Short communication Microsatellite based ovine parentage testing to identify the source responsible for the killing of an endangered species Fernando Rendo, Mikel Iriondo, Carmen Manzano, Andone Estonba * Department of Genetics, Physical Anthropology and Animal Physiology, Faculty of Science and Technology, University of the Basque Country, E-48940 Bilbao, Spain 1. Introduction The use of microsatellite markers for individual identification and kinship determination, such as biological maternity/paternity, is well established in humans and domestic animals [1–5]. In humans, these data are often used for the resolution of forensic cases, but the application of microsatellites in animal forensic studies is scarce [6–10]. In the present study, we report a forensic case in which we employed an ovine microsatellite panel in order to identify the source of poisoning which lead to the death of a protected species of scavenger birds. The Egyptian vulture (Neophron percnopterus) is a scavenger bird from south-western Europe, northern Africa and southern Asia, which is listed as an endangered species [11]. In the Basque Country (northern Spain), where this study was conducted, no more than 38 nestling couples of this species have been observed [12]. The Iberian Egyptian vulture population has decreased over the last few decades, mainly due to the use of agricultural poisons [13]. In 2007, in the province of Gipuzkoa (Basque Country), the body of an Egyptian vulture presenting signs of poisoning was found next to the remains of a Latxa Blond Face sheep breed lamb. Toxicological analysis carried out by the Wildlife Forensic Laboratory (LFVS, Madrid, Spain) identified the poison as being Aldicarb, a substance which was present in the remains of both the vulture and the lamb. Aldicarb is a carbamate commonly used as a pesticide in agriculture. It is also the most frequently used poison in northern Spain related to the illegal control of predators in hunting properties [14]. Most scavenger bird deaths seem to be accidental and related to this activity, but during the last 5 years, a significant increase in intentional poisoning of vulture species has been observed [14]. The reason for this seems to be that during moments of food scarcity, these species can behave, not as scavengers, but also as predators of live domestic animals. Nevertheless, the number of attacks on livestock is low and in the majority of cases, involves animals which are sick, immobilized or newborn [15]. This investigation by the Gipuzkoa Environmen- tal Prosecution was based on interviews and inspections of the different breeders in the region carried out by wildlife technicians from the Gipuzkoa Provincial Council. All available evidence pointed to the owner of the lamb as the putative suspect responsible for the poisoning, i.e. the shepherd whose flock grazes in the same place where the lamb remains were found. In order to obtain additional evidence to substantiate this hypothesis, the wildlife technicians requested our collaboration. Since there was no forensic evidence which directly linked the suspect and the vulture or poisoned lamb, we aimed to obtain indirect evidence which would identify the parents of the lamb among the individuals of the flock through microsatellite based parentage testing. Microsatellite markers can be multiplexed into a single PCR (polymerase chain reaction), creating an easy and cost-effective panel which can identify the parents of an individual, even when a high degree of genetic kinship between the members of the population is expected, as occurs in a flock. Forensic Science International: Genetics 5 (2011) 333–335 ARTICLE INFO Article history: Received 25 September 2009 Received in revised form 16 September 2010 Accepted 20 September 2010 Keywords: Animal forensic genetics Parentage analysis Ovine microsatellites Wildlife protection ABSTRACT In this study, we present an animal forensic genetics case in which we applied ovine microsatellite based parentage testing to the resolution of an act against protected wildlife. The aim was to identify the origin of the animal responsible for the death of an Egyptian vulture (Neophron percnopterus) that had been poisoned through consumption of a Latxa Blond Face breed lamb. Thus, we carried out a 22 microsatellite based parentage test in order to identify the parents of the lamb in the flock which grazes in the same place where the vulture’s remains were found. Multiple parentage analysis revealed two possible parents, one ewe and one ram, with a combined paternity/maternity index (PI) higher than 9.09E9 and a likelihood (W) value of 99.9999%, assuming 50% probability a priori. This result confirmed the flock of origin of the poisoned lamb and shows that the microsatellite panel described herein is a potentially useful tool for the resolution of animal forensic cases. ß 2010 Elsevier Ireland Ltd. All rights reserved. * Corresponding author. Tel.: +34 94 601 5517; fax: +34 94 601 3145. E-mail address: andone.estonba@ehu.es (A. Estonba). Contents lists available at ScienceDirect Forensic Science International: Genetics journal homepage: www.elsevier.com/locate/fsig 1872-4973/$ – see front matter ß 2010 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.fsigen.2010.09.009