The Euscorpius Tergestinus (CL Koch, 1837) Complex in Italy

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Zoologischer Anzeiger 244 (2005) 97–113 www.elsevier.de/jcz

The Euscorpius tergestinus (C.L. Koch, 1837) complex in Italy: Biometrics of sympatric hidden species (Scorpiones: Euscorpiidae) Valerio Vignolia,Ã, Nicola Salomonea, Tancredi Carusob, Fabio Berninia aDepartment of Evolutionary Biology, University of Siena, Via A. Moro 2, 53100 Siena, Italy bDepartment of Environmental Science, University of Siena, Via Mattioli 4, 53100 Siena, Italy

Received20 October 2004; receivedin revisedform 29 April 2005; accepted5 May 2005

Corresponding editor: Andrew Parker

Abstract

This work stems from the results of a recent phylogenetic investigation on the Euscorpius carpathicus species complex from the Italian peninsula (Salomone et al. 2004. Phylogenetic relationships between the sibling species Euscorpius tergestinus and E. sicanus (Scorpiones, Euscorpiidae) as inferred from mitochondrial and nuclear sequence data. In: Proceedings of the16th Congress of Arachnology, August 2–7, 2004, Ghent University, Belgium, 268pp.; Salomone et al. in prep.). Molecular investigation produced interesting and unexpected findings on the scorpion Euscorpius tergestinus (C.L. Koch, 1837). Both nuclear and mitochondrial sequence data provided evidence of substantial genetic differentiation in specimens identified as Euscorpius tergestinus according to recent taxonomical changes (Fet andSoleglad2002. Morphology analysis supports presence of more than one species in the ‘‘ Euscorpius carpathicus’’ complex (Scorpiones: Euscorpiidae). Euscorpius 3, 51pp.). These specimens clearly belong to two well- differentiated evolutionary lineages. Molecular results highlighted the need for a new morphological investigation. The present study undertook the morphological analysis of specimens belonging to both genotypes with the aim of identifying morphological characteristics able to discriminate between the two taxa. The analysis of trichobothria patterns, morphometric ratios, granulation patterns andthe observation of the pectinal sensilla confirm the difficulty in distinguishing these two genotypes and the high polymorphism of the subgenus Euscorpius Thorell, 1876. The length of pedipalp segments and dorsal patellar spurs (DPS), as well as femur leg granulation, are the main diagnostic characters; other ratios together with body color also help to distinguish the different genotypes. This study confirms the presence in Italy of two different cryptic species belonging to the ‘‘Euscorpius tergestinus’’ complex. Euscorpius tergestinus is a reddish, slender euscorpiid with a large dorsal patellar spine (DPS). A darker and generally squat phenotype with a short DPS, which corresponds to Euscorpius carpathicus concinnus sensu Caporiacco (1950), is elevatedto the species level: Euscorpius concinnus (C.L. Koch, 1837). These two species are sympatric in several Italian regions, and their distribution pattern is possibly determined by intraguild predaction interaction. r 2005 Elsevier GmbH. All rights reserved.

Keywords: Scorpions; Morphology; Biometrics; Multivariate analysis; ANOVA

ÃCorresponding author. Tel.: +39 0577 234393. E-mail address: [email protected] (V. Vignoli).

