Article. Rhithrogena sartorii, a new mayfly species (Ephemeroptera: Heptageniidae) from North Africa

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Zootaxa 3139: 63 68 (2011) www.mapress.com/zootaxa/ Copyright 2011 Magnolia Press Article ISSN 1175-5326 (print edition) ZOOTAXA ISSN 1175-5334 (online edition) Rhithrogena sartorii, a new mayfly species (Ephemeroptera: Heptageniidae) from North Africa SONIA ZRELLI 1,3, MICHEL SARTORI², MUSTAPHA BEJAOUI 1 & MONCEF BOUMAIZA 1 1 Unit of Hydrobiology, Laboratory of Environment Biomonitoring (LBE), Faculty of Sciences of Bizerta, 7021, Jarzouna. Tunisia. E-mails: zr_sonia@yahoo.fr, moncef_boumaiza@yahoo.com, bejaouimustapha@yahoo.fr 2 Museum of Zoology, Palais de Rumine, Place Riponne 6, CH-1005 Lausanne, Switzerland. E-mail: michel.sartori@vd.ch 3 Corresponding author. E-mail: zr_sonia@yahoo.fr Abstract A new species, Rhithrogena sartorii Zrelli & Boumaiza, sp. nov. is described in subimaginal, egg and nymphal stages from material collected in Northern Tunisia. This species shows affinities with members of the insularis species group. Features distinguishing the new species from other North African species, as well as from Rhithrogena insularis (from Corsica) and Rhithrogena nuragica (from Sardinia) are discussed. Keys words: Mayflies, taxonomy, morphology, egg, Rhithrogeninae, Khroumirie, Tunisia Introduction Recently, the generic situation in Heptageniidae of the world has been discussed by Webb and McCafferty (2008). The genus Rhithrogena Eaton, 1881 belongs to the subfamily Rhithrogeninae and includes ca 150 species mostly distributed in Palearctic region (Barber-James et al, 2008), with a high degree of endemism in mountain areas and on islands. Larvae of the genus are easily recognized by the shape of the first pair of gills which meet or overlap ventrally to form a friction disk (Webb & McCafferty, 2008). In North Africa, this genus is represented by 5 species: R. ourika Thomas & Mohati, 1985, R. ayadi Dakki & Thomas, 1986, R. ryszardi Thomas et al., 1987, R. giudicelliorum Thomas & Bouzidi, 1987 and R. mariae Vitte, 1991. Up to now, only R. mariae is known at the nymphal stage and belongs to the semicolorata group; R. ryszardi belongs to the germanica group and the three other species have not been assigned to any group (Sartori & Hughes, 2007). In Tunisia, the genus Rhithrogena is mentioned in ecological papers (Boumaiza & Thomas, 1986), checklist on aquatics insects (Thomas, 1998) or hydrobiological contributions (Kraiem, 1986). However, no specific identification of this material has been proposed so far. In the present study, an investigation of the streams in Northern Tunisia (Zrelli et al, 2011) resulted in the discovery of several Rhithrogena populations. After careful examination, it appeared that these populations belonged to a new species. Mature nymphs, eggs, and subimagos are described. The new species presents affinities with the insularis species group, i.e. R. insularis Esben-Petersen, 1913 from Corsica and R. nuragica Belfiore, 1987 from Sardinia. Materiel and methods The study sites are located in Khroumirie in Northern Tunisia (Fig. 1). The mayflies were preserved in ethanol (70%), some were dissected under the stereo microscope and were mounted on slides in Canadian balsam after a short stay in Creosote solution. The male and female subimagos were obtained by rearing in the laboratory. Unfortunately, no adults were obtained. Accepted by F.F. Salles: 17 Nov. 2011; published: 21 Dec. 2011 63

