Anais da Academia Brasileira de Ciências (2019) 91(1): e20180047 (Annals of the Brazilian Academy of Sciences) Printed version ISSN 0001-3765 / Online version ISSN 1678-2690 http://dx.doi.org/10.1590/0001-3765201920180047 www.scielo.br/aabc | www.fb.com/aabcjournal Redescription of immatures of Dasyhelea flavifronsGuérin-Méneville (Culicomorpha: Ceratopogonidae) and new contribution to the knowledge of its larval habitats FLORENTINA DÍAZ1, CAROLINA MANGUDO2, 3, RAQUEL M. GLEISER4, 5 and MARÍA M. RONDEROS1 1Centro de Estudios Parasitológicos y de Vectores/CEPAVE, Facultad de Ciencias Naturales y Museo/UNLP, Consejo Nacional de Investigaciones Científicas y Técnicas/CONICET, Boulevard 120, s/n, e/61 y 62 La Plata, Buenos Aires, Argentina 2Instituto de Investigaciones en Energía No Convencional/INENCO, Universidad Nacional de Salta, Av. Bolivia, 5150, A4400FVY Salta, Argentina 3Instituto de Investigaciones en Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado, 751, Orán, 4530 Salta, Argentina 4Universidad Nacional de Córdoba, CONICET, Instituto Multidisciplinario de Biología Vegetal, Centro de Relevamiento y Evaluación de Recursos Agrícolas y Naturales/CREAN, Av. Valparaíso, s/n, CC 509, 5000 Córdoba, Argentina 5Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales, Cátedra de Ecología, Av. Vélez Sársfield, 299, Córdoba, Argentina Manuscript received on January 15, 2018; accepted for publication on May 11, 2018 How to cite: DÍAZ F, MANGUDO C, GLEISER RM AND RONDEROS MM. 2019. Redescription of immatures of Dasyhelea flavifrons Guérin-Méneville (Culicomorpha: Ceratopogonidae) and new contribution to the knowledge of its larval habitats. An Acad Bras Cienc 91: e20180047. DOI 10.1590/0001-3765201920180047. Abstract: The fourth instar larva and pupa of Dasyhelea flavifrons Guérin-Méneville are redescribed, illustrated, and photomicrographed using binocular, phase-contrast, and scanning electron microscopy. Comparisons with the American species of the grisea group were made. The immatures were collected by using a siphon bottle in tree-holes and from water collected in dead snail shells in Salta Province, Argentina, transported to the laboratory and there bred to the emergence of the adults. Details on larval habitats are given. These are the first records from Argentina and in gastrotelmata. Key words: Ceratopogonidae, Dasyhelea flavifrons, gastrotelmata, immatures, tree holes. INTRODUCTION Adult females feed on nectar from flowers and pollen. Moreover, some are important pollinators The biting midges Dasyhelea Kieffer are common of commercial trees such as cacao (Theobroma and mainly diurnal dipterans occurring in all zoogeographical regions, except Antarctica. The cacao) and rubber (Hevea brasiliensis) (Borkent larvae of Dasyhelea develop mostly in shallow and Spinelli 2007, Córdoba et al. 2013). aquatic and semiaquatic habitats, and in general Taxonomically, the recognition of subgenera feed on detritus and algae, though a few species and/or groups of species have been proposed for feed in dead insects (Borkent and Spinelli 2007). all regions of the world. Borkent (2016) listed 77 species for the Neotropics, of which only 12 are Correspondence to: Florentina Díaz known also from their immatures. Subsequently, E-mail: [email protected] / [email protected] Grogan et al. (2016) described 13 new species ORCid: https://orcid.org/0000-0002-5991-6756 from Guadeloupe, and Díaz et al. (2017) described BIOLOGICAL SCIENCES An Acad Bras Cienc (2019) 91(1) FLORENTINA DÍAZ et al. IMMATURES OF Dasyhelea flavifrons two species from Peru and Brazil, all based on Winnertz, but also incompletely. The aim of this adults. Recently, Díaz et al. (2018) described two work is to provide the full redescription of these species, including their immature, from the Yungas immature in accordance with modern standards ecoregion, Argentina. and to present the first records from Argentina and The phytotelmata are aquatic in gastrotelmata as larval habitat. microenvironments formed by the accumulation of water in any part of the body of plants (e.g. MATERIALS AND METHODS leaves, flowers, stems, trunks, tree holes and STUDY AREA artificial containers). The gastrotelmata are aquatic microhabitats formed by the collection of water San Ramón de la Nueva Orán (hereafter Orán), (usually from rainfall) in the shells of dead snails is a city established in northwestern Argentina (Janetzky et al. 1995). In spite of their small size, near the border with Bolivia (23º08’ S, 64º20’ W, snail shells retain water longer than any other natural elevation 337 m). The region has a subtropical container, providing a relatively permanent habitat climate, with an average summer temperature of (Lounibos 1980). The communities of organisms 27.7 °C and winter temperature of 16.4 °C, and that live in these microenvironments are simple a mean annual rainfall of 1,000 mm, occurring and based on detritus; the macroinvertebrates most mostly from October to April, the warmer months. frequently found are insects, immature stages of The study area has been described