1. Introduction demonstrate that although populations of E. tergestinus from Italy are morphologically very similar, they do The systematics of the genus Euscorpius Thorell, 1876 indeed represent different evolutionary lineages. DNA (Scorpiones: Euscorpiidae) has undergone several taxo- analysis led to the fascinating discovery of unexpected, nomic changes since 1999, after the first molecular hidden or cryptic species, as previously found for other investigation (Gantenbein et al. 1999). Euscorpius animal groups (Trewick 1998; Taylor et al. 1998; Lessios (Euscorpius) carpathicus (Linnaeus, 1767) is now con- et al. 1999; Colborn et al. 2001; Wellborn andCothran sidered a complex of at least 7 sibling species. Euscorpius 2004), including arachnids such as pseudoscorpions carpathicus in sensu stricto remains validbut only for (Zeh andZeh 1994 ; Wilcox et al. 1997). We here the populations of the type locality in Romania (Fet and focus on these two different genotypes of ‘‘E. tergesti- Sissom 2000; Fet andSoleglad2002 ). Some of these nus’’. The extensive intraspecific polymorphism exhib- species are characterizedby localizeddistributions such itedby E. tergestinus has been highlightedby several as the Balearic E. balearicus Caporiacco, 1950, the authors (Huber et al. 2001; Fet andSoleglad2002 ); Crimean E. tauricus (C.L. Koch, 1837) andthe Greek however, we notedthat in general, upon closer E. koschewnikowi Birula, 1900, which were elevatedto examination, our sequencedspecimens were easy to the species level after both molecular andmorphological distinguish through qualitative characters such as color analyses (Gantenbein et al. 2001; Fet 2002; Fet and andgeneral morphology. We aimedto morphologically Soleglad2002 ). Euscorpius hadzii Caporiacco, 1950, is distinguish the two different genotypes with the typical considered a Balkan species with a widespread geo- E. tergestinus trichobothrial pattern. We report mor- graphic distribution (Fet andSoleglad2002 ). Two other phological data on all specimens included in the ‘‘new’’ species with a relatively wide distribution molecular analysis (Salomone et al. in prep.). Note that area are E. sicanus (C.L. Koch, 1837) and E. tergestinus some non-sequencedspecimens belonging to the same (C.L. Koch, 1837); they are the only known taxa populations of the sequencedspecimens were included belonging to the ‘‘carpathicus’’ complex that are present in this study in order to equilibrate the data set. in Italy. The geographical distribution of E. sicanus on the Italian Peninsula is more southern, including Sicily andSardinia ( Fet et al. 2003), while E. tergestinus is 2. Materials andmethods widely distributed from north to south (the exact geographic range is not yet well defined) and in the countries bordering northern Italy (Fet andSoleglad 2.1. Materials 2002; Vignoli andCrucitti 2003 ). The high phenotypic A total of 167 specimens, all collectedin Italy, were variability within these scorpions leddifferent authors analyzed(see AppendixA). The complete collection of (Koch 1837, 1841; Caporiacco 1950) to classify several scorpions labeled‘‘ Euscorpius carpathicus’’ within the different species and subspecies. Recent studies (Fet and Societa` Romana di Scienze Naturali (69 specimens) was Soleglad2002 ), basedon the morphological re-exam- included in this study together with some of Capor- ination of a large number of specimens, conclude that 9 ‘‘old’’ taxa actually belong to the same species and iacco’s specimens preservedin MZUF (12 specimens). The samples are preservedin the private collection of the are synonymous with E. tergestinus (Fet andSoleglad first author in the Dipartimento di Biologia Evolutiva, 2002). These taxonomical changes are basedon the University of Siena, Italy, with exclusion of the MZUF presence of a ‘‘standard’’ character, i.e. the trichobo- specimens. thrial formula of the pedipalp patella external aspect, with eb ¼ 4=4. The high morphological variability within andbetween populations was attributedto 2.2. Morphological analysis intraspecific polymorphism. A recent study based on nuclear and mitochondrial Starting from the specimens that were analyzedin the sequence data on the ‘‘E. carpathicus’’ complex from the molecular study, 8 adult specimens were randomly Italian peninsula confirmedthat E. sicanus is a well- selected to obtain a total of 10 adult individuals differentiated evolutionary lineage (Fet et al. 2003). (5 males and5 females) for each genetic group With respect to specimens determined on the basis of the (see Appendix A and Table 1). The selection criteria trichobothrial pattern, such as E. tergestinus (eb ¼ 4=4), were age and sampling locality. Only adults and, when genetic data clearly identified two deeply divergent possible, specimens from the same population as the lineages (Salomone et al. in prep.). The observed sequenced one were considered. We analyzed both the sequence divergence between these two assemblages patella trichobothria series (Pv, Pe) andpectinal tooth was large enough to be comparable to that separating counts of all the sequenced‘‘ tergestinus’’ specimens. All other Euscorpius species, e.g. E. flavicaudis (De Geer, morphometric ratios andcarination quotients were 1787) and E. italicus (Herbst, 1800). These results determined for the selected material. The conventions ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 99 Pe total Pv+Pe eb a (C.L. Koch, 1837) complex et est em esb eb LRLRLRLRLRLRL R L R Euscorpius tergestinus LRLR VVZC code Locality m/f Dp Pv Pe 260310304101 Poggibonsi106 (SI), Tuscany102 Brenna (SI), Tuscany46 Siena (SI), Tuscany Barberino95 del Mugello (FI),89 Lucca Tuscany (LU), Tuscany Montaione m (FI), Tuscany m Elba243 Island, 7 Tuscany 8 Bologna38 (BO), 8 Emilia m Romagna Pavia219 8 (PV), Lombardy 886 8 m 8 8172 Siena f (SI), 8 f 8 Tuscany f 8 662 Rome 9 6 (RM), Latium 9114 Rieti 7 6 (RI), 7 7 Latium 9 6246C f 8 8 Rome 4 7 (RM), 7 Latium112 Rome 6 4 (RM), 8 f Latium 9 4 8 8 7 6 4 Carapelle 6 Alvisio 8 4 (AQ), 8 Rome 8 7 Abruzzo (RM), Rome 4 7 4 Latium (RM), 6 Latium 6 4 6 6 9 4 7 4 f Rome 4 7 2 (RM), 7 Latium 6 8 f m 4 8 2 4 4 2 4 7 4 6 4 8 2 f 8 4 4 7 4 f 4 7 4 7 m 2 6 4 9 4 4 7 4 4 7 9 4 4 2 6 4 7 9 8 2 4 9 4 4 m 7 8 4 4 m 4 3 4 7 9 9 2 8 3 4 2 9 6 2 4 8 4 4 4 9 6 8 9 m 4 2 6 24 2 9 6 2 4 8 4 4 24 9 6 8 9 4 5 4 8 4 6 4 4 24 2 9 3 4 6 4 9 6 4 24 4 4 8 4 6 4 4 2 9 24 2 6 32 4 9 5 4 4 4 9 4 32 4 4 4 4 6 2 4 4 32 6 4 23 24 4 4 9 4 4 4 4 4 32 6 4 4 4 6 4 4 25 6 33 24 24 4 2 24 3 4 4 4 2 4 4 2 6 4 2 32 25 4 4 25 32 4 24 4 2 4 4 2 4 2 4 23 4 5 34 32 2 4 32 4 2 32 4 4 2 4 3 23 2 4 25 33 4 2 33 4 32 4 4 4 2 4 4 2 4 24 32 4 4 4 4 2 4 4 2 4 4 33 2 31 4 24 4 4 24 4 4 24 2 4 32 4 24 4 4 4 24 4 24 25 4 24 4 24 33 4 33 4 24 33 4 24 32 4 23 33 24 33 24 33 4 33 33 23 33 24 24 33 32 33 24 33 32 33 33 33 Trichobothrial counts, sex andDp of 20 selectedspecimens belonging to Table 1. Black type 246 Volterra (PI), Tuscany m 8 8 8 8 6 6 4 4 4 4 2 2 4 4 4 4 24 24 32 32 Redtype 96 Sistiana (TS), Friuli V. Giulia f 5 5 9 9 6 6 4 4 4 4 2 2 3 4 4 4 23 24 32 33 ARTICLE IN PRESS 100 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 of Vachon (1974) were followedfor trichobothrial to test univariate differences between genetic groups and notations. All measurements are in millimeters (mm) sex for all variables (Underwood 1997). A logarithmic andfollow Stahnke (1970) and Sissom (1990). Biometric transformation was usedwhen datalackedthe assump- measurements were taken with a micrometric ocular lens tion of variance homogeneity. The 2-way ANOVA was appliedto an optical WildHeerbrugg stereomicroscope. also appliedto the following indices obtainedfrom the We followedthe methodusedby Fet andSoleglad ratio between the main biometric variables: pedipalp (2002); 26 measurements of each specimen were taken chela width/pedipalp chela length (ChelW/ChelL); fifth andsubsequently usedfor statistical analysis. Only the metasomal segment length/fifth metasomal segment measurement of pedipalp chela depth followed Sissom width (metasoma V length/metasoma V width); dorsal (1990). Granulation andcarination, of metasoma, patellar spine (DPS) length/pedipalp patella length; pedipalps and legs, were also observed with a stereo- pedipalp femur length/pedipalp femur width (femur microscope. To analyze the variation of carinae we length/femur width). employedthe ‘‘carinae development quotient’’ ( Fet and Soleglad2002 ) in order to provide a general distinction. The chelal carinae configuration follows the diagram of 3. Morphological results SolegladandSissom (2001: Fig. 54) . The pectinal tooth shape was investigatedandthe right pecten of four To avoidconfusion, insteadof considering the specimens was cut andanalyzedunder a scanning numerous known subspecies, we chose to divide speci- electron microscope (SEM). Scanning electron micro- mens into two distinct morphotypes according to their graphs were preparedfrom selectedspecimens storedin phenotypic aspect. 75% ethyl alcohol. They were ultrasonically cleaned ‘‘Red’’ morphotype: Medium–large euscorpiid with after chloroform treatment in order to remove the carapace length of 5.4–6.1 mm. Orange–brown body cerotegument. Finally, they were air-dried, placed on with yellowish chelicerae, legs andtelson. All analyzed aluminum stubs andcoatedwith 20 nm Au–Pdusing a specimens are similar in color. The elongatedchelas give Balzers MED 010 sputter-coater before observation this morphotype a slender appearance (Fig. 1b). under a Philips XL 20 SEM. Tubercles on leg femur are crenulatedon both dorsal Key to museum and collection acronyms: VVZC andventral surfaces ( Figs. 2b andd ). collection – V. Vignoli, Dipartimento di Biologia ‘‘Black’’ morphotype: Medium-sized euscorpiid with Evolutiva, University of Siena, Italy; VF collection – short pedipalp segments (Fig. 1a). Considerable poly- private collection of V. Fet; MZUF – Museo Zoologico morphism is evident in both the color and the size of the ‘‘La Specola’’, Sezione del Museo di Storia Naturale, sexually mature specimens analyzed. The carapace University of Florence, Italy. length ranges from 3.9 to 5.8 mm. Dark brown–blackish Abbreviations: DPS – dorsal patellar spur; Pv – patella body with clear maculation pattern. Legs, telson and ventral; Pe – patella external; m – male; f – female; Dp – chelicerae are yellowish andshow the same pattern. The pectinal teeth; L – left; R – right; et – external terminal; extremities of the chela fingers are reddish or pale est – external subterminal; em – external median; esb – brown. Granulation on leg femurs is present but weak external suprabasal; eb – external basal-a; eb – external a (Figs. 2a andc ). basal.

3.1. Comparisons between ‘‘red’’ and ‘‘black’’ 2.3. Biometrics morphotypes

Two factors were analyzedin a crosseddesign:genetic 3.1.1. Trichobothria pattern andnumber group (2 fixedlevels: 1 ¼ black type and2 ¼ redtype) Type C, major additive neobothriotaxic on patella. andsex (2 fixedlevels: males andfemales). A multi- All specimens have trichobothria statistical ranges variate matrix was constructedwith the 20 selected typical of the Euscorpius subgenera (ventral chela (V): individuals as objects and 27 biometrical measurements 4/4; ventral patella (Pv)46). The trichobothria of the as variables, following Fet andSoleglad(2002) , with the patella ventral surfaces range from 8 to 9, with higher addition of the DPS length. After normalization, the values in the ‘‘red’’ morphotype (9/9) than in the data set was analyzed using principal component ‘‘black’’ one (see Table 1). The patella external region analysis (PCA) ordination to detect the main morpho- shows the typical number of the recently re-described logical group (ChatfieldandCollins 1980 ). A variable E. tergestinus. Patella external formula: eb ¼ 4=4, discriminant analysis was performed using the eigen- eba ¼ 4=4, esb ¼ 2=2, em ¼ 4=4, est ¼ 4=4, et ¼ 527 vector scores that indicate the weight of each variable on (6). Even the pattern of patellar external trichobothria is the first 2 principal components (PCs). A two-way identical (see Fig. 60 in Fet andSoleglad2002 ). crossedanalysis of variance (2-way ANOVA) was used Pedipalps were all carefully examined, and the em series ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 101

