AMERICAN MUSEUM NOVITATES Number 3915, 19 pp. December 18, 2018 A new species of Eumanota Edwards (Diptera: Mycetophilidae: Manotine) from Colombia: evidence for a pseudogondwanan pattern DALTON S. AMORIM,1 SARAH S. OLIVEIRA,2 AND ANDREA C. HENAO-SEPtJLVEDA3 ABSTRACT Eumanota wolffae, sp. nov., is described from the high Andean forests of Colombia. This is the first Neotropical species of the clade of non-Manota genera of the mycetophilid subfamily Manotinae—to date known entirely from the Oriental and the northwestern Australasian regions, and in Baltic amber. With the other species of Eumanota Edwards, this species shares, among other features, a largely developed third maxillary palpomere, projecting beyond the base of fourth palpomere, with a wide sensorial pit, a flat inner face, and the last palpomere at least 4x the length of the fourth palpomere. The systematic position of the Colombian species within the Manotinae is addressed. The biogeographic significance of Eumanota in the Neo¬ tropical region is discussed, interpreted here as a concurrent incidence of a circumtropical pattern and Gondwanan distributions, in other words, a particular pattern of biogeographic pseudocongruence, referred to here as a “pseudogondwanan pattern.” This pattern is associated with an early Cenozoic tropical biota over Laurasian terranes that expanded its distribution to the south (in the Americas, Africa, and Australasia), followed by large-scale extinction of Nearctic and Palearctic representatives due to global cooling in the Neogene. A discussion is provided about the correlation between these patterns and evidence of Late Cretaceous-Paleo¬ gene tropical floras over Laurasian terranes. 1 Faculdade de Filosofia, Ciencias e Letras de Ribeirao Preto, Universidade de Sao Paulo, Sao Paulo, Brazil. 2 Instituto de Ciencias Biologicas, Universidade Federal de Goias, Goiania, Brazil. 3 Grupo de Entomologia, Universidad de Antioquia, Medellin, Colombia. Copyright © American Museum of Natural History 2018 ISSN 0003-0082 2 AMERICAN MUSEUM NOVITATES NO. 3915 INTRODUCTION The Mycetophilidae subfamily Manotinae includes the Recent genera Manota Williston, Paramanota Tuomikoski, Promanota Tuomikoski, and Eumanota Edwards. All four genera have been shown to comprise a monophyletic group, the last three genera in a clade sister to Manota (Hippa et al., 2005, 2016; Sevdk et al, 2013). Taxonomic knowledge of the world manotines has remarkably increased over the past 20 years. In the late 1990s there were less than 30 species of Manota described worldwide, two spe¬ cies of Eumanota, one species of Paramanota, and one of Promanota. We now have nearly 300 species of Manota (Kurina et al., 2018) and the number of species in the other genera have risen to 10 in Eumanota, nine in Paramanota, and two in Promanota (Hippa et al., 2017). The Creta¬ ceous species described in Alavamanota Blagoderov and Arillo—A. hispanica Blagoderov and Arillo (2002), from Spain, and A. burmitina Blagoderov and Grimaldi (2004), from Myanmar— do not belong in a clade with Eumanota, Paramanota, and Promanota (Hippa et al., 2005). The bulk of the diversity of the non-Manota genera of Manotinae is in the Oriental Region, with only three non-Oriental species in this group of genera: Eumanota malukensis Soli (2002), from the Molucca Islands, E. jani Papp (2004) from Papua New Guinea, and Paramanota gran- daeva Hippa (2010) from the Baltic amber. Fly diversity in Colombia is uniquely rich. There are tropical elements connecting Central America along the Pacific coast to the Choco region, tropical elements with Amazonian con¬ nections east to the Andes, and the unique high-mountain Andean forests, as well as the paramo ecosystems. The high Andean forests are located at the top of the northern range of the Andes mountain system. The early stages of the Andean orogeny are Early Jurassic in age (Ramos, 2009), but the northernmost range of the Andes was formed as late