Mapping the stranger: Genetic diversity of Manila clam in European coastal lagoons

Manila clam Ruditapes philippinarum - synonym Venerupis philippinarum (Adams and Reeve, 1850) is one of the most successful marine invaders worldwide and represents almost 20% of worldwide mollusc total production. Originally distributed in the Indo-Pacific region, the species was introduced for aquaculture and fisheries in North America, Polynesia, the US Virgin Islands, as well as in Atlantic and Mediterranean coasts of Europe. Due to the commercial value of the species, population genetics and stock composition are of primary importance both in natural and invaded environments. Moreover, molecular genetics can provide valuable data in invasion biology, as, for example, the structure of the exploited populations, the origin of source populations and the routes of invasions. Furthermore, molecular data might be useful for the tracking of seafood products, as requested by the European Commission for European seafood products (see EC laws n deg 178/2002, 509/2006, 510/2006 and their improvements). Despite all these considerations genetic data are lacking, especially for introduced populations, and should be urgently provided. In a framework of an European network of researchers involved in Manila clam studies, clams were collected in coastal lagoons and estuaries in both Mediterranean (Northern Adriatic Sea, Italy) and Atlantic (Spain and Portugal) coasts of Europe. Both mitochondrial and nuclear molecular markers were applied to investigate the genetic structure of populations, specifically by the direct sequencing of 16S rDNA gene fragments and by the genotyping of 7 microsatellite markers previously used in native populations. Results demonstrated the occurrence of multiple haplotypes for 16S rDNA, even if one main haplotype was identified in the European analyzed populations. However, few sequences were available in Genbank to make comparisons with native populations. Concerning the microsatellite data, the genotyping of Manila clam populations showed low genetic diversity among them, considering the Fst values, and a lack of genetic structuring when analysed by Structure software. Both these data suggest the absence of geographic differentiation among the introduced populations of Manila clam, as a result of introgression due to translocations and other human activities. The levels of genetic diversity within introduced populations were comparable to those described from native regions, suggesting that multiple introduction events and mixed source populations can counterbalance the loss of genetic diversity caused by the founder effect. However, it must be remarked that microsatellites showed a strong occurrence of null alleles when tested by Micro-checker, as previously observed in other bivalves, suggesting that the problem is differently distributed among Manila clam populations or specific loci, thus microsatellites should be carefully checked before used in extensive population studies. In the near future, population genetics and stock identification of this species on a large scale should be assessed using new powerful markers such as those obtained by NGS (Next Generation Sequencing) methods. In conclusion, this research represents the very first extensive genetic investigation on Manila clam's European populations, defining the levels of genetic diversity that will help to inform management practices.