Moth Flies (Diptera, Psychodidae) Living in the Dark of Caves in the Dinaric Karst

Zootaxa 4845 (2): 275–282 ISSN 1175-5326 (print edition) https://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2020 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4845.2.8 http://zoobank.org/urn:lsid:zoobank.org:pub:748422C9-347E-4830-99A8-4D4309107E23

Moth flies (Diptera, Psychodidae) living in the dark of caves in the Dinaric Karst

RÜDIGER WAGNER1* & TONĆI RADA2 1University of Kassel, FB 10, Biology-Zoology, Heinrich-Plett-Straße 40, D-34132 Kassel, Germany Parkstraße 65, D-36110 Schlitz, Germany. [email protected]; https://orcid.org/0000-0002-2024-1827 2Speleološko Društvo “Špiljar”, Zajdenica Tehničke Kulture Grada Splita, 21000 Split, Varardinska 53, Croatia [email protected]; https://orcid.org/0000-0001-5287-6336 *Corresponding author

Abstract

Seoda cavernicola sp. nov. and Psychoda glamocensis sp. nov., are new species and cave dwellers from Bosnia and Hercegovina, and Croatia. Adults of S. cavernicola are pale and small; the eye bridge is reduced, ommatidia irregularly arranged, epandrium with a pair of setose excrescences. The eye bridge of P. glamocensis is likewise reduced with 2 or 3 irregularly ordered facet rows, palpus segments of some individuals are malformed; its closest relative is Psychoda alticola Vaillant based on the morphology of male and female genitalia as well as on COI barcodes.

Key words Psychodidae, troglobites, new species, Balkan peninsula

Introduction

The systematic research on cave dwelling animals has the best development worldwide on the Balkan Peninsula; the greatest amount of troglophilic animals is known from the Dinaric Karst. Among the cave living species found there are representatives of fish, amphibia, insects, millipeds, centipeds, spiders, crustaceans and other groups (e.g. Sket 2016). Organisms found in caves are classified troglobites (cave dwellers), troglophiles (species that sometimes occur outside of caves), and trogloxenes (species that cannot live permanently in caves). Life in caves affects species ecology, morphology, and physiology, and has further implications on evolutionary processes. Insect collections from caves usually contain few Psychodidae, at most species typically live near the entrance of the caverns with permanent contact to the surface environment (troglophiles, trogloxenes). They use caves as refugium against sunlight, high air temperature, low moisture, and to avoid predators. Thus, representatives of all extant psychodid subfamilies (Phlebotominae, Bruchomyiinae, Trichomyiinae, Sycoracinae, Psychodinae) have been reported from caves on almost all continents (e.g. Bravo et al. 2008, Bravo & Barata 2012a, 2012b, Khadri Shahar et al. 2011, Sarà 1950, 1962, Polseela et al. 2011, Quate 1962, 2000, Vaillant 1966). Usually most Psychodidae from collections in caves belong, besides Phlebotominae, to the subfamily Psycho- dinae, tribus Psychodini, genus Psychoda (sensu lato); ecologically these species are assumed to be euryecious. Larvae feed on feces of bats, birds, and any other decomposing organic material (biofilms, plant and animal cadav- ers). Adults of these species are without particular adaptation to permanent life in caves. Below we describe new species of the genera Seoda Enderlein and Psychoda (s.l.) Latreille, that show remark- able morphological variation that are probably adaptations to life in caves.

Material and methods

Specimens were collected with hand net in caves at high altitudes in Croatia and Bosnia and Hercegovina. Adults were put in situ in 80% ethanol and used for morphological and genetic studies. For morphological study, wings of specimens were dissected and kept in clove oil; the body was macerated in

Accepted by D. Cordeiro: 12 Aug. 2020; published: 2 Sept. 2020 275 10% KOH for 12-18 hrs, washed in acetic acid and put with drops of acetic acid to the clove oil with wings. After 1 day the acid evaporated and head, thorax abdomen and wings were arranged in drops of Canada balsam under 4 separate coverslips. Inspection and line drawing were made with a Leitz binocular microscope MZ 125 and Leitz MZ 20 EB with drawing mirror attached. Morphological terminology has changed and follows Kvifte & Wagner (2017).

