taxonID	type	description	language	source
3345934FFF9A6C41E7CA0B7EFD0C457B.taxon	description	(Herrich-Schäffer) * Lethocerus cordofanus Mayr MNHN Lethocerus delpontei De Carlo LEBIP Lethocerus maximus De Carlo LEBIP Limnogeton expansum MACN / MNHN Montandon Limnogeton fieberi Mayr MNHN / MRAC Limnogeton hedenborgi (Stål) MNHN / MRAC	en	Ribeiro, José Ricardo I., Ohba, Shin-Ya, Pluot-Sigwalt, Dominique, Stefanello, Fabiano, Bu, Wenjun, Meyin-A-Ebong, Solange E., Guilbert, Eric (2018): Phylogenetic analysis and revision of subfamily classification of Belostomatidae genera (Insecta: Heteroptera: Nepomorpha). Zoological Journal of the Linnean Society 182: 319-359
3345934FFF9A6C40E7CA08DBFDE942FF.taxon	description	NEPIDAE Curicta borellii Montandon LEBIP Curicta cf. pelleranoi LEBIP Curicta volxemi (Montandon) * LEBIP Laccotrephes japonensis (Scott) LEBIP Laccotrephes pfeiferiae (Ferrari) LEBIP Laccotrephes sp. * LEBIP Nepa cinerea Linnaeus LEBIP Nepa hoffmanni Esaki * LEBIP Ranatra brevicauda Montandon LEBIP Ranatra chinensis Mayr * LEBIP Ranatra heydeni Montandon * LEBIP Ranatra robusta Montandon LEBIP Ranatra sattleri De Carlo LEBIP	en	Ribeiro, José Ricardo I., Ohba, Shin-Ya, Pluot-Sigwalt, Dominique, Stefanello, Fabiano, Bu, Wenjun, Meyin-A-Ebong, Solange E., Guilbert, Eric (2018): Phylogenetic analysis and revision of subfamily classification of Belostomatidae genera (Insecta: Heteroptera: Nepomorpha). Zoological Journal of the Linnean Society 182: 319-359
3345934FFF9A6C40E7CA08DBFDE942FF.taxon	materials_examined	CHINA: Chekiang Province; JAPAN: Kumamoto, Tokyo, Alpes de Nikko; LAOS: Baudan BRAZIL: Rio de Janeiro; Rio Grande do Sul SOMALIA: Glohar BRAZIL: Rio Grande do Sul FRENCH GUYANA: Route de Pointe Combi REPUBLIC OF THE CONGO: Brazzaville; CAMEROON: Batouri, Nyaounderé; TANGANYIKA: Mlingano EGYPT; SUDAN; CAMEROON: Garoua SENEGAL: Kolda; EGYPT; REPUBLIC OF THE CONGO; CAMEROON: Yaoundé; UGANDA: Victoria Nyanza EGYPT; ETHIOPIA; CENTRAL AFRICAN REPUBLIC: Lamaboké; DEMOCRATIC REPUBLIC OF THE CONGO: Molindi River; REPUBLIC OF THE CONGO: Brazzaville; ZAMBIA: Muliba BRAZIL: Amazonas, Mato Grosso; FRENCH GUIANA: Piste Coralie BRAZIL: Rio Grande do Sul BRAZIL: Mato Grosso do Sul BRAZIL: Mato Grosso do Sul JAPAN: Kumamoto CHINA: Tianjin (Wuqing Country) CHINA: Tianjin (Wuqing Country) FRANCE: Vayral JAPAN: Hyogo BRAZIL: Mato Grosso do Sul JAPAN: Kumamoto BRAZIL: Mato Grosso do Sul BRAZIL: Mato Grosso do Sul; Rio Grande do Sul BRAZIL: Mato Grosso do Sul † List of specimen depositories is based on Arnett, Samuelson & Nishida (1993), except for DZRJ, LEBIP and MNRJ. DZRJ, Departamento de Zoologia, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; LEBIP, Laboratório de Estudos da Biodiversidade do Pampa, São Gabriel, Universidade Federal do Pampa, Rio Grande do Sul, Brazil; MNRJ, Museu Nacional, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil. such material was examined at 5 kV, without a coat of gold – palladium. DNA PREPARATION Insect specimens collected in the field were placed directly in 95 – 100 % ethanol and stored at − 20 ° C until processing. To amplify genes, genomic DNA was extracted from a single foreleg and associated muscles using a modified ethanol precipitation / resuspension protocol (Bender, Spierer & Hogness 1983) or the DNEasy tissue kit (Qiagen Inc.). In some cases, multiple individuals from the same species were extracted. All belostomatid vouchers were stored in 99.5 % ethanol pro-analysis (PA) at − 20 ° C deposited at the LEBIP (Laboratório de Estudos da Biodiversidade do Pampa, Universidade Federal do Pampa, Rio Grande do Sul, Brazil), except for W. rhomboides, which was dried and is deposited at the MNHN (Museum National d’Histoire Naturelle, Paris, France). For amplification of genes from W. rhomboides, genomic DNA was extracted following protocols adapted from those of Gilbert et al. (2007), Thomsen et al. (2009) and Lis, Ziaja & Lis (2011), which allowed us to recover amplifiable DNA from dried museum specimens that were up