scabiei in wild lagomorphs in Andalusia (southern Spain). Variable Categories p-value OR 95%CI Species Iberian hare 0.016 3.2 (1.2–8.3) European wild rabbit a a Age Adult 0.034 1.8 (1.0–3.1) Young 0.786 0.8 (0.2–2.9) Subadult a a Geographical area Western 0.047 2.9 (1.0–8.2) Central 0.100 2.2 (0.9–5.7) Eastern a a a Reference Category. S. Castro-Scholten et al. Research in Veterinary Science 177 (2024) 105344 5 (species, age, and geographical area) as risk factors potentially associated with S. scabiei exposure (Table 2). Six explanatory variables were excluded from the GEE analysis due to collinearity (Table S3). Seropositivity in Iberian hares was significantly higher compared to European wild rabbits (p =0.016). Additionally, significantly higher seropositivity was found in adult animals (20.7%; 86/415) compared to young (5.4%; 2/37) and subadult individuals (4.4%; 6/135). The prevalence of anti-S. scabiei antibodies increased significantly in western Andalusia (27.2%; 56/206) compared to central (15.3%; 27/177) and eastern (5.6%; 12/213) regions. 4. Discussion The present large survey provides new epidemiological information regarding the distribution of sarcoptic mange among wildlife in Mediterranean ecosystems. The overall seroprevalence found in this study (15.9%) indicates a moderate exposure of wild lagomorph populations to S. scabiei. Specifically, the seroprevalence found in European wild rabbits (11.8%) is consistent with the 13.0% detected in the only serosurvey carried out in this species to date (Mill´ an et al., 2012). To the best of author’s knowledge, this is the first epidemiological study conducted on this parasite in hare species worldwide. The seroprevalence detected in Iberian hare (29.1%) indicates a high circulation of S. scabiei in their populations in southern Spain. It should be noted that none of the animals sampled in the present study showed macroscopic lesions compatible with sarcoptic mange, and outbreaks of sarcoptic mange were not observed by the gamekeepers in the hunting grounds sampled. Taking into account the observed seroprevalences in both species, these findings may reflect different situations, including: i) the presence of undetected skin lesions during external inspection, ii) self-limiting infestation (Mill´ an et al., 2013) and/or iii) clinically asymptomatic infestations as previously suggested (Mill´ an et al., 2012). Nevertheless, further studies are needed to clarify this aspect. Significantly higher seropositivity was found in Iberian hares compared to wild rabbits, indicating that either both species are not equally exposed to S. scabiei or present differences in susceptibility to infection. In this respect, while various outbreaks of sarcoptic mange have been reported in wild rabbits (Mill´ an, 2010; Navarro-Gonzalez et al., 2010), only one clinical case has been described in hares to date (Cardells et al., 2021). Additionally, besides species, seropositivity to S. scabiei in wild lagomorphs was also age-related. There was a significantly higher seroprevalence observed in adults compared to subadults and young animals. This difference may be indicative of the cumulative likelihood of re-exposure to the parasite throughout the lifetime and/or the prolonged persistence of antibodies. Moreover, although anti- S. scabiei antibodies were not detectable after eight weeks in experimentally infested wild rabbits that recovered from the disease (Mill´ an et al., 2013), the antibody response was shown to be stronger and longer after re-infestation (Casais et al., 2014). In natural conditions, wild lagomorphs may be regularly exposed to low doses of the parasite, leading to the production of long-lasting antibodies without actually developing clinical disease. Seropositive animals were found in all the hunting seasons, including young or subadult animals in each of them. At least one positive animal was also detected in 38.0% and 71.4% of the hunting grounds and hunting areas sampled, respectively. These findings