Taxonomy

Seoda cavernicola sp. nov. (Figs 1–9)

Etymology. The name refers to the species’ preference for caves (caverna Latin). Diagnosis. Seoda cavernicola sp. nov. is distinguished from congeners by the pale body, the eye bridge of only 2–3 irregularly ordered facet rows, scape short, pedicel globular, and 1st flagellomere elongate, ventral surface of the subepandrial plate with 2 large setose excrescences; genitalia with robust outgrowths of gonocoxites, distiphallus lobes bowed out. Material. Holotype, 1♂, Croatia, Seget Gornji, Jama I. u. Gospinu Gaju, 17 February 2019; paratypes, 2♂, collected at the type locality with the holotype; (one paratype with wings destroyed). Further material (all paratypes): 2♂, 3♀, Croatia, Zelovo, Jama na Vranjinoj Glavici, 19 May 2019; 2♂, Croatia, Radošić, Baračeva jama, 31 March 2019; 1♂, Croatia, Zelovo, Elezova jama 19 May 2019; all leg. T. Rada. Holotype and 1 paratype (slide mounted) and remaining material in ethanol deposited in the Natural History Museum & Zoo, Split, Croatia. 1 paratype from Seget Gornji, and 1 paratype from Radošić (slide mounted) in the collection of the senior author. Description. Male: Specimens pale, whitish. Eyes reniform, eyebridge reduced with only 2 or 3 irregular facet rows, interocular suture wide U-shaped, distance between the eyes 2 to 2.5 facet diameter (figs 1–3). Antennae (fig. 4) of all specimens incomplete; scape pipe-shaped 2x longer than wide, pedicel globular 0.5x as long as the pedicel. First flagellomere almost symmetric elongate widest at middle, subsequent flagellomeres asymmetric bottle-shaped with basal bulb and long neck. Ascoids probably lost, 2 circular holes at about middle of segments may indicate the presence of ascoids. Antennal flagellum broken, length of remaining antennal articles: 40-26-73-64-58-62-; absolute length: 0.105-0.0685-0.1921-0.1684-0.1526-0.1631- mm. Basal palpus segment (fig. 5) slightly longer than the scape. Four palpus segments, terminal segment flexible; relative length: 30-58-55-95; absolute length: 0.08-0.15- 0.14-0.25 mm. Wing (fig. 6) pale translucent, length 2.30 mm, width 0.74 mm; length/width 3.11. Sc short terminates in wing; radial fork distal of medial fork, at level of Cu tip; no crossveins r4-r5 or r-m. Thorax without specific features. Abdomen with 8 segments and inverted genitalia; hypandrium equally thin, in the middle with slighter sclero- tization, with fine setae in that area. Gonocoxite tubular, about 2x longer than wide, gonostylus bent dorsal about as long as the gonocoxite, basally wider. From the basal inner edges of the gonocoxites outgrowths (gonocoxal apodemes/parameres?) arise that are half as long as the gonocoxites and enclose the dorsal lobes of the basiphallus and the basal part of the distiphallus sclerites. The dorsal parameral bridge consists of two median prolongations of the gonocoxites, converging in the middle forming a fissure or keel. Basiphallus dorsoventrally flattened, bilobed from the middle on. On dorsal side of the bifurcation thin lobes arise directed dorsally converging towards the fissure where they are jointed to the dorsal parameral bridge. The ventral lobes of the basiphallus sclerite are apically jointed to claw-shaped laterally bent distiphallus sclerites. At the positions of the joints between basiphallus and distiphallus lobes the aedeagus is jointed to (enclosed by) the large outgrowth of the gonocoxites. Epandrium basally wider with two sharp corners, lateral and hind edges almost straight; basally with 2 small foramina. On the ventral surface of the epandrium is a pair of large setose excrescences, a well-defined subepandrial plate is not discernable. Epandrial processes almost straight in the distal part on inner side with few stronger setae in line, distally with 8 apically serrate retinacula. The hypoproct is large, rhomboid, apically setose; epiproct small setose triangular. Remarks: Small size compared with species of the genera mentioned below and pale appearance of the type specimens are interpreted as adaptation to life in caves. The species shares large apodemes of the gonocoxites and

