The SISBIOTA-Diptera Brazilian Network: A long-term survey of Diptera from unexplored Brazilian Western Arc of Amazon, Cerrado, and Pantanal

ABSTRACT The SISBIOTA-BRASIL was a three-year multimillion-dollar research program of the Brazilian government to document plants and animals in endangered/understudied areas and biomes in Brazil. Distributional patterns and the historical events that generated them are extensively unknown regarding Brazilian fauna and flora. This deficiency hinders the development of conservation policies and the understanding of evolutionary processes. Conservation decisions depend on precise knowledge of the taxonomy and geographic distribution of species. Given such a premise, we proposed to research the diversity of Diptera of the Brazilian western arc of Amazon, Cerrado, and Pantanal in the states of Mato Grosso, Mato Grosso do Sul, and Rondônia. Three important biomes of the South American continent characterize these Brazilian states: Amazon forest, Cerrado (Brazilian Savannah), and Pantanal. Besides their ecological relevance, these biomes historically lack intensive entomological surveys. Therefore, they are much underrepresented in the Brazilian natural history collections and in the scientific literature, which is further aggravated by the fact that these areas are being exponentially and rapidly converted to commercial lands. Our project involved over 90 collaborators from 24 different Brazilian institutions and one from Colombia among researchers, postdocs, graduate and undergraduate students, and technicians. We processed and analyzed nearly 300,000 specimens from ~60 families of Diptera collected with a large variety of methods in the sampled areas. Here, we provide a detailed overview of the genera and species diversity of 41 families treated. Our results point to a total of 2,130 species and 514 genera compiled and identified for the three states altogether, with an increase of 41% and 29% in the numbers of species and genera known for the three states combined, respectively. Overall, the 10 most species-rich families were Tachinidae, Cecidomyiidae, Tabanidae, Psychodidae, Sarcophagidae, Stratiomyidae, Bombyliidae, Syrphidae, Tephritidae, and Asilidae. The 10 most diverse in the number of genera were Tachinidae, Stratiomyidae, Asilidae, Mycetophilidae, Syrphidae, Tabanidae, Muscidae, Dolichopodidae, Sarcophagidae, and Chloropidae. So far, 111 scientific papers were published regarding taxonomic, phylogenetic, and biogeographical aspects of the studied families, with the description of 101 new species and three new genera. We expect that additional publications will result from this investigation because several specimens are now curated and being researched by specialists.

Geometric morphometrics as a tool to understand biogeographical and evolutionary patterns in crane fly genus Ischnotoma Skuse (Diptera, Tipulidae).

Background: The geometric morphometric analysis is applied for the first time for the family Tipulidae to distinguish evolutionary and biogeographical patterns on Ischnotoma species from Neotropical and Australian regions. We included 45 recognized species of the genus, representing its three subgenera I. (Icriomastax), I. (Ischnotoma), and I. (Neotipula). This paper aims to test if the three subgenera are recoverable using this set of morphometric data. Methods: Twenty-two landmarks were selected on the wing, mostly located on the radial and medial veins. A Regression Analysis, Principal Components Analysis (PCA), a Shape Coordinates PCA and a Canonical Variate Analysis (CVA) were used to test the variations among individuals, wing shape and groups. Results: For all analyses, the species of Ischnotoma (Neotipula) has a strong dissociation from the remaining species and the CVA shows a complete separation of the three subgenera. This study represents the first insight for a new assessment of Ischnotoma and the first step to giving a possible new status for I. (Neotipula).

Form and function of labral fan-ray microtrichia and posterior proleg barbs in Neotropical black flies larvae (Diptera: Simuliidae).

The microtrichia of the labral fan primary rays, considered as the finest structure of the larval filter mechanism in Simuliidae, and the posterior proleg hooks, a structure used for substrate attachment, were describe using Scanning Electron Microscopy for the first time in 14 Neotropical species of black flies. Four patterns of primary rays microtrichia and two types of hooks barbs disposition were found. The present study provides new morphological information for Neotropical black fly species, compares it with previous information for worldwide species. Also, shows that the presence of posterior proleg hooks barbs is common among black fly species, that these barbs are not related with the larval habitat or with the primary ray microtrichia pattern, and that flow conditions may influence the last-instar larvae microtrichial patterns, at least in most of the Neotropical species studied here.