indicate a wide endemic circulation of the parasite among wild lagomorph populations in southern Spain. Nevertheless, the significantly higher seroprevalence detected in western Andalusia, and the two clusters found in this region and central Andalusia revealed that the distribution of the parasite in these species was not homogeneous across the study area (Fig. 1). The transmission dynamics of sarcoptic mange in wildlife has been associated with several factors, such as environmental conditions and animal density (Iacopelli et al., 2020; Mill´ an et al., 2012; Pence and Ueckermann, 2002). Consistent with this, the western region of Andalusia exhibits higher mean humidity, average annual rainfall, less-extreme mean temperatures, and abundance of wild lagomorphs than those observed in central and eastern regions (Agencia Estatal de Meteorología (AEMET), 2023; CAPMA. Consejería de Agricultura Pesca y Medio Ambiente, 2013). These conditions not only favour greater parasite survival in the environment but also increases the number of direct contacts between individuals, thus facilitating the circulation of S. scabiei among their populations (Arlian, 1989; Mill´ an et al., 2012). In conclusion, the results obtained indicate a widespread, endemic, and heterogeneous circulation of S. scabiei in wild lagomorphs within the Spanish Mediterranean ecosystems of southern Spain. The seroprevalence values detected, particularly in Iberian hare populations, underscore the importance of implementing integrated surveillance programs for S. scabiei focused on wild lagomorphs and other sympatric species involved in the transmission and maintenance of this multi-host parasite. Further studies are required to evaluate the differences between wild lagomorph species and to assess the impact of S. scabiei in their populations in the Iberian Mediterranean ecosystems. Supplementary data to this article can be found online at https://doi. org/10.1016/j.rvsc.2024.105344. Ethical statement This study did not involve purposeful killing of animals. All samples were collected from legally hunted animals during the hunting seasons or by passive surveillance under Spanish and Andalusian legislation. Thus, no ethical approval was necessary. CRediT authorship contribution statement Sabrina Castro-Scholten: Writing – review & editing, Writing – original draft, Methodology, Investigation, Formal analysis, Data curation, Conceptualization. Ignacio García-Bocanegra: Writing – review & editing, Writing – original draft, Visualization, Validation, Supervision, Resources, Project administration, Methodology, Investigation, Funding acquisition, Formal analysis, Data curation, Conceptualization. Natalia Iglesias: Writing – review & editing, Visualization, Validation, Methodology, Formal analysis. Rosa Casais: Writing – review & editing, Visualization, Validation, Resources, Methodology, Formal analysis. David Cano-Terriza: Writing – review & editing, Visualization, Validation, Investigation, Data curation, Conceptualization. D´ ebora Jim´ enez-Martín: Writing – review & editing, Visualization, Validation, Methodology, Investigation, Formal analysis. Jesús Barbero-Moyano: Writing – review & editing, Visualization, Validation, Methodology, Investigation, Data curation. Leonor Camacho-Sillero: Writing – review & editing, Visualization, Validation, Formal analysis, Data curation. Carlos Rouco: Writing – review & editing, Visualization, Validation, Methodology, Funding acquisition, Data curation, Conceptualization. Javier Caballero-G´ omez: Writing – review & editing, Writing – original draft, Visualization, Validation, Supervision, Methodology, Investigation, Formal analysis, Data curation, Conceptualization. 