276 · Zootaxa 4845 (2) © 2020 Magnolia Press WAGNER & RADA a bilobed basiphallus sclerite with Jungiella Vaillant, Vaillantodes Wagner, Seoda Enderlein, and Panimerus Eaton; size and shape of the distiphallus sclerites are similar to Seoda. The mode of connection of the dorsal parameral bridge and the aedeagus is unique. Usually this is by a separated Y- or U-shaped sclerite, the furca, that appears as an individual sclerite jointed with the basiphallus sclerite and dorsally coadunate in the middle with the parameral bridge. The presence of large outgrowth of the gonocoxites is similar to Jungiella Vaillant. In contrast to Jungiella, Vaillantodes, Seoda, and Panimerus which possess a separate furca the dorsal lobes in S. cavernicola sp. nov. arise from the basiphallus sclerite, they are not separated from or jointed to it. However, the task in the mode of operation is the same as for the other species: to support the back and forth movement of the aedeagus and its components. Another striking feature is the pair of setose outgrowth on the ventral surface of the epandrium; a subepandrial plate is not recognizable; however, the setose outgrowths are in the position of the subepandrial plate and therefore are interpreted as part of this. They are positioned exactly dorsal of the outgrowth of the gonocoxites.

FIGURES 1–9. Seoda cavernicola. 1. Eyebridge of holotype. 2–3. Eyebridges of 2♂ paratypes. 4. Scape, pedicel, 1st and 2nd flagellomeres. 5. Basal palpus segment. 6. Wing. 7. Hypandrium, styles and aedeagus ventral view. 8. Epandrium, epandrial processes and proctiger, ventral view. 9. Basiphallus sclerite and dorsal parameral bridge with ‘furca’, ventral view. Scale: 1.0 mm (6), 0.1 mm (1–4, 7–9), 0.05 mm (5).

Etymology. Named after Glamoč, a town and municipality located in Canton 10 of the Federation of Bosnia and Herzegovina. Diagnosis. Psychoda glamocensis sp. nov. is distinguished from its congeners by the reduced eyebridge, gonostyli with a very long seta in the basal third and the subtriangular aedeagus with the axe-shaped sclerite. Type material. Holotype 1♂, Bosnia and Hercegovina, Golubnjača kod Skucana, Skucani, Glamoč (43˚58’31 .9’’N/016˚54’43.6’’E, 1.005 m a.s.l.). July 2018; paratypes, 8♂, 1♀ same as holotype; 8♂, 2♀, (paratypes), Croa- tia, Golubinka pod Kraljevcom, Radošić (43˚35’45.2’’ N/ 016˚20’32’’E, 233 m asl.) 15 February 2019; 49♂, 32♀, (paratypes), Croatia, Radošić, Golubinka pod Kraljevkom Pit, 8 October 2018. Further material. 36♂, 5♀, Croatia, Ercegovici, Rušića jama, 6 April 2019; 49♂, 8♀, Croatia Trilj, Bakuša 5 June 2019; 1♂ Croatia, Voštane, Golubinka u Ljutu, 24 February 2019; 42♂, 2♀, Croatia, Labin Dalmatinski, Velika Zečica Donji, 13 March 2019; 5♂, 2♀, Croatia, Zelovo, Elezova jama, 19 May 2019; 12♂, 5♀, Croatia, Unešić, Dželalijina Golubinka, 10 March 2019; all leg. T. Rada; 1♂ Austria, Lower Austria, Lunz am See, emergence trap at Schreierbach, 12 October 1972 leg. Malicky; 1st report from Austria (in collection RW). Holotype and paratypes and the mentioned further material are deposited in the Natural History Museum & Zoo, Split, Croatia. Three paratypes (2♂, 1♀) remain in the collection of the senior author. Description. Male: Eyes reniform, eyebridge with 2 or 3 rows of irregularly arranged facets, interocular suture wide U-shaped, distance between the eyes 2–2.5 facet diameter (figs 10–12). Antenna (fig. 14–15) with scape, pedicel and 13 flagellomeres; scape short, wider than long, pedicel spherical, flagellomeres bottle-shaped, with remarkably long necks, flagellomeres 1–10 with a pair of Y-shaped ascoids about as long as a flagellomere, apical flagellomeres 11–13 reduced in size, 12 and 13 with a subapical lobe and a short stiff bristle. Relative length of scape, pedicel and flagellomeres: 49-28-88-83-82-81-77-76-72-66-69-57-25-15-12; absolute length: 0.13-0.07- 0.23-0.22-0.22-0.21-0.20-0.20-0.19-0.17-0.18-0.15-0.07-0.04-0.03 mm. Four palpus segments, terminal segment flexible; relative length: 49-49-46-55; absolute length: 0.13-0.13-0.12-0.14 mm. Mouth parts with 4 acute apical teeth (fig. 16).

