Published by Associazione Teriologica Italiana Volume 28 (1): 104–106, 2017 Hystrix, the Italian Journal of Mammalogy Available online at: http://www.italian-journal-of-mammalogy.it doi:10.4404/hystrix–11897 Short Note The phylogeography of Crocidura suaveolens from southern Italy reveals the absence of an endemic lineage and supports a Trans-Adriatic connection with the Balkanic refugium Riccardo Castiglia 1,* , Flavia Annesi 1 , Giovanni Amori 2 , Emanuela Solano 1 , Gaetano Aloise 3 1 Department of Biology and Biotechnology “Charles Darwin”, University of Rome “La Sapienza”, Via Borelli 50, 00161 Rome, Italy 2 National Research Council, Institute of Ecosystem Study, Largo Tonolli 50, 28922 Verbania Pallanza, Italy 3 Museo di Storia Naturale della Calabria e Orto Botanico, University of Calabria, Via Savinio s.n., 87036 Rende, Italy Keywords: white-toothed shrew mtDNA Pleistocene refugium taxonomy Article history: Received: 30 May 2016 Accepted: 27 October 2016 Acknowledgements The authors thank Paolo Colangelo and Emiliano Mori for helpful sug- gestions on the manuscript. Thanks are extended to Laura Barbieri for her work in laboratory. R. Castiglia received grants from Università di Roma “La Sapienza” and PRIN (2012). Abstract A molecular phylogeographic study using a fragment of the mitochondrial gene for cytochrome b (cytb) was performed on the lesser white-toothed shrew, Crocidura suaveolens, from seven localit- ies in central and southern Italy. Comparison with cytb European haplotypes revealed the absence of endemic lineages in the region, in contrast to what has been observed for many other Italian ter- restrial vertebrates. Indeed all the Italian specimens results nested with Balkanic conspecific within an Italo-Balkan clade. Historical demography of this clade showed a scenario of expansion which preceded the LGM. This evidence of glacial persistence indicates a certain flexibility of the classic models of Pleistocene biogeography. Southern European peninsulas (Italy, Balkans and Iberia) played an important role as refugial areas during Pleistocene cold periods, shap- ing the distribution of genetic variation of autochthonous European species (Hewitt, 2000). The lesser white-toothed shrew Crocidura sua- veolens (Soricomorpha, Soricidae) is widely distributed in the Palearc- tic, extending from the Atlantic coast of Spain eastwards through Europe and Asia to Siberia. The phylogeography of the species has been thoroughly studied (Dubey et al., 2006, 2007) and it reveals the presence of ten parapatric clades across the species range. In Europe two divergent mtDNA lineages, can be found, the first derived from the Iberian refugium and the second of Italo-Balkan origin (Fig. 1). The latter lineage comprises haplotypes from Switzerland, Austria, Hungary, Bulgaria, Greece and localities in northern and central Italy (Dubey et al., 2007). It is now well established that the southernmost part of the Italian Peninsula represents a “hotspot” of intraspecific diversity for many terrestrial vertebrates, with the presence of endemic genetic lineages (Amori et al., 2009; Vega et al., 2010; Bertolino et al., 2015). The most likely source of historical barriers causing this biogeographic pattern are the glacio-eustatic sea level oscillations throughout the Pleistocene, with consequent insularization of southern Italy during the multiple in- terglacial transgressions. Among mammals, species rank has been pro- posed for some of these southern Italian populations (Sciurus vulgaris, Grill et al., 2009; Myodes glareolus, Colangelo et al., 2010; Microtus savii “species group” Castiglia et al., 2008 and Bezerra et al., 2016). For this reason, a genetic study of C. suaveolens from the southern part of the Italian refugium was carried out to provide a complete pic- ture of its genetic diversity and to determine if it belongs to the Italo- Balkan clade, as previously suggested by Dubey et al. (2007), or if it represents an additional endemic lineage for southern Italy. * Corresponding author Email address: riccardo.castiglia@uniroma1.it (Riccardo Castiglia) Figure 1 – Distribution of Crocidura suaveolens in Europe (top left, orange). The small red circles indicate the 7 localities from central and southern Italy studied in this work. The large circles indicate the localities belonging to the Iberian (black) and to the Italo-Balkan clade (white) studied in previous works (Dubey et al., 2006, 2007). Twelve lesser white-toothed shrews from seven localities in central- southern Italy were analyzed (Fig. 1 and Tab. 1). Procedures for DNA extraction, amplification and sequencing followed Castiglia et al. (2007). A fragment (997 bp) of the mitochondrial gene for cyto- chrome b (cytb) was sequenced in six individuals (cro1, 3, 11–13, 19), using a combination of the universal primers L14723, L15408, H15553 and H15915. For the remaining specimens it was possible to sequence only a smaller fragment (476 bp) with primers L15408 and H15915. To place the southern Italian haplotypes in the European phylogenetic Hystrix, the Italian Journal of Mammalogy ISSN 1825-5272 22nd February 2016 ©cbe2016 Associazione Teriologica Italiana doi:10.4404/hystrix–11897