BRIEF RESEARCH REPORT published: 01 April 2022 doi: 10.3389/fvets.2022.830157 Ivermectin Plasma Concentration in Iberian Ibex (Capra pyrenaica) Following Oral Administration: A Pilot Study Barbara Moroni 1*† , José Enrique Granados Torres 2† , Jorge Ramón López-Olvera 3† , José Espinosa Cerrato 4† , Arián Ráez Bravo 3 , Gregorio Mentaberre 5 , Paulino Fandos 6† , Marco Pazzi 7† , Monica Romagnoli 7† , Giulia Gardini 1 , Luca Rossi 1 , Marta Valldeperes 3† , Edited by: J. Alberto Montoya-Alonso, Emmanuel Serrano 3 , Blanca Ramos 2† and Rosangela Odore 1† University of Las Palmas de Gran 1 Department of Veterinary Sciences, University of Turin, Turin, Italy, 2 Parque Nacional y Parque Natural de Sierra Nevada Canaria, Spain Carretera Antigua Sierra Nevada, Granada, Spain, 3 Wildlife Ecology & Health Group (WE&H) and Servei d’Ecopatologia de Reviewed by: Fauna Salvatge (SEFaS), Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Vincenzo Veneziano, Barcelona, Barcelona, Spain, 4 Departamento Sanidad Animal, Instituto de Ganadería de Montaña (CSIC-ULE), Facultad de University of Naples Federico II, Italy Veterinaria, León, Spain, 5 Wildlife Ecology & Health Group and Departament de Ciència Animal, Escola Tècnica Superior Cristóbal Briceño, d’Enginyeria Agraria (ETSEA), Universitat de Lleida (UdL), Lleida, Spain, 6 Agencia de Medio Ambiente y Agua, Sevilla, Spain, 7 University of Chile, Chile Department of Chemistry, University of Turin, Turin, Italy Juan Alberto Corbera, University of Las Palmas de Gran Canaria, Spain Sarcoptic mange is considered the main driver of demographic declines occurred in the *Correspondence: last decades in Iberian ibex (Capra pyrenaica) populations. Mass treatment campaigns Barbara Moroni by administration of in-feed acaricides are used as a measure to mitigate the impact barbara_moroni@yahoo.it of mange in the affected populations. However, there are no data on ivermectin (IVM) † ORCID: Barbara Moroni pharmacokinetics in this wild caprine, and the treatment through medicated feed is orcid.org/0000-0003-1478-8436 not endorsed by evidence on its effectiveness. The aim of this study is to determine José Enrique Granados Torres the pharmacokinetic profile of IVM in plasma samples of ibexes after the experimental orcid.org/0000-0002-9787-9896 Jorge Ramón López Olvera oral administration of IVM, using high performance liquid chromatography (HPLC) with orcid.org/0000-0002-2999-3451 automated solid phase extraction and fluorescence detection. A dose of 500 µg of IVM José Espinosa Cerrato orcid.org/0000-0002-9036-1402 per body weight was orally administered in a feed bolus to nine healthy adult ibexes Paulino Fandos (seven males and two females). Blood samples were collected by jugular venipuncture orcid.org/0000-0002-9607-8931 into heparin-coated tubes at day 1, 2, 3, 4, 7, 10, 15, and 45 post-administration Marco Pazzi orcid.org/0000-0002-9763-8785 (dpa). The highest plasma concentration of IVM (Cmax = 3.4 ng/ml) was detected 24 h Monica Romagnoli after the oral administration (T1), followed by a rapid decrease during the first week orcid.org/0000-0003-1119-4229 Marta Valldeperes post-administration. Our results reveal that plasma IVM concentration drops drastically orcid.org/0000-0002-6178-3147 within 5 days of ingestion, questioning the effectiveness of a single in-feed dose of this Blanca Ramos drug to control sarcoptic mange. To the best of our knowledge, this is the first study on orcid.org/0000-0003-1417-2455 Rosangela Odore plasma availability of oral IVM in ibexes and in any wild ungulate species. orcid.org/0000-0001-8529-2003 Keywords: ivermectin, sarcoptic mange, wildlife, Iberian ibex, treatment, medicated feed Specialty section: This article was submitted to Parasitology, a section of the journal INTRODUCTION Frontiers in Veterinary Science Iberian ibex (Capra pyrenaica) is a wild ungulate native to the mountain ranges of the Iberian Received: 06 December 2021 Peninsula and has recently been reintroduced in the French side of the Pyrenees. This caprine Accepted: 09 March 2022 currently enjoys a favorable conservation status (“least concern” according to the IUCN), which Published: 01 April 2022 allows sustainable hunting as a valuable game