Two new species of Tanytarsus van der Wulp (Diptera: Chironomidae) from Costa Rica.

Tanytarsus is a highly diverse genus of Chironomidae (Diptera), distributed worldwide. In the Neotropics, 84 species are known, but only five have been recorded from Central America to date. In the present article, two new species of the Tanytarsus ortoni species group are described and illustrated based on male specimens collected from Costa Rica, T. braini sp. nov. and T. costarica sp. nov.

Effects of Bacillus thuringiensis var. israelensis on the Black Fly Communities (Diptera, Simuliidae) in Tropical Streams.

Bacillus thuringiensis var. israelensis (Bti) Berliner, 1911 is widely used in the biological control of black fly and mosquito populations. The objective of this study was to evaluate the effects of Bti on the black fly communities in streams in the Atlantic Forest domain. The study was carried out in eight streams of Serra do Mar, in the municipality of Ubatuba, São Paulo. Some parts of the streams in this locality have received applications of Bti fortnightly for more than 25 years by the sanitary agency of the region. In each stream, two sections were sampled, with and without application of Bti (June 2015 and 2016). Black flies were collected and identified to the species level in the laboratory with 1382 larvae being identified, distributed in six species. Of this total, 73% of the individuals were found in sections where Bti was not applied. There was a difference in the abundance of simulids between the sections with and without Bti application, being more abundant in the latter. We measured the head capsule of individuals of the most abundant species, Simulium pertinax Kollar 1832, using a stereoscopic microscope with millimeter lens. These measurements demonstrated that there was a difference between the sections regarding the age structure of Simuliidae populations. In the sections without Bti application, there was a higher proportion of larvae in the last instar, while in the Bti-treated sections, smaller instars were predominant, possibly due to constant recolonization processes.

Austrocnephia, new genus, for five species of 'Paracnephia' (Diptera: Simuliidae), with a key to Australian black fly genera.

A segregate of the so-called Australian 'Paracnephia' (Diptera: Simuliidae) is assigned to a new genus, Austrocnephia. The taxon is fully diagnosed and a key to constituent species presented. Two species-groups are recognized: the aurantiaca species-group, comprised of A. aurantiaca (Tonnoir 1925) and A. strenua (Mackerras Mackerras 1950), and the tonnoiri species-group, comprised of A. fuscoflava (Mackerras Mackerras 1948), A. orientalis (Mackerras Mackerras 1950) and A. tonnoiri (Drummond 1931). Both species-groups are diagnosed and the included species fully redescribed. Detailed locality data is given, as is information about biology, when known. Brief comments are offered about the historical biogeography of Austrocnephia. A key to Australian simuliid genera is also provided.

New synonymy to Simulium ( Inaequalium ) diversibranchium Lutz, with comments on the Inaequale species-group (Diptera, Simuliidae)

ABSTRACT Simulium (Inaequalium) Coscarón and Wygodzinsky, 1984 is a homogeneous subgenus widely spread on the Neotropical Region, which the adults can only be reliably identified from linked-reared specimens. Despite many revisionary works only a single character of the pupal gill separates Simulium diversibranchium Lutz, 1910 from S. mariavulcanoae Coscarón and Wygodzinsky, 1984. Specimens of both species show that S. mariavulcanoae is a junior synonym of S. diversibranchium. The relationship between S. diversibranchium and S. subnigrum Lutz, 1910 needs further investigation. Some features of the pupal gill often used as diagnosis in Simulium (Inaequalium) species, such as the direction of gill filaments or the height of the bifurcation of the secondary or tertiary branches of the pupal gill are highly polymorphic, making its use as diagnostic character not reliable.

Comments on the association of immatures of Hemerodromia (Diptera, Empididae) and Simulium (Diptera, Simuliidae), and first record of this association in the Atlantic Forest (Brazil).