Fig. 1. Euscorpius tergestinus (C.L. Koch, 1837): ‘‘black’’ morphotype (VVZC Eut516: a, c, e) and‘‘red’’morphotype (VVZC Eut243: b, d, f). (a, b) Dorsal view of adult female; (c, d) dorsal aspect of right patella; (e, f) dorsal aspect of pedipalp chela.

seemedto differ between the two morphotypes. While and in some cases between different pedipalps of the the disposition of the four trichobothria in ‘‘red’’ same scorpion, only the distance between trichobothria specimens is more compact, that of ‘‘black’’ specimens 3 and4 differssomewhat; however, the general is more ‘‘open’’. The angle formedbetween trichobo- pattern in the studied material is always as described thria 1–2 and3–4 is larger in the ‘‘black’’ than in the above. Other differences in trichobothria patterns are ‘‘red’’ specimens. Although the single position of each the position of d and i on the pedipalp femur. In the trichobothria couldchange from specimen to specimen, ‘‘red’’ specimens d is more distant from i, while the ARTICLE IN PRESS 102 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113

Fig. 2. Euscorpius tergestinus (C.L. Koch, 1837): ‘‘black’’ morphotype (VVZC Eut304: a, c; Eut102: e, g) and‘‘red’’morphotype (VVZC Eut96: b, d, h; Eut243: f). (a, b) lateral view of leg femur ( Â 30); (c, d) detail of the dorsal aspect of leg femur ( Â 90); (e, f) microstructure of peg sensilla on tooth ( Â 3200); g, h. detail of peg sensilla ( Â 6400).

position of e on the dorsoexternal carina is similar in 3.1.2. Morphometric ratios both. Even the position of trichobothria V4 is different: Our morphometric dataset indicated that some ratios are it is situatedin a dimple on the ventral external carina representative while others are not, due to the high (V1) in ‘‘black’’ specimens, whereas it is on the external polymorphism of ‘‘black’’ specimens. The ratio obtained surface of the other morphotype. between the total length andwidthof chela (chela length/ ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 103 chela width) statistically highlights the slender appearance Table 2. Eigenvalues, percentage (%) variation andcumula- of the ‘‘red’’ type. This is evident only when females and tive percentage (cum. %) variation for the first 4 principal males are considered separately. The exceptional develop- components (PCs) of the multivariate data set ment of the DPS spine in ‘‘red’’ specimens is clear (Fig. 1d), PC Eigenvalues % Variation Cum. % variation without having to exclude either sex, in the plot that takes into account the ratio DPS/patella length (Fig. 4d). 1 19.65 72.8 72.8 2 1.83 6.8 79.6 3.1.3. Pectinal tooth count andshape 3 1.67 6.2 85.7 As shown in Table 1, this character does not help to 4 1.10 4.1 89.8 distinguish our morphotypes. Pectinal tooth counts vary between specimens, and sexual dimorphism is evident. Values range from 5/5 to 7/7 in females, while they are (85) (Figs. 2a–d). The ventral carination of the same more constant, predominantly 8/8, in males. The shape segment is also stronger in the ‘‘red’’ specimens. of pectines sensilla of the ‘‘black’’ specimens (VVZC Eut102-260) is similar to that of the ‘‘red’’ morphotypes (VVZC Eut96-243) and is identical to the drawing of 4. Statistical results ‘‘E. carpathicus’’ in Bonacina (1980) (Figs. 2e–h). SEM In the PCA analysis the first 2 components account images of the pectines sensilla did not reveal morpho- for 79.6% of cumulative variance (Table 2). The first logical differences, confirming that these characters are component (PC1) distinguishes morphotype 1 (‘‘black’’) only useful at higher taxonomically levels (Valle 1975). from morphotype 2 (‘‘red’’) with some overlap (Fig. 3). Eigenvector coefficients reveal that, comparedto in- 3.1.4. Carinae development: metasoma, pedipalps and dividuals with high positive scores, individuals with high legs negative values along PC1 all have higher biometric Results show that metasoma carination is more values; PC1 therefore generally separates the 2 genetic developed in the ‘‘red’’ morphotypes (carinae develop- groups according to general dimensions, although some ment quotient: 375) than in the ‘‘black’’ morphotypes individuals lie in an intermediate position. PC2 reveals (carinae development quotient: 223). Moreover, while some contrasts between several variables: pedipalp the lateral keel of the first segment is never present in the ‘‘black’’ specimens, it is always present, though some- patella, femur andchela length, pedipalp length (femur+patella+chela), DPS length andcarapace times faint andsmooth, in the other morphotype. length increase with positive scores along PC2. Other Another finding of metasoma observations was the lack variables, on the other hand, in particular femur and of the inferior median carina in the first 3 segments palm width, palm depth, and the width of all metasomal and, in some cases, also in the fourth segment. In the segments, increase with negative scores. Because the 2 ‘‘red’’ type this carina is always present in the fourth morphotypes are also separatedalong PC2, this second segment andsometimes also in the thirdmetasomal axis should be considered an index of form in which the segment. Observation of pedipalp carinae development revealedthat carination is more markedand‘‘orga- features of morphotype 2 are more elongatedthan those of morphotype 1. The 2-way factorial ANOVA on each nized’’ in ‘‘red’’ morphotypes than in ‘‘black’’ morpho- variable of the same data set confirmed multivariate types (Figs. 1dandf ), andis similar in all analyzed results. Because the major source of variation in the ‘‘red’’ specimens. The most apparent differences in PCA was explainedby general dimensions, in all cases pedipalp carination between the 2 morphotypes are: the average value of a variable was significantly greater on the femur, dorsal carinae are granulate in the ‘‘black’’ in morphotype 2 (‘‘red’’) than in morphotype 1 types but crenulate in the ‘‘red’’ types; granules on the (‘‘black’’). Therefore, as an example, only the tests ventral surface of the latter are also larger andmore numerous. The patella of the 2 morphotypes are not for 3 of the analyzedvariables are reportedin Table 3. ANOVA on the indices of form also yielded significant very different (Figs. 1c andd ). Most of the chelal carinae results (Table 3), confirming several differences in the are similar; however, carinae D3 andD4 are rough and morphology of the two genetic groups (Fig. 4). weakly granulate in the ‘‘black’’ types andsmooth, like in the dorsal intercarinal spaces, in the ‘‘red’’ types (Figs. 1e andf ). The ‘‘black’’ specimens are more polymorphic; some carinae are less developed, and 5. Discussion granules are often unequal in size andirregularly distributed. In addition, we considered the dorsal 5.1. Taxonomy carination of leg femurs. Each leg was analyzed, and results show that carination is much stronger in the From a taxonomic standpoint, the 2 studied pheno- ‘‘red’’ specimens (171) than in the ‘‘black’’ specimens types belong to different described subspecies. The most ARTICLE IN PRESS 104 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113

3.0 2f 2.5 2.0 2m 2f 1.5 2m 1.0 2f 2m 0.5 2f 2f 2m 1f 1m 0 1f PC2 -0.5 2m 1f 1m -1.0 1f 1m 1m -1.5

-2.0 1f -2.5 1m -3.0 -6 -4 -2 0 2 4 6 8 10 PC1 Fig. 3. Principal component analysis plot. Codes: 1 ¼ morphotype 1 (‘‘black’’); 2 ¼ morphotype 2 (‘‘red’’); f ¼ female; m ¼ male.

Table 3. Two-ways ANOVA table

Mean7SD Morphotype Sex

Variable/index 1 2 FpFp

Carapace length 4.970.7 5.770.2 20.2 *** 3.55 n.s. Chela length 8.271.4 10.370.5 23.9 *** 2.81 n.s. DPS 0.3370.07 0.7270.09 119.5 *** 0.32 n.s. Chela length/chela width 2.570.2 2.970.1 42.63 *** 2.20 n.s. Femur length/femur width 2.470.5 2.970.3 9.09 ** 0.26 n.s. Metasoma V length/metasoma V width 2.870.5 3.270.1 7.16 * 0.02 n.s. DPS/patella length 0.0870.01 0.1570.02 162.7 *** 2.62 n.s.