Egg procedure for Scanning Electronic Microscope study follows Ubero-Pascal & Puig (2007). Eggs were mounted, sputter coated with gold-palladium and finally examined with JSM-6300F scanning electron microscope with working voltages of 5 KV. The holotype and some of the paratypes are housed in the Museum of Zoology, Lausanne, Switzerland (MZL). Other paratypes are deposited in the Collection of the Laboratory of Environment Biomonitoring (LBE), Unit of Hydrobiology,Faculty of Sciences of Bizerta, Tunisia. FIGURE 1. Location of the Rhithrogena sartorii sp. nov. sampling sites in Northern Tunisia: 1. Ennour. 2. Rennagha. 3. Berbeg. 4. Ghezala. Rhithrogena sartorii Zrelli & Boumaiza, sp. nov. (Figs. 2 19) Material examined. Holotype: 1 nymph (preserved in ethanol), Ennour, 36.80072N 8.65871E 418 m., 28 IV 2010, S. Zrelli (MZL). Paratypes: same data as the holotype: 30 nymphs, 1 male subimago, 2 females subimagos (MZL, LBE). Other material examined. Same locality 24 V 2005, 1 nymph, 30 IV 2006, 1 nymph; Berbeg, 36.74968N 8.69693E, 558 m., 21 V 2005, 1 nymph; Rennagha, 36.85997N 8.72096E, 58 m. 29 VII 2005, 1 nymph; Ghezala, 36.64313N 8.69852E, 229m, 24 V 2005, 1 nymph. Description. Nymph. Body length of final instar, excluding caudal filaments, up to 4.5 mm and 7.5 mm for male and female nymphs respectively. Cerci, 5.0 7.3 mm. Color: body brownishwith visible tracheationonthe wholebody. 64 Zootaxa 3139 2011 Magnolia Press ZRELLI ET AL.

Head dark brown, subtrapezoidal with convex sides and rounded hind corners (Fig. 2). Labrum narrow, with distinct anterior teeth (Fig. 3s), 9 11 comb-shaped bristles on fore margin of maxilla with 6 7 teeth (Fig. 4). Glossae subovoidal with rounded apex (Fig. 5s). FIGURES 2 10. Rhithrogena sartorii sp. nov., larva: 2. Head, dorsal. 3s. Labrum, dorsal face 4. Comb-shaped setae from maxillary lacinia. 5s. Labium (dorsal view). 6. Hind femur. 7s. Bristles from the margin and the central part of hind femur. 8. First abdominal sternite. 9. Posterior margin of tergum V. 10a. Gill I. 10b. Gill II. 10c. Gill VII. R. insularis, 3i. 5i. 7i. R. nuragica, 3n. 5n. 7n. NEW RHITHROGENA SPECIES FROM TUNISIA Zootaxa 3139 2011 Magnolia Press 65

FIGURES 11 13. Rhithrogena sartorii sp. nov., male subimago: 11. Styliger plate and gonopods in ventral view. 12. Penis. 13. Titillator, lateral view. FIGURES 14 16. Rhithrogena sartorii sp. nov., egg: 14. General view. 15. Detail of the micropyle. 16. Polar cap. Dorsal surface of femora of all legs with few spines and an ovoid dark spot well visible on the center of the median pale area (Fig. 6). Bristles on hind femora very short with diverging margins (Fig. 7s). Tarsal claws with two or three denticles. Lateral sclerites of the first abdominal sternite rectangular with their anterior margin perpendicular to the body axis (Fig. 8). Posterior margin of abdominal tergites regularly dentate in central part, with few supplementary microdenticles (Fig. 9). All gill lamellae crenulated (10a, 10b and 10c). Plica on the first gill triangular with slightly concave apex (Fig. 10a). Male subimago. Body length 9.0 mm, forewing 8.0 mm. Cerci 9.0 mm. General color brown except the thorax, lighter. Eyes darkish brown. Sterna lighter than terga. Fore legs brown, femora with an elongated dark spot. Mid- and hind legs lighter, yellowish brown, and femora with a dark spot more elongated. Styliger plate and for- 66 Zootaxa 3139 2011 Magnolia Press ZRELLI ET AL.

