The Grotta dei Pipistrelli is located in the Pantalica area (Sortino, Syracuse, southeastern Sicily; Fig. 1A), awarded in 2005 as UNESCO World Heritage Site for its history, archaeology, speleology and landscape. The area is characterized by a nature plateau, deeply engraved by the quarries of the Anapo River valley and the Calcinara stream. The site consists of natural and semi-natural habitats along with arable land, which are essential habitats for the invertebrate and vertebrate communities.

The karst cavity opens on a rocky wall overhanging the Calcinara stream, about 10 m from the watercourse (Fig. 2A), in the Miocene “Calcari di Siracusa” formation (Grasso and Lentini, 1982 and Schilirò et al., 2002). The cavity has a sub-horizontal development with a 7.3% WE average slope and it has been explored for about 260 m, between the entrance of the cave (234 m a.s.l.), to the left of the Calcinara stream, and the ending point (253 m a.s.l.), where a thick layer of detritus blocks the duct (Fig. 1B). At the entrance, the cave has a large opening that quickly shrinks into a funnel that enters into a first room. A gallery links the room to a large hall called “Sala del Guano”, with a 15 m high vault consisting of three domes, which currently hosts a huge bat colony. The cavity continues with a series of galleries and small dome halls; it is interrupted in a large terminal hall, where it ends in a duct enclosed by coarse detritus (Schilirò, 2007).

The Grotta dei Pipistrelli is an important bat cave in Sicily since it hosts the largest colonies of bats and it represents the biggest nursery roost of the region (Ferrante et al., 2018; Spena et al., 2013). In 2012, a monitoring programme started to identify the species, estimate the number of specimens and their seasonality. To date, the Grotta dei Pipistrelli is the only cave of Sicily systematically monitored (Spena et al., 2013). The maximum number of occurrences (taking into account all the species together) was recorded in summer (over 9000 in July 2013) and the presence of numerous juveniles in summer suggests that colonies from this cave are primarily reproductive colonies (Ferrante et al., 2018, Spena et al., 2013 and Spena et al., 2017).

In 1996, it was noticed that the walls of the talus (particularly the right wall) had several ossiferous breccia (Figs. 2B–D) containing many small vertebrate remains (birds, rodents, insectivores and mostly bats). The ground close to the right wall also contained osteological evidences of small vertebrate and some bone remains of large mammals. Sediments of a gallery yielded an elephant tusk portion, attributable to Palaeoloxodon mnaidriensis (Adams, 1874). From 2014, several campaigns on fossil remains collection were conducted by the authors (particularly JDM, RG, MTS).

The analysed material is presently deposited in the Department of Biological, Geological and Environmental Sciences of the Catania University, pending the final collocation and cataloguing, in agreement with the Natural Reserve “Pantalica, Valle dell’Anapo e Torrente Cava Grande”.

Order Chiroptera Blumenbach, 1779

Indeterminate Chiroptera

Material: a very encrusted upper molar and 6 mandibular fragments without teeth, from the late Pleistocene breccia; 3 maxillary fragments and 7 mandible fragments without teeth, and 2 left mandibular rami, one with a deciduous tooth and one without teeth, from Holocene soil (Fig. 4A).

Description and remarks. The upper molar belongs to a large bat species, but no diagnostic trait has been observed. The maxillary and mandible fragments without teeth can be referred to at least 8 individuals of medium-small size species, while the two mandibular rami probably belong to two immature individuals of medium-large species (Table 1 and Table 2).

Family Rhinolophidae Gray, 1825

Genus Rhinolophus Lacépède, 1799

Rhinolophus ferrumequinum (Schreber, 1774). Greater horseshoe bat.

Material: 1 cranium fragment, 1 maxillary, 1 mandible fragment, 2 lower teeth, and a distal humerus, from the late Pleistocene breccia; 1 encrusted splancnocranium, 3 maxillaries, 1 upper tooth, 3 mandibles of which one fragmented, and 2 distal humeri, from the Holocene soil (Figs. 4B-C).

Description and remarks. These fossil remains have the typical morphology of the genus Rhinolophus: cranium with prefrontal swelling, premaxillaries not fused with maxillary bones, two upper premolars of which the anterior one is very small and displaced in the labial margin of the tooth-row, ascending ramus of mandible short and large with the coronoid process slightly higher than the articular one, three lower premolars of which the intermediate is very small and displaced in the labial margin of the tooth-row, lower molars of nyctalodontic type, distal epiphysis of humerus with epicondyle, trochlea, inner ridge and lateral ridge separate by grooves, and styloid process long and thin (Felten et al., 1973, Gaisler, 2001a, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986). The large size (Table 3) enables a definite attribution to the greater horseshoe bat. The remains of R. ferrumequinum can be referred to at least eight individuals (three from the late Pleistocene breccia and five from the Holocene soil; Table 1 and Table 2).

Rhinolophus ferrumequinum is a species of Mediterranean affinities, with preference for warm and open areas with trees and shrubs (vegetation mosaics), in limestone areas close to wetlands. The summer retreats of the species are found mainly in buildings, but also in rock crevices, hollow trees, caves and mine galleries. Hibernation takes place in natural and artificial cavities with temperatures of about 7–12 ̊C (Gaisler, 2001a, Horáček et al., 2000, Lanza, 2012 and Lanza and Agnelli, 2002).

Rhinolophus ferrumequinum occurs from North Africa and southern Europe through Southwest Asia, the Caucasus, Iran, Afghanistan, Pakistan and the Himalayas to south-eastern China, Korea, and Japan (Gaisler, 2001a, Horáček et al., 2000 and Piraccini, 2016). In Italy, this species occurs in all regions (Agnelli et al., 2006 and Lanza, 2012). In Sicily and the surrounding islands, R. ferrumequinum is the most frequent species (Agnelli et al., 2008) and its occurrence has been recorded since the nineteenth century (Doderlein, 1872, Galvagni, 1837 and Minà Palumbo, 1868).