as the Pleistocene (Hoorn et al., 2010). These environments are subject to extreme climatic conditions, including intense solar radiation, strong winds, extensive cloudiness, and strong temperature fluctuations (with changes from “strong winter” to “intense summer” in a single day) (Rangel-Ch., 2000, 2005). No doubt a result of these physical regimes, the high Andean forests host a particular set of very localized, endemic species (Rangel-Ch., 2000). These ecosystems currently suffer drastic impact caused by anthropic activities. Due to the very restrict species ranges, this impact is rapidly leading to a great loss of biodiversity (Herzog et al., 2011). Taxonomic and ecological studies of invertebrates inhabiting the paramos are scarce or nonexistent (for an overview, see Wolff et al., 2016). In what concerns Diptera diversity, the high Andean forest faunas combine some tropical Mesoamerican and Amazonian elements, but most frequently austral temperate and Nearctic components (Amorim and Pires, 1996; Oliveira et al., 2007; Amorim, 2009), producing not only a species-rich fauna but a phylogeneti- cally diverse one as well. Recent collecting in the high Andean forests of Colombia with Malaise traps produced an amazing diversity of fungus gnats. Among rather rare rangomaramids, ditomyiids, and various genera of mycetophilids, we found three males and a female of an undescribed species of Eumanota. This is the first non-Manota species of Manotinae known from the Americas. This 2018 AMORIM ET AL.: NEW SPECIES OF EUMANOTA EDWARDS 3 paper describes and illustrates this species, discusses the position of the species among the manotines, and discusses the evolutionary significance of the presence of this species in the Neotropical region. Material and Methods All specimens of the type series were collected with Malaise traps in the high Andean forest from the paramos of two sites, one in the municipality of Envigado and the other in Yarumal— both belonging to the Central Cordillera in the Department of Antioquia, Colombia. The male holotype and single male and female paratypes were deposited at the Coleccion de Entomologia of the University of Antioquia (CEUA), and one male paratype was deposited at the Museu de Zoologia of the University of Sao Paulo (MZUSP). The male paratype housed at the CEUA was stored in 95% ethanol at -20°C for molecular studies. The male paratype housed at MZUSP had the wings removed and was mounted on a per¬ manent slide; the remainder of the specimen was treated with 10% KOH for about 10 minutes at 40°C to soften internal tissues. The specimen was then kept for 10-15 minutes in 100% glacial acetic acid, and mounted using Euparal on the same slide as the wing. Digital images were made with specimens in gel ethanol or after slide mounting. Photos were taken using a Leica DC 500 camera on a Leica M16 stereomicroscope. Stacking used Helicon Focus 6.7.1. Final photos were treated using Adobe Photoshop CC. Initial drawings of the male terminalia were made using a camera lucida on a compound microscope, vectorized with Adobe Illustra¬ tor CC. Photos of the complex male terminalia were taken in many different levels and stacking was made for particular ranges of focus, so individual inner sclerites could be visualized with¬ out blending with sclerites at other focus levels. Morphological homology and terminology mainly follows Cumming and Wood (2009), Soli (1997), and Hippa et al. (2005), except for wing venation, which follows Cumming and Wood (2017), based on wing vein homology interpretation of Wootton and Ennos (1989) and Saigusa (2006). We reran Hippa et al’s (2005) phylogenetic analysis of the relationships among manotine genera only to verify the position of the new species in Eumanota, using the same dataset and all their terminals and Eumanota woljfae. Analyses were performed using TNT (Goloboff et al., 2008), with all characters unordered and equal weight and implied weighting with