Checklist of the dipterofauna (Insecta) from Roraima, Brazil, with special reference to the Brazilian Ecological Station of Maracá

Roraima is a Brazilian state located in the northern portion of the Amazon basin, with few studies regarding its biodiversity. The Ecological Station of Maracá (Brazil, state of Roraima) harbors the third largest Brazilian pluvial island and is composed of a transitional landscape of savanna and Amazon rainforest components. Despite its ecological importance and strategic localization, few studies covered the dipterofauna of this locality. An updated checklist addressing 41 families of true flies (Diptera) occurring in Roraima is presented based on the literature and the specimens collected during a field expedition that occurred in 2015. This checklist brings several improvements such as new records of 165 taxa to the state of Roraima, 29 taxa to Brazil, and 259 morphotypes, mostly likely representing undescribed species

Genetic and evolutionary analysis in species of the family Calliphoridae (Diptera: Brachycera: Calyptratae)

Orientadores: Ana Maria Lima de Azeredo-Espin, Nilson Ivo Tonin ZanchinTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A superfamília Oestroidea (Diptera:Brachycera:Calyptratae), com +13.000 espécies descritas, compreende um dos grupos mais numerosos e ecologicamente diversos da ordem Diptera. O grupo possui grande interesse para atividades humanas por englobar espécies de importância médica, veterinária e forense, muitas das quais compõem a família Calliphoridae. Apesar do grande número de estudos disponíveis, as relações evolutivas no grupo, o qual é composto predominantemente por linhagens de rápida diversificação e radiação, ainda são controversas e pouco compreendidas, encorajando a caracterização de novos marcadores moleculares para análises de filogenia molecular. Neste contexto, esta tese foi desenvolvida e organizada em três capítulos descrevendo estudos genéticoevolutivos em espécies da superfamília Oestroidea, com ênfase em Calliphoridae. O primeiro capítulo trata da caracterização e avaliação do segundo espaçador transcrito interno (ITS2) do DNA ribossomal como um marcador molecular para análises filogenéticas em Calliphoridae, incorporando informações tanto da sequência primária quanto da estrutura secundária adquirida pela região. A análise do ITS2 revelou um padrão hierarquicamente organizado das distâncias genéticas nos níveis de espécies, gêneros e subfamílias, enquanto pouca variação intra-específica foi encontrada. As árvores inferidas recuperaram muitas das relações comumente aceitas entre os táxons amostrados, sendo que a inclusão da informação estrutural nas análises resultou na recuperação de topologias mais confiáveis. Sendo assim, o potencial da região ITS2 como um marcador molecular para análises evolutivas na família Calliphoridae foi confirmado e seu uso em análises de maior escala, incluindo marcadores de diferentes naturezas de evolução, encorajado. O segundo capítulo da tese descreve a caracterização in vitro da estrutura secundária adquirida pelo ITS2, através de padrões de digestão enzimática e análise dos fragmentos gerados, em espécies representantes das três superfamílias de Calyptratae: Glossina morsitans, Musca domestica e Cochliomyia hominivorax. A análise do padrão de fragmentos gerados pelas enzimas RNAse I, A, T1 e V1, quando mapeados na estrutura secundária predita in silico, corroborou muitos dos domínios inicialmente preditos pelo método computacional, ressaltando a importância e confiabilidade desses métodos na predição de estruturas secundárias. O terceiro capítulo da tese descreve análises de filogenia molecular na superfamília Oestroidea, com ênfase na amostragem de espécies de Calliphoridae, utilizando quatro marcadores moleculares, dois nucleares (ITS2 e 28S) e dois mitocondriais (COI e 16). As análises, que incluíram uma extensa avaliação dos efeitos de diferentes estratégias de particionamento dos dados em análises de inferência Bayesiana (por conformação estrutural e posição no códon), revelaram a existência de dois clados principais em Oestroidea: Tachinidae + Mesembrinellinae e Oestridae + Rhiniinae + Sarcophagidae + Calliphoridae (definida em senso estrito). O status de família recentemente atribuído à Rhiniinae foi encontrado, enquanto há também evidências para sugerir o mesmo para a subfamília