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Genetic Identification of Native Populations of Mediterranean Brown Trout Salmo Trutta L. Complex (Osteichthyes: Salmonidae) in Central Italy

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Genetic Identification of Native Populations of Mediterranean Brown Trout Salmo Trutta L. Complex (Osteichthyes: Salmonidae) in Central Italy The European Zoological Journal ISSN: (Print) 2475-0263 (Online) Journal homepage: https://www.tandfonline.com/loi/tizo21 Genetic identification of native populations of Mediterranean brown trout Salmo trutta L. complex (Osteichthyes: Salmonidae) in central Italy A. R. Rossi, G. Petrosino, V. Milana, M. Martinoli, A. Rakaj & L. Tancioni To cite this article: A. R. Rossi, G. Petrosino, V. Milana, M. Martinoli, A. Rakaj & L. Tancioni (2019) Genetic identification of native populations of Mediterranean brown trout Salmotrutta L. complex (Osteichthyes: Salmonidae) in central Italy, The European Zoological Journal, 86:1, 424-431, DOI: 10.1080/24750263.2019.1686077 To link to this article: https://doi.org/10.1080/24750263.2019.1686077 © 2019 The Author(s). Published by Informa View supplementary material UK Limited, trading as Taylor & Francis Group. Published online: 12 Nov 2019. Submit your article to this journal Article views: 92 View related articles View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=tizo21 The European Zoological Journal, 2019, 424–431 Vol. 86, No. 1, https://doi.org/10.1080/24750263.2019.1686077 Genetic identification of native populations of Mediterranean brown trout Salmo trutta L. complex (Osteichthyes: Salmonidae) in central Italy A. R. ROSSI1, G. PETROSINO1, V. MILANA1, M. MARTINOLI2, A. RAKAJ2,& L. TANCIONI2* 1Department of Biology and Biotechnology C. Darwin, University of Rome “La Sapienza”, Rome, Italy, and 2Experimental Ecology and Aquaculture Laboratory, Department of Biology, University of Rome “Tor Vergata”, Rome, Italy (Received 30 July 2019; accepted 23 October 2019) Abstract Italian native populations of Mediterranean brown trout belong to the Salmo trutta complex. This species complex includes many mitochondrial lineages and phenotypes that have caused taxonomic controversies over time. The spatial distribution and the genetic diversity of these fishes are threatened by habitat destruction, global warming and, mainly, by the introduction of domestic trout of Atlantic origin. Indeed allochthonous trouts were massively restocked in Italian rivers for a century and they admixed with native populations. In order to identify residual native populations of Mediterranean brown trout, a genetic analysis of specimens collected within Latium region, on the Tyrrhenian slope of central Italy, was undertaken. To this purpose, 210 trout specimens were collected from six different rivers and analyzed for the identification of their nuclear (LDH- C1* RFLP) and mitochondrial (Control Region sequences) genotypes. Genetic characterization with these molecular markers allowed a quantitative estimate of allochthonous genotypes, which are present in all brown trout populations of the six sites, even if not equally distributed across the sampling area. At least three populations, inhabiting diverse lotic ecosystems (mountain, hilly and coastal streams respectively), are characterized by a high percentage of native nuclear allele *100 at locus LDH-C1* and typical Mediterranean haplotypes (of AD and ME lineages), which can be considered as different management units (MUs). This finding highlighted the aquatic ecosystems of the Latium Region as an important hotspot of salmonid biodiversity within the Italian peninsula, with important implications from a conservation perspective. Keywords: Biodiversity conservation, salmonids, hybridization, restocking, threatened fish Introduction different impact of paleoclimatic events (i.e. glaciations) Salmonids include freshwater and anadromous fishes on the isolation of ancestral populations into refuges widespread in the Northern hemisphere (Nelson et al. and subsequent secondary contact (Bernatchez 2001; 2016). In the Mediterranean area, the Italian peninsula Cortey et al. 2004, 2009). Since the last century, the represents a hotspot of salmonid biodiversity, being original distribution of these fishes has been altered by inhabited by a huge number of native taxa. Among the massive restocking with domestic brown trout these, a number of fishes of the genus Salmo, commonly mainly of Atlantic origin, that was translocated for known as trout, are included in the national and inter- recreational fishery and mixed with natural populations national red list (Bianco et al. 2013)andpartlyin (Splendiani et al. 2016b; Meraner & Gandolfi 2018). annexe II of the European Union “Habitat Directive” The coupling of these