The earliest fossil records of R. ferrumequinum come from some Southwest Europe localities referred to the late early Pleistocene (middle early Biharian, late Villafranchian; Galán et al., 2016a, Salari et al., 2013 and Tata and Kotsakis, 2005). Indeed, many Pliocene or earliest Pleistocene specimens assigned in the past to R. ferrumequinum or R. cf. ferrumequinum have been ascribed to other species of the group (Topál, 1979 and Wołoszyn, 1987). Some Pliocene or earliest Pleistocene remains recently assigned to the same taxa (e.g., Agustí et al., 2011 and Jin et al., 2000) are reported in preliminary lists and their attribution needs to be confirmed. Fossil remains of this species have been collected in several sites from the middle Pleistocene in central and southern Europe and in North Africa, and in many sites from the late Pleistocene and Holocene from Portugal and England to Japan and South to Maghreb and Near East (Gaisler, 2001a, Salari, 2009, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Kotsakis, 2017, Salari and Silvestri, 2015, Salari et al., 2013 and Tata and Kotsakis, 2005, and references therein; Bañuls-Cardona et al., 2017, Fagoaga et al., 2017, Galán et al., 2016b, Galán et al., 2018a, Lebreton et al., 2016 and Pereswiet-Soltan, 2016). In Sicily and the surrounding islands, R. ferrumequinum remains come from upper and lower layers of the Grotta di Spinagallo, dating back early middle Pleistocene (Kotsakis and Petronio, 1980 and Tata and Kotsakis, 2005).

Rhinolophus euryale Blasius, 1853. Mediterranean horseshoe bat.

Material: 1 splancnocranium and 2 distal humeri, from the late Pleistocene breccia; 3 maxillaries of which one fragmented, 1 subintact mandible, 2 humeri and 3 distal portions of humerus, from the Holocene soil (Figs. 4D-E).

Description and remarks. This material has the distinctive shape of the Rhinolophus genus (see above) and the dimensions of intermediate size species (Felten et al., 1973, Krapp, 2001, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986). We excluded R. birzebbugensis, an extinct species distributed in the Mediterranean regions between Pliocene and late early Pleistocene, for chronological reasons and for its larger size than the other species (Salari et al., 2013 and Storch, 1974). Furthermore, in all the specimens referred to the middle-sized rhinolophid species, the anterior upper premolar is very small, with a single cusp, clearly displaced in the labial margin of the tooth-row, compressed between C and P4; the anterior lower premolar is oval in occlusal view, with coronal area not wider than half of p4; the p3 (alveolus) is clearly displaced in the labial margin of the tooth-row, compressed between p2 and p4, and the length p2–p4 is less than 1.75 mm. According to Lanza (1959), Krapp (2001), Kryštufek and Dulić (2001) and Popov and Ivanova (2002), these morphological and morphometric features permit to exclude R. blasii.

Some morphological features of the above listed remains enable a certain attribution to R. euryale and not to R. mehelyi: the preorbital bridge is strong (Lanza, 2012), the upper canine is relatively robust, with trapezoidal occlusal outline and with a clear longitudinal groove in labial view, the upper molars have distal a concave edge and the labial cingulum is very prominent, and the mental foramen of the mandible is placed under p2 (Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986); the styloid process of the humerus is relatively long and thin, and in lateral view slightly dorsally inclined (Felten et al., 1973). The sizes fall within the variability of living species (Table 3). Fossil remains of R. euryale can be attributed to at least five individuals (two from the late Pleistocene breccia and three from the Holocene soil; Table 1 and Table 2).

Rhinolophus euryale is a species of strict Mediterranean affinities and it prefers warm areas with abundant shrubs or forests (deciduous), in areas rich in limestone caves and close to water environments. During the summer, the roosts are located in caves mainly in warmer parts of the cavities; in winter it finds refuge in caves and mine galleries, preferably with temperatures around 10–12 ̊C (Gaisler, 2001b, Horáček et al., 2000, Lanza, 2012 and Lanza and Agnelli, 2002).

Rhinolophus euryale is distributed in the Mediterranean area from Northwest Africa over most Mediterranean islands (except for the Balearics and Crete), throughout the Iberian Peninsula, southern France, the Balkan Peninsula, Slovakia, Hungary, Romania and in the Near East (Gaisler, 2001b, Horáček et al., 2000 and Juste and Alcaldé, 2016). In Italy, the Mediterranean horseshoe bat occurs throughout the territory with the probable exception of some northern regions (Agnelli et al., 2006 and Lanza, 2012). In Sicily and the surrounding islands, only a few reports from the 1950s are known, but its presence has been confirmed in recent times (Agnelli et al., 2008).

Bat fossils referred to Rhinolophus ex gr. R. euryale are known in eastern Europe since the early Pliocene (late Ruscinian), and in Italy from late early Pleistocene (middle early Biharian, late Villafranchian; Salari et al., 2013 and Tata and Kotsakis, 2005). Fossil remains of the Mediterranean horseshoe bat come from several middle and late Pleistocene and Holocene localities of the Mediterranean basin, Poland and Near East (Gaisler, 2001b, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Kotsakis, 2017 and Salari and Silvestri, 2015, Salari et al., 2013; Tata and Kotsakis, 2005, and references therein; Fagoaga et al., 2017 and Galán et al., 2018a). It also occurs in the late Pleistocene and Holocene layers of Ghar Dalam Cave, Malta (Storch, 1970 and Storch, 1974).

Rhinolophus mehelyi Matschie, 1901. Méhely's horseshoe bat.

Material: 1 maxillary and 2 distal portions of humerus, from the late Pleistocene breccia; 1 fragmented splancnocranium, 6 maxillaries mostly fragmented, 3 mandibles and 4 mandible fragments, an intact humerus and 9 distal epiphyses of humerus, from the Holocene soil (Figs. 4F-G).

Description and remarks. The morphology and dimensions of these fossil remains are typical of the middle-sized rhinolophid species (Felten et al., 1973, Krapp, 2001, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986). For the morphological and morphometric features that allow excluding Rhinolophus blasii and R. birzebbugensis, see above. Some morphological features enable a reliable attribution to R. mehelyi and not to R. euryale: the preorbital bridge is very thin (Lanza, 2012); the upper canine is robust, without longitudinal groove in labial view, the upper molars have a slightly concave distal edge and accentuated labial cingula, the mental foramen of the mandible is placed under the anterior part of p2; the p4 is robust and subrectangular with a pyramidal cusp, the cingulum shows two convexities separated by a shallow concavity in labial view (Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986); the styloid process of humerus is long and thin, and it is parallel to the axis of the humerus (Felten et al., 1973; Salari et al., 2013). The dimensions of the material assigned to R. mehelyi agree with the variability of the living species (Table 3). Rhinolophus mehelyi fossils can be referred to at least ten individuals (two from the late Pleistocene breccia and eight from the Holocene soil; Table 1 and Table 2).