Mesembrinellinae, como proposto anteriormente por outros autores. As diferentes estratégias de particionamento do conjunto de dados amostrados resultaram em diferenças discretas em termos de topologia, comprimentos de ramo e suporte geral das filogenias inferidas. Embora o resultado geral indique uma melhor resolução das análises quando do uso de combinações de partições e modelos mais complexas, as mesmas podem ocasionar também um aumento considerável na incerteza associada às análisesAbstract: The Oestroidea superfamily (Diptera: Brachycera: Calyptratae), with +13,000 described species, comprises one of the most numerous and ecologically diverse groups in the Diptera order. The group is actually of great interest for human activities since it includes species of medical, veterinary and forensic importance, most of them included in the Calliphoridae family. Despite the existence of several studies addressing the issue, evolutionary relationships in Oestroidea, a group mainly composed of rapidly diverged lineages, remains contentious and poorly understood, encouraging the characterization of new molecular markers for phylogenetic inference analyses. In this context, this thesis was developed and organized in three chapters describing genetic and evolutionary studies in species of the Oestroidea superfamily, with emphasis in Calliphoridae. The first chapter deals with the characterization and evaluation of the second internal transcribed spacer region (ITS2) of the ribosomal DNA cluster as a molecular marker for phylogenetic inference in Calliphoridae, including information of both primary sequence and secondary structure. The analyses revealed an hierarchically organized pattern of genetic distances in the specific, generic and subfamilial level, while little intraspecific variation was detected. Inferred trees were able to recover most of the commonly accepted relationships among the sampled taxa, with the consideration of structural information resulting in better supported topologies. Thereby, the potential of the ITS2 region as a molecular marker for phylogenetic inference in the Calliphoridae family was corroborated and its use in larger scale analyses, including other markers with different evolutionary patterns, encouraged. Chapter II describes the in vitro characterization of the secondary structure of the ITS2 region, through patterns of enzymatic digestion and analysis of the generated fragments, in representative species of the three superfamilies of the Calyptratae clade: Glossina morsitans, Musca domestica and Cochliomyia hominivorax. Analyses of the patterns of the fragments generated by enzymatic digestions with the RNAses I, A, T1 and V1, when mapped in the in silico predicted secondary structure, corroborated the folding of most of the domains predicted by computational methods, highlighting the importance and reliability of these methods in secondary structure prediction. Chapter III describes molecular phylogenetic analyses in the Oestroidea superfamily, with emphasis on the Calliphoridae family, using four different molecular markers, two nuclear (ITS2 and 28S) and two mitochondrial (COI and 16S) regions. The analyses, which included a comprehensive evaluation of the effects of different data partitioning strategies in a Bayesian framework (by structural conformation and codon position), revealed the existence of two main clades in Oestroidea: Tachinidae+Mesembrinellinae and Oestridae+Rhiniinae+Sarcophagidae+Calliphoridae (defined in a strict sense). The recently attributed family status to Rhiniinae was confirmed, and there are evidence to also suggest the same for Mesembrinellinae, as previously pointed out by other studies. The different data partitioning strategies used in the sampled dataset resulted in small differences in terms of inferred topologies, estimated branch lengths and average support. Although the overall results indicate a significant increase in phylogeny resolution when more complex and parameter-rich models / partitions combinations are used, they can also lead to an increased uncertainty in the phylogenetic estimation processDoutoradoGenetica Animal e EvoluçãoSoutor em Genética e Biologia Molecula

Morfometria geométrica identifica diferenças fenotípicas em asas de linhagens de Mesembrinella bellardiana aldrich, 1922 (Diptera: Mesembrinellidae) da amazônia e mata atlântica

