Abstract INTRODUCTION: Despite their widespread usage, synthetic insecticides and larvicides are harmful for controlling disease-causing mosquitoes owing to the development of resistance. The leaves of Eugenia astringens, Myrrhinium atropurpureum, and Neomitranthes obscura were collected from Marambaia and Grumari restingas. The safety and larvicidal efficacy of their extracts were tested against Aedes (Stegomyia) aegypti L. and Simulium (Chirostilbia) pertinax Kollar. METHODS: The dry leaves were subjected to static maceration extraction using 90% methanol. A. aegypti and S. pertinax larvae were exposed to 7.5, 12.5, and 25.0 µL/mL of the extracts (n= 30). The larvicidal activity after 24 h and 48 h, and the mortality, were determined. The median lethal concentration (CL50) was estimated by a Finney's probit model. RESULTS: M. atropurpureum and E. astringens extracts exhibited the strongest larvicidal effects against A. aegypti. M. atropurpureum extracts (25 µL/mL) caused mortalities of over 50% and 100% after 24 h and 48 h, respectively (CL50 = 11.10 and 9.68 ppm, respectively). E. astringens extracts (25 µL/mL) caused mortalities of 50% and 63.33% after 24 h and 48 h, respectively. High concentrations of N. obscura extracts induced a maximum mortality of 46.66% in A. aegypti larvae after 48 h (CL50= 25 ppm). The larvae of S. pertinax showed 100% mortality following exposure to all the plant extracts at all the tested concentrations after 24 h. CONCLUSIONS: The extracts of M. atropurpuerum exhibited the strongest larvicidal activity against A. aegypti. The larvae of S. pertinax were sensitive to all the extracts at all the tested concentrations.

ABSTRACT Biotic factors such as predation, although important drivers of the black fly community, are rarely investigated in the literature. This study aimed to test the hypothesis that the patterns of co-occurrence of black fly larvae and its potential predators is not random and that there is a correlation between its frequencies and Simuliidae larvae abundances. Larvae were sampled from two localities in the Pedra Branca State Park, Rio de Janeiro, Brazil, during the dry season in june 2018. We collected Simulium pertinax Kollar, 1832, Simulium subpallidum Lutz, 1910, Simulium (Inaequalium) sp., Simulium (Psaroniocompsa) sp. and Simulium (Trichodagmia) sp. The predators families present were Perlidae, Hydropsychidae, Leptoceridae, Libellulidae and Chironomidae. The null models showed that species co-occurred significantly more than expected by chance. The canonical correlation analyses for the Vargem Grande and Pau da Fome areas area showed a significant relationship between black fly abundances and predator abundances. In Vargem Grande the abundance of Chironomidae showed a highly significant positive correlation to S. (Psaroniocompsa) sp., while Hydropsychidae was significantly correlated to Simulium sp. On the other hand, in Pau da Fome Libellulidae was significantly correlated to S. (Psaroniocompsa) sp. and Simulium sp. and Chironomidae correlated significantly to Simulium sp. The result is consistent with what would be expected of organisms that exercise mutual population regulation, although other factors than biotic interactions may be causing these patterns. However the lack of mechanistic evidences on the effect of biotic interactions on black fly populations pose a challenge on the understanding of these patterns.

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.

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.

In this paper, the overall morphological differences between populations of Simulium subpallidumLutz, 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.

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.

Este estudo teve como objetivo avaliar, nos criadouros de simulídeos, a influência da velocidade da água sobre a ocorrência de suas larvas e estrutura de suas taxocenoses. Para obtenção da amostragem foram utilizados "quadrats" de madeira distribuídos aleatoriamente no rio Caputera e um de seus tributários. Foram encontradas quatro espécies: Simulium incrustatum sl. Lutz, 1910; Simulium (Inaequalium) sp. ; Simulium pertinax sl. Kollar, 1832 e Simulium subpallidum sl. Lutz 1909. S. (Inaequalium) sp. teve uma distribuição quase constante ao longo do gradiente de correnteza. S. pertinax se mostrou fortemente associada às velocidades mais altas de correnteza, enquanto que S. subpallidum se mostrou associado às velocidades mais baixas.