k = 3, 10000 trees retained in memory, random seed 0, 1000 replicates, and TBR saving 10 trees for interaction. Eumanota Edwards, 1933 Eumanota Edwards, 1933: 231. Type species: Eumanota leucura Edwards, 1933, by original designa¬ tion. Soli, 2002: 46-52, Papp, 2004: 228-232, Hippa et al., 2005: 410-414. Amended Diagnosis (modified from Soli, 2002): A row of strong bristles around hind margin of eyes. Three ocelli in straight line, median ocellus about half the diameter of lateral ones. Four palpomeres visible; palpomere 1 lost or fused to palpomere 2; palpomere 3 large, 4 AMERICAN MUSEUM NOVITATES NO. 3915 triangular, projected beyond insertion of palpomere 4, with a deep, conspicuous sensory pit; palpomere 5 from 4-6x longer than palpomere 4. Scutum densely clothed with minute setae, no defined acrostichals, a single row of dorsocentrals on posterior half, some stronger supra- alars. Basisternum shieldlike, with setae at least laterally Anepisternum setose; mesepimeron bare; laterotergite ovate, bulging, setose; mediotergite, metepisternum, and metepimeron bare. Wing membrane either clear or darkened at distal half, at least posterior half with dorsal mac- rotrichia on membrane. C extends well beyond apex of R5. Sc typically short, ending in bR; Rx shorter or only slightly longer than r-m; r-m longitudinal. M1+2 less than half r-m length, basal most part of Mx weakly sclerotized. M4 separated from CuA at base of wing. Tibiae and tarso- meres with trichia arranged in regular rows. Spurs well developed. Comments. Some adjustment of the original diagnosis was necessary to include this new Neotropical species. Most features referred to as diagnostic for the Oriental/Australasian spe¬ cies of Eumanota (but not all) are also present in the Neotropical species (exceptions are dis¬ cussed below). In E. wolffae, sp. nov., the hind tibia does not have a sensory groove and the female clearly has a two segmented cercus. Eumanota wolffae, sp. nov. Figures 1-5 Zoobank: urn:lsid:zoobank.org:pub:61CC009E-0407-48FD-8E10-D7B06CAB80A2. Material examined: Holotype male, Colombia, Antioquia, Yarumal, Vereda El Respaldo, Farm La Maruja, 6°55' 05"N, 75°24'48"W, Malaise trap, Bosque, 2,300 m, 15.Dec.2016, C.H.- Sepulveda and J. Medina cols. (CEUA) (in ethanol). Paratypes. 1 male, same data as for holo¬ type (CEUA); 1 female, Colombia, Antioquia, Envigado, Loma del Escobero, Parcelacion Nubarrones, 6°13'97"N, 75°55'54"W, Malaise trap, Bosque, 29.Jan.-ll.Feb.2017, M. Wolff and C. H.-Sepulveda cols. (CEUA) (in ethanol); 1 male, Colombia, Antioquia, Envigado, Loma del Escobero, Parcelacion Nubarrones, 06°08'23"N, 75°33'14"W, Malaise trap, Bosque, 25.Sept.-01. Oct.2017, M. Wolff and C. H-Sepulveda cols. (MZUSP) (on a slide-mount). Diagnosis. Maculation on distal third of wing and over distal part of CuA (fig. 2E). Body mostly brown, with light brown to cream areas on sides of scutum, anterior half of thoracic pleura, coxae, and sternites 1-6. Female terminalia with two-segmented cercus (fig. 5A, B). Each gonocoxite with two distal projections, one lateral and one ventral, distal lateral projection with a megaseta on inner face; gonostylus small, well sclerotized, slightly asymmetric, tapering to apex, with an irregular indentation near tip, and a pair of straight megasetae on ventral face; gonocoxal bridge with setae; T9 with a pair of well-developed megasetae on inner face, tergite 9 elongate trapezoid; cercus projecting from distal margin of tergite 9 (figs. 3 A, 4C). Description. Male: Wing length, 3.2 mm. Head (fig. 1A-D). Frons, face, and clypeus whitish yellow, occiput whitish yellow ventrally, darker toward vertex, dark brown at area of ocelli. Anten¬ nal scape and pedicel whitish yellow, flagellum light brownish yellow on first three flagellomeres, light brown on more distal flagellomeres. A crown of 8-10 black, long postocular bristles at each 2018 AMORIM ET AL.: NEW SPECIES OF EUMANOTA EDWARDS 5 FIGURE 1. Eumanota