species. Besides its biological and ecological value, Frontiers in Veterinary Science | www.frontiersin.org 1 April 2022 | Volume 9 | Article 830157 Moroni et al. Ivermectin Plasma Concentration in Iberian Ibex ibex trophy hunting and the associated related tourism represent feed. In domestic ruminants, interspecies and intra-species a significant source of revenue for remote and disadvantaged variability in IVM pharmacokinetics has been observed. In rural communities in Spain (1). As for the majority of large domestic goat, a greater plasma clearance of IVM leads herbivores in Europe, the hunting quotas are established by to lower plasma concentrations than in cattle and sheep, public conservation agencies, based on the estimated population regardless of the route of drug administration (14). After size, although hunting is banned in specific protected areas such oral administration, IVM bioavailability is about four times as National Parks (2, 3). greater and plasma concentrations can be detected longer in Since the late eighties of the twentieth century, free-ranging sheep than in goats (15). Moreover, IVM pharmacokinetics Iberian ibex populations suffered from epidemic waves of may depend on sex and age, and the presence of rumen sarcoptic mange, a contagious skin disease caused by the digesta significantly influences the systemic availability of the burrowing mite Sarcoptes scabiei. After the epidemics, mange has drug (16). remained endemic in all the populations affected. Devastating The main findings related to domestic ruminants and demographic effects have been recorded in naïve herds, including the paucity of data concerning wild species recommend mortality rates over 90% (4, 5). Sarcoptic mange is now caution when extrapolating treatment protocols (e.g., dose considered the main driver of short to medium term population and formulation) from one species to another and, as a declines in this wild ruminant (6). consequence, extended pharmacokinetic data in target animal Control of sarcoptic mange epizootics in wildlife is species are much-needed to avoid drug misuse. Therefore, challenging. Amongst other measures like non–intervention the aim of the present experimental study is to begin filling (laissez-faire), massive lethal control, and selective culling of the existing knowledge gaps by exploring, for the first time, clinically-affected animals (7), individual and mass treatments the pharmacokinetic profile of a single oral dose of IVM in with acaricides have been proposed and empirically implemented Iberian ibexes. at local scale with unknown success (8). However, beyond ethical considerations, treating free-ranging wildlife has disadvantages, such as the impracticability of drug administration on a MATERIALS AND METHODS large population scale, and the potential environmental contamination with drug residues (9, 10). In Spain, the lack Animals of shared protocols among regions to tackle the population Nine adult healthy Iberian ibexes (seven males and two females, decline caused by sarcoptic mange in exposed wild ruminants age ranging 1–5 years) were captured in Sierra Nevada Natural has promoted the empirical use of acaricides massively Space as part of the regular management of the species and administered through medicated mixture feed (6). Nevertheless, transferred to the “Iberian ibex stock reservoir El Toril”, in ′ ′ ′ ′ no studies support the actual effectiveness of this common Dílar, Granada (37◦ 02 -7◦ 03 N, 3◦ 22 -3◦ 33 W), southern Spain practice. In fact, essential gaps in knowledge need to be filled (17). The ibexes were kept isolated from the rest of the before considering oral mass treatments as a possible control reservoir ibex population during all the study period. Average measure for the management of sarcoptic mange in free-ranging weight was 19 kg for females and 26 kg for males. The study Iberian ibex and other susceptible wild ruminants (11, 12). was approved by the Ethics on Animal Welfare Committee Among the knowledge gaps to fill before implementing of the University of Jaén and authorized by the Dirección any in-field treatment, the pharmacokinetics of orally- General de Producción Agrícola y Ganadera of the Consejería administered