Mehely's horseshoe bat is a species of strict Mediterranean affinities, with apparently similar habits to those of R. euryale. It prefers groves and terrains covered by tree-shrub vegetation. Both summer and winter roosts are found in caves and galleries located in limestone areas next to water, with temperatures of 15–24 ̊C in the nursery, and about 11–13 ̊C in winter (Gaisler, 2001c, Horáček et al., 2000, Lanza, 2012 and Lanza and Agnelli, 2002).

Rhinolophus mehelyi is largely restricted to the Mediterranean, and it has a discontinuous distribution from North Africa and southern Europe (southern Iberian Peninsula, the Balkans and perhaps in southern France) through Asia Minor, Anatolia, to Transcaucasia, Iran and possibly Afghanistan (Alcaldé et al., 2016; Gaisler, 2001c and Horáček et al., 2000). In the entire range of its distribution, this cave-dwelling bat that is particularly sensitive to human disturbance is today considered Vulnerable (Alcaldé et al., 2016): it has declined in numbers in the Iberian Peninsula (where it is relatively more abundant), close to extinction in France and possibly already extinct in Corsica, Croatia and Israel (Alcaldé et al., 2016; Mucedda et al., 2009). In Italy, its present distribution is limited to Sardinia, Sicily and Apulia (Agnelli et al., 2006, Agnelli et al., 2008, Dondini et al., 2014, Lanza, 2012 and Mucedda et al., 2009). In Sicily, R. mehelyi seems to occur only in the Grotta di Calafarina (Portopalo di Capo Passero, Syracuse; Mucedda et al., 2009 and Mucedda et al., 2015), while previous reports, including the Grotta dei Pipistrelli (Caruso, 1982 and Caruso and Grasso, 1996), should be confirmed.

This species is known with certainty from the middle Pleistocene in Spain and Italy, since the Pliocene and early Pleistocene chronosubspecies R. mehelyi birzebuggensis was recognized as a valid species, R. birzebuggensis (see Salari et al., 2013; Tata and Kotsakis, 2005). During the late Pleistocene and Holocene R. mehelyi is reported in Spain, France, Italy, former Yugoslavia, Bulgaria, Armenia, Azerbaijan (Gaisler, 2001c, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011 and Salari and Silvestri, 2015; Tata and Kotsakis, 2005, and references therein; Sevilla, 2016). Rhinolophus mehelyi fossils are recorded also from the upper layer of the Grotta di Spinagallo (Sicily), early middle Pleistocene (Kotsakis and Petronio, 1980 and Tata and Kotsakis, 2005) and the late Pleistocene layer of Ghar Dalam Cave, Malta (Storch, 1974).

Rhinolophus euryale Blasius, 1853 vel Rhinolophus mehelyi Matschie, 1901

Material: 2 mandibular fragments, from the late Pleistocene breccia; 2 cranium fragments, 2 maxillaries fragmented, and a mandible branch, from the Holocene soil (Table 1).

Description and remarks. The discriminant morphological features are not appreciable in the few remains. Moreover, as the variability in the range of size between R. euryale and R. mehelyi is overlapping, this material could not be attributed with certainty to either of the two species.

Fossil remains related to R. euryale vel mehelyi are reported in several middle and late Pleistocene and Holocene sites of the Western Palaearctic (Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011 and Salari and Silvestri, 2015, and references therein; Bañuls-Cardona et al., 2017, Lebreton et al., 2016, López-García et al., 2014 and López-García et al., 2015), including Vallone Inferno, Holocene (López-García et al., 2013a, as Rhinolophus sp.) in Sicily.

Family Vespertilionidae Gray, 1821

Genus Myotis Kaup, 1829

Myotis myotis (Borkhausen, 1797). Greater mouse-eared bat.

Material: 2 crania, 3 splancnocrania, 3 fragmented maxillaries, 14 mandibles of which 2 intact, 1 upper and 2 lower teeth, 5 humeri and 1 distal epiphysis of humerus, from the late Pleistocene breccia; 6 splancnocrania of which one fragmented, 14 maxillaries mostly fragmented, 1 upper tooth, 54 mandibles mostly fragmented, and 6 humeri and 3 distal portions of humerus, from the Holocene soil (Figs. 5A-C).

Description and remarks. These fossil remains have the typical shape and size of the large Myotis species: dorsal cranium outline straight in side-view, prefrontal swelling absent, premaxillaries fused with maxillary bones but separated from each other, three upper premolars of which the intermediate one is smaller than the others, ascending ramus of mandible wide and with a tall and strong coronoid process, a low articular process and a long and thin angular process, three lower premolars of which the intermediate is a little smaller, lower molars of myotodontic type, m1–m2 with trigonid and talonid of equal length, third molar with reduced talonid, distal epiphysis of humerus with the epicondyle reduced, trochlea, inner and outer ridges separated by grooves, styloid process strongly reduced (Felten et al., 1973, Güttinger et al., 2001, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988, Sevilla and Lopez-Martinez, 1986 and Topál and Ruedi, 2001). In our samples of large Myotis, the upper third molar displays aboral reduction and the posterior lower premolar is subrectangular. These morphological features, along with chronological reasons, enable to exclude M. ghardalamensis, an extinct species from late early Pleistocene of Ghar Dalam Cave, Malta (Storch, 1974 and Topál and Ruedi, 2001). As known from the living species, Myotis myotisis larger in size, but the dimensions of large M. blythii and small M. myotis overlap (Arlettaz et al., 1991, Bachanek and Postawa, 2010, Benda and Horáček, 1995, Ghazali, 2009, Güttinger et al., 2001, Ruedi et al., 1990 and Topál and Ruedi, 2001). The only certain morphological difference between the teeth of M. myotis and M. blythii occurs on the talonid of m3, which is more reduced in M. myotis (see Mein, 1975, Sevilla García, 1988 and Topál and Tusnadi, 1963). Therefore, the talonid reduction index (TRI = talonid width/tooth length) was calculated and the specimens with the TRI close to or less than 0.45 were assigned to M. myotis, and those with TRI close to or more than 0.50 to M. blythii (see Mein, 1975). Although Galán et al. (2018b) showed some overlap also for this index in the extant Iberian specimens of the two species, in our samples of the large Myotis from the Grotta dei Pipistrelli, both extant and fossil, there are no overlaps between the two species (Table 4; Fig. 6). The dimensions of other cranial and dental measurements support these attributions (Table 4), although the sizes of the bone remains are slightly smaller than that of M. myotis roosting today in the Grotta dei Pipistrelli. Concerning the humeri of identical morphology of distal epiphysis (Felten et al., 1973), discrimination between the two species was made on the intact specimens, ascribing only the longest ones to M. myotis and all the others to M. blythii (Figs. 5C–E). On this basis, it was then possible to discriminate the distal fragments of the humerus, attributing to M. myotis those with breadth of distal epiphysis equal to or greater than 4.4 mm, to M. blythii those with breadth of distal epiphysis equal to or less than 4.0 mm, with all the remaining material being assigned to M. myotis vel blythii. According to the above criteria, M. myotis remains can be referred to at least 38 individuals (ten from the late Pleistocene breccia and twenty-eight from the Holocene soil; Table 1 and Table 2).