The value of the F-statistic is reportedtogether with the p-level of signiﬁcance for factors Morphotype andSex. For the factor Morphotype, mean values (7SD) are reported for each variable pooling males and females data. Data are expressed in mm. Codes are: n.s. ¼ not signiﬁcant; Ãpo0:05; ÃÃpo0:01; ÃÃÃpo0:001; 1 ¼ morphotype (‘‘black’’); 2 ¼ morphotype (‘‘red’’)

recent taxonomical revision of the Euscorpius carpathi- sary to determine the taxonomical position of our cus complex (Fet andSoleglad2002 ) considers 9 ‘‘black’’ types. The study of Caporiacco’s material different old taxonomical entities as synonyms of provided a better understanding of his described E. tergestinus (C.L. Koch, 1837). Three of these were subspecies andallowedthe comparison of ‘‘black’’ described by C.L. Koch: Scorpius aquilejensis (Koch specimens with Caporiacco’s specimens. Our black 1837), S. concinnus (Koch 1837)andS. niciensis (1841). phenotypes clearly correspondto E. c. concinnus re- Four subspecies were described by Caporiacco (1950): described in the manuscript on Italian euscorpiids by E. carpathicus apuanus, E. c. corsicanus, E. c. picenus and Caporiacco (1950). The author considered this scorpion E. c. oglasae. The 2 other taxa are: S. tergestinus var. as the most widely distributed taxon, with diagnostic austriacus (Ferrari, 1872) and E. c. mesotrichus Hadzˇ i, characters such as medium size, dark color and an 1929; since the latter is a homonym of E. italicus oligotrichous trichobothrial pattern. Other morphologi- mesotrichus Hadzˇ i, 1929, this taxon cannot be used. The cal aspects are also identical: the pectinal tooth count, reason for this attribution of synonymy was the 7/7 in females and8/8 in males, Pe ¼ 24 andPv ¼ 8. insufﬁciency of diagnostic morphological characters While comparing 4 of Caporacco’s specimens from andthe presence of one character, external pedipalp central Italy (Emilia Romagna, Umbria), it was possible patella series eb ¼ 4=4, in all the above-mentioned to determine other ratios and characteristics that were taxa. Our ‘‘red’’ types undoubtedly coincides with the not taken into account by Caporiacco. The DPS is short E. tergestinus neotype accurately re-described by Fet andthe pedipalp chela is stout, while chela carinae D3 andSoleglad(2002) . Comparative analysis was neces- andD4 are rough andthe leg femur granulation is weak. ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 105

Max 75% Max 75% Min 25% Min 25% Mediana Mediana

Chela length / Chela width Metasoma V length / Metasoma V width 3.2 4.0

3.6 3.0

3.2 2.8 2.8

2.6 2.4

2.4 2.0

2.2 1.6 12 12

Max 75% Max 75% Min 25% Min 25% Mediana Mediana

Femur length / Femur width DPS / Patella length 4.2 0.20

0.18 3.6

0.16 3.0 0.14 2.4 0.12 1.8 0.10

1.2 0.08

0.6 0.06 12 12 Fig. 4. Average7SD for the 4 indices of form calculated for morphotypes 1 (‘‘black’’) and 2 (‘‘red’’) (including males and females).