ceps brown, posterior margin deeply incised in the middle (Fig. 11). Forceps uniformly brown, 3 segmented with a well defined basal segment, the 1st segment the longest, the 2nd and the 3rd equal in length. Genitalia as in Fig. 12. Titillators with apical margin bearing 3 4 teeth with 3 subapical teeth visible on their surface (Fig. 13). Female subimago. Body length 7.3 8.0 mm, forewings 5.7 7.0 mm. General color yellowish brown. Thorax brown, abdomen with color pattern similar to male. All legs with elongated spots on the dorsal side of femora. Eggs. Length: 180 200 µm, width: 150 170 µm. General shape ovoid (Fig. 14). Polar cap at one pole with numerous adhesive elements (KCTs). Micropyle with well developed margin (Fig. 15). Chorionic surface with several KCTs and with microgranules irregularly arranged (Fig. 16). Male and female imagos unknown Etymology. The species is named to honour Dr. Michel Sartori (Museum of zoology, Lausanne, Switzerland), who is a long-time investigator of the genus Rhithrogena. Affinities. Rhithrogena sartorii sp. nov. seems to belong to the insularis-group which included up to now only R. insularis Esben-Petersen 1913 and R. nuragica Belfiore 1987. According to Belfiore (1987) and Belfiore et al. (1992) the nymphs of both species shows a unique combination of characters: 1) lateral sclerites of the first abdominal sternite with anterior margin perpendicular to the body axis; 2) all gills crenulated; 3) a dark spot on the dorsal side of femora; 4) plica on the first gill triangular; and 5) the presence of well-developed tubercules on egg chorion. First 3 characters are also shared by species of the hybrida/hercynia species group, but the last two clearly differ from it. In the present state of our knowledge, R. sartorii is morphologically close to the species of the R. insularis group, but genetic data based on both mitochondrial and nuclear markers are congruent and place R. sartorii in a cluster quite distant from them (L. Vuataz, comm. pers.). When compared to the species found in Morroco, R. sartorii clearly differs from R. mariae and R. ryszardi which belong to the semicolorata and the germanica species group respectively, which gills II VII are not crenulated. It also differs from R. ourika, R. ayadi and R. giudicelliorum, mainly by the shape of the male titillators. The main diagnostic nymphal characters are described and compared with the two insularis-group species in Table 1. The nymph of the new species can be distinguished especially by the number of comb-shaped bristles and the number of teeth on the comb-shaped bristles at the tip of lacinia. The shape of the chorionic tubercles is somewhat intermediate between those of R. insularis and R. nuragica. KCTs lack the finger-like processes found in R. insularis, and the arrangement of the KCTs is less regular than in R. nuragica (Belfiore, 1987). TABLE 1. Some descriptive characters of Rhithrogena sartorii sp. nov. compared with two related species: R. nuragica and R. insularis. Character Rhithrogena sartorii sp. nov. Rhithrogena nuragica Belfiore 1987 Rhithrogena insularis Esben-Peterson 1913 Mean body length (mm) 6 7,5 6,8 Plica on the first gill Triangular with slightly Subtriangular Subtriangular concave apex Form of the second gill Rectangular Rhomboïdal Rhomboïdal Glossae Subovoidal Subtriangular and narrow Subovoidal and wide Emargination of the Labrum Smooth pronounced smooth Number of comb-shaped setae on the 9 11 8 9 7 8 maxilla Number of teeth on comb-shaped setae 6 7 9 10 9 11 of the maxilla Dark spot of the femora Well visible Hardly visible Visible Length of bristles on dorsal side of hind Very short Short Long femora Shape of setae on dorsal side of hind femora Divergent Divergent Narrow Ecology. Nymphs of the new species were collected in four different sites in strong current of streams and riffles of rivers with cobble substrates. Streams ranged in width from 4 m (Berbeg) to 10 m (Ennour), and water depth NEW RHITHROGENA SPECIES FROM TUNISIA Zootaxa 3139 2011 Magnolia Press 67