Myotis myotis is a thermophilous species of Mediterranean affinities. It prefers temperate and warm places in plains and in hills, it is frequent in forest environments with sparse undergrowth and also in open and semi-desert environments, these last not far from woody areas. In spring, it searches refuge in buildings, in underground natural or artificial environments and, more rarely, in tree hollows; it normally winters in caves and galleries both natural and artificial, with temperatures of 2–12 ̊C and high humidity (85–100%) (Arlettaz, 1996, Güttinger et al., 2001, Horáček et al., 2000, Lanza, 2012 and Lanza and Agnelli, 2002).

The recent range of greater mouse-eared bat includes eastern, southern and central Europe, reaching southern England and southern Sweden, and the northern Mediterranean basin and Near East (Coroiu et al., 2016, Güttinger et al., 2001 and Horáček et al., 2000). In Italy, M. myotis occurs in all regions (Agnelli et al., 2006 and Lanza, 2012), even if the specimens from Sardinia should be ascribed to M. punicus (see Agnelli et al., 2006). In Sicily, the greater mouse-eared bat was recorded since the nineteenth century (Doderlein, 1872, Galvagni, 1837, Minà Palumbo, 1868 and Monticelli, 1886; all the records as Vespertilio murinus Schreber, 1774, an ancient synonym for M. myotis). It is, after R. ferrumequinum and Pipistrellus kuhlii (Kuhl, 1817), the third frequent species (Agnelli et al., 2008).

According to Arlettaz et al. (1997), in Europe the sibling species, M. myotis and M. blythii, would have appeared for first time one to the West and the other to the East and they would have progressively widened the distribution area toward the East and West, respectively. Myotis blythii would have reached southwestern Europe only in the late Holocene, whereas M. myotis would have reached eastern Europe in the late Pleistocene (Arlettaz et al., 1997). However, this hypothesis must be revised and adequately updated by the palaeontological record. As for M. myotis, the earliest records come from the late early Pleistocene of Spain (Galán et al., 2016a; Sevilla, 2014; Sevilla Garcia, 1988), it is recorded from several middle Pleistocene sites of France (Mein, 1975, as Myotis cf. myotis; Desclaux, 1992 and Sevilla and Chaline, 2004) and Italy (Kotsakis and Barisone, 2008), and reaches European Turkey (Tracia, eastern Europe) in the same timeframe (Lindenau, 2005). Thus, the expansion of its distribution towards East occurred earlier than previously assumed. The expansion to the North (e.g., Poland; Wołoszyn, 1987), instead, took place in the late Pleistocene. Indeed, fossil remains attributed to the greater mouse-eared bat occur in many late Pleistocene and Holocene sites of Portugal, Spain, France, Belgium, Germany, Switzerland, Italy, Austria, Poland, former Czechoslovakia, former Yugoslavia, Hungary, Romania, Ukraine (Güttinger et al., 2001, Hutterer et al., 2012, Salari, 2009, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Silvestri, 2015 and Tata and Kotsakis, 2005, and references therein).

Myotis blythii (Tomes, 1857). Lesser mouse-eared bat.

Material: 1 splancnocranium, 3 maxillary fragments and 2 mandible fragments, from the late Pleistocene breccia; 4 maxillary fragments, 18 mandible of which 8 intact, 1 humerus and 2 distal portions of humerus, from the Holocene soil (Figs. 5D-E).

Description and remarks. This material shows the typical morphology and size of the large Myotis species (Felten et al., 1973, Güttinger et al., 2001, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988, Sevilla and Lopez-Martinez, 1986 and Topál and Ruedi, 2001). For the exclusion of M. ghardalamensis and for the morphological and morphometric features that allow the distinction between M. blythii and M. myotis, see above and Table 4. Additional comparisons were done with M. punicus, recently added to the Sicilian bat fauna (Bogdanowicz et al., 2015 and Fulco and Lo Valvo, 2015), although it seems to be based on a single female specimen (Bogdanowicz, in litteris). This species was originally described as a subspecies of M. blythii, for the greater width of M2 compared to European M. blythii populations (Felten, in Felten et al., 1977). Myotis punicus occurs in the Mediterranean part of the Maghreb (from western Morocco to eastern Tunisia), Libya, Corsica, Sardinia and Malta (Agnelli et al., 2006, Benda et al., 2014, Felten et al., 1977 and Lanza, 2012). The Sardinia and Corsica populations differ significantly in some craniometric features from the one from Maghreb (Evin et al., 2008) and they are currently genetically isolated between them, despite their ability to fly and the narrowness of Bonifacio strait (Biollaz et al., 2010). In our samples, the second upper molars are less wide (2.65–2.90 mm) than the range given for M. punicus (2.97–3.36 mm; Felten et al., 1977). Fossil remains of M. blythii can be attributed to at least 16 individuals (three from the late Pleistocene breccia and thirteen from the Holocene soil; Table 1 and Table 2).

