A new species of Bungona in Turkey (Ephemeroptera, Baetidae): an unexpected biogeographic pattern within a pantropical complex of mayflies Sroka, Pavel Godunko, Roman J. Rutschmann, Sereina Angeli, Kamila B. Frederico F. Salles, Gattolliat, Jean-Luc Zoosystematics and Evolution 2019 2019-01-25 95 1 1 13 http://zoobank.org/78B55194-D8FC-422C-A0C1-5715400FEEAE Sroka, Godunko & Gattolliat Sroka, Godunko & Gattolliat 2019 Insecta Baetidae Bungona CoL Animalia Bungona (Chopralla) pontica Ephemeroptera 0 1 Arthropoda species pontica sp. n. Chopralla  Type material.   Holotype.Male mature larva (IE CAS), TURKEY, Dipsiz OenueStream, Gemicilervillage, 500 m upstream from the village, in forest near Inebolu-Ayancikroad, 50 m a.s.l., 41°57.641'N, 33°53.026'E; 06.vii.2011, Sroka & Godunko leg. [locality code: TUR11/52].   Paratypes.2 mature male larvae (IE CAS: 1 larva in EtOH with some body parts mounted on a slide: mouthparts, legs, gills, tergum X, paraprocts, cerci; 1 larva dried and gilded as a SEM sample), same data as holotype; 1 mature male larva (IE CAS: in EtOH), TURKEY, IlisiStream, Inebolu-Ayancikroad, Yakaoerenvillage, vicinity of Abana town, 50 m a.s.l., 41°56.244'N, 34°13.360'E; 06.vii.2011, Sroka & Godunko leg. [locality code: TUR11/53]; 2 female larvae (MZL: 1 larva in EtOH: GBIFCH00272819 [FREDIE SR24E11] and 1 larva on a slide GBIFCH00272820 [FREDIE SR24E12]), same data as holotype.  Diagnosis. The prostheca of right mandible simple (not bifid) with several minute denticles apically; setae on the dorsal margin of the femur reaching 1/4 of the femur width; the surface of pronotum without tubercles. A detailed comparison with related species is presented in the Discussion.  External morphology of the larva. Body length approx. 4.5 mm-4.7 mm ( n= 2). Length of cerci ca 1.5-2.0 mm (0.3 xbody length), paracercus equal in length to cerci (Fig. 1A).   Figure 1. Bungona (Chopralla) ponticasp. n., habitus. ADorsal. BLateral.   Head.Labrum (Fig. 2A) ca 1.3 xwider than long, broadly rounded distally, with shallow medial emargination. Dorsal surface of labrum (Fig. 2A, right) with one long seta submedially (sI in Fig. 2A), apicolateral arc of three slightly shorter setae (sII in Fig. 2A); and with few short hair-like setae scattered on surface. Dense row of short branched setae present along anterior margin of labrum, longer setae anteromedially. Ventral surface of labrum (Fig. 2A, left) with group of fine hair-like setae near anterior margin. Hypopharynx with trilobed lingua apically, slightly longer than superlingua. Distal parts of lingua and superlingua covered with short, hair-like setae. Right mandible (Fig. 2C) with two partially fused incisor groups (outer incisor group (oig) in Fig. 2C, and inner incisor group (iig) in Fig. 2C), each equipped with three denticles. Right prostheca (prs in Fig. 2C) simple, not bifid, with several minute denticles apically. Numerous short setae present between prostheca and mola. Left mandible (Fig. 2B) with two mostly fused incisor groups, outer incisor group with four denticles (oig) in Fig. 2B, and inner incisor group with three denticles (iig) in Fig. 2B. Left prostheca (prs in Fig. 2B) robust, with about three short rounded denticles and comb-shaped structure apically. Numerous short setae present between prostheca and mola. Maxilla (Fig. 2H) with two-segmented maxillary palp. Segment II 1.8 xlonger than segment I, narrowing distally, and pointed at apex. Labium (Figs 2D-2G) with glossa slightly longer than paraglossa, inner margin of glossa with row of setae increasing in length apically. Second row of shorter setae present submarginally on ventral surface of glossa. Outer margin of glossa mostly without setation except for subapical part.   Figure 2. Bungona (Chopralla) ponticasp. n., mouthparts. ALabrum (right side dorsal, left side ventral). BIncisors of left mandible (dorsal, same scale bar for Band C). CIncisors of right mandible (dorsal). DGlossa and paraglossa (dorsal, same scale bar for D-G). ELabial palp (dorsal). FGlossa and paraglossa (ventral). GLabial palp (ventral). HMaxilla. Abbreviations: oig-outer incisor group, iig-inner incisor group, prs-prostheca, sI-submedial seta, sII-apicolateral arc of setae. Paraglossa along outer margin with row of setae, increasing in length apically. Groups of similar setae in subapical region present on both, dorsal and ventral surface. Along inner margin, short rows consisting of ca five setae submarginally also present on both, dorsal and ventral surface. Labial palp with segment I slightly longer than segments II and III combined. Segment I equipped with sparse short hair-like setae. Segment II with ca four stout setae in central part of dorsal surface, not expanded distoventrally. Segment III quadrangular, slightly distally expanded, with numerous setae on ventral surface, increasing in length and thickness distally.   