First cladistic analysis of Toxotarsinae (Diptera: Calliphoridae), with insights on the evolution of the group and on the transformation series of some historically controversial characters.

Toxotarsinae is a subfamily of Calliphoridae (Diptera: Calyptratae: Oestroidea) with 11 species currently distributed in three genera: Neta Shannon, 1926, Sarconesia Bigot, 1857, and Toxotarsus Macquart, 1851. All known species are endemic to South America, mostly restricted to areas with cooler temperatures-like high elevation areas of the Andean Cordillera-and lowlands and coastal regions in the subtropical and temperate parts of the continent. The classification of the subfamily has been somewhat controversial, with eleven, mostly monotypic nominal genera established to accommodate its species, mostly due to distinct interpretations of certain controversial characters by different authors. To provide a sound basis for a generic classification, we performed the first study aimed to understand the transformation series of those controversial characters and character states under a phylogenetic framework. Our maximum parsimony analyses, under both equal and implied weighting of characters and including all currently known species in the subfamily, retrieved two main clades of Toxotarsinae: the roraima group, composed of Sarconesia roraima (Townsend, 1935), Neta chilensis (Walker, 1836), and S. magellanica (Le Guillou, 1842), and the splendida group, composed of the remaining species of the subfamily. The position of S. magellanica varied, but the species was mostly recovered within the roraima group except in the analyses under implied weighting with k values between 1 and 6. Our results, which include an explicit interpretation of the transformation series associated with some characters and character states commonly used in Toxotarsinae systematics, suggest that both Sarconesia and Neta, along with other genera proposed in the past for species of the subfamily such as Sarconesiomima Lopes & Albuquerque, 1955, Chlorobrachycoma Townsend, 1918, Sarconesiopsis Townsend, 1918, and Roraimomusca Townsend, 1935, previously suggested to be synonyms of Sarconesia, do not properly reflect the evolution of lineages in the subfamily and should not be considered as valid nominal genera. Toxotarsus was the only genus consistently recovered as monophyletic and, based on the phylogenetic relationships recovered herein, we suggest including all known Toxotarsinae within this genus, with the roraima and splendida clades treated as species-groups. Finally, we discuss the characters commonly used to delimit the taxa subordinated to Toxotarsinae, such as the length of the first flagellomere, the position of the rays of the arista relative to the pedicel, the extension of the plumosity of the arista, and the number of postsutural acrostichal setae.

Vertical stratification of insect abundance and species richness in an Amazonian tropical forest.

Tropical forests are among the most biodiverse biomes on the planet. Nevertheless, quantifying the abundance and species richness within megadiverse groups is a significant challenge. We designed a study to address this challenge by documenting the variability of the insect fauna across a vertical canopy gradient in a Central Amazonian tropical forest. Insects were sampled over two weeks using 6-m Gressitt-style Malaise traps set at five heights (0 m-32 m-8 m intervals) on a metal tower in a tropical forest north of Manaus, Brazil. The traps contained 37,778 specimens of 18 orders of insects. Using simulation approaches and nonparametric analyses, we interpreted the abundance and richness of insects along this gradient. Diptera, Hymenoptera, and Coleoptera had their greatest abundance at the ground level, whereas Lepidoptera and Hemiptera were more abundant in the upper levels of the canopy. We identified species of 38 of the 56 families of Diptera, finding that 527 out of 856 species (61.6%) were not sampled at the ground level. Mycetophilidae, Tipulidae, and Phoridae were significantly more diverse and/or abundant at the ground level, while Tachinidae, Dolichopodidae, and Lauxaniidae were more diverse or abundant at upper levels. Our study suggests the need for a careful discussion of strategies of tropical forest conservation based on a much more complete understanding of the three-dimensional distribution of its insect diversity.

The Tree Cricket Neoxabea brevipes Rehn, 1913 (Orthoptera: Gryllidae: Oecanthinae) from the Brazilian southern Atlantic Forest: morphology, bioacoustics and cytogenetics.

In this paper we improve the morphological description of the tree cricket Neoxabea brevipes Rehn, 1913 and a new occurrence record for the species at the São Francisco de Paula National Forest Conservation Unit, State of Rio Grande do Sul, southern Brazil. Morphological characterization is provided together with illustrations emphasizing the most significant diagnostic features, such as the metanotal gland, abdominal specializations and phallic sclerites. The calling song was field recorded, comprising a continuous trill with dominant frequency of 3.6 kHz (19.7°C). The karyotype was described for the first time to Neoxabea Kirby, 1906, including diploid number of 2n=26, with two sexual chromosomes.

Two new species of Huascaromusca Townsend from Colombia, and a new combination for Giovanella carvalhoi (Diptera: Mesembrinellidae).