The angle between trichobothria 1 and4 of the em series mens (VVZC Lucca, Eut106; Bologna, Eut95) are is slightly variable, particularly the position of number totally black, chelicerae and telson included, except for 4, but all 8 patterns are more ‘‘open’’ than the narrow the dark yellow tarsus and the slightly reddish apix of pattern of E. tergestinus. It wouldbe unwise to consider chelae fingers. The specimen from Elba Island(VVZC this difference diagnostic; intraspecific variability of the Eut46) is totally dark reddish brown, except for the position (andnumber) of trichobothria is not rare. This lighter-coloredlegs that show no maculation. Very kindof polymorphism was observedfor the same series similar is the specimen from Pavia (VVZC Eut89). The (em) in euscorpiids such as E. hadzii Caporiacco, 1950 unnamedspecimens from Elba Island(Tuscany Archi- by Fet andSoleglad(2002) and S. niciensis C.L. Koch, pelago) belonging to an eb ¼ 4=4 morphotype were 1837 by Lacroix (1991). The color may be compared notedby Valle (1975) andconsideredas E. tergestinus with that of the most recent of Caporiacco’s specimens (Fet et al. 2003). They show all characters of the ‘‘black’’ from Umbria, but not with that of specimens from phenotype. Scorpius aquilejensis (C.L. Koch, 1837), Emilia Romagna, which are too oldandhave lost their which was described on the basis of only one specimen pigmentation. As explainedabove, this qualitative andrepresentedin Koch’s famous paper, ‘‘Die Arachni- character is variable but similar between specimens den’’ (1837), as a ‘‘yellow’’ Euscorpius with crenulate collectedin central andsouthern Tuscany. Two speci- metasomal keels andventral patellar series 8/8. Our ARTICLE IN PRESS 106 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 dataset contains 1 unusual ‘‘yellow’’ specimen (Eut278) late; moreover, the granulation of both ventral and collectedin Rome; given that molecular andmorpho- dorsal metasomal carinae is sparse, as in our analyzed logical aspects of this specimen are the same as those of E. tergestinus specimens. As we do not have this our ‘‘red’’ type ( ¼ E. tergestinus), 1 hypothesis is that phenotype in our molecular dataset we cannot come to S. aquilejensis was defined on the basis of 1 anomalous any taxonomical conclusions; we therefore follow the specimen which was light in color andlackedsome taxonomy proposedby Fet andSoleglad(2002) , which trichobothria on the patellar ventral series. Caporiacco considers E. c. apuanus synonymous with E. tergestinus. (1950) analyzedspecimens from several localities in the During this study we analyzed some specimens from vicinity of Trieste (locus typicus), andgave the ventral Montecristo Island, a small island situated in the central patella series (9/9) andthe external patella (24/24) as Tyrrhenian Sea; E. c. oglasae was defined as an diagnostic trichobothrial characters. These values, oligotrichous euscorpiidsimilar to the subspecies ( E. c. together with the color description and geographic corsicanus) described by the same author in Corsica, correspondence (we also have 1 specimen from Sistiana, France, but lighter in color (Caporiacco 1950). As we Trieste), indicate that our ‘‘red’’ morphotype corre- only have juveniles, andespecially because we lack sponds even to S. aquilejensis (E. c. aquilejensis sensu molecular data, we do not discuss their taxonomy but Caporiacco 1950). We also analyzed an additional follow Fet andSoleglad(2002) . Finally, during the specimen from Trieste (Nabresina) deposited in MZUF. investigation of the material preservedin MZUF, a This specimen shows a polytrichous trichobothrial unique specimen from Monte Pisano, Pisa (Tuscany), pattern (Pe: eb ¼ 4=4, eba ¼ 4=4, esb ¼ 2=2, em ¼ 4=4, which Caporiacco (1950, p. 186) considered similar to est ¼ 4=4, et ¼ 8 À 8; Pv ¼ 10/11) andprovidesevi- E. c. canestrinii (Fanzago 1872), was foundand dence of the high polymorphism of the complex; besides, analyzed. The damaged specimen presents typical the geographic position of Trieste is a crucial point for characteristics of a ‘‘pale-colored’’ E. tergestinus,such dispersal between the Balkans and the Italian Peninsula. as the pedipalp patella trichobothria formula (eb ¼ 4=4, No type specimens of this taxon are available for study eba ¼ 4=4, esb ¼ 2=2, em ¼ 4=4, est ¼ 4=4, et ¼ 6=6; (Fet andSoleglad2002 ); although it remains an Pv ¼ 9/9), rather than those of the ‘‘E. sicanus’’ complex enigmatic entity, we agree with the authors that it to which this Sardinian endemic morphotype is ascribed shouldbe consideredsynonymous with E. tergestinus. (Vachon 1978; Fet et al. 2003). In keeping with the Euscorpius c. picenus was described by Caporiacco results of this morphological andstatistical analysis, (1950) as a Euscorpius subspecies extremely similar to molecular results (Salomone et al. in prep.), indicate E. c. concinnus, but with a higher trichobothrial number that the ‘‘red’’ and ‘‘black’’ morphotypes belong to the (Pv ¼ 9/9) anddarker color. We analyzed3 specimens E. tergestinus complex andmay be distinguishedas from Abruzzo (Caramanico negli Abruzzi) of Capor- separate forms. Although multivariate analysis revealed iacco’s material. Two show the same morphology as the high polymorphism of the ‘‘black’’ morphotype, the E. tergestinus, while 1 has the features of our ‘‘black’’ 2 morphotypes are certainly distinct and belong to 2 type (pedipalp chela and DPS short, Pv ¼ 8/8). As different morphological groups (Fig. 3). The 2-way previously notedby Fet andSoleglad(2002) , the factorial ANOVA analysis confirmedmultivariate specimens extremely similar to E. tergestinus have a results and provides further indication of morphological different metasomal carination, which is less granulate. differentiation between the two studied types. The To avoid confusion we decided to consider E. c. picenus plots that provide the best evidence of differentiation synonymous with E. tergestinus,asinFet andSoleglad (Figs. 4a–c) mainly highlight the more slender structure (2002). The last subspecies to be examinedand (chela length/chela width; femur length/femur width; comparedwas E. c. apuanus Caporiacco, 1950. This metasoma V length/metasoma V width) of the ‘‘red’’ was considered a taxonomical entity principally on the type with respect to that of the ‘‘black’’ type. The DPS/ basis of its yellow color; the author also gave other patella length ratio only highlights the shape of a morphological characteristics, but they are insufficient specific structure, i.e. the DPS spine (Fig. 4d). The large to distinguish it from other taxa, which are very DPS spine, the markedleg granulation andthe similar. To distinguish this subspecies from the other elongatedchelae, especially in females, are the most ‘‘yellow’’ euscorpiids, Caporiacco highlighted the fainter important features distinguishing the ‘‘red’’ morphotype granulation of the leg femurs. Two females from from the ‘‘black’’ morphotype. Although the color Levigliani, in the Apuan Alps, were studied. Their pattern is considered in this paper, this parameter is general aspect is similar to that of E. tergestinus, highly qualitative andcannot be consideredalone. with elongatedpedipalpchelae, granulation of the We can split the Italian ‘‘E. tergestinus’’ complex into pedipalp patella and femora, long total length and 2 validspecies: E. tergestinus s.s. (C.L. Koch, 1837) and similar pigmentation; nevertheless, they also show some the ‘‘black’’ type which corresponds to E. c. concinnus intermediate characters. The dorsal surface of the sensu Caporiacco (1950). Basedon our comparative pedipalp chela and D1–D4 carinae are weakly granu- morphological analysis, which is sustainedby molecular ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 107 results (Salomone et al. 2004 in prep.), we elevate this locality is not known, we maintain this name as valid taxon to the species level. andfix neotype for E. concinnus here. Euscorpius concinnus (C.L. Koch, 1837) Euscorpius tergestinus: Fet andSissom 2000: 372 ; Fet (Figs. 1a, c ande ; Table 4) andSoleglad2002: 16–24 . Scorpius concinnus C.L. Koch, 1837: 105-103, pl. CVI, Type material: Holotype: 1 female (lost), type locality Fig. 246. unknown. Neotype: Adult female (VVZC Eut516) References (selected): collectedin the locality of Ponte a Bozzone Scorpius concinnus: C.L. Koch, 1850: 86; Pavesi, 1876: (N 43121001.200,E11123010.100), Castelnuovo Berardenga 430. (SI), Tuscany, central Italy; under tree bark, pine wood, Euscorpius carpathicus concinnus: Caporiacco 1950: 273 m a.s.l., 13 October 2003 (V. Vignoli & F. Cicco- 190–194. Note: Caporiacco (1950 p. 194) usedthis nardi coll.). Type specimen will be deposited in MZUF. subspecies name since it did fit Koch’s description Diagnosis: Medium-sized euscorpiid with general illustration. Since Koch’s holotype is lost andthe type squat aspect (Fig. 1a; Table 4). Blackish–dark brown body, with reddish chela and pale brown legs and chelicerae. The pedipalp patellar external trichobothria Table 4. Meristic data for the neotype (female), and an adult number andpattern are typical of the ‘‘ tergestinus’’ male (VVZC Eut304) of Euscorpius concinnus complex (Fet andSoleglad2002 ). The dorsal patellar spur (DPS) is quite developed, as is granulation on leg Neotype Male femurs (Figs. 1c, 2a andc ). Pectinal tooth: males 8–8; females 7–7. Total Length 28.7 29.9 Description of neotype: Color. Carapace, mesosoma, Carapace Length 4.2 4.5 metasoma andtelson vesicle all brown ( Fig. 1a). Posterior width 4.2 4.6 Pedipalp femur is the same color as the body, while Metasoma Length 9.3 11.4 the patella andchelae are more reddish.The extremities of chela fingers are paler. The chelicerae are pale brown, Segment I Length 1.2 1.4 with maculate pigmentation, andare similar to the Width 1.5 1.8 telson aculeus. Carapace. Characterizedby a slight, Segment II Length 1.4 1.7 uniform roughness. Mesosoma. All tergites are slightly Width 1.4 1.6 rough, like the carapace, anddonot show any carinae. Segment III Length 1.6 2.0 Ventral side is yellow–brown with smooth sternites. Width 1.4 1.5 Stigmata are small andhardlyvisible. Metasoma. Generally squat structure. Intercarinal spaces of