candidate acaricides in the target hosts are de Agricultura, Pesca y Medio Ambiente of the Junta de crucial. Pharmacokinetic is the keystone to establish effective Andalucía (Ref: SA/SIS/MD/ps/October 25, 2012). The Sierra drugs, dosages, vehicles, frequency of administration, number Nevada National and Natural Park Administration also approved of feeding points according to surface area and population, and this study. uptake rates of medicated feed that should be reached in order to obtain a mass effect limiting the impact of the disease under field conditions. Difficulties in recruiting, maintaining and repeatedly Pharmacotherapy and Blood Sample handling Iberian ibexes have, so far, understandably limited the Collection necessary trials. After an acclimation period of at least 1 month in the In captive wild ruminants, the macrocyclic lactone ivermectin experimental facilities, an ad hoc formulated bolus containing (IVM) has been used against S. scabiei infection at dose rates 0.5 mg of IVM per kg of body weight was orally administered of 200–400 µg/kg, and repeated subcutaneous administrations to each ibex. For this, specialized wildlife operators physically were necessary to eliminate severe clinical signs (8). Oral immobilized, blindfolded, and administered the boluses to the administration has also been described in wild free-ranging ibexes using commercial applicators. Blood samples (20 ml) were animals (6, 8, 13); nevertheless, the plasma concentration collected from the jugular vein into heparin-coated vacutainers in treated individuals, and the drug plasma therapeutic just prior to drug administration (time 0) and 1 (T1), 2 (T2), 3 concentrations against sarcoptic mange were unknown, as well (T3), 4 (T4), 7 (T5), 10 (T6), 15 (T7), and 45 (T8) days after the as the proportion of individuals having access to medicated bolus administration (dpa). Plasma was obtained immediately after the blood collection by centrifugation at 2,000 g for 20 mins Abbreviations: IVM, ivermectin; HPLC, high pressure liquid chromatography. and stored in 2 ml containers at −80◦ C until analyzed. Frontiers in Veterinary Science | www.frontiersin.org 2 April 2022 | Volume 9 | Article 830157 Moroni et al. Ivermectin Plasma Concentration in Iberian Ibex Chemical Extraction, Derivatization and DISCUSSION HPLC With Fluorescence Detection IVM was analyzed in the plasma samples by high performance To the best of our knowledge, this is the first study investigating liquid chromatography (HPLC) with automated solid phase the pharmacokinetic profile of orally administered IVM in extraction and fluorescence detection following a validated Iberian ibex and in any wild ruminant species. The controlled method previously described (18). Briefly, 1 ml plasma aliquot conditions of the experiment will allow the use of the was mixed with 5 ng internal standard abamectin and vortexed. results as reference for future experimental scenarios and/or The solution was mixed with 1 ml acetonitrile/water solution field studies. Sarcoptic mange has been pharmacologically (4:1) for 30 mins and then centrifuged at 17,000 g at room treated in captive wildlife, and some experiences in free- temperature for 5 mins. The resultant supernatant was applied ranging wildlife populations have also been carried out, mostly at 10 mg/ml to a cartridge (Strata-X 33 µm Polymeric Reversed empirically (8, 10, 23). Phase, Phenomenex, Torrance, CA, US), previously activated However, the complete eradication of mange from a with 1 ml methanol and 1 ml of water. Elution was performed population in endemic areas through pharmacological control with 1.3 ml methanol. The dried residue was dissolved in strategies has been never achieved and so far deemed as 100 µl N-methylmidazole solution in acetonitrile (1:2) and unrealistic or inconclusive (10). derivatized with 150 µl trifluoroacetic anhydride solution in Literature is poorly informative on the efficacy of different acetonitrile (1:3). doses of orally administered macrocyclic lactones for the A 100-µl aliquot of the derivatized samples was injected treatment of sarcoptic mange in wild ruminants (8). Hence, the into the chromatograph. The mobile phase consisted of acetic IVM dose in