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. Data availability The data that support the findings of this study are available from the authors upon reasonable request. Acknowledgements We gratefully acknowledge the help of the personal of the S. Castro-Scholten et al. Research in Veterinary Science 177 (2024) 105344 6 Epidemiological Surveillance Program in Wildlife (Regional Government of Andalusia) an all the members of GISAZ research group in the collection of samples and epidemiological information. This work has benefited from the financial aid of research grants funded by the Spanish Ministry of Science and Innovation (project LagoHealth; REF: PID2019- 111080RB-C21 and LAGOMED project REF: PRIMAS2-11-PCI2019- 103698) and by the University of C´ ordoba, Spain (REF: UCO-FEDER- 1264967; Sub-modality 2.4. “UCOLIDERA”). This research was also supported by CIBER-Consorcio Centro de Investigaci´ on Biom´ edica en Red (CB 2021), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovaci´ on and Uni´ on Europea-Next Generation EU. S. Castro-Scholten, J. Barbero-Moyano and D. Jim´ enez-Martín are supported by an FPU grant from the Spanish Ministry of Universities (FPU19/06026, FPU20/ 00180 and FPU22/03649, respectively). We also wish to thank Dr. Javier Mill´ an for gratefully giving us sera from the 17 scabies-free wildcaught rabbits used to calculate the cut-off point of the ELISA. Funding for open access charge: Universidad de C´ ordoba / CBUA. References Agencia Estatal de Meteorología (AEMET), 2023. Visor del Atlas clim´ atico de la Península y Baleares. Retrieved from. https://www.aemet.es/es/serviciosclimatico s/datosclimatologicos/atlas_climatico/visor_atlas_climatico (Accessed 8 th January 2023). Arlian, L.G., 1989. Biology, host relations, and epidemiology of Sarcoptes scabiei. Annu. Rev. Entomol. 34 (1), 139–159. Bomstein, S., Wallgren, P., 1997. Serodiagnosis of sarcoptic mange in pigs. Vet. Rec. 141 (1), 8–12. CAPMA. Consejería de Agricultura Pesca y Medio Ambiente, 2013. Programa de Vigilancia Epidemiol´ ogica de la Fauna Silvestre. Retrieved from: https://www.junt adeandalucia.es/medioambiente/portal/landing-page-%C3%ADndice/-/asset _publisher/zX2ouZa4r1Rf/content/programa-de-vigilancia-epidemiol-c3-b3gica-de- la-fauna-silvestre-en-andaluc-c3-ada-pve-1/20151?categoryVal=(Accessed 05 th January 2024). Cardells, J., Lizana, V., Martí-Marco, A., Lavín, S., Velarde, R., Rossi, L., Moroni, B., 2021. First description of sarcoptic mange in an Iberian hare (Lepus granatensis). CRPVBD 1, 100021. Casais, R., Prieto Martín, M., Balseiro Morales, A., Solano, P., Parra, F., 2007. Identification and heterologous expression of a Sarcoptes scabiei cDNA encoding a structural antigen with immunodiagnostic potential. Vet. Res. 38, 435–450. Casais, R., Dalton, K.P., Mill´ an, J., Balseiro, A., Oleaga, ´ A., Solano, P., Goyache, F., Prieto, J.M., Parra, F., 2014. Primary and secondary experimental infestation of rabbits (Oryctolagus cuniculus) with Sarcoptes scabiei from a wild rabbit: factors determining resistance to reinfestation. Vet. Parasitol. 203, 173–183. Dohoo, I.R., Martin, W., Stryhn, H.E., 2003. Veterinary Epidemiologic Research. University of Prince Edward Island, Charlottetown, P.E.I. Escobar, L.E., Carver, S., Cross, P.C., Rossi, L., Almberg, E.S., Yabsley, M.J., Niedringhaus, K.D., Van Wick, P., Dominguez-Villegas, E., Gakuya, F., Xie, Y., Angelone, S., Gort´ azar, C., Astorga, F., 2022. Sarcoptic mange: an emerging panzootic in wildlife. Transbound. Emerg. Dis. 69, 927–942. Fern´ andez-Mor´ an, J., G´ omez, S., Ballesteros, F., Quir´ os, P., Benito, J., Feliu, C., Nieto, J., 1997. Epizootiology of sarcoptic mange in a population of cantabrian chamois (Rupicapra pyrenaica parava) in northwestern Spain. Vet. Parasitol. 73 (1–2), 163–171. Flux, J.E., 1971. Validity of the Kidney Fat Index For Estimating the Condition of Hares: A Discussion, pp. 238–244. Fürstenberg, M.H., 1861. Die Kr¨ atzmilben der Menschen und Thiere. Engelmann. Gonz´ alez-Candela, M., L´ eon-Vizcaíno, L., Cubero-Pablo, M.J., 2004. Population effects of sarcoptic mange in barbary sheep (Ammotragus lervia) from sierra Espu˜ na Regional Park, Spain. J. Wildl. Dis. 40 (3), 456–465. Gort´ azar, C., Ferroglio, E., H¨ ofle, U., Fr¨ olich, K., Vicente, J., 2007. Diseases shared between wildlife and livestock: a European perspective. Eur. J. Wildl. Res. 53, 241–256. Hollanders, W., Vercruysse, J., Raes, S., Bornstein, S., 1997. Evaluation of an enzymelinked immunosorbent assay (ELISA) for the serological diagnosis of sarcoptic mange in swine. Vet. Parasitol. 