New species of Sycorax (Diptera: Psychodidae) from the Brazilian Amazon

. Moth flies of the subfamily Sycoracinae have been associated with anurans. Females of some species have been found feeding on the blood of these vertebrates. Here we describe a new species of the genus Sycorax Curtis from a preserved site of Amazon Forest in the city of Manaus, Amazonas, Brazil, with one male specimen collected in the dorsum of the frog Anomaloglossus stepheni (Martins).


INTRODUCTION
Among the Psychodidae, the subfamily Sycoracinae comprehend a group of moth flies which adult of some species have been collected in association with anurans (Desportes, 1942;Bravo & Salazar-Valenzuela, 2009;Curler et al., 2015;Ježek et al., 2015).These associations showed evidence of females feeding on blood of these vertebrates and males with an aggregation behaviour, possibly to mate.This subfamily has 55 current species described (Bravo et al., 2023;Faé et al., 2023), distributed in four genera: Sycorax Curtis, 1839, with 43 known species, Parasycorax Duckhouse, 1972, with six described species, Falsosycorax Bravo & Araújo, 2023, with five species, and the monospecific Aposycorax Duckhouse, 1972.In the Neotropics, 12 species of Sycorax have been described (Bravo et al., 2023).Here we describe a new species from central Amazonia and discuss its possible association with the frog Anomaloglossus stepheni (Martins, 1989).

MATERIAL AND METHODS
All specimens were collected in the grids used for long term ecological studies of two forested areas of the municipality of Manaus, Amazonas, Brazil: Reserva Florestal Adolpho Ducke and Fazenda Experimental da UFAM.The codes L3 3500 and NS01/100 indicate specific points on those grids which coordinates are 02°56′28.4″S,59°56′10.4″Wand 02°39′40.9″S,60°03′49.8″W,respectively.Specimens were collected using Malaise (one male and one female) and CDC light traps (three males and one female), except for one male that was manually collected while resting on top of the frog Anomaloglossus stepheni.Specimens were preserved in alcohol 70%.They were mounted using the following methodology: diaphanization in KOH 10% at 50℃ for approximately 3 hours, and immersion in the following solutions for 10 minutes each: water, 10% acetic acid, distilled water, 70% alcohol, absolute alcohol, clove oil and 5 minutes in butyl acetate, and finally slide mounted using Canadian balsam.Photomicrographs were captured using a Leica DM5500 B optical microscope, with Leica DFC295 camera and the z-stacking function included in Leica Application Suite LAS V3.6 digital imaging software.The specimens will be deposited at the Entomological Collection of the Instituto Nacional de Pesquisas da Amazônia (INPA), Manaus, Amazonas, Brazil, and at the Entomological Collection Prof. Johann Becker of the Museu de Zoologia da Universidade Estadual de Feira de Santana, Feira de Santana, Bahia, Brazil (MZFS).General morphology follows Cumming & Wood (2017) with specifications for the family of Kvifte & Wagner (2017).Etymology: The epithet 'manauara' is a noun in apposition, referring to the gentilic used for people born in the city of Manaus.Female.Similar to male, except for the following: Head (Fig. 15) 1.25 times broader than long; eyes rounded, discrete groove close to insertion of the antenna, interocular distance 10 times diameter of ommatidium.Palp segment 2 with a cluster of Newstead sensilla on medial surface.Cibarium with one pair or inconspicuous spines.Antennae with one ascoid inserted on inner side on flagellomeres I-IV, ascoids more discrete than in males.Wing (Fig. 16) similar to that of male, except for length 1.69 mm, 3.18 times longer than width, R₁ reaching wing margin beyond level of R₂₊₃ fork, R₂ converging to R₁ on basal third then diverging from it apically, M₃ surpassing level of R₂₊₃ fork, M₄ reaching wing margin at level of M₁₊₂ fork.Tergite 8 and sternite 8 without bristles.Sternite 8 with two pilose lobes (Fig. 17); tergite 9 sub-rectangular and pilose; cercus longer than broad in dorso-ventral view; hypoproct with apex rounded and micropilose.Sternite 9 (genital fork) well sclerotized, 2 times longer than greatest width (Fig. 18); spermathecal duct slightly sinuous, terminating in sclerotized button, which gives way to an amorphous tenuous sac (Fig. 18).