Wing length 3.1 mm, width 1.1 mm. C basally with 2 swellings; Sc short, a setose area, terminates in wing. R1 terminates in the distal half of the wing; division of Rs with fork R2/ R3+4 at level of Sc tip. R5 terminates short behind wing tip; radial and medial fork complete; medial fork basal of radial fork. CuA basally broader; in the basal part of the wing all veins are broken, here is fold that enables specimens to hold the wing in roof shape at rest. Cross veins not detected. Thorax without specific features. Abdomen with 8 segments, genitalia (figs 22–26) inverted; hypandrium broad, slightly broader in the midst. Gonocoxites short tubular about 1.5 times longer than wide, probably without setae. Dorsal parameral bridge (fig. 25) of gonocoxites thin in the middle, laterally widened; ventral bridge of parameres are blade shaped sclerites with sharp bent tips that do not meet in the middle. Gonostyli elongate triangular. almost straight; behind the basal swelling with a remarkably long and strong seta, about as long as the gonostylus. Epandrium (fig. 23) with 1 median foramen; edges roundish. basal edge deeply curved, apical edge almost straight. Subepandial plate increasingly wide apically with a broad median connection to the proctiger and to lateral plates connecting with the bases of the basally swollen epandrial appendages. Proctiger not clearly discernable; the epandrial processes clearly longer than the epandrium, strongly bent, apically with a sharp tip beside the basis of the retinaculum. Aedeagus consists of an asymmetric bilobed basal sclerite, lobes apically curved inwards; the distal sclerite is shorter than the basal sclerite, asymmetric, tomahawk shaped. Female: in most respects similar to male but poorly preserved. Differences are minor in the antenna and wing venation. Eye bridge fig. 13, mouth parts fig. 17 and 18. Relative length of antennal articles: 34-24-57-51-53-48-52-48-42-; absolute length: 0,09-0,06-0,15-0,13-0,14- 0,13-0,14-0,13-0,11- mm, apical segments missing. Four palpus segments, terminal segment flexible; relative length: 30-28-28-34; absolute length: 0.08-0.07-0.07-0.09 mm. Mouth parts with 4 acute apical teeth. Wing length approximately 2.20 mm. Genitalia (figs. 20, 21) with a bilobed subgenital plate with an elongate tubular structure, laterally accompanied by a pair of sclerites that appear to be superficially similar to the parameral complex of the male. Basally is a com- plicatedly folded sclerite of probably specific shape.

278 · Zootaxa 4845 (2) © 2020 Magnolia Press WAGNER & RADA FIGURES 10–21. Psychoda glamocensis. 10. Eyebridge of male holotype. 11–12. Eyebridges of male paratypes. 13. Same of female paratype. 14. Flagellomeres 10–13. 15. 2nd flagellomere with ascoid. 16. Male mouth parts.17–18. Female mouth parts. 19. Wing. 20. Female subgenital plate. 21. Internal genital sclerites, female. Scale: 1.0 mm (19), 0.1 mm (10–15, 20–21), 0.05 mm (16–18).