Myotis blythii is a thermophilous and troglophilous species of Mediterranean affinities. The biology of the lesser mouse-eared bat seems to be overall very similar to that of the greater mouse-eared bat. However, it differs significantly in the diet and, consequently, the areas of foraging are different (Arlettaz, 1996). Myotis blythii prefers to hunt in areas more or less rich in grasslands, both primary (steppes, grasslands) and anthropogenic (meadows, pastures), avoiding the arid and naked areas, or with degraded or freshly cut grass, and any type of woodland and forest; hibernacula are found in hypogean cavities usually with a temperature of 4–14 ̊C (Horáček et al., 2000, Lanza, 2012, Lanza and Agnelli, 2002 and Topál and Ruedi, 2001).

The recent range of the species includes southern Europe, the southern part of Central Europe, parts of southwestern Asia and east to China and Mongolia (Horáček et al., 2000 and Juste and Paunović, 2016; Topál and Ruedi, 2001). In Italy, this species occurs in all regions, except for Sardinia (Agnelli et al., 2006 and Lanza, 2012). In Sicily, its occurrence is reported since the nineteenth century (Monticelli, 1886; as Vespertilio oxygnathus Monticelli, 1865, ancient synonym for M. blythii) and today it is recorded in few localities (Agnelli et al., 2008).

The earliest fossil records referred to Myotis cf. M. blythii and M. blythii longicaninus Popov, 2004 come from some eastern European sites referred to late Pliocene (late Ruscinian; Popov, 2004 and Topál, 1983). As mentioned above, it seems that in Europe the distribution of M. blythii has subsequently progressively expanded towards the west. After the late Pliocene, there are further evidences from eastern Europe, it occurs in Italy in the earliest and late early Pleistocene (Salari et al., 2013 and Tata and Kotsakis, 2005), in Austria in the middle Pleistocene (Rabeder, 1972 and Rabeder, 1973, as M. oxygnathus) and it reached the Iberian Peninsula in the late Pleistocene (Galán et al., 2016b, as M. oxygnathus in Appendix B, 2018a; Fagoaga et al., 2017), although, since middle Pleistocene, fossil remains attributed to M. myotis vel blythii are recorded both in southern France (Sevilla and Chaline, 2011) and in Spain (Sevilla García, 1988). In this case too, the expansion of its distribution area towards the west seems to occur earlier than hypothesized by Arlettaz et al. (1997). Myotis blythii fossils occur in several middle Pleistocene localities and in many late Pleistocene and Holocene sites in southern and eastern Europe and the Near East (Salari, 2009, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Kotsakis, 2017, Salari and Silvestri, 2015, Salari et al., 2013, Tata and Kotsakis, 2005 and Topál and Ruedi, 2001, and references therein). The species is reported also from some late Pleistocene and Holocene sites of Morocco (e.g., López-García et al., 2013b and Ouahbi et al., 2003) and Malta (Storch, 1970 and Storch, 1974), but these finds should be attributed to M. punicus.

Myotis myotis (Borkhausen, 1797) vel Myotis blythii (Tomes, 1857)

Material: 1 fragmented splancnocranium without teeth, 4 maxillary fragments, 40 mandible fragments mostly without teeth, 34 upper and 21 lower teeth, and 45 distal epiphyses of humerus, from the late Pleistocene breccia; 1 splancnocranium without teeth, 17 maxillary fragments mostly without teeth, 94 mandible fragments mostly without teeth, 3 lower teeth, and 74 distal epiphyses of humerus, from the Holocene soil.

Description and remarks. The overlap in the range of size variability between the two species and any evident discriminant morphological feature does not allow us to attribute the material to M. myotis or to M. blythii. Among the distal portions of the humerus, six specimens from the late Pleistocene breccia and seven from the Holocene soil do not have the distal epiphysis fused and therefore they belong to immature individuals. Thus, the remains of M. myotis vel blythii can be referred to at least 82 individuals (28 from the late Pleistocene breccia and 54 from Holocene soil; Table 1 and Table 2).

Fossil remains attributed to M. myotis vel blythii are reported in many middle and late Pleistocene and Holocene sites of Western Palaearctic (Salari, 2009, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011 and Salari and Silvestri, 2015, and references therein; Bañuls-Cardona et al., 2017, Galán et al., 2016b, López-García et al., 2014 and López-García et al., 2015), including the Grotta di San Teodoro, late Pleistocene (Mangano et al., 2005, as Myotis sp.) and Vallone Inferno, Holocene (López-García et al., 2013a) in Sicily.

Myotis capaccinii (Bonaparte, 1837). Long-fingered bat.

Material: 1 lower canine and 3 distal humeri from the late Pleistocene breccia and 2 distal humeri, from the Holocene soil (Fig. 7A).

Description and remarks. Morphology and dimensions (Table 5) agree with those of the living M. capaccinii: lower canine is very similar to those of the great-sized Myotis, but with more reduced size, it shows a single three-sided cusp and, unlike M. mystacinus, the coronal area is slightly more elongated and without accessory cusp (Lanza, 1959 and Spitzenberger and Helversen, 2001); the distal epiphysis of humerus shows the epicondyle with reduced trochlea, inner and outer ridges separated by grooves and the styloid process moderately developed (Felten et al., 1973). These fossil remains can be referred to at least four individuals (two from the late Pleistocene breccia and two from the Holocene soil; Table 1 and Table 2).

Myotis capaccinii is a species of strict Mediterranean affinities; it inhabits wooded or bushy karst areas associated with rivers or ponds, in which it may also prey upon water larvae and small fish. It is typically throglophilous, refuging throughout the year in both natural and artificial caves and cavities; the winter shelters are with temperatures between 4 and 15 ̊C and relative humidity of 90–100% (Horáček et al., 2000, Lanza, 2012 and Lanza and Agnelli, 2002; Spitzenberger and Helversen, 2001).

The current range of distribution includes the Mediterranean basin, south-western Asia from Turkey to Iran (Horáček et al., 2000, Paunović, 2016 and Spitzenberger and Helversen, 2001); in Italy, this species occurs in all regions (Agnelli et al., 2006 and Lanza, 2012). In Sicily and surrounding islands, Bonaparte (1837, as Vespertilio capaccinii) established the species on Sicilian specimens; in the nineteenth century it was also recorded by Minà Palumbo (1868) and Doderlein (1872, as Comastes capaccini Fitzinger, 1870, a synonym for M. capaccinii), but its current occurrences are limited to a few caves (Agnelli et al., 2008).