Larvae of Empididae (Diptera) prey on black fly immatures and its pupae can be collected from pupal cases of Simuliidae (Diptera). The aim of our work was to report the second record of association between immatures of Empididae and Simuliidae in the Neotropical Region and the first for the Atlantic Forest (Rio de Janeiro, Brazil). We collected 4982 pupae and exuviae of Simulium Latreille, (Diptera, Simuliidae) and found three with a pupa of Hemerodromia Meigen (Diptera, Empididae) inside. This shows that the use of black flies cocoons by dance flies occurs at extremely low frequencies, which might explain why this association is so rarely recorded. Our results are relevant for a better comprehension of the predator-prey relationship between these families.

FAMILY BLEPHARICERIDAE.

Blephariceridae are poorly known in Colombia, being reasonably well sampled only in Antioquia Department. In the present catalogue, five species of Limonicola Lutz and eight species of Paltostoma Schiner are reported for the country.

FAMILY CORETHRELLIDAE.

Corethrellidae are poorly known in Colombia, with few specimens deposited in collections, and only nine named species of Corethrella Coquillett are reported for the country. An additional six undescribed species have been recently collected, indicating that the number of recorded species in Colombia is underestimated.

Water pollution and distribution of the black fly (Diptera: Simuliidae) in the Atlantic Forest, Brazil.

Black flies have medical importance because some species are vectors of the unenocerciasis and Mansonelosis, nevertheless, their ecology and potential use as bioindicators is still poorly studied in the Neotropical Region. In Brazil, bioindicators use is strongly focused in a multimetrical ecological index approach; this way, we investigated the black fly spatial distribution, in relation to abiotic factors correlated to water quality, to provide baseline information for their utilization as standalone indicators of lotic systems integrity. We have tested the hypothesis that environmental changes related to urbanization, lead to decreased abundance and loss in the number of species of the black fly fauna. The sampling was conducted in 10 urban and 10 preserved streams during the dry season (August-September) of 2012, in the mountainous region of Teres6polis, State of Rio de Janeiro, Brazil. The streams were characterized for their environmental integrity conditions and physico-chemical properties of water. In each stream, five different rapid points were sampled in a section of 50 meters, 10 meters apart from each other. The black flies were sampled with a kick-net sampler on rocky substrates. The material was separated and the larvae were sorted in morphotypes, and later, the final instar specimens were dissected and identified with the help of taxonomical literature at species level. A total abundance of 488 larvae from nine species were collected, 5 (1.02 %) in extremely impacted streams, 470 (96.31 %) in intermediate streams and 13 (2.66 %) in preserved streams. The visual evaluation (HII) differed in relation to the water physico-chemical evaluation, in which more variation in the characterization of the sampling sites was observed. In Canonical Correspondence Analysis Simulium subpallidum, S. inscrustatum and S. pertinax were significantly associated with intermediate values of most of the variables, and then to intermediate impacted sites. On the other hand, Lutzsimulium hirticosta, S. subnigrzm and Simulium sp. A were associated to low values of chemical variables, and then to more preserved sites. Most studies on the bioindicator potential of Simuliidae have suggested an approach to agricultural impacts, while our results, on the other hand, showed that Simuliidae species were present in streams with intermediate urban pollution impacts, but absent in heavily impacted sites. Thus, our data suggested that some species are associated to more pristine breeding sites, such as L. hirticosta and Simulium sp. A, while others may be good bioindicators of moderately impacted streams, such as S. nertinax. S. subnigrum and S. subpallidum.