DNA Barcoding. The availability of a DNA barcode permits comparison with other Psychoda species. The two sequences (ZFMK-TIS-2617210 consensus sequence (reversed), ZFMK-TIS-2617212 consensus sequence (reversed)) indicate the closest relation with specimens of Psychoda alticola Vaillant 1973, from the Norwegian Finnmark (Kvifte & Andersen 2012). Sequences from Alpine specimens are presently not available. Remarks. The most striking feature of both new species is the reduction of the eyes probably due to life in caves The number of facets and facet rows varies extremely and differs even between specimens. In Psychoda s. lat. species the eye bridge usually consists of 4 parallel regular rows but in Ps. glamocensis sp. nov. the number of facet rows is reduced to mostly 2, with irregular arranged facets. Eyes are widely separate in both sexes so that the shape of the eyebridge varies between individuals. All specimens were collected deep in the caves, at Golubnjača kod Skucana at a depth of 30 m at permanent darkness, and at Golubinka pod Kraljevcom 25m deep at semidark-

MOTH FLIES FROM DINARIC KARST CAVES Zootaxa 4845 (2) © 2020 Magnolia Press · 279 ness. The malformation of palpus segments in several specimens is probably due to in-breeding in small populations in different caves. Number of teeth on mouthparts is as well variable and differs between specimens. This indicates that populations live for a long time in the caves. Long term low temperatures and enduring darkness are probable preconditions to develop morphological changes of the eyes and palpus segments. The antenna appears longer compared with the head diameter and the long seta on the gonostyli are further indications that antennae and setae may play major roles as tactile instruments to detect and locate conspecifics or avoid predators.

FIGURES 22–26. Psychoda glamocensis. 22. Hypandrium, gonostyli and aedeagus, holotype. 23. Epandrium, subepandrial plate and bases of epandrial processes. 24. Right epandrial process with retinaculum. 25. Left gonocoxite, parameral bridges and position of distiphallus sclerite, holotype. 26. Basiphallus and distiphallus sclerites and periaedeagal membrane. Scale: 0.1 mm (22–26).

The sex ratio in collections from all localities is unusual for Psychoda species; light trap collections often contain exclusively females. However, in Golubnjača kod Skucana cave 90%, in the Golubinka pod Kraljevcom cave 80%, and in Golubinka pod Kraljevkom cave 60% males were collected. The first two caves give shelter to

280 · Zootaxa 4845 (2) © 2020 Magnolia Press WAGNER & RADA wild pigeon (Columba livia Gmelin); probably the larvae of Psychoda glamocensis sp. nov. feed on their feces and remains; bats that are usually further inhabitants of caves may provide further organic material. The additional male from Austria was found as undetermined but close to P. alticola in the collection of the senior author; identification was finally easy based on the reduced eye bridge and male genitalia. The locality Sch- reierbach is a permanent cold stream (probably a very large rheocrene) with constant water temperature of about 8°C all year round. Here P. glamocensis was collected together with specimens that are doubtless P. alticola; both species seem to prefer cold conditions that occur in the Alps at higher elevation, but as well in cold springs, and caves at lower altitudes.

Discussion

The environment in caves is unique with almost permanently darkness and reduced seasonal changes of temperature and many other variables. Food supply originates to some extent from outside the cave: feces of birds, bats and other organisms that temporarily use caves. Both new species demonstrate similar morphological features interpreted as adaptation to life in caves. Per- manent darkness may affect size reduction of the eyes and irregular organization of facets in the eye bridge. Other morphological changes in specimens of P. glamocensis are in addition interpreted as effects of inbreeding in small populations. The high percentage of males is surprising. Usually females predominate by far in most collections of Psy- choda. From light trap collections (pers. observ.) it is known that in summer among thousands of females only few males occur. An approximately balanced sex ratio occurs in spring and autumn, with suboptimal conditions of nutri- tion and temperature and small populations. Thus, the high number of males collected may as well be an adaptation to the continuing awkward life in caves at high elevations in the Balkans.

Acknowledgements

We thank Mr. Siniša Šolak, experienced guide from Glamoč area for guiding Tonći Rada and co-workers to the entrance of the Golubnjača Kod Skukana pit. Many thanks to Björn Rulik (Zoologisches Forschungsmuseum A. Koenig, Bonn, Germany) for executing a COI analysis following the precepts used in the German Barcode of Life (GBOL) project. The recently late Phil Withers kindly revised linguistics of the text. Thanks to two anonymous reviewer for their helpful comments.

References

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