A sistemática do grupo Mesembrinellidae apresenta algumas controvérsias. Um exemplo em nível de espécie é Mesembrinella bellardiana Aldrich, 1922. Guimarães (1977) propôs a divisão da linhagem específica em duas subespécies: M. bellardiana bellardiana (Aldrich, 1922), com ocorrência restrita ao sul da América do Sul (predominantemente na Mata Atlântica); e M. bellardiana fuscicosta (Séguy, 1925), restrita ao norte da América do Sul (predominantemente na Amazônia e Andes). Bonatto (2001) não encontrou diferenças significativas na genitália masculina destas linhagens, não adotando assim a proposta de Guimarães (1977). Posteriormente, Marinho et al. (2017), através da análise de sequências dos genes COI e ITS2, sugeriram que a espécie nominal M. bellardiana seria composta provavelmente por duas linhagens evolutivas distintas, correspondendo às subespécies propostas anteriormente. Assim, este trabalho buscou encontrar diferenças nas formas das asas entre as linhagens evolutivas identificadas em Marinho et al. (2017). Através de morfometria geométrica (ADAMS et al., 2013), buscou-se encontrar padrões na venação de asas que possam embasar as evidências de divergência fenotípica entre linhagens e verificar se é possível sua identificação

Distribuição e registros de ocorrência de Calliphoridae e mesembrinellidae (diptera: calyptratae) no bioma pampa e no Rio Grande do Sul, Brasil

O presente trabalho propõe a realização de uma compilação dos dados de registros de ocorrência e distribuição de espécies das famílias Calliphoridae e Mesembrinellidae em localidades do estado do Rio Grande do Sul, Brasil, bem como nas áreas do Pampa associadas também à Argentina e ao Uruguai

The First Phylogenetic Study Of Mesembrinellidae (diptera: Oestroidea) Based On Molecular Data: Clades And Congruence With Morphological Characters

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)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.332134152Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2012/23200-2]CNPq [309240/2013-1]FAPESP [2003/10274-9]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

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

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 necrosaprophagy and obligate parasitism. 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 d(N)/d(S) rate to infer possible sites under selection. 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. 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 approach4CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP477335/2009-8; 150441/2016-9 307502/2011-22008/58106-0; 2014/13933-8; 2009/51723-7; 2009/13463-3; 2014/01600-4; 2012/19987-7; 2012/23200-

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

Distributed under Creative Commons CC-BY 4.0 Evolution of genes involved in feeding preference and metabolic processes in Calliphoridae (Diptera: Calyptratae)

ABSTRACT 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 necrosaprophagy 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 d N /d S 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)

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. © 2015 Sociedade Brasileira de Entomologia

12,500+ and counting: biodiversity of the Brazilian Pampa

Knowledge on biodiversity is fundamental for conservation strategies. The Brazilian Pampa region, located in subtropical southern Brazil, is neglected in terms of conservation, and knowledge of its biodiversity is fragmented. We aim to answer the question: how many, and which, species occur in the Brazilian Pampa? In a collaborative effort, we built species lists for plants, animals, bacteria, and fungi that occur in the Brazilian Pampa. We included information on distribution patterns, main habitat types, and conservation status. Our study resulted in referenced lists totaling 12,503 species (12,854 taxa, when considering infraspecific taxonomic categories [or units]). Vascular plants amount to 3,642 species (including 165 Pteridophytes), while algae have 2,046 species (2,378 taxa) and bryophytes 316 species (318 taxa). Fungi (incl. lichenized fungi) contains 1,141 species (1,144 taxa). Animals total 5,358 species (5,372 taxa). Among the latter, vertebrates comprise 1,136 species, while invertebrates are represented by 4,222 species. Our data indicate that, according to current knowledge, the Pampa holds approximately 9% of the Brazilian biodiversity in an area of little more than 2% of Brazil’s total land. The proportion of species restricted to the Brazilian Pampa is low (with few groups as exceptions), as it is part of a larger grassland ecoregion and in a transitional climatic setting. Our study yielded considerably higher species numbers than previously known for many species groups; for some, it provides the first published compilation. Further efforts are needed to increase knowledge in the Pampa and other regions of Brazil. Considering the strategic importance of biodiversity and its conservation, appropriate government policies are needed to fund studies on biodiversity, create accessible and constantly updated biodiversity databases, and consider biodiversity in school curricula and other outreach activities