wolffae male paratype: A, frontal view of head and thorax; B, head, frontal view; C, head, dorsal view; D, Head, lateral view side around eyes. Midocellus present, about half the width of lateral ocelli, ocelli in line, lateral ocelli closer to eye margin than its own width. No eye bridge, ommatotrichia present. Antennal scape and pedicel with a row of distal short, brown setae; first flagellomere 1.6x longer than wide, no black or stronger setae on any flagellomere. Fronto-clypeus 2.8x higher than wide, densely covered with blackish short setae, setation longer along ventral margin. Labella quite elongate, whitish yellow. Maxillary palpus with a small basal palpomere (either palpomere 1+2 or pal- pomere 2), palpomere 3 well developed, brownish, projecting over base of palpomere 4, a large, rounded sensory pit on inner basal fifth of palpomere; palpomere 4 whitish, distal two-thirds wider than basal third; palpomere 5 whitish, about 4x longer than palpomere 4. Thorax (fig. 2A-C): Scutum largely brown, with a pair of whitish-yellowish bands along lateral margins. Scutellum brown dorsally, whitish-yellow along posterior margin. Basisternum well developed, shieldlike, with some lateral setae. Antepronotum, postpronotum, prosternum, and proepisternum whitish, proepimeron, dorsal part of anepisternum and of mesepimeron, metepisternum, and metepimeron light brownish yellow; ventral end of anepisternum, katepi- sternum, and ventral part of mesepimeron yellowish brown; laterotergite and mediotergite light 6 AMERICAN MUSEUM NOVITATES NO. 3915 CuA FIGURE 2. Eumanota wolffae, male paratype: A, thorax, dorsal view; B, thorax pleural, lateral view; C, thorax, posterior view; D, wing tip; E, full wing. Rsl, first sector of the radial sector. 2018 AMORIM ET AL.: NEW SPECIES OF EUMANOTA EDWARDS 7 brown. Scutum with a regular row of moderately strong dorsocentrals, besides a very irregular row of long black supraalars, and a few prescutellars; no defined acrosticals. Anterior parapsidal suture short but well marked, ending outside row of dorsocentrals. Antepronotum and proepi- sternum with long, black setae; anepisternum with scattered short, brownish setae and a pair of longer setae on dorsoposterior margin; laterotergite with about 18 long, brownish setae; katepisternum, mesepimeron, metepisternum, and mediotergite bare. Mesepimeron not reach¬ ing ventral margin of thorax. Legs: Foretibia with an anteroapical depression furred with setulae, no regular comb of setae. Front coxa whitish yellow, mid- and hind coxae light brownish yellow, with darker lon¬ gitudinal areas. Front femur, tibia, and tarsus light brown; mid femur, tibia, and tarsus light yellowish brown, femur with a darker area ventrobasally, tarsus darker to apex; hind femur mostly light brown, lighter dorsodistally, tibia light brownish yellow, tarsus light brown. Front coxa entirely covered with short setae on anterior and lateral faces. Tibial spurs 1:2:2, mid and hind tibial spurs of about same length, about 3.8x longer than width of tibial apex. Tarsal claws with a long basal tooth. Wing (fig. 2D-E): Membrane quite translucent on basal 3/5, distal 2/5 grayish brown, distal end of CuA also with a grayish-brown area, as well as membrane around M1+2 and distal end of r-m. Entire membrane densely covered with macrotrichia. All anterior and posterior veins with dorsal setae, including Sc, except for first sector of Rs. Sc clearly ending in bR. Rs originat¬ ing at 0.6x wing length, transverse. Rx quite long, about 1.4x r-m length, R5 curved posteriorly on distal fourth, joining C in an acute angle, C extending beyond R5, almost reaching Mx; r-m quite longitudinal, elongate, about 2.5x M1+2 length, anterior end of r-m apart from bR, hence br cell much wider medially; bM long, 1.7x r-m length, originating at level of h, very close to wing base; M1+2 weakly sclerotized, short, 2x length of first sector of Rs; basal tenth of Mx dark, with setae, but not fully