Sycorax manauara
Discussion: The sinuous shape of genital filaments of the aedeagus seen in the new species is unique, although it resembles those of the five Falsosycorax species, which are known from the Andes of Ecuador and Colombia.However, the new species lacks the median dorsal process of the aedeagus, that can be found in these five species, and lacks, too, the two or three long basal spines in the gonostylus present in the Andean species.Instead, it has two spiniform setae, not developed as spines.A long subapical seta, near de apical spine, is present in most species of Sycorax, but it is absent in the new species, as well as in F. andicola (Young, 1979) andF. trispinosa (Young, 1979).A long bristle in the gonocoxite as described to the new species is only observed in three other species of the genus, Sycorax bravoi Santos, Ferreira & Falqueto, 2011, Sycorax longispinosa Bravo, 2007, andSycorax malayensis Quate, 1962.
The number and disposition of spines on the gonostyli is an important character for Sycoracinae species diagnosis.Along with Sycorax manauara sp.nov., other nine Neotropical species of Sycorax (S. longispinosa, S. confusa Bravo, Rocha & Santos, 2010, S. bahiensis Bravo, 2003, S. cariacicaensis Santos & Bravo, 2009, S. tuberculata Santos, Bravo & Falqueto, 2013, S. assimilis Barretto, 1956, S. espiritosantensis Santos & Bravo, 2009, S. bravoi and S. canaanensis Santos, Bravo & Falqueto, 2013) have also only an apical spine on each gonostylus, but their distally bifid aedeagus are not developed in long genital filaments as seen in the new species.Sycorax longispinosa and S. confusa are also easily differentiated from the new species by the long and strong setae near the paramere apex, measuring twice the length of the paramere.In the new species the paramere has two long setae near its apex, one on the dorsal and one on the ventral surface of the paramere, both shorter than the length of the paramere.Sycorax canaanensis have one long and one short setae near the apex of the paramere, but both on the dorsal surface.The other six neotropical species of Sycorax have only a single long setae or no long setae at all on the paramere.Also, the overall shape of the paramere is unique for all these species.
Although we did not test for the identification of DNA samples of vertebrate blood in the female specimens, one male of the new species was collected in the dorsum of the frog Anomaloglossus stepheni.A single male specimen does not confirm an association but given our current knowledge, it is expected that species of Sycoracinae can be associated with anurans.Bravo & Salazar-Valenzuela (2009) reported the presence of many males (in majority) and females of Falsosycorax wampukrum (Bravo & Salazar-Valenzuela, 2009) on the dorsum of the frog Atelopus sp. in Ecuador and suggested the behavior of aggregation of males to mate.Ježek et al. (2015) reported aggregation of males of two species of Sycorax on two frog species in Brunei and Curler et al. (2015) showed that the females of Aposycorax chilensis (Tonnoir, 1929) prefer to feed on the blood of species of frogs.We believe that males of Sycorax manauara sp.nov.may have a similar behavior and encourages new field expeditions to confirm and better understand what would be the first association in Brazil between a Sycoracinae and an anuran species.
The new species can be differentiated from all other Sycorax by the combination of the following characteristics: gonocoxite with long bristle inserted subapically, on internal surface; gonostyle with apical spine and without subapical setae; paramere with an external convex and robust main structure, bearing one dorsal and one ventral subapical long setae, and a medial hooked branch; aedeagus bifid, developed in two long and sinuous genital filaments.
AUTHORS' CONTRIBUTIONS: DPC: Conceptualization, Visualization, writing -original draft; ACR, DPC, FB: Investigation, Resouces; DPC, FB: formal analysis, writing -review & editing.All authors actively participated in the discussion of the results, they reviewed and approved the final version of the paper.CONFLICTS OF INTEREST: Authors declare there are no conflicts of interest.FUNDING INFORMATION: Fieldwork had financial support from CNPq (Universal 461.573/2014-8 and 429.132/2016-6) and Excellence Program in Basic and Applied Health Research (PROEP FIOCRUZ FAPEAM 001/2014).Images were acquired on equipment provided by the program CAPES PROEQUIPAMENTOS from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).DPC and FB receive a research grant from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (PCI Program 300891/2022-9 and process number 305855/2019-0, respectively).