Water pollution and distribution of the black fly (Diptera: Simuliidae) in the Atlantic Forest, Brazil / Contaminación del agua y distribución de la mosca negra (Díptera: Simuliidae) en el bosque Atlán&tico, Brasil

Abstract:Black flies have medical importance because some species are vectors of the Onchocerciasis and Mansonelosis, nevertheless, their ecology and potential use as bioindicators is still poorly studied in the Neotropical Region. In Brazil, bioindicators use is strongly focused in a multimetrical ecological index approach; this way, we investigated the black fly spatial distribution, in relation to abiotic factors correlated to water quality, to provide baseline information for their utilization as standalone indicators of lotic systems integrity. We have tested the hypothesis that environmental changes related to urbanization, lead to decreased abundance and loss in the number of species of the black fly fauna. The sampling was conducted in 10 urban and 10 preserved streams during the dry season (August-September) of 2012, in the mountainous region of Teresópolis, State of Rio de Janeiro, Brazil. The streams were characterized for their environmental integrity conditions and physico-chemical properties of water. In each stream, five different rapid points were sampled in a section of 50 meters, 10 meters apart from each other. The black flies were sampled with a kick-net sampler on rocky substrates. The material was separated and the larvae were sorted in morphotypes, and later, the final instar specimens were dissected and identified with the help of taxonomical literature at species level. A total abundance of 488 larvae from nine species were collected, 5 (1.02 %) in extremely impacted streams, 470 (96.31 %) in intermediate streams and 13 (2.66 %) in preserved streams. The visual evaluation (HII) differed in rela&tion to the water physico-chemical evaluation, in which more variation in the characterization of the sampling sites was observed. In Canonical Correspondence Analysis Simulium subpallidum, S. inscrustatumand S. pertinaxwere significantly associated with intermediate values of most of the variables, and then to intermediate impacted sites. On the other hand, Lutzsimulium hirticosta, S. subnigrumand Simuliumsp. A were associated to low values of chemical variables, and then to more preserved sites. Most studies on the bioindicator potential of Simuliidae have suggested an approach to agricultural impacts, while our results, on the other hand, showed that Simuliidae species were present in streams with intermediate urban pollution impacts, but absent in heavily impacted sites. Thus, our data suggested that some species are associated to more pristine breeding sites, such as L. hirticostaand Simuliumsp. A, while others may be good bioindicators of moderately impacted streams, such as S. pertinax, S. subnigrumand S. subpallidum.

Resumen:Aunque las moscas negras tienen importancia médica debido a que algunas de sus especies son vectores de la Oncocercosis y Mansonelosis, su ecología y su potencial como bioindicadores es aún poco estudiado en la Región Neotropical. En Brasil el uso de bioindicadores se centra principalmente en un índice ecológico multimétrico, en este estudio investigamos la distribución espacial de las moscas negras y su relación con los factores abióticos, para su utilización como indicadores de la integridad de los sis&temas lóticos. En este sentido, hemos probado la hipótesis de que los cambios ambientales relacionados con la urba&nización conducen a la disminución de la abundancia y la pérdida de especies de la mosca negra. El muestreo se llevó a cabo en la región montañosa del Estado de Río de Janei&ro, Brasil, en 10 zonas urbanas y 10 arroyos conservados durante la estación seca (agosto-septiembre) de 2012. Los arroyos se caracterizaron por sus condiciones de integridad del ambiente y las propiedades físico-químicas de agua. Las moscas negras se muestrearon en sustratos rocosos con un kick-net sampler.El material se separó y las larvas se ordenaron por morfotipos y más tarde los especímenes en último estadio se disectaron e identificaron con la ayuda de la literatura taxonómica a nivel de especie. Una abundancia total de 488 larvas de nueve especies fueron recolectadas, 5 (1.02 %) en arroyos muy afectados, 470 (96.31 %) en intermedios y 13 (2.66 %) en conservados. La evaluación visual (HII) difiere de la físico-química del agua, en la que se observó una mayor variación. En el Análisis Correspon&dencia Canónica, Simulium subpallidum, S. inscrustatumy S. pertinaxestaban asociadas significativamente con sitios de impacto intermedio, para la mayoría de las variables. Por otro lado, Lutzsimulium hirticosta, S. subnigrumy Simuliumsp. A se asocian con valores bajos de las varia&bles químicas, relacionado a sitios más conservados. La mayoría de los estudios sobre el potencial bioindicador de Simuliidae tienden a enfocarse en los impactos agrícolas, mientras que nuestros resultados, por otro lado, sugieren que las especies de Simuliidae están presentes en arroyos con impactos intermedios de contaminación urbana, pero ausentes en los sitios fuertemente afectados. Por lo tanto, nuestros datos sugieren que algunas especies tales como L. hirticostay Simuliumsp. A están asociadas a lugares de reproducción más prístinos, mientras que otras pueden ser bioindicadores de arroyos moderadamente afectados, como S. pertinax, S. subnigrumy S. subpallidum.