sclerotized as a vein; M2 slightly curved anteriorly on basal third, with a quite unique shape; M4 originating close to wing base, actually disconnected from other veins (with a very faint connection to CuA at base), with a gradual, discrete posterior curve along its length, with a slightly stronger curve at tip; CuA well sclerotized, with a stronger curve before level of proximal tip of r-m; CuP slightly sclerotized on basal half, entirely absent on distal half; no trace of AY. Abdomen: Tergite 1 dark brown, tergites 2-5 dark brown with some yellowish-brown basal areas, tergite 6 brown; sternites 1-6 yellowish; segments 7 and 8 brown. Tergite 8 slender, wide, extending laterally toward ventral side of abdomen, sternite 8 triangular and elongate, with seven distal setae. Terminalia (figs. 3A, B, 4A-D): Terminalia slightly elongate, gonocoxites (gc) at least 2x as long as wide, mesial fusion reaching distal margin with no suture, mediodistally with a short posterior incision with indented margin; distal end of gonocoxites with a distal lateral projec¬ tion (glp) extending beyond insertion of gonostylus (gs) and an additional more mesial oblique projection (gmp) as long as lateral ones; gonocoxites with a large modified seta directed inward (gci) at inner face of tip of each lateral projection, besides a regular, elongate distal seta at tip of external face of lateral projection (gcd) and additional shorter setae on external face. Gono- AMERICAN MUSEUM NOVITATES NO. 3915 B FIGURE 3. Eumanota wolffae, male paratype: A, ventral view; B, dorsal view. Abbreviations: aap, aedeagal apodeme; ae, aedeagus; ce, cercus; gcb, gonocoxal bridge; gcd, gonocoxal distal setae on lateral projection; gci, gonocoxal inner seta on latera projection; glp, gonocoxite lateral projection; gmp, gonocoxitemesal pro¬ jection; gs, gonostylus; pa, paramere; pap, parameral apodeme; S8, S10, sternites 8, 10; T8, tergite 8; T9, tergite 9; T9s, differentiated seta of tergite 9. stylus well sclerotized, slightly asymmetrical, shorter than gonocoxite projections, displaced to a quite dorsal position at terminalia, bearing two long, strong straight setae at ventral face, distal end with irregular indentation. Aedeagus (ae) present as a long, straight, well-sclerotized medial sclerite with two pairs of distal lateral projections and an additional medial projection bearing gonopore at tip, aedeagal apodeme (aap) reaching anterior end of terminalia; paramere (pa) with a pair of anterior apodemes, each with a short lateral, curved extension (pap), medi¬ ally fused on distal half dorsal to aedeagus, with a slightly asymmetrical pair of distal projec¬ tions curved outward extending beyond tip of aedeagus, and a pair of short, well-sclerotized elongate projections adjacent to aedeagus. Tergite 9 (T9) trapezoid, elongate, wider at base than distally, with curved distal margin, bearing at inner face a pair of distal, long, differentiated setae (T9s). Gonocoxal bridge (gcb) with setae at its distal and inner margins, gonocoxal apodemes elongate, close together anteriorly. Cerci (ce) projecting beyond distal margin of tergite 9, covered with elongate setae and elongate microtrichia, sternite 10 (S10) quite well developed, triangular, with long, straight setae. Female (paratype): As male, except as follows. Wing: Length, 3.5 mm. Thorax: Pleural sclerites light brown. Abdomen: Segments light brown, segments 2-5 with yellowish-brown areas, all tergites and sternites setose. Tergite 7 much wider than long, sternite 7 slightly pro¬ jected medially over base of tergite 8. Terminalia (fig. 5A, B): Sternite 8 elongate, with a pair of wide bare, basal lobes, each with a short distal projection with some few dark setae. Tergite 8 wide, short, with a row of setae along posterior margin, a laterodistal lobe quite ventrally bearing some few setae. Tergite 9 extending posteriorly at each lateral end, with some few setae. 