MICROSATELLITE LETTERS Development and characterization of thirteen microsatellite markers for the longbill spearfish (Tetrapturus pfluegeri) A. M. Bernard • A. W. S. Hilsdorf • A. F. Amorim • R. R. Domingues • M. S. Shivji Received: 5 August 2013 / Accepted: 17 September 2013 Ó Springer Science+Business Media Dordrecht 2013 Abstract The longbill spearfish (Tetrapturus pfluegeri; Istiophoridae) is an epipelagic billfish species broadly distributed in the Atlantic Ocean and exploited by multi- national fisheries. No data exist on the population structure or genetic diversity of this apex predator, despite a strong need for this information to inform international conser- vation and management efforts. Thirteen microsatellite loci, comprising mostly tri- and tetra-nucleotide repeats, were isolated and characterized for this species by geno- typing individuals (n = 29–42) obtained from western North and South Atlantic fisheries. The average number of alleles ranged from 3 to 16 and the expected heterozygosity ranged from 0.35 to 0.86. This suite of markers provides the first population genetic nuclear resources for the longbill spearfish, allowing initiation of studies for assessing the population structure and demography of this poorly understood species. Keywords Billfish genetics Á Billfish conservation and management Á Istiophoridae Á STR The longbill spearfish (Tetrapturus pfluegeri) is a widely distributed (40°N–35°S) epipelagic billfish species ende- mic to the Atlantic Ocean and capable of extensive migratory movements (Collette et al. 2011). It is caught primarily as by-catch in commercial fisheries targeting other pelagic teleosts. Currently, this species falls under the management jurisdiction of the International Commission for the Conservation of Atlantic Tunas (ICCAT), but almost nothing is known regarding its stock status and population trajectory to inform management efforts. Fur- thermore, recent work suggests that a large number of species misidentifications are occurring between the mor- phologically similar longbill spearfish, white marlin (Ka- jikia albida), and roundscale spearfish (Tetrapturus georgii) (Beerkircher et al. 2009; Bernard et al. 2013), further hindering conservation and management of the longbill spearfish. To inform conservation and manage- ment efforts for this enigmatic apex predator, we have developed 13 microsatellite markers for the longbill spearfish, which may be used to delineate genetic man- agement units and assess both the contemporary and his- torical demographic history of this species. Genomic DNA from five longbill spearfish individuals was extracted using the DNeasy Blood and Tissue Kit (Qiagen Inc.) and provided to Genoscreen, France (www. geneoscreen.fr) for commercial microsatellite library pro- duction. Microsatellite library isolation was performed via 454 GS-FLX Titanium pyrosequencing (Roche Diagnos- tics) of enriched DNA libraries as described in Malausa et al. (2011). Enrichment was performed using eight DNA probes: TG, TC, AAC, AAG, AGG, ACG, ACAT, and Electronic supplementary material The online version of this article (doi:10.1007/s12686-013-0038-y) contains supplementary material, which is available to authorized users. A. M. Bernard Á M. S. Shivji (&) Guy Harvey Research Institute, Oceanographic Center, Nova Southeastern University, 8000 North Ocean Drive, Dania Beach, FL 33004, USA e-mail: mahmood@nova.edu A. W. S. Hilsdorf (&) Á R. R. Domingues Unit of Biotechnology, University of Mogi das Cruzes, PO Box 411, Mogi das Cruzes, SP 08701-970, Brazil e-mail: wagner@umc.br A. F. Amorim Fisheries Institute, Age ˆncia Paulista de Tecnologia dos Agronego ´cios, Av. Bartolomeu de Gusma ˜o, 192, Santos, SP 11030-906, Brazil 123 Conservation Genet Resour DOI 10.1007/s12686-013-0038-y