Evidence of phenotypic plasticity of larvae of Simulium subpallidum Lutz in different streams from the Brazilian Cerrado

In this paper, the overall morphological differences between populations of Simulium subpallidum Lutz, 1909 are studied. Several studies found in the literature point to a relationship between the labral fans and body size and the habitat where blackfly larvae occur. However, other characteristics potentially related to the microhabitat, such as abdominal hook circlet morphology, which is used for larvae to fix themselves in the substratum, and thoracic prolegs morphology, which help larvae move in the substratum, were analyzed in three different populations of S. subpallidum, one of which occupied a faster flow. The results suggest phenotypic plasticity in S. subpallidum and a tendency toward larger structures in faster flows.

Seasonal variation in black fly (Diptera: Simuliidae) taxocenoses from the Brazilian Savannah (Tocantins, Brazil).

We investigated the spatial and temporal distribution of blackfly larvae on a mesohabitat scale atfour different breeding sites in the Brazilian savannah "Cerrado." A total of 2,385 larvae of 12 different species were collected, and three of the sampling sites revealedspecies richness and diversity peaks in April. A principal components analysis was performed to reduce abiotic data into a smaller set of independent variables, such as precipitation and river size, and subsequent linear regressions showed that Simulium subpallidum was positively correlated with precipitation. This species, along with S. nigrimanum, was negatively correlated withriver size, whereasS. perflavum was negatively correlated with precipitation. Beta diversity was negatively correlated withprecipitation, and black fly taxocenoses were markedly different between wet and dry seasons, as shown through nonmetric multidimensional scaling, suggesting that precipitation playsa major role in structuring these communities, probably because of enhanced drift and the dendritic nature of lotic metacommunities.

On the enigmatic Heterostomus curvipalpis Bigot, 1857, with a description of the pupa (Diptera, Brachycera).

The previously unknown pupa of Heterostomus curvipalpis Bigot is described. The morphology of the pupa of Heterostomus is compared with the pupae of Xylophagomorpha, Tabanomorpha and Stratiomyomorpha families, based on five characters, and its phylogenetic position is discussed. Currently placed in Xylophagidae, we conclude that the pupa of Heterostomus shares most characters with the pupae of Pelecorhynchidae.

Simulium (Chirostilbia) brunnescens (Diptera: Simuliidae) - new species from the Brazilian cerrado, Manso Dam, state of Mato Grosso, Brazil.

A new species of Simuliidae, Simulium (Chirostilbia) brunnescens, was discovered at Chapada dos Guimarães, state of Mato Grosso, Brazil, and nearby municipalities (Paranatinga, Rosário do Oeste and Nobres). This species is described here based on the adults, pupae and larvae. This species is closely related to Simulium (C.) subpallidum Lutz, but could be differentiated in all stages: females, leg colour pattern and frontal dilatation size; males, gonostyle shape; pupae, number of gill filaments; larvae, body size and colour, postgenal cleft, ratio between antenna and stalk of labral fan.

Simulium (Chirostilbia) brunnescens (Diptera: Simuliidae) - new species from the Brazilian cerrado, Manso Dam, state of Mato Grosso, Brazil

A new species of Simuliidae, Simulium (Chirostilbia) brunnescens, was discovered at Chapada dos Guimarães, state of Mato Grosso, Brazil, and nearby municipalities (Paranatinga, Rosário do Oeste and Nobres). This species is described here based on the adults, pupae and larvae. This species is closely related to Simulium (C.) subpallidum Lutz, but could be differentiated in all stages: females, leg colour pattern and frontal dilatation size; males, gonostyle shape; pupae, number of gill filaments; larvae, body size and colour, postgenal cleft, ratio between antenna and stalk of labral fan.