Thorax.Colour whitish with distinct dark brown pattern (Fig. 1A, B). Surface of pronotum with short minute scales and without any protuberance (Fig. 6A). Legs whitish, tarsi slightly darker (Fig. 1B). Scales abundant on surface of femora, tibiae, and tarsi (Fig. 4A). Femur in all leg pairs with dorsal and ventral margin subparallel, ca 4 xlonger than wide. Dorsal margin with sparse row of 8 or 9 long, apically rounded setae, slightly widened apically (Fig. 3A, C, E). Length of setae ca 0.25 xfemur width. Occasional short setae present along anterior margin of femur. Tibia with patella-tibial suture (middle and hind leg; pts in Figs 3D, F, 4C, D) which is absent on foreleg (Fig. 3B, 4B). Position of patella-tibial suture at middle of tibia length in hind leg, and slightly more distally in middle leg (Fig. 3D, F). Length of row of long setae on anterior surface of tibia extending for ca 0.5 xlength of tibia in all legs. Width of row of long setae on posterior surface of tibia extends ca 0.5 xwidth of tibia in fore- and middle leg. In hind leg, row of long setae on posterior surface of tibia running parallel to outer margin of tibia, for same distance as row of setae on anterior surface. Angle between rows of setae on anterior and posterior margin of tibia more acute on hind leg compared to fore- and middle leg. (Fig. 3B, D, F). Short, bluntly pointed setae situated along inner margin of tibia.   Figure 3. Bungona (Chopralla) ponticasp. n., legs. AForeleg (dorsal, same scale bar for A, C, E). BBasal part of fore tibia (dorsal, same scale bar for B, D, F). CMiddle leg (dorsal). DBasal part of middle tibia (dorsal). EHind leg (dorsal). FBasal part of hind tibia (dorsal). Abbreviations: pts-patella-tibial suture. Tarsi equipped with several rows of long hair-like setae along outer margin. Most regular row apparent on anterior surface, and accompanied by more irregular rows posteriorly. Length of rows of setae reaching ca 0.5 xlength of tarsus in all legs. Occasional short spine-like setae present along inner margin of tarsus. Claws equipped with two rows of 3 or 4 flattened denticles, subapical striations, and minute subapical setae (Fig. 4E, F). Hind wing pads vestigial (Fig. 5B).   Figure 4. Bungona (Chopralla) ponticasp. n., legs. AScales on surface of legs. BForetibia (dorsal). CMiddle tibia (dorsal). DHind tibia (dorsal). EClaw. FDetail of claw apex. Abbreviations: s-scale, ss-scale socket, pts-patella-tibial suture.   Figure 5. Bungona (Chopralla) ponticasp. n., thorax and abdomen. ASetae on abdominal sterna IV, V and VI. BPart of metathorax with vestigial hind wing pad. CSurface and posterior margin of abdominal terga II, V and VIII. DParaproct. EAbdominal tergum X. FGills.   Abdomen.Colour pale whitish with dark brown pattern (Fig. 1A). Tergite I pale, with dark stripe along posterior margin. Tergites II-VI mostly dark, with tiny paired pale dots submedially and several larger pale areas medially, submedially, and laterally. Tergites VII-VIII mostly pale, darker stripes along posterior margins. Tergites IX-X darker, with pale area anteriorly on tergite IX. Sternites pale whitish, with darker longitudinal stripes sublaterally (Fig. 1B). Tergites equipped with numerous elongate scales, scale bases and short hair-like setae on surface (Fig. 5C). Posterior margin of tergites bear triangular spines (Fig. 5C); limited to lateral side on tergite I, larger and more elongated spines on tergites II-VII generally with median spines shorter than lateral, tergites VIII and IX similar to previous ones except central spines more reduced or completely absent. Spines on posterior margin of tergite X in two groups laterally and further two groups submedially (Fig. 5E). Sternites also equipped with scales and scale bases occasionally scattered over the surface. Posterior margins of sternites IV to IX with triangular spines, very reduced on sternite IV, more distinct on posterior segments; spines absent in segments I-III. Row of conspicuous long setae present on sternites IV-VI (row of shorter setae also present on sternite III; Fig. 5A). Gills (Fig. 5F) present on segments I-VII, slightly asymmetrical, with indistinct tracheation, apically pointed, margins occasionally bearing short setae. Brownish line medially on dorsal surface, parallel with medial trachea, not distinguishable on gills I and VII. Paraprocts (Fig. 5D) with six pronounced marginal spines sometimes accompanied with 1-2 smaller ones. Surface equipped with sparse scales and scale bases. Posterolateral extension with few small marginal spines, absent in some specimens. Caudal filaments (Fig. 1A, B) whitish with dark rings on segment margins. Distal margin of each segment equipped with pointed spines and scales. Outer margin of cerci bears enlarged spines on every second segment. Secondary swimming setae present.  