in more Diptera being dominant (Campos et al. 2011). The detail in Mangudo et al. (2015). Briefly, Orán is knowledge of Ceratopogonidae from Argentina located in the pedemontane floor of the Yungas that breed in these microenvironments is poor; only subtropical montane moist forest (Brown et al. 7 of the 31 genera registered (Borkent and Spinelli 2001), which remains mostly to the East and North 2007) have been associated with phytothelmata: of the city. Besides urbanization, other human Atrichopogon Kieffer, Forcipomyia Meigen, related ecological modifications include industrial Dasyhelea Kieffer,Culiciodes Latreille, Stilobezzia development, agriculture and forestry (Brown et al. Kieffer, Bezzia Kieffer, and Palpomyia Meigen 2001). The city is characterized by a densely built (Campos et al. 2011). central area surrounded by suburban areas with During a sampling program focused on the bigger gardens and more trees; throughout the city collection and study of Diptera that was carried out most buildings are low. in Salta province between 2011 and 2012, larvae ENTOMOLOGICAL SAMPLING and pupae were collected from tree holes and dead snail shells, and reared to adults. Based on Larvae, larval exuviae and pupae were collected the characters of adult males, they were identified between February and March in 2011 and 2012, as Dasyhelea flavifrons. This is considered a wide as a part of a larger study on mosquito (Diptera: spread Holarctic arboreal species, whose larvae Culicidae) larval habitats. Samples were collected have been reported in Europe as terrestrial and from tree holes using a siphon bottle (Müller and inhabiting sap flows, mushrooms and tree holes Marcondes 2006, Mangudo et al. 2010) and from (Dominiak and Szadziewski 2010). However, the snail shells by overturning and examining the original description and the different redescriptions water content from each shell in a white plastic of immatures are incomplete, except for the tray, from which larvae and pupae were collected contribution of Keilin (1921), who described all with a pipette. Tree holes were located in the city stages of this species under the name D. obscura and in yunga forest patches to the north and east of An Acad Bras Cienc (2019) 91(1) e20180047 2 | 14 FLORENTINA DÍAZ et al. IMMATURES OF Dasyhelea flavifrons town, while snail shells were collected from forest various tree species); Borkent and Grogan 2009: 11 patches only. For details on tree holes selection see (in Nearctic catalog; distribution), Borkent 2016: Mangudo et al. (2015, 2018) and for snail shell 65 (in online World catalog); Grogan et al. 2016: sampling see Mangudo et al. (2017). 208 (in review of Guadeloupe records, key). Larvae were preserved in ethanol 80% and Dasyhelea oppressa Thomsen 1935: 285 (New pupae were kept alive in the laboratory isolated in York); Waugh and Wirth 1976: 230 (in revision plastic vials (2 mm) holding water from the larval of eastern United States Dasyhelea; distribution, habitat and containing a piece of humid filter paper, larval habitats – tree holes, sap of oak, elm); Graves to maintain the humidity inside the vials, until adult and Graves 1985: 88 (USA: North Carolina, larval emergence. Emerged adults were maintained alive habitats – shelf fungi); Wilkening et al. 1985: 519 for 24 hours to ensure the development of their final (Florida records); Hribar and Grogan 2005: 231 pigmentation. Adults and their respective exuviae (Monroe County, Florida records); Szadziewski and were stored in vials containing 80% ethanol. Dominiak 2006: 142 (as synonym of C. flavifrons Larval, pupal exuviae and adults were mounted in Guérin-Méneville); Borkent and Grogan 2009: 11 Canada balsam following the technique described (in Nearctic catalog; distribution; as synonym of by Borkent and Spinelli (2007). The technique of D. flavifrons). Ronderos et al. (2000, 2008) was followed to prepare REDESCRIPTION OF THE FOURTH INSTAR LARVA larvae for scanning electron microscopy (SEM). A (FIGS. 1-3, 5) (N=3) camera lucida was used to make illustrations with pen and ink. Photomicrographs were taken with a Total length 4.5 mm. Color in life whitish with the Micrometrics SE Premium digital camera, through head capsule brown, short, wide, tapering to apex a Nikon Eclipse E200 microscope. For larval terms (Fig. 1c, d); chaetotaxy as in Fig. 1b, c. HL 0.29- see Díaz et al. (2018) and for pupae see Borkent 0.30 (0.295, n=3) mm; HW 0.215-0.220 (0.216, (2014). n=3) mm; HR 0.255-0.257 (0.256, n=3); SGW The plates were made in TIFF format in Adobe 0.17-0.19 (0.18, n=3) mm; SGR 1.21 (1.16-1.26, Photoshop version14. The material studied is n=3). Antenna cylindrical (Figs. 1b-d, 2a). Labrum deposited in the División Entomología, Museo de (Fig. 1c, d) 0.80 times longer than wide; palatum La Plata (MLPA), La Plata, Argentina. (Figs. 1b, 2a, 3a) with four pairs of sensillae campaniformia (Figs. 1d, 3b, c), posterior of them RESULTS three pairs of sensillae coeloconica (Figs. 1b, d, 2a, b, 3b, c): mesal one serrate, others simple; messors Dasyhelea flavifrons (Guérin-Méneville) (FIGS.