seg- Segment IV Length 2.0 2.4 ments I–IV are smooth; only ventral intercarinal space Width 1.3 1.3 of segment V presents slightly enlargedgranules. Segment V Length 3.1 3.9 Segment I: dorsal and dorsal lateral carinae partially Width 1.2 1.3 granulate; lateral, inferior lateral andinferior median Telson Length 3.4 4.5 obsolete. Segment II: similar to segment I. Segment III: dorsal carina weakly granulate, inferior lateral and Vesicle Length 2.0 3.2 inferior median smooth but evident; other carinae are Width 1.2 1.8 absent. Segment IV: similar to segment III. Segment V: Depth 1.2 1.9 dorsal lateral weakly granulate; lateral carina obsolete; Aculeus Length 1.4 1.1 inferior lateral andmedian, weakly granulate. Telson. Pedipalp Length 13.9 15.1 Squat shape andsmooth surface. Vesicle with 8 long Femur Length 3.3 3.6 ventral setae, and2 long setae in the region between Width 1.3 1.4 vesicle andaculeus. Aculeus with strong curvature. Pectines. Pectinal tooth count: 7/7. Middle lamellae: 5/5. Patella Length 3.5 3.7 The peg sensilla shows the typical shape of the Width 1.5 1.6 ‘‘carpathicus complex’’ drawn by Bonacina (1980) for Chela Length 7.1 7.7 E. carpathicus. Length: 1.9; width: 1.0. Genital oper- Palm Length 3.4 3.7 culum. Separatedmost of length. Length: 0.6; width:1.7. Width 2.7 3.4 Sternum. Pentagonal shape. Length: 1.4; width: 1.5. Depth 1.8 2.3 Chelicerae. Movable finger: ventral edge typically smooth with brush-like setae on the entire inner part; Movable finger Length 4.1 4.6 dorsal edge: two subdistal denticles. Fixed finger: Pectinal teeth 7/7 8/9 standard configuration. Pedipalps. The general structure Measurements are in millimeters. is stocky. Femur. Dorsal carinae granulate, ventral ARTICLE IN PRESS 108 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 crenulate andintercarinal spaces partially granulate. Trichobothria d and i are situatedat the same distance on the femur. Patella. Dorsal carinae are granulate (Fig. 1c) while the ventral carinae are weakly granulate, like the intercarinal spaces. DPS short: 0.28 mm. Ventral patellar spine reduced to a little tubercle. Trichobothria pattern, type C neobothriotaxic (major additive) (Vachon, 1974); Patella external (Pe) formula: et ¼ 7=7, est ¼ 4=4, em ¼ 4=4, esb ¼ 2=1, eba ¼ 4=4, eb ¼ 4=4; patella ventral (Pv): 8/9. Chela. Carinae granulation: dorsal carinae D1 andD4 are weakly granulate; D3 andD5 are rough. Dorsal intercarinal surface rough. Ventral carina V1 andV2 are weakly granulate; V2 smooth. External (E) carina weakly granulate. Dorsal surface between carina D3 andD4 is rough ( Fig. 1e). Trichobothria V4 is situatedon the ventral carina V1. Legs. Two pedal spurs without spines. Tarsus: median ventral row with 8–9 stout spinules (leg III); tarsal setae (adjacent to the median ventral spinule row), flanking pairs: 3; terminal pairs: 2. Basitarsus: 2–3 small proventral spinules on legs I–II; 1 spinule on leg III, absent on leg IV. Male. Specimen similar to the neotype in color and body granulation. Total body length: 29.9 mm; carapace length: 4.5 mm. Telson vesicle swollen andaculeus more strongly curvedthan female one. Pectinal tooth count: 8/9. DPS: 0.32 mm. Fig. 5. Map indicating sampling sites of investigated specimens Materials examined: Besides the material from VVZC of Euscorpius tergestinus (D)andEuscorpius concinnus (*). andthe specimens from the Societa` Romana di Scienze Naturali, the first author analyzedsome of Caporiacco’s specimens preservedin MZUF labeledas E. c. concinnus with yellowish legs andvesicle, andlarger than the other (C.L. K.) andas E. carpathicus (see Appendix A). specimens; the carapace length of these large specimens Variations: The diagnostic characters are reasonably is: 4.9 (VVZC Eut512), 5.0 (VVZC Eut515), 5.3 (VVZC constant in all examinedpopulations. Although we Eut513). notedthe variability of the carinae on metasomal Distribution andecology : Euscorpius concinnus has a segments, especially of the inferior median carinae, this wide geographic distribution (Fig. 5); E. c. concinnus characteristic was not considered in the morphological was considered 1 of the most common and widely discussion because it has already been widely documen- distributed ‘‘carpathicus’’ subspecies in Italy (Caporiac- ted( SolegladandSissom 2001 ; Fet andSoleglad2002 ). co, 1950). We analyzedspecimens belonging to different The metasomal ventral granulation of three specimens populations collectedin an area of more than 650 km from three close localities (Barberino del Mugello, (Bergamo-Salerno) throughout the Italian Peninsula, Bologna, Lucca) is different despite the molecular which comprises 9 regions (Lombardy, Liguria, Friuli V. similarity (Salomone et al. in prep.). The granulation Giulia, Emilia Romagna, Tuscany, Marche, Umbria, of the dorsal surface between carinae D3 and D4 of the Latium and Campania). Their distribution extends from pedipalp chela is another variable character. The dorsal the Tyrrhenian coast to the Adriatic coast, and includes surface of chela in two specimens, one from Emilia insular populations such as those of Palmaiola andElba Romagna (VVZC Bologna, Eut95) andthe other from Island(Tuscany Archipelago). This species seems to be Latium (VVZC Cori, Eut105), is smoother than usual. tolerant (eurytopic) of a wide range of habitats and Body color and total length vary among different altitudes, from sea level to 1500 m, where they have populations. The southern populations, from Monte principally corticolous andlapidicolous habitus. Never- Semprevisa andNational Park of Circeo (Latium), are theless, a preference for natural rather than anthropo- entirely black (vesicle, legs, tarsus andbasitarsus genic habitats was observed. included). Two males from eastern Tuscany (Castel- l’Azzara (GR) andArezzo) are similar; the specimen 5.2. Ecology from Arezzo (VVZC Eut142), with a carapace length of 6.2 mm, is the largest studied specimen (total length: Cryptic taxa can not only show small different mor- 45 mm). Specimens from the Marche region are brown, phological features but also ecological andbehavioral ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 109 differences (Colborn et al. 2001; Muster et al. 2004; and E. sicanus (C.L. Koch, 1837) (larger) and Wellborn andCothran 2004 ); these couldbe the result E. concinnus (smaller) in central Tuscany. The unusual of an adaptation, given that the phenotypic similarity recovery of E. tergestinus under bark in a pine wood of of similar species also implies limitedscope to occupy Sistiana, near Trieste, insteadof in a rural construction different habitats (Wellborn andCothran 2004 ). couldbe dueto the presence of E. italicus in the same We observedecological differences between the 2 similar region; the latter, larger species is known to colonize species, which are sympatric but not syntopic; anthropogenic habitats (Braunwalder 2001; Crucitti E. concinnus is more common in natural habitats, such et al. 1998a, b). as woods, whereas E. tergestinus was prevalently found We do not exclude behavioral differences between in anthropogenic habitats. We suppose that the dis- these cryptic taxa, andmating tests will help determine tribution of these 2 euscorpiids in different microhabi- whether there are stable reproductive barriers and, if so, tats (not really different habitats as Wellborn and whether they are pre- or post-zygotic. Cothran (2004) confirm) couldbe the result of strong interspecific interaction (intraguildpredation interaction) insteadof ‘‘anthropotolerance’’ (i.e. major or minor tolerance to anthropogenic habitats (Crucitti et al. 6. Conclusions 1998a, b)) or ‘‘anthropophily’’ (Vachon 1983; Lacroix 1990). Intraguildpredation interaction is thought to The most important outcome of our study, based on greatly affect scorpion distribution patterns (Polis the observation of 167 specimens, is that in Italy there andMcCormick 1987 ), andthis competition could are at least 2 different valid species belonging to the promote their coexistence. Size differences between ‘‘E. tergestinus’’ complex. Euscorpius tergestinus is a sympatric scorpion species seem to be important in slender, reddish euscorpiid with a big dorsal patellar determining this interaction, and larger species occupy spine. Euscorpius concinnus is generally darker and squat the most favorable microhabitats (Polis andMcCormick (see Table 5 for a summary of characters usedto 1987). Wall crevices protect specimens from extremes in distinguish the 2 taxa). Moreover, the latter species is temperature (Benton 1992) andpredators, andensure characterizedby a heavy metasoma, relatively short relatively constant humidity during the dry Mediterra- pedipalp segments, a short dorsal patellar spine (DPS), nean summer; these are all reasons why man-made andweak granulation on leg femurs. The 2 taxa are environments couldbe consideredfavorable habitats. sympatric in several Italian regions (Lombardy, Friuli The susceptibility to climatic conditions, such as V. Giulia, Latium, Tuscany, Umbria). Intraguildpre- precipitation, which are correlatedto relative humidity dation interaction could be the principal cause of the and appear to determine the distribution of euscorpiids specific distribution in different microhabitats, establish- (Fet et al. 2001), sustains this hypothesis. Euscorpius ing an equilibrium that makes coexistence of these concinnus and E. tergestinus were never foundtogether; cryptic species possible. The distribution of E. tergesti- the larger species (E. tergestinus) was present in nus, previously limitedto northern Italy ( Fet and anthropogenic habitats, while the smaller (E. concinnus) Soleglad2002 ), has been extended to include Siena occupiednatural habitats. The same distribution pattern andRome, both in central Italy. has been observedin Italy for other pairs of sympatric Further studies will be useful to complete knowledge euscorpiids, such as E. italicus (Herbst 1800) (larger) of this polymorphic scorpion group in Italy. In and E. concinnus (smaller) in northeast Tuscany particular, relationships among the endemic subspecies (V. Vignoli andJ. Ove Rein pers. obs.) andLatium, of Montecristo Island, E. c. oglasae Caporiacco, 1950,