The oldest fossils of the long-fingered bat are known in the late early Pleistocene (middle early Biharian, late Villafranchian) in a few Mediterranean localities (Adrover, 1967, Salari et al., 2013 and Storch, 1974). It is found in some middle and late Pleistocene and Holocene sites from central and southern Italy and other circum-Mediterranean localities, invariably represented by few remains, whereas in northern Italy and in the remaining western Palaearctic it is rarely reported (Salari, 2009, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Kotsakis, 2017 and Salari and Silvestri, 2015; Salari et al., 2013, Spitzenberger and Helversen, 2001 and Tata and Kotsakis, 2005, and references therein). Myotis capaccinii fossils occur also in the lower layer of the Grotta di Spinagallo (Sicily), early middle Pleistocene (Kotsakis and Petronio, 1980 and Tata and Kotsakis, 2005) and in the late early Pleistocene layer of Ghar Dalam Cave, Malta (Storch, 1974).

Myotis cf. M. mystacinus (Kuhl, 1817). Whiskered bat.

Material: a distal humerus, from the Holocene soil (Fig. 7B).

Description and remarks. In the distal epiphysis of humerus, the epicondyle is reduced, with trochlea, inner and outer ridges separated by grooves; the styloid process is strongly reduced but still present and in external view does not overtop the trochlea. According to Felten et al. (1973), these morphologic features and the very small size (Table 5) enable the attribution to M. mystacinus.

According to numerous authors (e.g., Agnelli et al., 2006, Benda and Tsytsulina, 2000, Benda et al., 2016 and Helversen et al., 2001), this taxon is a morphogroup with several cryptic species. In effect, since the 70s of last century, Myotis brandtii (Eversmann, 1845), previously considered a subspecies of M. mystacinus, was identified as a separate species (Agnelli et al., 2006, Hanák, 1970, Horáček et al., 2000, Lanza, 2012, Simmons, 2005 and Tupinier, 2001). Recently, the modern bioacoustic and mostly the molecular biology techniques enabled the identification of new species belonging to the M. mystacinus group, such as M. aurascens Kusjakin, 1935 and M. alcathoe Helversen and Heller, 2001 (Benda and Tsytsulina, 2000 and Helversen et al., 2001), but the lack of morphometric data on their humeri did not permit adequate taxonomic assessment of them.

The geographical distribution of these species is poorly known. Myotis aurascens is not part of the Italian bat fauna (Lanza, 2012 and Tsytsulina et al., 2012), but Benda and Paunović (2016) seem to have a different view. On the other side, both M. brandtii and M. alcathoe have been reported also from continental Italy (Agnelli et al., 2006, De Pasquale and Galimberti, 2014 and Lanza, 2012), but not from Sicily and the surrounding islands (Agnelli et al., 2008, Fulco and Lo Valvo, 2015 and Lanza, 2012). For these reasons, this distal epiphysis of humerus was tentatively attributed to Myotis cf. M. mystacinus.

Myotis mystacinus is a nemoral species, preferring forests or open areas close to wetlands and frequently it is observed in parks and gardens in towns, preferably near rivers or lakes. Hibernation takes place in natural and artificial cavities, with temperatures of 2–8 ̊C and high relative humidity, ranging from 80 to 100%. (Horáček et al., 2000, Lanza, 2012, Lanza and Agnelli, 2002 and Tupinier and Aellen, 2001).

Today's range of distribution includes Europe (except Iceland), northwestern Africa and a great part of Asia to Japan (Horáček et al., 2000 and Tupinier and Aellen, 2001), but the species distribution is likely to be reconsidered in the light of the recent identification of new species of the M. mystacinus group. In Italy, the whiskered bat probably occurs in all regions (Agnelli et al., 2006 and Lanza, 2012). In Sicily, the species was dubiously reported since the nineteenth century (Doderlein, 1872) and it is considered part of the Sicilian bat fauna (Fulco and Lo Valvo, 2015, Lanza, 1959 and Lanza, 2012), even if there are no recent reports (Agnelli et al., 2006 and Agnelli et al., 2008).

Fossil remains attributed to Myotis cf. M. mystacinus are known since the late Pliocene (MN 16a biounit, early Villanyian; Horáček, 1976 and Sabol et al., 2004) and to M. mystacinus since the early Pleistocene (Topál, 1963), while those referred to M. brandtii would be known since the late middle Pleistocene (Tupinier, 2001). Bone remains referred to M. mystacinus come from several middle and late Pleistocene and Holocene sites in Portugal, France, Belgium, Germany, Switzerland, Austria, Poland, former Czechoslovakia, Hungary, Bulgaria, Ukraine (Salari, 2010, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011 and Tupinier and Aellen, 2001, and references therein; Hutterer et al., 2012, López-García et al., 2017a and Sevilla, 2016). By contrast, in Italy there are very few reports: Myotis capaccinii vel mystacinus is recorded at Covolo di Trene, latest Pleistocene (Bon et al., 1991), M. mystacinus s.l. (= sensu lato) comes from Grotta Breuil, late Pleistocene (Salari and Di Canzio, 2009 and Salari and Kotsakis, 2011) and Caverna delle Arene Candide, latest Pleistocene (Salari, 2010). Finally, M. mystacinus is reported from upper layer of the Grotta di Spinagallo (Sicily), in the early middle Pleistocene (Kotsakis and Petronio, 1980 and Tata and Kotsakis, 2005). However, it cannot be ruled out that some of the aforementioned reports (particularly those of the last century) can be referred to M. brandtii, M. aurascens or M. alcathoe.

Genus Plecotus Geoffroy, 1818

Plecotus cf. P. auritus (Linnaeus, 1758). Brown long-eared bat.

Material: 2 distal epiphyses of humerus (Fig. 7C), referable to two different individuals, from the late Pleistocene breccia (Table 1 and Table 2).

Description and remarks. The distal epiphysis of humerus is wide (2.8 mm, Table 5) and displays the epicondyle with reduced trochlea, inner and outer ridges separated by grooves, the styloid process is moderately developed and does not overtop the trochlea in external view; in inner view, the transition from the epitroclea to the styloid process is angular, as in P. auritus, and not roughly straight, as in Plecotus austriacus (Fischer, 1829) (Felten et al., 1973).

