Description of a new species of Wormaldia from Sardinia and a new Drusus species from the Western Balkans (Trichoptera, Philopotamidae, Limnephilidae) Author Vitecek, Simon Author Previsic, Ana Author Kucinic, Mladen Author Balint, Miklos Author Keresztes, Lujza Author Waringer, Johann Author Pauls, Steffen U. Author Malicky, Hans Author Graf, Wolfram text ZooKeys 2015 496 85 103 http://dx.doi.org/10.3897/zookeys.496.9169 journal article http://dx.doi.org/10.3897/zookeys.496.9169 1313-2970-496-85 DE6EE3CABEFB4D20A41151743759BCB8 Taxon classification Animalia Trichoptera Limnephilidae Drusus crenophylax Graf & Vitecek sp. n. Material. Holotype. 1 male: Bosnia and Herzegovina, Cvrcka river; 44°32.932'N 17°23.562'E ; 393 m a.s.l.; 01.10.2014; leg. Dejan Dmitrovic , Goran Sukalo ; specimen identifier: fDsp4501M. Paratypes: 2 females: Bosnia and Herzegovina, Spring of Cvrcka river, Vilenjska vrela; 44°33.003'N , 17°23.580'E ; 456 m a.s.l.; 12.09.2012; leg. Dejan Dmitrovic ; specimen identifiers: fDsp3401F, fDsp3402F. 4 males, 3 females, 19 larvae: Bosnia and Herzegovina, Spring of Cvrcka river, Vilenjska vrela; 44°33.003'N , 17°23.580'E 456 m a.s.l.; 12.09.2012; leg. Dejan Dmitrovic , Goran Sukalo ; specimen identifiers for 3 larvae: fDsp4502L, fDsp4503L, fDsp4504L. Holotype and paratypes currently in coll. W. Graf, will deposited in the Biologiezentrum des Oberoesterreichischen Landesmuseums, Linz, Austria. Type locality. Bosnia and Herzegovina, Republika Srpska, Cvrcka River. Diagnosis. Males of the new species are most similar to Drusus discophorus Radovanovic and Drusus vernonensis Malicky, but exhibit (1) subtriangular superior appendages in lateral view, (2) subtriangular, low tip of the intermediate appendage in lateral view, and (3) simple, rounded tips of intermediate appendages in caudal view. Drusus discophorus males have suboval superior appendages and a high round tip of the intermediate appendage in lateral view; Drusus vernonensis males have round superior appendages in lateral view and trilobate tips of intermediate appendages in caudal view. Females of the new species show the reduced median lobe of the vaginal sclerite and high base of the lateral lobe of segment IX as typical for Balkan Drusinae , and are most similar to Drusus vernonensis , but exhibit (1) a sharp dorsal notch of segment X in lateral view, and (2) segment X with 2 round median lobes in dorsal view. Drusus vernonensis females have a rounded dorsal outline of segment X and lack the median lobes of segment X. Larvae of the new species are most similar to Drusus klapaleki Marinkovic-Gospodnetic and Drusus serbicus Marinkovic-Gospodnetic , but exhibit (1) a semicircular area dorsomedially on the pronotum anterior the pronotal ridge void of white recumbent setae, (2) lateral gills, and (3) a subtriangular pronotal ridge in lateral view. Larvae of Drusus klapaleki have white recumbent setae covering the whole pronotum, and larvae of Drusus serbicus lack lateral gills and have an annular pronotal ridge. Description . Adults. Habitus dark; sclerites and tergites brown; cephalic and thoracic setal areas pale; cephalic, thoracic and abdominal setation blond; legs light brown to fawn, proximally darker; haustellum and intersegmental integument pale, whitish. Wings smoky, with dark setae. Male maxillary palp 3-segmented. Forewing length 11-13.2 mm, spur formula 1 -3- 3 in males; forewing length 13-14.5 mm, spur formula 1 -3- 3 in females. Male genitalia (Fig. 3 A-E ). Tergite VIII dark brown, in dorsal view cranially distinctly incised, with lighter areas around fused alveoli; setation concentrated at laterocranial borders of spinate areas; spinate area as two +/- triangular laterocaudal lobes medially connected by a band of spines, embracing a medial, indent less sclerotized area (translucent in cleared specimens) with scarce spines. Ninth abdominal segment (IX) ventrally wider than dorsally in caudal view; in lateral view medially with a sharp caudad protrusion and a ventral protrusion, embracing the base of the inferior appendices. Superior appendages in lateral view subtriangular, somewhat Y-shaped with a shorter dorsal and a longer ventral protrusion separated by a slight indentation. Intermediate appendages in lateral view blocky with 2 tips, the proximal sharp, the distal high, rounded, rough; in dorsal view the tips parallel, extending laterally: a bar-shaped, laterally rounded distal tip and a sharp proximal tip, separated by a rounded excision with round edges; in caudal view approximately triangular, tips rounded. Inferior appendages (gonopods sensu Snodgrass 1935 ) in lateral view proximally wide, medially slightly constricted with a slight dorsal triangular protrusion, curved dorsadly in the slender posterior third; in dorsal, ventral and caudal view proximal part laterad, distal part approximately straight in dorsoventral plane, curved dorsad; in caudal view tips distinctly slender; setal alveoli fused, creating a rugged, less sclerotized ventral area. Parameres simple, with a distinct medial thorn-like spine and 2 proximal spines in the proximal half. Figure 3. Genitalia of Drusus crenophylax sp. n. A-E male genitalia: A right lateral view B paramere in right lateral view C ventral view D caudal view E dorsal view F-I female genitalia: F right lateral view G ventral view H caudal view I dorsal view. Scale bar: 1 mm. Female genitalia (Fig. 3 F-I ). Segment IX setation abundant, concentrated in the caudal half; lateral lobe of segment IX membraneous, in lateral view oblique triangular, the ventral edge about twice as long as the dorsal edge, with a dorsal sclerotized setose part protruding