not exceeding 65 cm at all collecting sites. The very low mineralization of water (<0.15 PSU) can explain the absence of the new species in the other streams of Northern Tunisia which are more mineralized. Acknowledgments We are most grateful to Laurent Vuataz (Lausanne) for the rich comparative material of the insularis-group and to our colleagues Dr. Sadok Boulaaba and Dr. Jean-Luc Gattolliat for their help. Alain Thomas (Toulouse) and an anonymous reviewer are thanked for their comments. Thanks are extended to Mme Geneviève L Eplattenier for her technical help and to the staff of the Electronic Microscope Center (CME) of Lausanne University for their help and advices. The first author is grateful to all the members of Museum of Zoology in Lausanne and all the members of Laboratory Hydrobiology of Faculty of Sciences in Bizerta for their encouragement and advises. References Barber-James, H.M., Gattolliat, J.L., Sartori, M. & Hubbard, M.D. (2008) Global diversity of mayflies (Ephemeroptera, Insecta) in freshwater. Hydrobiologia, 595, 339 350. Belfiore, C. (1987) Heptageniidae from Corsica and Sardinia. Rhithrogena nuragica n. sp., R. eatoni Esben Petersen 1912, and R. insularis Esben Petersen 1913 (Ephemeroptera). Annales de Limnologie, 23 (2), 87 94. Belfiore, C., Picariello, O., Scillitani, G. & Cretella, M. (1992) Genetic divergence between two insular species of the genus Rhithrogena (Ephemeroptera, Heptageniidae). Fragmenta Entomologica, 23, 235 242. Boumaïza, M. & Thomas, A.G.B. (1986) Répartition et écologie des Ephéméroptères de Tunisie (1ère partie) (Insecta, Ephemeroptera). Archives de l Institut Pasteur de Tunis, 63 (4), 567 599. Dakki, M. & Thomas, A.G.B. (1986) Rhithrogena ayadi n. sp., Éphéméroptère nouveau du Moyen Atlas marocain (Heptageniidae). Annales de Limnologie, 22 (1), 27 29. Kraiem M. (1986) Contribution à l'étude hydrobiologique de trois cours d'eau du Nord-Ouest de la Tunisie. Présentation, physico-chimie et aperçu faunistique. Bulletin Mensuel de la Société Linnéenne de Lyon, 55, 96 104. Sartori, M. & Hughes, SJ. (2007) Description of a peculiar Rhithrogena nymph from the Iberian Peninsula (Ephemeroptera, Heptageniidae). Limnética, 26 (2), 435 440. Thomas, A.G.B., Vitte, B. & Soldán, T. (1987) Rhithrogena ryszardi n. sp., Éphéméroptère nouveau du Moyen Atlas (Maroc) et redescription de Rh. soteria Navás, 1917 (Heptageniidae). Annales de Limnologie, 23 (3), 169 177. Thomas, A.G.B. & Mohati, A. (1985) Rhithrogena ourika n. sp., Éphéméroptère nouveau du Haut Atlas marocain (Heptageniidae). Annales de Limnologie, 21 (2), 145 148. Thomas, A.G.B. & Bouzidi, A. (1986) Trois Éphéméroptères nouveaux du Haut Atlas marocain (Heptageniidae, Baetidae, Leptophlebiidae). Bulletin de la Société d'histoire Naturelle de Toulouse, 122, 7 10. Thomas, A.G. B. (1998) A provisional checklist of the mayflies of North Africa (Ephemeroptera). Bulletin de la Société d'histoire Naturelle de Toulouse, 134, 13 20. Ubero-Pascal, N. & Puig, M.A. (2007) Egg morphology update based on new chorionic data of Potamanthus luteus (Linnaeus), Ephemera danica Müller and Oligoneuriella rhenana (Imhoff) (Insecta, Ephemeroptera) obtained by scanning electron microscopy. Zootaxa, 1465, 15 29. Vitte, B. (1991) Rhithrogena mariae n. sp. Éphéméroptère nouveau du Rif marocain (Ephemeroptera, Heptageniidae). Nouvelle Revue d'entomologie (N.S.), 8 (1), 89 96. Webb, J.M. & McCafferty, W.P. (2008) Heptageniidae of the world. Part II. Key to the genera. Canadian Journal of Arthropod Identification, 7, 1 55. Zrelli, S., Boumaiza, M., Bejaoui, M., Gattolliat, J.-L. & Sartori, M. (2011) New reports of mayflies (Insecta: Ephemeroptera) from Tunisia. Revue Suisse de Zoologie, 118 (1), 1 8. 68 Zootaxa 3139 2011 Magnolia Press ZRELLI ET AL.