The recent applications of modern bioacoustic and molecular biology techniques (combined with conventional approaches based on morphological and morphometric criteria), have allowed identification of new species of genus Plecotus, such as Plecotus macrobullaris Kuzjakin, 1965, Plecotus kolombatovici Dulić, 1980 and Plecotus gaisleri Benda, Kiefer, Hanák and Veith, 2004. In particular, P. kolombatovici and/or P. gaisleri were reported from central Mediterranean islands surrounding Sicily, such as Pantelleria and the Maltese Archipelago (Benda et al., 2004, Benda et al., 2014, Lanza, 2012 and Spitzenberger et al., 2006), but the lack of morphometric data on their humeri did not permit adequate taxonomic assessment of them.

Plecotus kolombatovici was originally described as a subspecies of P. austriacus, occurring on the Adriatic coast and islands of Croatia (Dulić, 1980). It is a Mediterranean endemic species with a fragmented distribution in the southern regions of the Balkans, Asia Minor and probably also Malta and Pantelleria (Lanza, 2012 and Spitzenberger et al., 2006). Plecotus gaisleri was originally described as a subspecies of P. teneriffae Barret Hamilton, 1907 (already subspecies of P. auritus, see Horáček et al., 2000), occurring in Cyrenaica (Libya) and Mediterranean northwestern Africa (Benda et al., 2004). It is endemic to the Mediterranean part of Africa and some adjacent islands, such as Pantelleria and Malta (Benda et al., 2004, Benda et al., 2014 and Lanza, 2012). Spitzenberger et al. (2006) ascribed the P. teneriffae population from the Mediterranean north-western Africa and adjacent islands to P. kolombatovici. For these reasons, our specimens were tentatively attributed to Plecotus cf. P. auritus.

Plecotus auritus is a boreal species, found typically in woodland, in evergreen boreal, mixed and deciduous forests, as well also in parks and gardens of villages and towns. During the milder seasons, the roosts are located in hollow trees and occasionally in rock fissures or in the roofs of buildings; during the winter it takes refuge in cellars and in hollow trees, as well as in caves and mines (Agnelli et al., 2006, Horáček et al., 2000 and Lanza, 2012). The brown long-eared bat is a species relatively resistant to cold temperatures; in caves, it preferably chooses the areas near the entrance. Hibernacula are usually located in very humid environments (humidity between 80 to 100%) and with quite low temperatures ranging between 2 and 5 ̊C, but it has been observed to survive for even a few days with lower temperatures, up to −3.5 ̊C (Lanza, 2012 and Lanza and Agnelli, 2002).

The species currently occurs throughout the whole of Europe, where it is widely distributed South of 65̊N, West of the Urals and North of the Caucasus. However, its distribution in the South is patchy (Horáček et al., 2000, Hutson et al., 2008a and Spitzenberger et al., 2006). In Italy, P. auritus has been reported in the northern and central regions and in Sardinia, where it is in sympatry with its congeneric P. sardus Mucedda, Kiefer, Pidinchedda and Veith, 2002 (Agnelli et al., 2006 and Lanza, 2012). In Sicily and the surrounding islands, its presence is debated. In the nineteenth century, the brown long-eared bat was reported by Galvagni (1837, as Vespertilio auritus, an ancient synonym for P. auritus), Sanvisente (1849, as Vespertilio Auritus), Minà Minà Palumbo (1868, along with P. brevimanus Jenyns, 1828, synonymized with P. auritus, see Simmons, 2005), Doderlein (1872, together to P. brevimanus) and Monticelli (1886). After the distinction of P. austriacus from the sibling species P. auritus in the mid-1950s, and new observations in the years 1960–1990, the above records were attributed to P. austriacus, known to surely occur also in Sicily and southern Italy (Agnelli et al., 2006, Agnelli et al., 2008, Lanza, 2012 and Sarà, 1999). However, Fulco et al. (2015) recently reported the capture of a young female of P. auritus, near Caronia (Messina).

Fossil remains referred to P. auritus come from several localities of central and eastern Europe, whereas in the circum-Mediterranean regions are quite rare (Salari and Di Canzio, 2009). Earlier material described as Plecotus aff. auritus is reported in the earliest Pleistocene (late Villanyan, middle Villafranchian) in Germany (Kowalski, 1962). Plecotus auritus vel austriacus is recorded in middle and late Pleistocene localities in Spain (Galán et al., 2016b, Laplana et al., 2013 and Sevilla García, 1988), France (Sevilla and Chaline, 2011) and Italy (Berto et al., 2016, López-García et al., 2014, López-García et al., 2015 and López-García et al., 2017b), while P. auritus vel macrobullaris is reported in the late Pleistocene of Transcaucasia (Sevilla, 2016). In the middle and late Pleistocene and Holocene P. auritus is recorded in Spain, France, Belgium, Germany, Austria, Poland, former Czechoslovakia, Hungary, Bulgaria, Ukraine, Russia and Turkey (Bañuls-Cardona et al., 2017, Hutterer et al., 2012, Pazonyi et al., 2014, Rosina, 2012 and Salari and Di Canzio, 2009 with references). In Italy, late Pleistocene and Holocene remains referred to P. auritus or P. auritus s.l. come from Zandobbio (Maviglia, 1954), Palmaria Island (Cigna et al., 1967), Grotta di Pozzo (Salari and Di Canzio, 2009), Caverna delle Arene Candide (Salari, 2010), Grotta dei Cocci (Salari, 2011) and Grotta Mora Cavorso (Salari and Silvestri, 2015). It is likely that several of the abovementioned remains actually belong to P. austriacus, P. macrobullaris or P. kolombatovici. Therefore, a systematic review of the fossil material assigned to the genus Plecotus is necessary.

Family Miniopteridae Dobson, 1875

Genus Minioperus Bonaparte, 1837

Miniopterus schreibersii (Kuhl, 1819). Schreiber's bat.

Material: 1 maxillary fragment, 21 mandibles mostly fragmented, 1 upper and 1 lower tooth, 2 humeri and 8 distal epiphyses of humeri from the late Pleistocene breccia; 1 splancnocranium, 4 fragmented maxillaries, 32 mandibles of which 5 are intact, and 4 humeri and 19 distal epiphyses of humeri from the Holocene soil (Figs. 7D-E).

