Vocalizations During Electroejaculation in Anaesthetized Adult and Young Pampas Deer (Ozotoceros bezoarticus) Males F Fumagalli 1 , JP Dami an 2 and R Ungerfeld 3 1 Cl  ınica Semiol  ogica, Facultad de Veterinaria, Universidad de la Repu ´blica, Montevideo, Uruguay; 2 Departamento de Biolog ıa Molecular y Celular, Facultad de Veterinaria, Universidad de la Rep ublica, Montevideo, Uruguay; 3 Departamento de Fisiolog ıa, Facultad de Veterinaria, Universidad de la Rep ublica, Montevideo, Uruguay Contents The aim of this study was to characterize the vocalizations produced during electroejaculation under general anaesthesia in pampas deer males and to determine whether the characteristics of those vocalizations differ in adult and young pampas deer males. Electroejaculation was applied to 13 adults (AM) and 13 young (YM) males under general anaesthesia. Vocalizations were digitally recorded, and the number and duration of vocalizations, the latency in relation to each voltage, the total time vocalizing, and the structure of the fundamental frequency (F 0 ) [initial frequency (F start ), maximal frequency (F max ), min- imal frequency (F min ) and final frequency (F end )] were analysed. No male vocalized with 0 V; the number of animals that vocalized increased at 2 and 3 V and increased again at 4, 5 and 6 V (p < 0.05). The latency time from the beginning of each series (each voltage) decreased until 4 V (p < 0.01). The number of vocalizations/voltage increased from 4 V (p < 0.05). The length of each vocalization and the total time during which animals vocalized were greater in YM than AM (p = 0.02 and p = 0.01, respectively). Similarly, the fundamental frequencies were higher in YM than AM (p ≤ 0.05). Overall, we concluded that the vocalizations emitted during electroejaculation in pampas deer under general anaesthesia are related to the voltage applied during the process. Young males vocalize more time, probably due to a greater sensibility to the electric stimulation. The differences in the characteristics of the vocalizations between adult and young males may be related to the anatomic differences in the neck of adult or young males. Introduction The pampas deer (Ozotoceros bezoarticus, Linnaeus, 1758) used to be a widespread species originally distributed in the open grasslands of South America (Jackson and Langguth 1987). However, habitat frag- mentation, agriculture development, competition with farmed animals and unregulated hunting led to the decrease in size and distribution of the species (Demar  ıa et al. 2003). This species is listed in Appendix I (see supporting information) of the Convention on Interna- tional Trade in Endangered Species of Wild Fauna and Flora (CITES 2014). In Uruguay, there are two wild populations, and there is also a third semicaptive group (Ungerfeld et al. 2008). This third population is allo- cated at the Estaci  on de Cr  ıa de Fauna Aut octona Cerro Pan de Az ucar (ECFA) (Maldonado, Uruguay; 34 ° 3 0 S, 54 ° 0 0 W) and consists of approximately 80 individuals. The cryopreservation of gametes and the development of gene banks are useful techniques to improve conservation status of endangered species (Garde et al. 2006). For this purpose, techniques for semen collection should be adequately adapted for each species. As artificial vagina can only be used in tame and trained individuals, electro- ejaculation (EE) is the most widely used technique for semen collection in wild animals. In deer, due to nervous temperament and thus, risks for operators, EE is conven- tionally used under general anaesthesia (Umapathy et al. 2007; Mart  ınez et al. 2008; Mart  ınez-Pastor et al. 2009). Electroejaculation produces some undesirable physio- logical responses. For example, in anaesthetized pampas deer, we previously observed increases in heart and pulse rates, as well as in creatine kinase, aspartate aminotrans- ferase and alkaline phosphatase concentrations, and a decrease in rectal temperature (Fumagalli et al. 2012). Responses to EE may also differ in relation to males’ age, as adult males had a greater increase in heart rate, but lower increases in alkaline phosphatase and creatine kinase concentrations. These negative responses have been associated with the occurrence of vocalizations in bulls (Falk et al. 2001; Whitlock et al. 2012) and rams (Dami an and Ungerfeld 2011), which are considered as reliable indicators of pain (Dami an and Ungerfeld 2010). Although we did not record vocalizations in anaesthe- tized domestic (rams, bucks) or wild (antelopes, muflon, dama deer) ruminants, strikingly we have previously observed that pampas deer males vocalize during EE even while they are maintained under general anaesthesia. At least under several other contexts, the increases in cortisol secretion, and the number, duration and characteristics (high-frequency) of vocalizations are reliable indicators for pain (Prunier et al. 2013). Considering all this information, a first aim of this study was to characterize the vocalizations produced during EE under general anaesthesia in pampas deer males. Another objective was to determine whether the characteristics of the vocalizations, as well as the cortisol and creatine kinase serum concentration, differ in adults and young pampas deer males during EE under general anaesthesia. Materials and Methods Animals and facilities The study was conducted from September to March (spring to autumn), at the ECFA, with 26 males. From Reprod Dom Anim doi: 10.1111/rda.12494 ISSN 0936–6768 © 2015 Blackwell Verlag GmbH