Zootaxa 4237 (3): 454–470 ISSN 1175-5326 (print edition) http://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2017 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4237.3.2 http://zoobank.org/urn:lsid:zoobank.org:pub:BC7FE3FF-4B54-41FB-B74D-0AAAB784CD3B New species of Endecous (Grylloidea, Phalangopsidae, Luzarinae) from the Iguaçu National Park (Brazil), including bioacoustics, cytogenetic and distribution data PEDRO G. B. SOUZA-DIAS1,5, NEUCIR SZINWELSKI2, MARCOS FIANCO2, ELLIOTT CENTENO DE OLIVEIRA3, FRANCISCO DE A. G. DE MELLO4 & EDISON ZEFA3 1Departamento de Zoologia, Instituto de Biociências, Universidade de São Paulo, rua do Matão, travessa 14, n. 101, 05508-900, Cidade Universitária, São Paulo, SP, Brazil 2Centro de Ciências Biológicas e da Saúde, Universidade Estadual do Oeste do Paraná, rua Universitária, n. 2069, 85819-110, Universitário, Cascavel, PR, Brazil 3Programa de Pós-Graduação em Biologia Animal, Departamento de Ecologia, Zoologia, Universidade Federal de Pelotas, rua Capão do Leão s/n, 96010-900, Pelotas, RS, Brazil 4Departamento de Zoologia, Instituto de Biociências de Botucatu, Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP). 18618-000, Botucatu, SP, Brazil 5Corresponding author. E-mail: [email protected] Abstract In this study we describe two new species of cavicolous-straminicolous crickets for southern Brazil. Endecous chape n. sp. and E. naipi n. sp. are sympatric crickets that inhabit the Atlantic Semideciduous Forest of the Iguaçu National Park and adjacent areas. The descriptions were based on morphological characters, mainly from male genitalia and tegmina of adult males. Furthermore, we describe the chromosomes and the calling song of one of the new species, E. chape n. sp., presenting a discussion about the morphology of the phallic complex of Endecous and a distribution map for the species of the genus. The type-material is deposited in the Museu de Zoologia da Universidade de São Paulo (MZSP), and in the Coleção de Insetos do Departamento de Zoologia (Zoology Department Insect Collection), Universidade Estadual Paulis- ta—UNESP—Botucatu campus. Key words: Insect, Orthoptera, calling song, chromosomes, taxonomy, cricket Resumo Neste estudo descrevemos duas novas espécies de grilos cavícolas-estraminícolas para o sul do Brasil. Endecous chape n. sp. e E. naipi n. sp. são grilos simpátricos habitantes da Floresta Estacional Semidecidual do Parque Nacional do Iguaçu e áreas próximas. As descrições basearam-se em caracteres morfológicos, sobretudo da genitália e das tégminas dos ma- chos. Além disso, descrevemos os cromossomos e o som de chamado de uma das espécies, E. chape n. sp., apresentamos uma discussão sobre a morfologia do complexo fálico de Endecous, e um mapa de distribuição das espécies do gênero. O material-tipo está depositado no Museu de Zoologia da Universidade de São Paulo (MZSP) e na Coleção de Insetos do Departamento de Zoologia da Universidade Estadual Paulista—UNESP—campus Botucatu. Palavras-chave: Inseto, Orthoptera, som de chamado, cromossomos, taxonomia, grilo Introduction Endecous Saussure (1878) is one of the largest Luzarinae genera in South America, with 14 known valid species grouped in three subgenera, occurring through Brazil, Argentina, Bolivia and Uruguay (Gorochov, 2014; Cigliano 454 Accepted by D. Rentz: 14 Dec. 2016; published: 1 Mar. 2017 et al., 2016). The genus is widely distributed in Brazil, where its 10 recognized species are spread throughout the most parts of the country (Souza-Dias et al., 2014; Zefa et al., 2014; Bolfarini & Bichuette, 2015). Species of Endecous present tegmina with a functional stridulatory apparatus, except one species. E. apterus Bolfarini & Souza-Dias, 2014 has lost its tegmina as a result of troglobiomorphosis (Souza-Dias et al. 2014). Until now, only E. alejomesai Zefa, 2010, E. betariensis de Mello & Pellegatti-Franco, 1998, and E. itatibensis Rehn, 1918 had their calling songs recorded. These songs are composed of phrases, and including species-specific song parameters (de Mello & Pellegatti-Franco, 1998; Zefa, 2006; Zefa et. al. 2010). Zefa (2006) observed that populations of E. itatibensis from different regions present intraspecific variations in the carrier frequency and rhythm of the emitted signals. The Phalangopsidae comprises 975 valid species (Cigliano et al., 2016). Studies on cytogenetics are known for only 17 species. The karyotypes vary from 2n=11 to 2n=21, and with the basic X0 sex system (Zefa et al., 2010). Regarding Endecous, six of the 14 described species were karyotyped, varying in number, chromosome morphology and sex system determination: E. onthophagus (Berg, 1891) and E. itatibensis have 2n=19, X0 (Cardoso et al., 1984; Mesa & Garcia-Novo, 1997), E. alejomesai, E. cavernicolus Costa-Lima, 1940, and E. betariensis have 2n=21, X0 (Piza, 1945; Zefa et al. 2010), and E. ubajarensis Zefa, 2014 2n=14, X1X20 (Zefa et al. 2014). Endecous is considered a cavicolous-troglobitic cricket: its species are active at night on leaf litter, while during daytime remain