this study (0.5 mg/kg) must be intended as an acid 0.2% in water/methanol/acetonitrile (10:40:50) pumped empiric trade-off between a limited number of relevant sources. at a flow rate of 1.4 ml/min through a column (Gemini Amongst them, Yeruham and colleagues (24) successfully treated C18, 150 x 4.6 mm, Phenomenex) with fluorescence detection. five ruminant species affected by sarcoptic mange in zoo gardens, Fluorescence detection (RF 2000 Fluorescence Detector, Dionex) including the Nubian ibex (Capra nubiana), a close relative of was performed at 365 nm excitation and 475 nm emission the Iberian ibex. Their protocol provided for the administration wavelength. Recovery rate was calculated for each sample of an oral dose of 0.2 mg/kg of IVM for three consecutive through internal standard recovery correction. days repeated three times at 2 week intervals. This protocol is clearly unfit for mass use under free-ranging conditions, since regular access of target wildlife to medicated feed is far from being guaranteed; nevertheless, the results are encouraging with Data Analysis respect to possible oral treatment options in Sarcoptes-exposed A Linear Mixed Model (LMM with a normal error distribution wild ruminants. Moreover, Leon-Vizcaino and colleagues (25) and identity link function) was fitted to explore the showed that 0.4 mg/kg of subcutaneously administered IVM pharmacokinetics of plasma IVM concentrations (ng/ml, log- were preferable to 0.2 mg/kg for the treatment of spontaneously transformed) in the study ibexes. Time since drug administration infested Iberian ibex, and Foreyt (26) reported the successful in- was included as fixed factors whereas ibex identity as a random feed use of IVM for 7 consecutive days at the dose of 1 mg/kg intercept term in the LMM (19). For the mixed models we used for the treatment of Rocky Mountain bighorns (Ovis canadensis) the library lme4 1.1-15 version (19), whereas the library MuMIn experimentally infested with Psoroptes mites. The 0.5 mg/kg IVM 1.43.6 version (20) was employed to assess the marginal and dose in our study took into account the above findings, the lower conditional contribution of the fixed and random terms (21). All antiparasitic efficacy of oral IVM when administered at the same the statistical analysis were performed with R Statistical Software dosage recommended via subcutaneous inoculation (27) and, 4.1.0 version (22). prospectively, the individual variability in medicated food intake by free-ranging wild ruminants. This study revealed that IVM plasma concentrations in RESULTS orally-treated Iberian ibexes drop drastically within four dpa. Not surprisingly, the oral route leads to lower IVM plasma The limit of quantification was 0.2 ng/ml. Although IVM was concentrations compared to those reached after a subcutaneous detected in plasma from dpa 1 to dpa 45, mean plasma IVM administration (28), likely due to the binding to the particulate concentration dropped from the initial 3.40 ng/ml in dpa 1 to phase of digesta (14, 29). Moreover, the binding of IVM to 0.63 ng/ml in dpa 4 (Figure 1A), and stabilized thereafter around plasma albumin and lipoproteins should be taken into account 0.25 ng/ml. This temporal decrease (β = −0.4, SE = 0.04, t especially in animals with chronic stages of sarcoptic mange = −11.11, p-value < 0.01) explained 61% of the plasma IVM usually showing emaciation and poor body condition, in which concentration variability in the ibexes of the study. Moreover, a decrease in plasma proteins (thus a higher free fraction of IVM the individual contribution to the observed patterns was 8% in the plasma), might be more likely associated to an unsuccessful (Figure 1B). treatment (14). The pharmacokinetic values of orally-administered IVM in The maximum drug concentration and bioavailability found the Iberian ibexes are reported in Table 1. No adverse response in Iberian ibex (3.40 ± ng/ml) were lower than those described in was observed throughout the study. goats (6.03 ± 0.95; 15.85 ± 5.29 ng/ml, respectively) after the oral Frontiers in Veterinary Science | www.frontiersin.org 3 April 2022 | Volume 9 | Article 830157 Moroni et al. Ivermectin Plasma Concentration in Iberian Ibex FIGURE 1 | (A) Time trend