Diversity and microdistribution of black fly (Diptera: Simuliidae) assemblages in the tropical savanna streams of the Brazilian cerrado

We describe the abiotic factors affecting the distribution of black flies at a microhabitat scale, rather than at the regional scale usually present in the literature on the Neotropics. Black fly larvae were sampled from the Tocantins River and three tributaries, located in the Brazilian savanna (state of Tocantins, Brazil) during six bi-monthly sampling periods from October 2004-August 2005. At each sampling site, 15 random quadrats (30 x 30 cm) were sampled each period and for each quadrat were determined mean water velocity, predominant substrate type (rocks, riffle litter or riparian vegetation) and depth detrended correspondence analysis (DCA) was used to determine associations with current velocity, whereas correspondence analysis (CA) was used to estimate site specific current velocity associations. Canonical correspondence analysis (CCA) was used to identify general microhabitat associations. The CCA showed that most species had a trend towards riffle litter, except for Simulium nigrimanum associated with rocky substrate and Simulium cuasiexiguum associated with riparian vegetation. The DCA showed a well defined pattern of water velocity associations. The CA revealed that the species showed different speed associations from one site to another, suggesting different competitive pressures resulting in the occurrence of different realized niches.

Diversity and microdistribution of black fly (Diptera: Simuliidae) assemblages in the tropical savanna streams of the Brazilian cerrado.

We describe the abiotic factors affecting the distribution of black flies at a microhabitat scale, rather than at the regional scale usually present in the literature on the Neotropics. Black fly larvae were sampled from the Tocantins River and three tributaries, located in the Brazilian savanna (state of Tocantins, Brazil) during six bi-monthly sampling periods from October 2004-August 2005. At each sampling site, 15 random quadrats (30 x 30 cm) were sampled each period and for each quadrat were determined mean water velocity, predominant substrate type (rocks, riffle litter or riparian vegetation) and depth detrended correspondence analysis (DCA) was used to determine associations with current velocity, whereas correspondence analysis (CA) was used to estimate site specific current velocity associations. Canonical correspondence analysis (CCA) was used to identify general microhabitat associations. The CCA showed that most species had a trend towards riffle litter, except for Simulium nigrimanum associated with rocky substrate and Simulium cuasiexiguum associated with riparian vegetation. The DCA showed a well defined pattern of water velocity associations. The CA revealed that the species showed different speed associations from one site to another, suggesting different competitive pressures resulting in the occurrence of different realized niches.

The phylogeny of Simulium (Chirostilbia) (Diptera: Simuliidae) and perspectives on the systematics of the genus in the Neotropical Region.

Recently, knowledge of Neotropical Simuliidae has been accumulating quickly. However, information about supra-specific relationships is scarce and diagnoses of Simulium subgenera are unsatisfactory. To investigate the relationships among Simulium (Chirostilbia) species and test the subgenus monophyly, we performed a cladistic analysis. The ingroup included all species of this subgenus and the outgroup included representatives of the 17 species groups of Neotropical Simulium and three Holarctic species. The study was based on a data matrix with 31 terminal taxa and 45 morphological characteristics of adult, pupa and larva. The phylogenetic analysis under equal weights resulted in eight most-parsimonious trees (length = 178, consistency index = 34, retention index = 67). The monophyly of the S. (Chirostilbia) was not supported in our analysis. The Simulium subpallidum species group was closer to Simulium (Psilopelmia) and Simulium (Ectemnaspis) than to the Simulium pertinax species group. Additionally, we describe the three-dimensional shape of the terminalia of male and female of Simulium (Chirostilbia) for the first time and provide comments about the taxonomic problems involving some species of the subgenus: Simulium acarayense, Simulium papaveroi, S. pertinax, Simulium serranum, Simulium striginotum and S. subpallidum.