inside caves, rock gullies, crevices, burrows, and general cavities, even in urban areas. Some species are generalists, with cavicolous and straminicolous populations, as E. itatibensis and E. aguassay Mews, 2008 and three species are troglobitic, as E. apterus, E. cavernicolus, and E. peruassuensis Bolfarini, 2015. For more information on the habitat occupation of Endecous see Souza–Dias et al. (2014), Zefa et al. (2014), and Bolfarini & Bichuette (2015), and references therein. Here we describe two new species of Endecous from the Iguaçu National Park (Brazil), and the calling song and chromosomes of one of the new species, with comments on the morphology of the phallic complex of the genus. Additionally, a distribution map of Endecous species is presented. Material and methods Taxonomy. Individuals of E. chape n. sp. and E. naipi n. sp. were collected at night on trails of the Iguaçu National Park, at the municipalities of Foz do Iguaçu (25°37'6.16"S, 54°28'56.34"W) and Céu Azul (25°9'20.96"S, 53°50'34.51"W), Paraná State. Individuals of E. chape n. sp. were also collected in the municipality of Concórdia, west of Santa Catarina State (27°15'38.50"S, 51°58'37.55"W). The national collecting permit was issued by the Instituto Chico Mendes de Conservação da Biodiversidade—ICMBio to PGBSD (SISBio 31497-1), and NS (SISBio 13543). The specimens were examined, described and compared using a Leica MZ-9.5 stereomicroscope. They were photographed immersed in 80% ethanol using a Leica MZ-16 stereomicroscope attached with a DFC-420 video camera. The same procedure was used for the male genitalia photographs. The software Leica Application Suite LAS v4.0 was used to take the images, and the software Helicon Focus 5.3 to stack them. Subsequently, the images were edited in a licensed Adobe Photoshop CS6 and GIMP (Gnu Image Manipulating Program) 2.8. The male phallic complexes were removed and treated with an aqueous solution of 10% KOH for 24h to remove muscular tissues, and stored in a small glass vial with 80% ethanol together with the respective specimen. The female copulatory papilla were removed and stored in a small glass vial with 80% ethanol together with the respective specimen. The genitalia terminology follows Desutter (1987), Desutter-Grandcolas (2003) and Souza- Dias (2015). Distribution map. The distribution map was built in the software QGIS 2.10. The biogeographic provinces followed the proposition of Morrone (2014), and the shapefiles were provided by Lowenberg-Neto (2014). The distribution data were obtained from Endecous’ previous studies, labels of studied specimens, field trips, and additional data provided by Dr. Marcio Bolfarini (Bolfarini & Bichuette, 2015). Bioacoustics. The calling songs was registered only for E. chape n. sp. Six males were recorded in laboratory with RR-US300 Panasonic tape recorder, and microphone placed about 10 to 20cm from the calling male. The room temperature was measured after each recording with an INCOTERM L-007/09 thermometer. The recorded songs were analyzed with Avisoft-SASlab Lite software, digitized at sampling frequency of NEW ENDECOUS FROM THE IGUAÇU NATIONAL PARK Zootaxa 4237 (3) © 2017 Magnolia Press · 455 22050 Hz, resolution 16 bit, fast Fourier transformation (FFT) – Length 256, frame 12.5%, window hamming and overlap 99.1%. We selected the best recordings of the calling song of each individual to analyze the phrase duration (i.e., the elapsed time from the first to the last pulse of a phrase); number of subphrases per phrase; subphrase period (elapsed time from the subphrase’s first pulse to the beginning of subsequent subphrase); number of pulses per subphrase; dominant frequency (highest intensity spectral component); and phrase amplitude. We consider a pulse as a train of sound cycles produced during inward movement of the forewings, and a phrase as a train of pulses. Cytogenetics. Cytological preparations were obtained only from E. chape n. sp., from the caeca of two males and three females, previously injected with a 0.05% aqueous solution of colchicine for 5h. The tissues were dipped in a hypotonic solution of sodium citrate (0.075 M) for 10 min prior to fixation in Carnoy I (3 parts of ethanol: 1 part of acetic acid). The cells were squashed in a drop of 45% acetic acid and stained with 0.5% lacto-acetic orcein. The chromosomes were photographed under a microscope Zeiss Lab. A1, and Axio Cam ERc5s. The chromosome terminology is according to Levan et al. (1964). Abbreviations. Male genitalia. Arc, ectophallic arc; Arm, pseudepiphallic arm; D.Ec.P., dorsal ectophallic projection; Ect.Ap., ectophallic apodeme; Ect.F., ectophallic fold; End.Ap., endophallic apodeme; End.Sc., endophallic sclerite; M.S., membranous sphere; PsP1, pseudepiphallic paramere 1; PsP2, pseudepiphallic paramere 2; V.E c.P., ventral ectophallic projection; Sc.A, sclerite A of the pseudepiphallic arms. General morphology. I, II, III, anterior, median, posterior (leg, tarsomere); DD, LL, dorsal disc, lateral lobe of pronotum; FW, forewing; F, femur; T, tibia.