of the mean plasma ivermectin concentration (ng/ml) in the nine Iberian ibexes orally administered with ivermectin from 1 (T1) to 45 (T8) days post-administration; (B) Differences among individual ibexes in the decrease of plasma ivermectin concentration throughout the study period. TABLE 1 | Ivermectin pharmacokinetic values in plasma of the experimental action of macrocyclic lactones is related to their plasma Iberian ibexes after the oral administration of a 0.5 mg/kg dose. concentrations, and more specifically to the bioavailability (34). Cmax (ng/ml) Tmax (days) AUC (days · ng/ml) Tlast (day) MRT (days) On the other hand, due to its high lipophilic nature, IVM is widely distributed within the body in all species, with highest 3.40 ± 1.2 1 21.27 45 30.80 concentrations achieved in liver and adipose tissue, where it tends to accumulate (14). This might explain the rapid drop of IVM Cmax, maximum concentration; Tmax, time for maximum concentration; AUC, Area Under Curve; Tlast, time of last detection of ivermectin; MRT, mean residence time. observed in the plasma of Iberian ibexes in this study, as well as the long MRT (30.8), pointing to a possible quick deposit in other tissues. Interestingly, it has been experimentally observed administration of an IVM dose of 0.2 mg/kg (30, 31). By contrast, that IVM is detectable in the skin of scabietic pigs for at least 10 the time to maximum concentration (1 day) was consistent with days after a single oral administration of 0.2 mg/kg (35). previous reports of oral administration in other medium-sized Therapeutic or effective threshold for IVM plasma ruminants, sheep (1.7 day), goat (2.8 days), and reindeer (2 days concentrations against sarcoptic mange has not been defined for oral mixture, 1 day for oral paste) (15, 18, 28). By comparing in any ruminant model. In cattle, the minimum IVM blood two different oral formulations in reindeer, Oksanen et al. (28) concentration for anthelmintic activity against nematodes ranges found a lower relative plasma availability for a paste than for from 0.5 to 1 ng/ml (36). By contrast, a higher therapeutic an oral liquid drench formulation, which could also partially threshold level (8 ng/ml) has been suggested after a subcutaneous explain the lower values observed in this study. The solid bolus injection of 0.6 mg/kg long-acting IVM against common bovine formulation used in this study does not appear in previous similar ticks (Boophilus spp) (37), showing that therapeutic blood trials, but we deemed it: i) better representative, than a liquid levels may depend, among others, on the animal species and drench, of the medicated pellet administration that is already related metabolism, as well as the target parasite. Moreover, empirically used in Iberian ibex (32); ii) better suited to provide the therapeutic efficacy and the associated threshold of IVM an accurate and precise IVM dosing than medicated pellets. plasma concentrations may vary depending on the severity of Several factors known to influence drug absorption and sarcoptic mange, with the most affected individuals being less bioavailability may explain the interindividual variability in IVM responsive to the therapeutic effect, thus requiring a higher plasma concentrations, observed in particular at T1-T4 (33). dosage to eliminate the mite and heal (30), as shown in other Nevertheless, as anticipated, the variability could be even greater animal species (31). under field conditions (e.g., mass-delivery treatment schemes The efficacy of IVM plasma concentrations against S. scabiei with delivery of medicated feed), thus the variability in the in Iberian ibex and other wild ungulate species should be IVM plasma concentrations reached should likely be greater further investigated, also considering that underdosing is a due to inter-individual variability in medicated food intake, with recognized driver for the development of resistance to endo and unpredictable consequences on IVM efficacy. The antiparasitic ectoparasiticides (8, 10, 38, 39). At present, resistance of S. scabiei Frontiers in Veterinary Science | www.frontiersin.org 4 April 2022 | Volume 9 | Article 830157 Moroni et al. Ivermectin Plasma Concentration in Iberian Ibex