Description and remarks. Both the morphology and size (Table 6) of the above fossil remains match with those of the living Schreiber's bat: cranium without prefrontal swelling and with dorsal outline strongly sinuous in side-view, premaxillaries fused with maxillary bones but separated from each other, three upper premolars of which the anterior greatly reduced, mandible with coronoid process higher than the articular one, and angular process long and robust, three lower premolars consecutively higher from the anterior to the posterior one, lower molars of nyctalodontic type, distal epiphysis of humerus with the medial epicondyle relatively narrow, trochlea separated from the inner ridge by a shallow groove, inner and lateral ridges separated one from each other by a deep groove, the styloid process long and narrow in anterior view (Felten et al., 1973, Lanza, 1959, Lanza, 2012, Salari, 2004, Sevilla García, 1988 and Sevilla and Lopez-Martinez, 1986). These M. schreibersii remains can be attributed to at least 32 individuals (twelve from the late Pleistocene breccia and twenty from the Holocene soil; Table 1 and Table 2).

Miniopterus schreibersii is a species of probable tropical origin, it is sub-cosmopolitan, but, within Europe, of strict Mediterranean affinities and typically troglophilous. It is found mainly linked to environments not anthropized or scarcely modified by man, clearly preferring karstic areas, but also occurring in a variety of both forested (especially deciduous woodlands) and open habitats (herbaceous formations, including steppe). The temperature and relative humidity of the caves in which they find refuge normally range between 11–19 ̊C and 57–91% in the summer roosts and between 4–12 ̊C and 70–98% in the case of winter roosts (Horáček et al., 2000, Lanza, 2012, Lanza and Agnelli, 2002 and Vincent et al., 2011).

Recent studies of bioacoustics and molecular biology have identified and/or established to species some subspecies or geographic varieties of M. schreibersii from Asia, sub-Saharan Africa and Oceania (e.g., Goodman et al., 2011 and Miller-Butterworth et al., 2005). Therefore, the current range of distribution seems to be restricted to the southern part of western Palearctic and, perhaps, Guinea, from south-western Europe and North and West Africa through Anatolia and the Near East to the Caucasus (Hutson et al., 2008b). In Italy, M. schreibersii occurs throughout the whole country (Agnelli et al., 2006 and Lanza, 2012). In Sicily, Schreiber's bat is currently distributed in the island and it is one of the most observed species (Agnelli et al., 2008). Its occurrence has been recorded since the nineteenth century (Doderlein, 1872).

Fossil remains attributed to M. schreibersii are known since the earliest Pleistocene (late Villanyan, middle Villafranchian) in various localities of central Europe and in Italy from late early Pleistocene (middle early Biharian, late Villafranchian; Salari et al., 2013 and Tata and Kotsakis, 2005). Schreiber's bat occurs in several sites from middle and late Pleistocene and Holocene. It disappeared from central Europe during the glacial phases, but it was present in the Mediterranean basin and expanded its distribution area further north during the interglacial phases and the Holocene (Salari, 2009, Salari, 2010, Salari, 2011, Salari and Di Canzio, 2009, Salari and Kotsakis, 2011, Salari and Kotsakis, 2017 and Salari and Silvestri, 2015, Salari et al., 2013, Tata and Kotsakis, 2005, and references therein; Bañuls-Cardona et al., 2017, Galán et al., 2016b and López-García et al., 2017a). Fossil remains of M. schreibersii occur also in Sicily and Malta, from the late early Pleistocene and late Pleistocene layers of Ghar Dalam Cave (Storch, 1974), lower and upper layers of the Grotta di Spinagallo, early middle Pleistocene (Kotsakis and Petronio, 1980 and Tata and Kotsakis, 2005) and the Holocene of Vallone Inferno (López-García et al., 2013a).

The chiropteran remains recovered from the Grotta dei Pipistrelli assemblages amount to 664 identified specimens referable to at least 208 individuals (Table 1 and Table 2). Nine taxa, divided into four genera and three families of the order Chiroptera, have been identified.

Nine taxa of bats divided into three families (Rhinolophidae, Vespertilionidae and Miniopteridae) and four genera (Rhinolophus, Myotis, Plecotus and Miniopterus) have been discovered in the fossil assemblages from the Grotta dei Pipistrelli in Pantalica. All the taxa identified are still part of the extant Italian bat fauna and today also occur in Sicily, with the probable exception of brown long-eared bat. The bony remains do not show major morphological or morphometric differences from other late Pleistocene, Holocene and extant populations of the same bat species.

Radiometric dating suggest that the bat remains were deposited during the late Pleistocene (MIS 2, around the LGM) and in two close but distinct moments of middle Holocene (Atlantic chronozone).

Taphonomic remarks show that all the three assemblages are autochthonous thanatocoenoses and only very few bone remains from the Holocene soil at the base of the wall were accumulated by predators. The Grotta dei Pipistrelli hosted bat colonies mainly during the winter semester, for hibernation, but the cave was also utilized as a nursery.

Fossil assemblages are very similar to each other and with it current and historical bat fauna of the Grotta dei Pipistrelli. Rhinolophus ferrumequinum, R. euryale, R. mehelyi, Myotis myotis, M. blythii, M. capaccinii, Plecotus cf. P. auritus and Miniopterus schreibersii occur in the late Pleistocene ossiferous breccia. In the first Holocene assemblage appear Myotis cf. M. mystacinus and lack Plecotus cf. P. auritus, while R. ferrumequinum, R. mehelyi, M. myotis, M. blythii and M. schreibersii occur in the most recent Holocene assemblage.

From this cave, the first Pleistocene record of R. euryale in Sicily, and of M. myotis and M. blythii in the central Mediterranean islands are reported. The Grotta dei Pipistrelli probably played a role of refuge area for some species currently distributed also in continental Europe and it is the southernmost limit of the brown long-eared bat distribution, even considering its occurrences in the Pleistocene and the recent record from Messina province.

The ecological needs of the species present at the Grotta dei Pipistrelli allow to outline a composite environment near the cave, with wooded areas interrupted by open spaces (grassland or steppe) and rocky areas, where the water supply provided by Anapo River and its tributaries was great enough to support a rich vegetation cover. The percentage variations of frequencies of the recognized taxa suggest that in the late Pleistocene there was a greater vegetation cover than today and that Sicily, in the central Mediterranean basin, kept temperate–warm climatic conditions even during the LGM, whereas during the middle Holocene there was a progressive increase of open spaces, probably connected with the deforestation by Neolithic humans.