to 10 years old. The specimens were pinned again and redeposited in the collection as vouchers. PCR, SEQUENCING AND ALIGNMENT Modified primers based on Simon et al. (1994), von Dohlen & Moran (1995) and Simon et al. (2006) were used to amplify parts of the mitochondrial gene COI (~ 730 bp) and 16 S rDNA (~ 480 bp). Nuclear 18 S sequences were amplified using the primers Ns 1 and Ns 2 a (Barker et al., 2002) (~ 600 bp) (see Table 2). Amplification was carried out in a 25 - µL volume reaction, with 5 µL of Taq and Load Mastermix, 0.5 µL of each primer at 25 µM and 2 µL of extraction product. For 16 S loci, all reactions were performed using HifiTaq DNA Polymerase – an enzyme mixture that greatly increases fidelity and amplification of genomic targets. For COI reactions, a ‘ step-up procedure’ was used: 2 min at 95 ° C, five amplification cycles to improve DNA stock of 1 min at 95 ° C, 15 min at 45 ° C and 1 min at 72 ° C; followed by 35 cycles of 40 s at 95 ° C, 1 min at 51 ° C and 1 min at 72 ° C, with a final extension at 72 ° C for 5 min. The protocol for the 16 S rDNA region was: 94 ° C for 3 min and 80 ° C for 20 min (hot start), followed by 35 cycles of 94 ° C for 1 min, 50 ° C for 1 min and 72 ° C for 2 min, with a final extension at 72 ° C for 7 min. The protocol for 18 S rDNA was 94 ° C for 2 min, 35 cycles of 94 ° C for 30 s, 49 ° C for 30 s and 72 ° C for 30 s, with a final extension at 72 ° C for 7 min. Negative controls (no template) were always run simultaneously with our PCR experiments. All reaction mixtures were discarded when any DNA appeared in the negative control. Sequences were sent to the Genoscope (Centre National de Séquençage, Evry). The sequences obtained were read and cleaned with CodonCode (CodonCode Corporation, 2002 – 2015). All sequences were aligned using MUSCLE (Edgard, 2004), and MAFFT v. 7 (Katoh & Standley, 2013) for non-coding sequences. In all regions, gaps were checked manually and treated as missing data. This procedure yielded approximately 1830 bp per taxon, although sequences for some taxa were not complete (Table 1). All Belostomatidae and Nepidae sequences are original accessions, except Abedus breviceps Stål, 1862, Curicta scorpio Stål, 1862 and Nepa apiculata Uhler, 1862, which were taken from GenBank. NCBI accession numbers and references are summarized in Table 3. PHYLOGENETIC ANALYSES	en	Ribeiro, José Ricardo I., Ohba, Shin-Ya, Pluot-Sigwalt, Dominique, Stefanello, Fabiano, Bu, Wenjun, Meyin-A-Ebong, Solange E., Guilbert, Eric (2018): Phylogenetic analysis and revision of subfamily classification of Belostomatidae genera (Insecta: Heteroptera: Nepomorpha). Zoological Journal of the Linnean Society 182: 319-359
3345934FFF906C55E4FA0D57FD4C435E.taxon	materials_examined	Type genus: Diplonychus Laporte, 1833 ported tree clades conflicting with all input trees was the response variable and datasets and quality of those data were used as factors, under the null hypothesis that the different dataset and the presence of missing data, with different methods of analysis used to infer phylogeny, did not affect the existence of phylogenetic signal and, in turn, the genus-level relationships in Belostomatidae. A failure to reject H 0 suggests strong congruence among such treatments. Abbreviations: d. f. = degree of freedom; SS = sum of squares; MS = mean of squares Diagnosis: Diplonychini differs from other belostomatine suprageneric groups by the frons rounded or curved in dorsal view (Fig. 9 D, G, H), as well as the following male genitalia features: (1) the transverse bridge of basal plate of male genitalia clearly jointed and entire (Fig. 16 B); (2) plate of phallotheca as long as ventral diverticulum. Likewise, females of Diplonychini differ from the others by the presence of an ampulla located at the basal part of spermatheca (Fig. 21 A – C). Distribution: Africa, Australia, East Indies, southern Asia (Estévez & Ribeiro, 2011). Description: Measurements. – Total length (from apex of head to apex of abdomen at