69 (1–2), 117–123. Iacopelli, F., Fanelli, A., Tizzani, P., Berriatua, E., Prieto, P., Martínez-Carrasco, C., Le´ on, L., Rossi, L., Candela, M.G., 2020. Spatio-temporal patterns of sarcoptic mange in red deer and Iberian ibex in a multi-host natural park. Res. Vet. Sci. 128, 224–229. Kulldorff, M., Huang, L., Pickle, L., Duczmal, L., 2006. An elliptic spatial scan statistic. Stat. Med. 25 (22), 3929–3943. Le´ on-Vizcaíno, L., Ruíz De Yb´ a˜ nez, M.R., Cubero, M.J., Ortíz, J.M., Espinosa, J., P´ erez, L., Sim´ on, M.A., Alonso, F., 1999. Sarcoptic mange in spanish ibex from Spain. J. Wildl. Dis. 35, 647–659. Menzano, A., Rambozzi, L., Rossi, L., 2007. A severe episode of wildlife-derived scabies in domestic goats in Italy. Small Rumin. Res. 70 (2–3), 154–158. Mill´ an, J., 2010. First description of sarcoptic mange in wild European rabbit (Oryctolagus cuniculus). Eur. J. Wildl. Res. 56 (3), 455–457. Mill´ an, J., Cas´ ais, R., Delibes-Mateos, M., Calvete, C., Rouco, C., Castro, F., Colomar, V., Casas-Díaz, E., Ramírez, E., Moreno, S., Prieto, J.M., Villafuerte, R., 2012. Widespread exposure to Sarcoptes scabiei in wild European rabbits (Oryctolagus cuniculus) in Spain. Vet. Parasitol. 183, 323–329. Mill´ an, J., Casais, R., Colomar, V., Bach, E., Prieto, J.M., Velarde, R., 2013. Experimental infection of wild-caught European rabbits (Oryctolagus cuniculus) with Sarcoptes scabiei from a naturally infected wild rabbit. Med. Vet. Entomol. 27 (2), 232–235. Morris, P., 1972. A review of mammalian age determination methods. Mammal Rev. 2, 69–104. N´ ajera, F., Crespo, E., García-Talens, A., Grande-G´ omez, R., Herrera-S´ anchez, F.J., Gentil, M., Cort´ es-García, C., Müller, E., Calero-Bernal, R., Revuelta, L., 2021. First description of Sarcoptic mange in a free-ranging European wildcat (Felis silvestris silvestris) from Spain. Animals 11, 2494. Navarro-Gonzalez, N., Serrano, E., Casas-Díaz, E., Velarde, R., Marco, I., Rossi, L., Lavín, S., 2010. Game restocking and the introduction of sarcoptic mange in wild rabbit in North-Eastern Spain. Anim. Conserv. 13 (6), 586–591. Oleaga, A., Balseiro, A., Gort´ azar, C., 2008a. Sarcoptic mange in two roe deer (Capreolus capreolus) from northern Spain. Eur. J. Wildl. Res. 54, 134–137. Oleaga, ´ A., Casais, R., Gonz´ alez-Quir´ os, P., Prieto, M., Gort´ azar, C., 2008b. Sarcoptic mange in red deer from Spain: improved surveillance or disease emergence? Vet. Parasitol. 154 (1–2), 103–113. Oleaga, A., Alasaad, S., Rossi, L., Casais, R., Vicente, J., Maione, S., Soriguer, R.C., Gort´ azar, C., 2013. Genetic epidemiology of Sarcoptes scabiei in the Iberian wolf in Asturias, Spain. Vet. Parasitol. 196, 453–459. Oleaga, A., García, A., Balseiro, A., Casais, R., Mata, E., Crespo, E., 2019. First description of sarcoptic mange in the endangered Iberian lynx (Lynx pardinus): clinical and epidemiological features. Eur. J. Wildl. Res. 65 (3), 40. Pence, D.B., Ueckermann, E., 2002. Sarcoptic manage in wildlife. Revue Sci. Techn. (Int. Off. Epizoot.). 21 (2), 385–398. Rossi, L., Tizzani, P., Rambozzi, L., Moroni, B., Meneguz, P.G., 2019. Sanitary emergencies at the wild/domestic caprines interface in Europe. Animals 9 (11), 922. Tarigan, S., Huntley, J.F., 2005. Failure to protect goats following vaccination with soluble proteins of Sarcoptes scabiei: evidence for a role for IgE antibody in protection. Vet. Parasitol. 133 (1), 101–109. Valldeperes, M., Moroni, B., Rossi, L., L´ opez-Olvera, J.R., Velarde, R., Molinar Min, A.R., Mentaberre, G., Serrano, E., Angelone, S., Lavín, S., Granados, J.E., 2021. First report of interspecific transmission of sarcoptic mange from Iberian ibex to wild boar. Parasit. Vectors 14, 481. S. Castro-Scholten et al.