January 29, 2011 | Veterinary Record Short Communications Short Communications J. Giménez, BSc, P. Gauffier, MSc, R. Esteban, MSc, P. Verborgh, MSc, CIRCE (Conservation, Information and Research on Cetaceans), Calle Cabeza de Manzaneda 3, Algeciras-Pelayo, 11390 Cádiz, Spain R. De Stephanis, PhD, Department of Conservation Biology, Estación Biológica de Doñana, CSIC (Consejo Superior de Investigaciones Científicas), Calle Americo Vespucio s/n, 41092 Isla de la Cartuja, Sevilla, Spain E-mail for correspondence: joan@circe.biz Provenance: not commissioned; externally peer reviewed Published Online First 4 January 2011 Veterinary Record (2011) 168, 101b doi: 10.1136/vr.c5284 Biopsy wound healing in long-finned pilot whales ( Globicephala melas ) J. Giménez, R. De Stephanis, P. Gauffier, R. Esteban, P. Verborgh CONSERVATION and management of natural systems often require biological research so that decisions can be based on scien- tific evidence (Tracy and Brussard 1996). Most biological research with application to conservation requires fieldwork, and fieldwork on animals involves a variety of techniques, ranging from benign, such as simple observation, to more invasive, such as blood sam- pling (Cuthill 1991). Invasive techniques can potentially interfere with the animal studied, however, and alter it in ways that are at odds with the goals of the conservation programme. It is essential, therefore, to balance the risks and benefits of research applied to con- servation. In the past two decades, various invasive techniques to obtain samples for genetic analysis, or analysis for stable isotopes or contaminants have become widely used, especially in protected species, for example, fish (Bernatchez and others 1991, Sheffer and others 1997), snakes (Gibbs and others 1998), birds (Haig and Avise 1996) and terrestrial mammals (McGowan and Davidson 1994, Taylor and others 1997, Comstock and others 2002). The case of cetaceans is also important, especially due to the fact that handling large mammals such as whales is particularly difficult in the field. As a result of techniques such as biopsy darting, several questions have been answered regarding social structure (de Stephanis and others 2008b), feeding ecology (Walker and others 1999, de Stephanis and others 2008a), population genetics (Engelhaupt and others 2009), contaminant loads (Fossi and others 2003) and evolutionary rela- tionships (Leduc and others 1999) in cetaceans. Although biopsy darting is an invasive procedure, it has become widely used in ceta- cean biology and ecology due to its value for management and con- servation purposes; however, there has also been increasing concern about the behavioural response of cetaceans to the technique. It is important to investigate the risks and benefits of biopsy darting, especially when working with protected species with serious prob- lems of conservation (de la Chenelière 1998). While undoubtedly helping to add much-needed information on cetacean populations, the long-term consequences of biopsy collection remain almost unknown. In order to determine whether there are potential risks that should be considered, it is necessary to evaluate the physical effects at the target site in addition to studying behavioural responses to biopsy darting (J. Giménez, R. De Stephanis, S. García-Tiscar, P. Gauffier, R. Esteban, L. Minvielle-Sebastia, P. Verborgh, unpublished observations). The International Whaling Commission (1991) rec- ommended combining biopsy studies with photo-identification techniques to monitor the potential long-term effects. The few stud- ies that have analysed wound healing in cetaceans (Weller and others 1997, Krützen and others 2002) reported no evident physiological complications arising from biopsies in bottlenose dolphins (Tursiops truncatus). However, it is necessary to evaluate biopsy wound healing in target species with different conservation statuses. This short communication describes a study to evaluate biopsy wound healing in long-finned pilot whales (Globicephala melas) in the Strait of Gibraltar. The total population of this species in the Strait of Gibraltar is approximately 213 individuals (Verborgh and oth- ers 2009). The population is subject to whale-watching activities, with more than eight boats whale watching, mainly between April and November (Carbó-Penche and others 2007), and intense mari- time traffic, with more than 105,000 vessels crossing the whales’ main spatial distribution each year (R. De Stephanis, P. Verborgh, S. Pérez, R. Esteban, C. Guinet, unpublished observations). Studies of the whales’ diet (de Stephanis and others 2008b), social structure (de Stephanis and others 2008a), spatial distribution (de Stephanis 2008, R. De Stephanis, P. Verborgh, S. Pérez, R. Esteban, C. Guinet, unpublished observations), genetics (Verborgh and others 2009) and population dynamics (Gauffier 2008, Verborgh and others 2009) have been conducted on this population in the past 10 years, and have led the Spanish Cetacean Society to propose that the population be included in the ‘vulnerable’ category of the Spanish Catalogue of Endangered Species. Skin and blubber biopsies were collected from long-finned pilot whales in the Strait of Gibraltar between 2004 and 2008 as part of long-term studies on molecular sex determination, diet, genetics and contaminant studies. Biopsy darting was performed using a 67 kg draw crossbow (Zasdar) from a distance ranging between 5 and 15 m from the animal. The dart (Fig 1) was aimed at the mid-lateral region, near the dorsal fin. A stop collar attached to the tip of the bolt prevented penetration deeper than the biopsy tip and caused the bolt to rebound upon impact with the whale’s body. The darts were designed to float on the surface of the sea, and were retrieved using a dip net. The biopsy tips were made of stainless steel and both the tips and the arrows were designed and manufactured by Finn Larsen of the Danish Institute for Fisheries Research, Charlottenlund, Denmark. Using biopsy arrows with tips 1.5 cm long and with an internal diameter of 0.6 cm, fitted with tooth-like barbs to hold sample material, biopsies including epi- dermis, dermis and blubber were collected. Biopsy sampling complied with the current laws of Spain and was carried out under the a permit issued by the Spanish Ministry of Environment. All the biopsy darts were sterilised before use, following the same protocol: the tips were brushed with commercial liquid soap, then rinsed and immersed in bleach, then rinsed in alcohol at 96 per cent, then burned to remove the alcohol, and finally stored in a sterile plastic bag until use. Biopsy dart- ing was attempted only when the sea condition was between 0 and 1 on the Douglas scale and the wind conditions were between 0 and 1 on the Beaufort scale. At each biopsy attempt, it was ensured that there were no whale calves present in the area before the crossbow was fired. The whales were sampled only if they were found while travelling (never when they were foraging, milling, resting or socialising), and were sampled independently of their size, excluding calves and juve- niles. Before the biopsy attempt, the dorsal fin of the whale to be sam- pled was photographed, and its identity was confirmed on-board the research vessel using a photo identification catalogue (Verborgh and oth- ers 2009) to avoid double sampling. After the sampling procedure, the target whale was photographed once more to confirm its identity and to group.bmj.com on February 3, 2011 - Published by veterinaryrecord.bmj.com Downloaded from