Table 5. Diagnostic characters distinguishing Euscorpius tergestinus from Euscorpius concinnus

Character Euscorpius tergestinus Euscorpius concinnus

Pedipalp patella ventral (Pv) trichobothria number 9/9 8/8 Dorsal patellar spine length Long Short Pedipalp chela length Long Short Chela carinae D3 andD4 Smooth Rough Color Orange, reddish Black, dark brown Legs femur dorsal granulation Crenulate Granulate Angle between trichobothria 1 and4 of em series (Pe) Narrow Large Metasoma and pedipalp granulation Emphasized, organized, crenulated Irregular, weakly granulate to granulate ARTICLE IN PRESS 110 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 andparticular morphotypes (specimens with Pe ¼ eba: Eut101), Barberino del Mugello (FI), under stone, beach 5/5, eb ¼ 4=4 andspecimens from the Apuan Alps, E. c. wood, 29 June 2002 (V. Vignoli coll.); 1 adult female apuanus Caporiacco, 1950), as well as their geographic (VVZC Eut102), Montaione (FI), under stone, pine distribution must be further investigated. We cannot wood, 27 June 2002 (J. Ove Rein coll.); 1 adult female exclude the presence of other cryptic species in Italy, (VVZC Eut106), Loc. Tereglio, Lucca (LU), under especially in the mountainous regions or on the islands, stone, beach wood, 02 May 2002 (A. Petrioli coll.); 1 where gene flow is absent or reduced due to natural adult male (VVZC Eut260), Poggibonsi (SI), under tree barriers. Last but not least, without molecular evidence bark, holly oak wood, 19 December 2002 (V. Vignoli & andmorphological congruence, it wouldnot have been N. Salomone coll.); 1 adult male (VVZC Eut304), Loc. possible to complete this study according to modern Ponte a Bozzone, Castelnuovo Berardenga, Siena (SI), scientific standards. under tree bark, pine wood, 05 May 2003 (V. Vignoli coll.); 1 adult male (VVZC Eut310), Brenna (SI), under tree trunk, deciduous wood, 03 May 2003 (F. Cicco- Acknowledgements nardi coll.); 1 adult female (VVZC Eut46), Marciana, Elba Island(LI), underchestnut tree bark, 620 m, 11 March 2001 (V. Vignoli coll.); 1 adult female (VVZC We are grateful to Gaia Pigino, Massimo Migliorini, Eut256), Castellina in Chianti (SI), under pine tree bark, Francesco Cicconardi, Jan Ove Rein, Andrea Petrioli, 08 December 2002 (V. Vignoli coll.); 1 adult female Marco Bastianini, Diego Facheris, Marco Colombo, (VVZC Eut265), Badia al Cerreto (SI), under bark, holly andMaria Vittoria Di Giovanni for collecting zoological material usedin this study.We also thank Pietro oak wood, 19 December 2002 (V. Vignoli & N. Salomone coll.); 1 adult female (VVZC Eut36), Castello Lupetti andEugenio Paccagnini for assistance with di Brolio, Gaiole in Chianti (SI), 01 October 2000 microscope images. Particular thanks go to Sarah (G. Manganelli coll.); 1 subadult female (VVZC Whitman for loaning specimens preservedin the Museo Eut245), Loc. Pecchiaiola, Volterra (PI), under tree Zoologico ‘‘La Specola’’ of Florence, andto Pierangelo bark, cypress wood, 10 November 2002 (V. Vignoli Crucitti for the generous loan of his entire euscorpiid coll.); 1 adult female (VVZC Eut311), Loc. Montalbuc- collection, preponderant for the results of this cio, Siena (SI), under chestnut tree bark, 07 May 2003 manuscript. Useful comments andsuggestions of Victor Fet (Marshall University) greatly improvedthe final (V. Vignoli coll.); 1 adult female (VVZC Eut266), Gambassi Terme (FI), under tree trunk, deciduous version of the manuscript. We also thank two anon- wood, 19 December 2002 (V. Vignoli & N. Salomone ymous reviewers for comments that improvedthe coll.); 1 adult female (VVZC Eut43), Marina di Campo, manuscript. Elba Island (LI), under stone, pine wood, 10 March 2001 (V. Vignoli coll.); Emilia Romagna: 1 adult female (VVZC Eut95), Loc. S. Luca, Bologna (BO), under tree Appendix A bark, holly oak wood, 16 March 2002 (V. Vignoli coll.); Liguria: 1 adult female (VVZC Eut88), Loc. Le Grazie, All specimens were collectedin Italy and,where not Porto Venere (SP), 12 June 2001 (D. Facheris, coll.), in otherwise indicated, the samples are preserved in the VF collection. Latium: 1 adult female (VVZC private collection of the first author in the Dipartimento Eut267), Campo di Montelanico, Monti Lepini (RM), di Biologia Evolutiva, University of Siena, Italy. Habitat under stone, beach wood, 900 m, 04 January 2003 indications are reported only when known. The follow- (A. Petrioli coll.); 1 adult female (VVZC Eut105), ing list of materials includes all the sequenced specimens Foresta di Cori, Cori (LT), 500 m, 28 May 2002 (Salomone et al. in prep.) with evidence of the typical (A. Petrioli coll.). ‘‘tergestinus’’ morphology. Euscorpius tergestinus (C.L. Several specimens (69) belonging to the zoological Koch, 1837): Tuscany: 1 adult female (VVZC Eut243), collection of the Societa` Romana di Scienze Naturali, Loc. Vico Alto, Siena (SI), house, 30 October 2002 (R. labeledas E. ‘‘carpathicus’’, andusedin a studyon the Dallai coll.); 1 adult female (VVZC Eut35), Siena (SI), Latium scorpiofauna (see Crucitti et al., 1998a for house, 09 October 2000 (G. Innocenti coll.), in VF detailed data), were morphologically reanalyzed. In collection. Latium: 1 subadult female (VVZC Eut278), particular, the study undertook the comparative bio- Rome (RM), house, 19 February 2003 (P. Crucitti coll.). metric analysis of the following 8 selectedadultspeci- Friuli V. Giulia: 1 adult female (VVZC Eut96), Sistiana mens. Euscorpius tergestinus (C.L. Koch, 1837): Latium: (TS), under tree bark, pine wood, 18 m, 11 March 2002 3 females (VVZC Eut219C-38C-86C) Antrodoco, Rieti (V. Vignoli coll.). Euscorpius concinnus (C.L. Koch, (RI); Via Leonina 7, Rome (RM); Via del Corso, Rome 1837): Lombardy: 1 female (VVZC Eut89), Brallo di (RM); 4 males (VVZC Eut/172C-112C-114C-246C), Via Pregola (PV), under stone, beach wood, 1500 m, 31 July dei Greci, Rome (RM); Via Margotta, Rome (RM); Via 2001 (D. Facheris coll.). Tuscany: 1 adult male (VVZC dei Panieri, Rome (RM); Via G. Giolitti, Rome (RM). ARTICLE IN PRESS V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 111