domestic brown trout with the (Nonnis Marzano et al. 2016). Their present distribu- native populations has caused introgressive hybridiza- tion is the result of the combination of historical natural tion and reduced the genetic integrity of native trout so colonization (Bianco 1990), and recent anthropogenic that in some geographic areas original populations have activities.Indeedthehistoryoftroutsofthe been almost completely admixed or replaced by alien Mediterranean area was geographically shaped by the trout (Splendiani et al. 2016b, 2019). This context, *Correspondence: L. Tancioni, Experimental Ecology and Aquaculture Laboratory, Department of Biology, University of Rome “Tor Vergata”, Via della Ricerca Scientifica 1, Rome 00133, Italy. Tel: +39 0672595975. Fax: +39 0672595965. Email: [email protected] © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Mediterranean brown trout 425 associated with the phenotypic plasticity and ecological populations with higher genetic integrity, and for use adaptation, has contributed to a confusing nomencla- in a parallel habitat modelling analysis (Martinoli ture picture and to “taxonomic inflation” (see Tougard et al. 2019). To this purpose, two molecular markers et al. 2018) of Italian native trout, with many morphae that proved to be diagnostic of lineages (Cortey & described as species (or subspecies) whose validity is García-Marín 2002) and geographic origin questionable. To date, it is still very difficult to delineate (McMeel et al. 2001) of brown trout were applied. boundaries and unequivocal correspondences between The mitochondrial marker (Control Region, CR) morphological species and evolutionary lineages identi- was used for lineage identification; the nuclear mar- fied by molecular markers; in order to overcome this ker (Lactate dehydrogenase C1, LDH-C1*), to dis- difficulty the use of Evolutionary Significative Units criminate specimens of European hatchery origin (ESUs) has been suggested (Zanetti et al. 2013). from those of Mediterranean native origin, and to However the problem is not completely solved, as identify their hybrids. Habitat Directive requires species names as labels of protection actions. Material and methods The main point of congruence about Italian trouts is that they are part of the Salmo trutta complex defined on A total of 210 specimens were collected from 6 a molecular basis (Sanz 2018), and include: trout that different rivers/locations (Table I) using electrofish- inhabit the Apennines and the western Alps (AD line- ing procedures. In order to minimize the risk of age) that likely corresponds to S. ghigii Pomini, 1940; underestimating the local population genetic varia- trout present in south eastern Sicily with AT native bility, associated with the sampling of individuals haplotypes, that should keep the name S. cettii belonging to the same family group (Hansen et al. Rafinesque, 1810, and whose distribution elsewhere in 1997), each sampling area was extended for more Italy still needs to be defined; trout from the Tyrrhenian than 20 times the wetted width of the bed of the area of the Italian Peninsula, Sardinia and north Sicily, streams, taking also into account the eventual pre- presently without a valid name, and characterized sence of small tributaries. by AD and ME haplotypes; S. fibreni (Zerunian & Specimens were anaesthetized with a 0.035% MS Gandolfi 1990)theendemictroutofthesmallkarstic 222 (Tricaine Methanesulfonate) solution. A small lake of Posta Fibreno (a split of the AD lineage); portion of the adipose fin was removed and fixed in S. marmoratus Cuvier, 1829 (MA lineage) present in 90% ethanol for genetic analysis. The procedures Italian and Slovenian rivers draining into the northern used for fish sampling were carried out in agreement Adriatic; S. carpio Linnaeus, 1758 the endemic trout of with relevant legislation (CEN EN 14011/2003 - the Garda Lake (AD and MA haplotypes) (Gratton Water quality - Sampling of fish with electricity), et al. 2014). In addition to these, allochthonous brown avoided animal sacrifice and permitted the live trout corresponding to Salmo trutta derived from north release of sampled specimens after data collection. European farmed stocks (mainly AT lineage), are pre- Fish sampling was authorized by the Direzione sent, and frequently hybridized with local native popu- Regionale Agricoltura, Promozione della Filiera lations (Bernatchez 2001; Nonnis Marzano et al. 2003; e della Cultura del Cibo, Caccia e Pesca of the Splendiani et al. 2006, 2016b, 2019; Gratton et al. Regione Lazio (Prot. n. 526425). 2014; Fabiani et al. 2018). Total genomic DNA was extracted from fin clips In the last decade a 30% decline of native trout according to Aljanabi and Martinez (1997). For populations,
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