against IVM has been described both, in vitro and in vivo, limited experimental studies in both domestic animals and captive to human model (40, 41). In the particular case of sustained oral wildlife (52–54). administration of IVM to free-ranging Iberian ibexes, resistance Nonetheless, and similarly to IVM, numerous gaps in to the drug could be also developed by generalist nematodes of knowledge must be addressed before the in-field use of these the digestive tract, eg the blood-feeding Haemonchus contortus, alternative drugs may be considered a valid option (10). that may be cross-transmitted with sympatric sheep and goats Recently, Mounsey et al. (55) highlighted the precarious balance (32, 42). This could result in negative herd health management between dosage, route of administration, ecotoxicity, drug issues at the wildlife/livestock interface (23). resistance and efficacy of macrocyclic lactones in the Australian Some limitations of the present study include the relatively wombats (Vombatus ursinus and Lasiorhinus latifrons), two low number of dosed individuals and the absence of time points highly susceptible wildlife species to sarcoptic mange, suggesting within the first 24 h post treatment, which would have provided that a better cooperation and a continuous debate between more precise information concerning IVM absorption rate and stakeholders, including veterinarians, wildlife carers and plasma maximum concentrations. researchers, should be encouraged to achieve best treatment The pharmacological treatment of sarcoptic mange options for wildlife. in wildlife is a complex and controversial management There is therefore a need to further investigate the measure, with absence or paucity of protocols regarding pharmacokinetics, pharmacodynamics, population target dosage, administration times, density of medicated baits per and environmental consequences of the administration of surface or animal population unit, and a lack of knowledge available macrocyclic lactones, isoxazolines and other candidate on the pharmacokinetics of macrocyclic lactones in most molecules, including long-acting formulations, before their in- wildlife species, on the chances for resistance to appear due to field delivery become a management option to control sarcoptic underdosage and on the potential environmental consequences mange in free-ranging wild ruminants. of the massive release of antiparasitic drugs in the environment (8, 10, 43–45). In Iberian ibex, traditional management has mostly relied in decreasing host density and/or the selective DATA AVAILABILITY STATEMENT culling of affected individuals (7, 46). However, short-term strategies including the non-selective mass administration The original contributions presented in the study are included of medicated feed to affected ibex populations has also been in the article/supplementary material, further inquiries can be tried, with little or no effect on the disease prevalence and directed to the corresponding author/s. demographic impact (6, 47, 48). The rapid drop of plasma IVM concentration in Iberian ibex after a single oral dose found in this study, and the lack of published results on therapeutic ETHICS STATEMENT trials with oral IVM in scabietic free-ranging individuals suggest This study complied with all Andalusian, Spanish, and European caution in promoting the mass delivery of IVM medicated legal requirements and guidelines regarding experimentation and pellets as a measure to efficiently control sarcoptic mange animal welfare. Handling procedures and sampling frequency at the individual and population level in this wild caprine. were designed to reduce stress and minimize the impact on The epidemiological effects of partially ineffective treatment the health of the subjects, as per European (2010/63/UE) and strategies could even be opposite, by lengthening the infective Spanish (R.D 53/2013) standards. The study was approved by stage of the individuals affected by sarcoptic mange and therefore the Ethics on Animal Welfare Committee of the University of increasing the dissemination and spread of the disease (25), Jaén and authorized by the Dirección General de Producción potentially to other mammal species too (12, 