Abruzzo: 1 male (VVZC Eut62C), Carapelle Alvisio, Eut182), Palmaiola Island(LI), 12 July 1969 (F. Giusti (AQ). coll.); 1 male (VVZC Eut261), Poggibonsi, Siena (SI), Additional material: Euscorpius tergestinus (C.L. under tree bark, oak wood, 19 December 2002 (V. Koch, 1837): Tuscany: 2 adult females (syntypes, MZUF Vignoli & N. Salomone coll.); 1 male (VVZC Eut237), 125/5938-5945), Monte Corchia, Vallecchia, Pietrasan- Marciana, Elba Island (LI), under tree bark, oak wood, ta, Levigliani, Apuan Alps, Lucca (LU), 03 August 1875 11 February 2001 (V. Vignoli coll.); 3 subadults (VVZC (Del Prete & G. Cavanna coll.); 1 female (VVZC Eut239-240-241), 1 female (VVZC Eut238), Marina di Eut521), Chiusi (FI), house, 10 August 1996 (F. Vignoli Campo, Elba Island (LI), under stone, pine wood, 10 coll.); 2 subadults (VVZC Eut218-219), Montecristo February 2001 (V. Vignoli coll.); 1 female (VVZC Island(LI), 13 July 1968 (L. Lazzeroni coll.); 1 female Eut200), Marina di Campo, ibidem, 22 August 1967 (VVZC Eut520), Siena, April 1996 (F. Vignoli coll.); 1 (F. Giusti coll.); 1 male (VVZC Eut99), Loc. Cercina, adult male (MZUF 305/9490), Monte Pisani, Pisa (PI), ? Florence (FI), on stone wall 14 June 2002 (V. Vignoli (Canara coll.). Abruzzo: 3 adult females (syntypes, coll.); 1 male (VVZC Eut179), Arezzo (AR), June 1998 MZUF 163/5997-6002-6009), Falde of Mt. Morone, (? coll.); 1 adult male (VVZC Eut246), Loc. Pecchiaiola, Caramanico negli Abruzzi, Pescara, 27 July 1876–04 Volterra (PI), under tree bark of cypres wood, 10 August 1878 (G. Cavanna coll.). Lombardy: 1 female November 2002 (V. Vignoli coll.). Abruzzo: 1 subadult (VVZC Eut522), Presezzo (BG), 11 November 1998 (VVZC Eut525), Venere, Aquila (AQ), under stone, (D. Facheris coll.); 1 female (VVZC Eut144), Mapello, 750 m, 08 June 1998 (V. Vignoli coll.). Campania: Bergamo (BG), 28 April 2002 (D. Facheris coll.); 1 male 2 adult females (VVZC Eut223), 700 m, Corleto (VVZC Eut338), Ambivivere (BG), 06 May 2003 Monforte, Salerno (SA), 16 June 1998 (C. Pignataro (D. Facheris coll.). Umbria: 1 female (VVZC Eut519), coll.); 1 female (VVZC Eut526), ibidem, 07 June 1998 Loc. S. Marco, Perugia (PG), 15 November 2003 (E. Di (C. Pignataro coll.). Marches: 2 females (VVZC Giovanni coll.); 1 female (VVZC Eut518), Coreiano, Eut512-513), ibidem, 15 August 2003 (E. Di Giovanni Perugia (PG), 11 November 2003 (Goretti coll.). coll.); 1 male (VVZC Eut515), Loc. Ponte Nativo, Latium: 1 female (VVZC Eut281C), Via Donizzetti, Roccafluvione, Ascoli Piceno (AP), neglectedhouse, 31 Rome, 10 February 1998, (G. Amori coll.). Friuli V. July 2003 (E. Di Giovanni coll.). Latium: 2 males Giulia: 1 subadult male (MZUF 187/6275), Nabresina, (VVZC Eut250-251), 1 female (VVZC Eut249), Loc. Trieste (TS), August 1879 (Paulucci coll.). Euscorpius Cerasella, National Park of Circeo, Latina (LT), under concinnus (C.L. Koch, 1837): Tuscany: 1 female (MZUF tree bark, oak wood, 13 November 2002 (F. Cicconardi 135/5699), Resceto, Massa Carrara, 500 m, 10 June coll.); 1 subadult (VVZC Eut143C), ibidem, humus 1879, (Del Prete coll.); 1 female (VVZC Eut147), under Quercus pubescens, 05 May 2002, (F. Cicconardi Barberino del Mugello (FI), under stone, deciduous coll.); 4 males and1 female (VVZC Eut506-510), Loc. wood, 25 June 2002, (V. Vignoli & J. Ove Rein coll.); 1 San Felice, ibidem, under tree bark, oak wood, 08 female (VVZC Eut524), Padule di Fucecchio, Pistoia, November 2003, (F. Cicconardi coll.); 1 male (VVZC pine wood, 05 March 1994 (V. Vignoli coll.); 1 female Eut230), Poggio Bustone, Mt. Reatini (RI), wood, 08 (VVZC Eut523), Brenna (SI), deciduous wood, 03 June August 1969 (F. Giusti coll.); 1 male (VVZC Eut228), 1996 (V. Vignoli coll.); 1 male (VVZC Eut257), Vallonina Bassa, Terminillo, (RI), 19 August 1967 (F. Castellina in Chianti (SI), under tree bark, pine wood, Giusti coll.); 1 female (VVZC Eut527), Mt. Semprevisa, 08 December 2002 (V. Vignoli coll.); 1 female (VVZC Bassiano, Mts. Lepini (RM), 1100 m, 19 March 2000 Eut303), Loc. Ponte a Bozzone, Castel Nuovo Berar- (M. Bastianini coll.); 1 female (VVZC Eut529), 1 denga (SI), under tree bark, pine wood, 05 May 2003 (V. subadult (VVZC Eut530), ibidem, 14 December 2003 Vignoli coll.); 2 males (VVZC Eut502-503), Colognole, (A. Petrioli coll.). Marches: 1 male (VVZC Eut199), Livorno (LI), holly oak wood, 09 November 2003, Sirolo, Ancona (AN), 31 August 1967 (B. Urbani coll.). (M. Migliorini coll.); 1 female (VVZC Eut289), Gaiole Friuli V. Giulia: 2 females (VVZC Eut226-227), Trieste in Chianti, Siena (SI), under stone, beach wood, 01 May (TS), 13 April 1967 (F. Giusti coll.); 1 subadult (VVZC 2003 (V. Vignoli coll.); 2 females (VVZC Eut290-291), Eut141), Doberdo` del Lago (TS), under tree bark, Radda in Chianti, Siena (SI), under tree bark, pine deciduous wood, 10 March 2002 (V. Vignoli coll.). wood, 01 May 2003 (V. Vignoli coll.); 1 female (VVZC Liguria: 1 female (VVZC Eut292), Pignone (SP), rock Eut288), Gaiole in Chianti, Siena, (SI), under stone, wall, 24 April 2003 (M. Colombo coll.); 1 female (VVZC beach wood, 01 May 2003 (V. Vignoli coll.); 1 female Eut90), Rio Maggiore (SP), 10 June 2001 (D. Facheris (VVZC Eut197), 04 May 1970, Loc. Fungaia, Siena (SI) coll.). Emilia Romagna: 1 female (MZUF 138/5653), (Gatti coll.); 1 male (VVZC Eut528), Castell’Azzara Modena, 1874 (Carruccio coll.), labeled as E. c. concinnus (GR), 01 November 2003 (M. Bastianini coll.); 1 female (C.L. K.); 1 male (MZUF 139/5621), Casinalbo, (VVZC Eut229), Montieri (GR), 29 April 1981 (F. Formigine, 1880 (Fiori coll.), labeledas E. c. concinnus Giusti coll.); 1 female (VVZC Eut210), Montalbuccio (C.L. K.). Umbria: 1 female (MZUF 158/5661), (SI), 19 August 1966 (C. Baroni coll.); 1 female (VVZC Marzano, Lippiano, Perugia, , VII.1925 (A. Andreini ARTICLE IN PRESS 112 V. Vignoli et al. / Zoologischer Anzeiger 244 (2005) 97–113 coll.), labeledas E. carpathicus; 1 male (MZUF 157/ biogeography of this genus. Biogeographica (Paris) 75, 5639), Lippiano, Perugia, 09.X.1934 (? coll.), labeledas 49–65. E. carpathicus. Gantenbein, B., Soleglad, M.E., Fet, V., 2001. Euscorpius balearicus Caporiacco, 1950, stat. nov.: molecular (allo- zymes andmtDNA) andmorphological datasupport the existence of an endemic scorpion species on the Balearic References Islands (Scorpiones: Euscorpiidae). Organisms Diversity Evolut. 1, 301–320. Benton, T.G., 1992. The ecology of the scorpion Euscorpius Huber, D., Gantenbein, B., Fet, V., Sherabon, B., 2001. flavicaudis in England. J. Zool. London 226, 351–368. Euscorpius carpathicus (L.) from Austria (Scorpiones: Eu- Bonacina, A., 1980. Sistematica specifica e sottospecifica del scorpiidae): phylogenetic position clarified by mitochondrial complesso ‘‘Euscorpius germanus’’ (Scorpiones, Chactidae). DNAanalysis.In:Fet,V.,Selden,P.A.(Eds.),Scorpions Riv. Mus. Civ. Sci. Nat. ‘‘Enrico Caffi’’ (Bergamo) 2, 2001. In Memoriam Gary A. Polis. British Arachnological 47–100. Society, Burnham Beeches, Bucks, pp. 273–278. Braunwalder, M.E., 2001. Scorpions of Switzerland: summary Koch, C.L., 1837. Die Arachniden. Nu¨rnberg, C. H. Zeh’sche of a faunistic survey. In: Fet, V., Selden, P.A. (Eds.), Buchhandlung 3 (6), 105–115. Scorpions 2001. In Memoriam Gary A. Polis. British Koch, C.L., 1841. Die Arachniden. Nu¨rnberg, C. H. Zeh’sche Arachnological Society, Burnham Beeches, Bucks, Buchhandlung 8, 1–114. pp. 279–286. Lacroix, J.B., 1990. Faune de France; Arachnida: Scorpionida. Caporiacco, L.Di., 1950. Le specie e sottospecie del genere 4e note. Genus Euscorpius Thorell, 1876. 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