rest): from 8.2 to 27.7 mm. General coloration. – Almost uniformly brown. External morphology. – Body ovate with wings usually covering abdomen. Frons rounded or curved (Fig. 9 D, G, H); vertex without median longitudinal carina (Fig. 9 D); antennae with segments 2 and 3 not flattened ventrally, with fourth segment similar to or slightly more bulbous than prolongations of segments 2 and 3 (Fig. 11 B); frontogenal suture slightly convergent and opened distally (Fig. 9 D). Pronotum without longitudinal median carina (e. g. Fig. 9 D); prosternal keel usually poorly elevated, except for some Appasus species; hemelytra with rounded pruinose area; clamp of clavus with its outer projection overlapping inner part, always far from the margin of hemelytra (Fig. 14 A); its outer carina with three rows of microtrichiae along its external margin, covering small portion in dorsal view (Fig. 13 A, B); tile-like microtrichiae rounded at apex, never toothed along the margin of its apex (Fig. 12 B); foretarsi with two segments, externally usually appearing one-segmented, with segment 1 conspicuous, with two symmetrical grooves; claws vestigial; hind trochanters carinated, with short hairs or bristles along outer margins. Pilosity developed, covering half of connexivum, slightly constricted between spiracles, extending posteriorly along about half of or almost entire genital operculum; pubescence of ventral laterotergites 3 and 4 not attaining entire external margin; air straps lanceolate, with somewhat uniform width along its extension. Male genitalia: Phallosoma fused to ventral diverticulum; arms of phallosoma well developed, extending nearly to apex of ventral diverticulum, enclosing ventral diverticulum in some Appasus (Fig. 16 E), somewhat laterally directed; orifice strongly developed, dorsally located on apex of phallosoma; ventral diverticulum contiguous, never bilobed, with its apex without ventroapical protuberance, not showing spines or tubercles in ventral view; transverse bridge of basal plate of male genitalia clearly jointed and entire (Fig. 16 B); plate of phallotheca somewhat developed, fused to or close to ventral diverticulum. Female genitalia: Operculum of females with two tufts of setae on apex; apex of second valvulae with an inconspicuous spine; basal part of spermatheca without distinct apodemes, clearly with the presence of an ampulla; median vagina area below spermatheca without pouch (Fig. 21 A – C). Taxonomic notes: In general, members of the new tribe Diplonychini share with W. rhomboides the prosternal carina poorly elevated (‘ prosternal keel rounded’ according to Estévez & Ribeiro, 2011: 51) (character 27: 0> 1) and with H. pelocoroides the surface of apex of ventral diverticulum (in ventral view) without spines or tubercles (character 89: 0> 1). Our findings support Diplonychini trib. nov. as the sister group of the tribe Belostomatini by the following unambiguous homoplastic synapomorphies: (1) clamp of clavus with its outer projection not flattened, overlapping the inner part (similar to Fig. 14 A) (character 31: 2> 3); and (2) outer carina of the clamp with three rows of microtrichiae along its external margin (character 32: 2> 1) (Fig. 13 A, B). The new tribe Diplonychini shares with other members of the subfamily Belostomatinae the foretarsi bearing both claws vestigial (character 44: 0> 3), phallosoma fused with ventral diverticulum (character 71: 0> 1), ventral diverticulum contiguous, never bilobed (character 85: 0> 1) (Fig. 16 E), and apex of second valvulae with a spine and / or protuberance (character 94: 0> 1). All morphological character states cited above are exclusive to this subfamily (Fig. 6).	en	Ribeiro, José Ricardo I., Ohba, Shin-Ya, Pluot-Sigwalt, Dominique, Stefanello, Fabiano, Bu, Wenjun, Meyin-A-Ebong, Solange E., Guilbert, Eric (2018): Phylogenetic analysis and revision of subfamily classification of Belostomatidae genera (Insecta: Heteroptera: Nepomorpha). Zoological Journal of the Linnean Society 182: 319-359