caudally; in dorsal and ventral view slender, projecting lateradly; in caudal view dorsal sclerotized setose part somewhat triangular. Segment X in lateral view with a proximal and a distal part, defined by a sharp dorsal notch; in dorsal view trapezoidal, with rounded shoulders, 2 small dorsal median lobes, and distally with 2 triangular, sharp-tipped lateral lobes, each with a lateral rounded setose and a small median rounded protrusion; ventrally unsclerotized, open. Supragenital plate in lateral view sinuously-edged quadrangular with a small, rounded dorsal protrusion, caudal line slightly indent; in ventral view quadrangular, in caudal view quadrangular, dorsally slightly wider than ventrally. Vulvar scale in lateral view triangular, rather straight, longer than the supragenital plate; in ventral view slender with 3 lobes: 2 lateral lobes, digitiform, roundly oval, straight; 1 median, short (reduced), of greater width than length: length approximately 1/6th of that of lateral lobes. Fifth instar larva (Fig. 4 A-I ). Head capsule hypognathous, finely granulated with a field of microspinules dorsal to each eye, dark brown dorsally, fading to yellow ventrally; 18 pairs of primary setae present: #1, 4, 6, 10, 12, 13 yellow and #6, 13 short, inconspicuous, the rest dark brown, long (Fig. 4A); antennae located on high carinae, each carina about as high as long, both strongly curved mediad (Fig. 4B); mandibles toothless. Pronotum dark brown, coarsely granulated; distinct medial ridge present, rounded, steeper anteriorly in lateral view; recumbent white setae present, but lacking in a semicircular area anterior the pronotal ridge (Fig. 4C); pronotal horn present. Mesonotum completely covered by 2 sclerites, dark brown, with darker apodemes; edges black; sa1 comprising 4-6 setae, sa2 and sa3 connected, comprising 28-34 setae in total on each sclerite (Fig. 4D). Metanotum with 3 pairs of sclerites: anteriomedian sclerites subtriangularly ovoid, dark brown with 11-19 setae; posteromedian sclerites rhomboid, pale brown, with 13-15 setae; lateral sclerites long, curved dorsally in lateral view, pale brown fading to yellow ventrally with a dark median spot and 21-25 setae (Fig. 4E). Legs yellow-light brown, dorsally and distally darker (Fig. 4 F-H ). Abdomen white (Fig. 4G), dorsal gills from II praesegmental position to VI praesegmental position, lateral gills from II praesegmental position to IV praesegmental position, ventral gills from II prasegmental position to VII postsegmental position; lateral line from last quarter of II to first quarter of VIII (Fig. 4I); abdomen I with 1 dorsal and 2 lateral protuberances, posterior sclerites absent on lateral protuberances, setal areas sa1-3 fused dorsally and ventrally (Fig. 4D, E), sternum bearing 2 setae with distinct basal plates; abdomen VIII with 2 long and 2-4 short posterodorsal setae on either side; abdomen IX with 1 posterodorsal seta on either side, dorsal sclerite IX semicircular, pale brown with 7 long and several shorter setae. Case simple, constructed of mineral particles. Figure 4. Larval characteristics of Drusus crenophylax sp. n. A head, frontal view B head, left lateral view C pronotum dorsal view D meso- and metathorax with abdominal segment I, dorsal view E abdominal segments I-V, left lateral view F left thoracic leg I, frontal view G left thoracic leg II; frontal view H left thoracic leg III, frontal view; bottom, gill and lateral line diagram, positions of gills are depicted as black circles, position of lateral line bold. Molecular species delimitation and larval affiliation. Analysis of the genetic distance of mtCOI between Drusus crenophylax sp. n. and the in the adult stage morphologically most similar species, Drusus discophorus and Drusus vernonensis , clearly supports the recognition of the new species. Uncorrected p-distances recorded in a fragment of the mtCOI gene (ranging from 2-8%; Fig. 5), agree with the interspecific distances commonly recorded in Limnephilidae (e.g., Graf et al. 2005 ; Kucinic et al. 2011a ; Previsic et al. 2014a , b ) and other caddisfly families (e.g., Hydropsychidae ; Pauls et al. 2010 ). Also, all haplotypes of Drusus crenophylax sp. n. adults were completely identical to another and those of undescribed Drusus -larvae collected at the locus typicus, enabling confident affiliation of larvae and adults of Drusus crenophylax sp. n. Figure 5. Distance matrix (lower left) and colour heat map (upper right) showing uncorrected inter- and intraspecific p-distances of the partial mtCOI sequence (541 bp) between Drusus crenophylax sp. n., Drusus vernonensis and Drusus discophorus . For detailed information on the haplotypes, see Table 1. Ecology and distribution. Drusinae species typically are members of crenal species communities, and mainly inhabit crenal sections of cold streams. Larval Drusus crenophylax were collected at eucrenal sections of the Cvrcka River (Fig. 6A, B) and behave as epilithic grazers, as indicated by mandible morphology ( Pauls et al. 2008 , Graf et al. 2009 ). Based on regional collection data, we assume that the species is a micro-endemic restricted to the watershed of the Cvrcka river. Figure 6. Habitat of Drusus crenophylax sp. n. at the type locality. A collection site of the larval paratypes B collection site of the male holotype. Etymology. The species epithet is a compound name, combining κρηνον ('well, spring, fountain' in Ancient Greek) and φυλαξ ('guard, keeper, protector' in Ancient Greek), terms that reflect the high degree of niche specificity of Drusus species, the majority of which inhabit crenal sections of streams ( Graf et al. 2008 ).