2018 AMORIM ET AL.: NEW SPECIES OF EUMANOTA EDWARDS 9 A B FIGURE 4. Eumanota wolffae, male paratype: A, ventral view at level of syngonocoxite; B, ventral view at level of parameres; C, ventral view at level of gonostylus and gonocoxal bridge; D, Ventral view at level of tergite 9. Abbreviations: gc, gonocoxite; for others, see figure 3. Tergite 10 short, slightly projected laterally, with some few, weaker setae. Sternite 10 elongate, wider at base, with rounded tip, covered with setae. First segment of cercus longer than second; second segment lobose, wider than long, apically rounded, setose. Etymology: The species epithet is named to honor of Marta Wolff, one of Colombia’s outstanding woman scientists, who has dedicated her career to the knowledge of the insect fauna of Colombia, particularly fly diversity. She also coordinates the larger project studying the Diptera fauna of the high Andean forests of Colombia, including the paramos, and took part in the hard fieldwork underlying the project. Biology: The project on the high Andean forests includes collecting fungi for to rear adult mycetophilids in the lab. Adults of E. wolffae, however, have so far not been obtained from fungi. There is no information, hence, on the biology of this species except the kind of environ¬ ment in which they were collected, the high Andean forests on Colombia (fig. 6). 10 AMERICAN MUSEUM NOVITATES NO. 3915 FIGURE 5. Eumanota wolffae, female paratype: A, lateral view; B, ventral view. Abbreviations: cel, ce2, cercus segments 1-2; S7-10, sternites 7-10; T7-10, tergites 7-10. Comments: Males and females of E. wolffae are identical to each other in general body color, including the wing pattern. The male terminalia of E. wolffae is very complex, and has some distinctive features compared to the Oriental/Australasian species of Eumanota. The gonocoxites are fused all the way to the distal margin of the syngonocoxites, with no medial suture except for a short, irregular distal incision (fig. 3A). The gonocoxites ventrally have a pair of more mesal projections at the distal margin and a pair of distal lateral projections both extending beyond the base of the gonostylus (fig. 4A, C). Each lateral projection of the gono- coxite bears a single, very distinctive distal megaseta on the inner face (figs. 3A, B, 4C), unique in the genus. The aedeagus is complex, with a long, straight apodeme that almost reaches the anterior end of the terminalia and a medial tubular distal structure bearing a terminal opening (figs. 3 A, 4A, B). The parameres have a pair of typical distal outward projections (figs. 3 A, 4B). The general shape of tergite 9 (fig. 4B, D) is very similar to that found in the Oriental species, but there is a pair of strong setae at the inner face of tergite 9, absent in other species of the genus. The well-sclerotized gonostylus (figs. 3A, B, 4C) has a unique shape, not only due to the two or three distal indentations, but to the pair of strong setae directed ventrally. DISCUSSION Phylogenetic Position of Eumanota wolffae in the Manotinae The genus Eumanota was originally described by Edwards (1933) based on two species from Borneo. Soli (2002) added three species to the genus, two from Thailand and one from the Maluku Islands. Papp (2004) described two species, one from Taiwan and one from Papua New Guinea, while Hippa et al. (2005) described one species from Myanmar and one species from Sumatra, Indonesia, referring to an additional species from Pahang, in Malaysia, known from a single female that they did not formally describe. Our phylogenetic analysis of the Hippa et al.’s (2005) dataset including E. wolffae showed that this Neotropical species groups with the remaining species of the genus. We had nine shortest trees under equal-weight analysis, in which all Hippa et al.’s (2005) main conclusions for the manotines were exactly recovered: Manotinae monophyletic, Manota sister of the clade of the remaining genera, Paramanota sister to (Promanota + Eumanota),