This study aimed to evaluate the influence of water velocity speed on the local distribution and taxocenosis structure of blackfly larvae. The larvae were collected from two adjacent streams located in the municipality of Angra dos Reis (RJ): Caputera River and one of its tributaries. Riffle litter patches were sampled randomly using a 30 x 30 cm quadrat. Four blackfly species were found: Simulium incrustatum s. l. Lutz, 1910; Simulium (Inaequalium) sp. ; Simulium pertinax s. l. Kollar, 1832 and Simulium subpallidum s. l. Lutz, 1909. Among these species, Simulium pertinax s. l. was clearly associated with higher water current speeds, while Simulium subpallidum s. l. showed association with lower water velocities, and Simulium (Inaequalium) sp. had a relatively constant distribution along the water current gradient.

A distribuição das taxocenoses de larvas de simulídeos (Diptera: Simuliidae) de diferentes altitudes no Parque Nacional de Itatiaia ( localizado no sudeste do estado do Rio de Janeiro, Brasil 44º34' - 44º42' W , 22º16' - 22º28' S ) foi investigada ao longo de quatro campanhas, cada uma em uma diferente estação do ano de 2003. Cada campanha foi composta de 60 amostras, obtidas a partir de "quadrats" de madeira de 30 x 30 cm, distribuídos através de quatro áreas: Taquaral, Véu da Noiva, Três Picos e Brejo da Lapa. Nas campanhas do inverno e da primavera, dois outros sítios, Alsene e Agulhas Negras, foram adicionados. Foram coletadas 3578 larvas e 292 pupas, sendo identificadas seis espécies: Simulium clavibranchium Lutz, S. subnigrum Lutz, S. rappae Py-Daniel, S. incrustatum Lutz, S. stellatum Gil-Azevedo, Figueiró & Maia-Herzog e Lutzsimulium pernigrum Lutz. As maiores abundância e diversidade foram encontradas no sítio de altura intermediária, o Brejo da Lapa. A análise de componentes principais (PCA) revelou que L. pernigrum, S. stellatum e S. incrustatum se mostraram associadas a rios de menor porte, com as duas primeiras espécies associadas a sítios com maior insolação e S. incrustatum com pouca insolação.

The distribution of the blackfly larval taxocenoses of different altitudes in the Itatiaia National Park (located in the southeast of Rio de Janeiro state, Brazil 44º34' - 44º42' W , 22º16' - 22º28' S) was investigated on four campaigns, one in each season of 2003. On each campaign 60 samples were collected, using 30 x 30 cm wooden quadrats, distributed at four sites: Taquaral, Véu da Noiva, Três Picos and Brejo da Lapa. The winter and spring campaigns included two additional sampling sites: Alsene and Agulhas Negras. A total of 3578 larvae and 292 pupae were sampled, representative of six species: Simulium clavibranchium Lutz, S. subnigrum Lutz, S. rappae Py-Daniel, S. incrustatum Lutz, S. stellatum Gil-Azevedo, Figueiró & Maia-Herzog and Lutzsimulium pernigrum Lutz 1910. The highest larvae abundance and diversity were reported at the intermediate altitude site, Brejo da Lapa. Principal Components Analysis (PCA) revealed that L. pernigrum, S. stellatum and S. incrustatum were associated with the smaller breeding sites, the first two species being associated with sunny sites, while S. incrustatum occurred in sites with little sunlight.

Esta nota apresenta um método alternativo para bioensaios com larvas de simulídeos, com que se espera proporcionar aos pesquisadores uma ferramenta mais simples, porém igualmente efetiva para a obtenção de dados referentes à eficácia de larvicidas.

This note presents an alternative method for bioassays using black fly larvae, with which we intend to provide a simpler but effective tool for accessing data on larvicide efficacy.