The Mesembrinellidae (Diptera: Oestroidea) are a group of exclusively Neotropical flies, with 38 recognized species distributed in nine genera. A total of 11 of these species, present in seven genera, were known to occur in Colombia. Huascaromusca obscura Wolff, sp. nov. and H. cordillera Wolff & Ramos-Pastrana, sp. nov. are herein described and their male terminalia are illustrated. Based on a prior molecular study of the family, Giovanella carvalhoi Wolff et al., 2013 is transferred to Huascaromusca Townsend. A key to all known species of Huascaromusca is provided.

The first phylogenetic study of Mesembrinellidae (Diptera: Oestroidea) based on molecular data: clades and congruence with morphological characters.

The Mesembrinellidae (Diptera: Oestroidea) comprise a small group of strictly Neotropical calyptrate flies, with 36 described species. The group has often been treated as a subfamily of Calliphoridae, but there is growing evidence that it corresponds to a distinct Oestroidea lineage. Internal relationships have so far been addressed based only on morphology, with results lacking resolution and support. This is the first molecular phylogeny for the group, which is based on the analyses of 80 terminal taxa (22 mesembrinellid and 28 outgroup species) and 5 molecular markers (ITS2, 28S, COI, COII and 16S). Maximum-parsimony, maximum-likelihood and Bayesian inference methods were used, the latter two with partitioning strategies considering codon position and secondary structure information. Results corroborate the Mesembrinellidae as a monophyletic lineage inside Oestroidea. Three clades were consistently recovered: (1) (Laneella + Mesembrinella patriciae); (2) (Mesembrinella (excluding M. patriciae)  + Eumesembrinella); and (3) (Huascaromusca + Giovanella). Re-examination of the female reproductive tract of M. patriciae revealed a Laneela-type spermatheca, which corroborates the position of the species recovered in the molecular phylogenetic analyses. Mesembrinella and Huascaromusca are in all cases paraphyletic with regards to Eumesembrinella and Giovanella, respectively. These latter two genera should, thus, be seen as subjective junior synonyms.

Evolution of genes involved in feeding preference and metabolic processes in Calliphoridae (Diptera: Calyptratae).

BACKGROUND: The genotype-phenotype interactions among traits governing feeding preference are of fundamental importance to behavioral genetics and evolutionary biology. The genetic basis of behavioral traits has been explored in different taxa using different approaches. However, the complex nature of the genetic mechanisms undergirding behavior is poorly understood. Here, we present an evolutionary study of candidate genes related to parasitism in Calliphoridae (Diptera: Calyptratae). Closely related species in this family exhibit distinct larval feeding habits, most notably necro-saprophagy and obligate parasitism. METHODS: To understand the genetic and molecular bases underlying these habits, expression levels of eight candidate genes for feeding behavior-Cyp6g2, foraging, glutamate dehydrogenase, Jonah65aiv, Malvolio, PGRP-SC2, RPS6-p70-protein kinase, and smooth-were measured in four species using qPCR. Moreover we used expression values and sequence information to reconstruct the relationship among species and the dN/dS rate to infer possible sites under selection. RESULTS: For most candidate genes, no statistically significant differences were observed, indicating a high degree of conservation in expression. However, Malvolio was differentially expressed between habits. Evolutionary analyses based on transcript levels and nucleotide sequences of Malvolio coding region suggest that transcript levels were correlated to feeding habit preferences among species, although deviations under a strictly neutral model were also observed in statistical tests. DISCUSSION: Malvolio was the only gene demonstrating a possible connection to feeding habit. Differences in gene expression may be involved in (or be a result of) the genetic regulation of Calliphoridae feeding habit. Our results are the first steps towards understanding the genetic basis and evolution of feeding behavior in Calliphoridae using a functional approach.

A survey of necrophagous blowflies (Diptera: Oestroidea) in the Amazonas-Negro interfluvial region (Brazilian Amazon)

ABSTRACT The fauna of blowflies (Calliphoridae and Mesembrinellidae) in three localities of primary Amazon forest coverage in the Amazonas-Negro interfluvial region was assessed. A total of 5066 blowflies were collected, with Chloroprocta idiodea being the most abundant species (66.3%). A difference in species richness between the localities ZF2 and Novo Airão was observed. Comparison among sampled sites revealed no considerable variation in fauna composition, except for the species Eumesembrinella benoisti (Séguy 1925) and Hemilucilia sp., whose occurrence was observed only in a single locality. Apparently, Amazon rivers are not efficient geographical barriers to influence the current composition of necrophagous blowfly assemblages. Also, most of the blowfly species did not show a noticeable specificity for any specific forest among the interfluvial areas of the ombrophilous forest. Finally, an updated checklist of necrophagous blowfly species of the Amazonas state in Brazil is presented.