49). Moreover, Agrícola y Ganadera of the Consejería de Agricultura, Pesca y aspects such as the proportion of the target population actually Medio Ambiente of the Junta de Andalucía (Ref: SA/SIS/MD/ps/ receiving the treatment, the access to the drug by non-target October 25, 2012). The Sierra Nevada Natural Park staff also species, and the environmental effects of the massive release approved this study. of IVM in the environment, are still in need to be thoroughly investigated and controlled (8, 10). A major concern is that IVM and its metabolites are mainly eliminated in the feces of treated AUTHOR CONTRIBUTIONS individuals (50), with potential impact of the residues on the biology and reproduction of dung invertebrate fauna (45, 50). BM, JG, JL-O, JE, AR, GM, PF, LR, MV, ES, BR, and RO In vitro and in vivo studies carried out with Sarcoptes conceptualized the project and agreed on the study design. RO mites var. suis showed that moxidectin concentration required prepared the IVM bolus. JG, JE, AR, and BR performed the IVM to kill 50% of mites was lower than that of IVM, and treatment and obtained blood samples from the ibexes. BM, GG, that a single dose of moxidectin or fluralaner were more MP, and MR performed the laboratory analysis and elaborated effective than two consecutive of IVM, suggesting that IVM the results. ES performed the statistical analysis. BM drafted the should be soon replaced in the treatment approach for original manuscript. JG, JL-O, JE, AR, GM, PF, LR, MV, ES, BR, sarcoptic mange (35, 51, 52). New long-acting oral and and RO contributed in shaping and revising the manuscript. topic isoxazolines are emerging as more efficient therapeutic All the authors have read and agreed to the final version of options against sarcoptic mange, as suggested by recent the manuscript. Frontiers in Veterinary Science | www.frontiersin.org 5 April 2022 | Volume 9 | Article 830157 Moroni et al. Ivermectin Plasma Concentration in Iberian Ibex FUNDING and ES by the Spanish Ministerio de Ciencia Innovación y Universidades (MICINN) through a Ramon y Cajal agreement This project was funded by the Consejería de Medio (RYC-2016-21120). GM is a Serra Húnter Fellow. Ambiente de la Junta de Andalucía (project 173/2009/M/00; 03/15/M/00; 861_11_M_00 and 2016/00014/M), and by ACKNOWLEDGMENTS the Spanish Ministerio de Economía y Competitividad (projects CGL2012-40043-C02-01, CGL2012-40043-C02- We are grateful to all the people involved in the 02, and CGL2016-80543-P). The authors’ research activities are experiment. Special thanks goes also to the park wardens partially supported by the Plan Andaluz de Investigación (RNM- and fieldworkers in the SNNS and, in particular, to 118 group). MV is supported by a FI-GENCAT Fellowship José López, Isidro Puga, Apolo Sánchez, and Antonio (2020_FI_B2_00049, co-financiated by Agència de Gestió Rodríguez for their professional and personal involvement in d’Ajuts Universitaris i de Recerca and European Social Fund) the study. REFERENCES 16. Steel JW. Pharmacokinetics and metabolism of avermectins in livestock. Vet Parasitol. (1993) 48:45–57. doi: 10.1016/0304-4017(93)90143-B 1. 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Valldeperes M, Granados JE, Pérez JM, Castro I, Bravo AR, Fandos P, Ráez Bravo, Mentaberre, Fandos, Pazzi, Romagnoli, Gardini, Rossi, Valldeperes, et al. How sensitive and specific is the visual diagnosis of sarcoptic mange Serrano, Ramos and Odore. This is an open-access article distributed under the in free - ranging Iberian ibexes? Parasit Vectors [Internet]. (2019) 21:1– terms of the Creative Commons Attribution License (CC BY). The use, distribution 7. doi: 10.1186/s13071-019-3665-7 or reproduction in other forums is permitted, provided the original author(s) and 47. Pérez JM, Giuseppe P, Dipartimento M, Torino U, Paolo L. Biology and the copyright owner(s) are credited and that the original publication in this journal management of sarcoptic mange in wild Caprinae populations. (2021) 51:82– is cited, in accordance with accepted academic practice. No use, distribution or 94. doi: 10.1111/mam.12213 reproduction is permitted which does not comply with these terms. 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