Etymology.  "Pontus"in Latin means "Black Sea" in reference to the geographical region where the type material of the new species was collected.  Habitat and ecology. Larvae were found in two slightly eutrophic small streams of different size, the Dipsiz Oenueand Ilisistreams. Both small streams flow in northern direction towards the Black Sea within shallow valleys in the westernmost part of the Pontic Mountains (Kuzey Anadolu Daglari). The slopes surrounding both valleys are relatively steep, formed by hills reaching up to 450 m a.s.l. (Fig. 7B) and are densely overgrown by the typical Northern Anatolian conifer and deciduous forests (Euxine-Colchic deciduous forests ecoregion). The Dipsiz Oenuestream at the type locality at 50 m a.s.l. is small, only approximately 0.8-1.5 m wide, and partly shaded by vegetation (Fig. 7A). The bottom consists of relatively coarse stony substratum, partly covered by detritus in the littoral region. The current velocity was approximately 0.5 m/s and the water temperature reached 18 °C (measured ca 5 cm below the water surface).   Figure 6.Difference in the arrangement of the posterior margin of pronotum between Bungona (Chopralla) ponticasp. n. ( A) and Bungona (Chopralla) liebenauae( B).   Figure 7. AType locality of Bungona (Chopralla) ponticasp. n. (Dipsiz Oenuestream near Gemicilervillage). BValley of Dipsiz Oenuestream approximately 400 m downstream from the type locality. The Ilisistream, at the collecting site, was up to 4-4.5 m wide, had a relatively high velocity current (up to 0.7 m/s), well-expressed stream discharge, and a bottom structure consisting of relatively coarse stony substratum with a low concentration of detritus. We can assume that the new species is probably very rare at the studied localities as well as in all Turkey. During extensive collecting trips in the Sinop Province in 2011 and 2017, only six larvae were found.  Bungona ponticasp. n. larvae co-occurred with mayfly larvae of  Epeorussp.,  Electrogenasp.,  Procloeon bifidum(Bengtsson, 1912),  Baetis fuscatus(Linnaeus, 1761), B. (Rhodobaetis) rhodani(Pictet, 1843),  B. vardarensisIkonomov, 1962,  Nigrobaetis digitatus(Bengtsson, 1912), and  Serratella ignita(Poda, 1761). Additional information on the species composition of the mayfly fauna within the Sinop and Kastamonu provinces was published by  Tanatmis(2004), Ertorun and Tanatmis(2004)and further east, in the rivers of the Trabzon Province by  Aydinli(2017). The presence of mature larvae at the beginning of July, indicates a flight period of B. (Ch.) pontican. sp. during the first half of the summer.  Molecular reconstruction. In total, 61 haplotypes were reconstructed, including 19 previously unknown haplotypes. The new sequences were deposited at GenBank (Acc. nos in Table 1). The cox1tree detected B. (Ch.) ponticasp. n. as a discrete lineage which is not nested within other Bungona (Chopralla)species (Fig. 9). Two other Bungona (Bungona)species, namely B. (B.) narillaand B. (B.) illiesi, formed a paraphyletic group. The  Cloeodesrepresentatives from South America (i.e.,  C. aymore,  C. barituensis,  C. ioachimi, and  C. itajara) together with  C. pseudogladiusfrom Madagascar formed a monophyletic clade. However, as the calculated branch support was very low, the phylogenetic relationships between species/genera remain mostly unsolved.   Figure 8.Distribution of Bungona (Chopralla)spp. Marked occurrence in Turkey encompass the position of both known localities of Bungona (Chopralla) ponticasp. n.   Figure 9.Molecular reconstruction including representative set of taxa of the  Cloeodes-complex (comprising Bungona (Chopralla) ponticasp. n.) and additional taxa of other lineages. Bayesian inference was used to reconstruct the tree based on the mitochondrial DNA barcoding gene cytochrome coxidase subunit 1. Bayesian posterior probabilities> 0.8 are indicated. Scale bar represents substitutions per site. Geographic origins of the specimens are indicated. Colours: green = Bungona (Bungona); blue = Bungona (Chopralla); purple = Bungona (Centroptella); yellow =  Cloeodes; white = other genera.