Change in susceptibility response of Aedes aegypti (Diptera: Culicidae) to organophosphate insecticide and Copaifera oleoresin.

The growth of resistance in vector mosquitoes to insecticides, especially the organophosphate Temephos can facilitate the transmission of various disease agents worldwide. Consequently, it arises a challenge to public health agencies, which is the urgency use of other possibilities as botanical insecticides. Such insecticides have specific properties against insects due to the plant's ability to synthesize products derived from its secondary metabolism. The diversity and complexity of active compounds of botanical insecticides can help reduce the selection of resistant individuals and consequently not change susceptibility. To corroborate this hypothesis, the aim of this study was to compare two populations of Aedes aegypti treated with Temephos and Copaifera oleoresin. Thus, Ae. aegypti larvae were exposed from (F1) up to tenth generation (F10) with sublethal doses (±LC25) of these products (Copaifera oleoresin: 40 mg/L and Temephos: 0.0030 mg/L). The triplicates and control groups were monitored every 48 hours and the surviving larvae were separated until the emergence of the adults. Each new population were then subjected to a series of concentrations (LC50 and LC95) of Temephos and Copaifera oleoresin to calculate the Resistance Ratio (RR) of each exposed generation. The population of Ae. aegypti exposed to Temephos had an increase in RR from 05 (considered low) to 13 (considered high). Those population exposed to Copaifera oleoresin, had no increasing in RR and continued susceptible to the oil in all generations. There was a significant difference in mortality between the generations exposed to the two products. The results presented here show that the change in the susceptibility status of Ae. aegypti population to Temephos was already expected. So, we believe that this work will be of great contribution to research related to mosquito control with plant products, and resistance to chemical insecticides.

Use of nontarget organism Chironomus sancticaroli to study the toxic effects of nanoatrazine.

Atrazine was banned by the European Union in 2004, but is still used in many countries. Agricultural research employing nanotechnology has been developed in order to reduce the impacts to the environment and nontarget organisms. Nanoatrazine was developed as a carrier system and have been considered efficient in weed control. However, its toxicity must be verified with nontarget organisms. In this context, the aim of the present study was to investigate ecotoxicological effects of solid lipid nanoparticles (empty and loaded with atrazine) and atrazine on Chironomus sancticaroli larvae, evaluating the endpoints: mortality, mentum deformity, development rate and biochemical biomarkers. The contaminant concentrations used were 2, 470, 950, and 1900 µg L-1 in acute (96 h) and 2 µg L-1 in subchronic (10 days) bioassays. An environmentally relevant concentration of atrazine (2 µg L-1) presented toxic and lethal effects towards the larvae. The nanoparticles loaded with atrazine showed toxic effects similar to free atrazine, causing mortality and biochemical alterations on the larvae. The nanoparticle without atrazine caused biochemical alterations and mortality, indicating a possible toxic effect of the formulation on the larvae. In the acute bioassay, most concentrations of nanoparticles loaded with atrazine were not dose dependent for the endpoint mortality. Only the atrazine concentration of 470 µg L-1 was statistically significant to endpoint mentum deformity. The atrazine and nanoparticles (with and without atrazine) did not affect larval development. The results indicate that Chironomus sancticaroli was sensitive to monitor nanoatrazine, presenting potential to be used in studies of toxicity of nanopesticides.

Accelerating the Morphogenetic Cycle of the Viral Vector Aedes aegypti Larvae for Faster Larvicidal Bioassays.

Any bioassay to test new chemically synthesized larvicides or phytolarvicides against Culicidae and more harmful mosquito species, such as Aedes aegypti and Aedes albopictus, which specifically transmit dengue, yellow fever, chikungunya viral fevers as well as Zika virus, or Anopheles gambiae, a vector for malaria and philariasis, requires thousands of well-developed larvae, preferably at the fourth instar stage. The natural morphogenetic cycle of Aedes spp., in the field or in the laboratory, may extend to 19 days at room temperature (e.g., 25°C) from the first permanent contact between viable eggs and water and the last stage of larval growth or metamorphosis into flying adults. Thus, accelerated sequential molting is desirable for swifter bioassays of larvicides. We achieved this goal in Aedes aegypti with very limited strategic and low-cost additions to food, such as coconut water, milk or its casein, yeast extract, and to a lesser extent, glycerol. The naturally rich coconut water was excellent for quickly attaining the population of instar IV larvae, the most advanced one before pupation, saving about a week, for subsequent larvicidal bioassays. Diluted milk, as another food source, allowed an even faster final ecdysis and adults are useful for mosquito taxonomical purpose.

Malathion insecticide resistance in Aedes aegypti: laboratory conditions and in situ experimental approach through adult entomological surveillance.

OBJECTIVE: In Brazil, the most common method of controlling outbreaks of arbovirus is by the use of chemical sprays, which kill the insect vector, Aedes aegypti. The main objective of this study was to evaluate the resistance of Ae. aegypti to the insecticide, malathion, in situ. The location of this study was the municipality of Foz do Iguaçu, in the state of Paraná, Brazil. METHODS: Ultra-low-volume (ULV) fogging equipment was used, by vehicle, to apply the insecticide in situ, and mosquito populations after treatment were compared with those of control areas. The resistance of strains collected from the municipality was compared to the Rockefeller strain under laboratory conditions. RESULTS: We found 220 adult female specimens and 7423 eggs of Ae. aegypti in the areas subjected to UBV treatment, whereas 245 adult females and 10 557 eggs were found in the control areas. The UBV treatment area showed no significant difference compared to the control area, for all the indices. Mortality of the Rockefeller colony varied more quickly when there were slight variations in malathion concentration than the Foz do Iguaçu population.

OBJECTIF: Au Brésil, la méthode la plus courante de lutte contre les épidémies d'arbovirus consiste à utiliser des pulvérisations chimiques qui tuent l'insecte vecteur, Aedes aegypti. L'objectif principal de cette étude était d'évaluer la résistance de Ae. aegypti à l'insecticide, le malathion, in situ. Le lieu de cette étude était la municipalité de Foz do Iguaçu, dans l'état du Paraná, au Brésil. MÉTHODES: Un équipement de brumisation à très faible volume (ULV) a été utilisé, par véhicule, pour appliquer l'insecticide in situ et les populations de moustiques après le traitement ont été comparées à celles des zones témoins. La résistance des souches collectées dans la municipalité a été comparée à la souche Rockefeller dans des conditions de laboratoire. RÉSULTATS: Nous avons trouvé 220 spécimens femelles adultes et 7.423 œufs d'Ae. aegypti dans les zones soumises au traitement ULV, alors que 245 femelles adultes et 10.557 œufs ont été trouvés dans les zones témoins. La zone de traitement ULV n'a montré aucune différence significative par rapport à la zone témoin, pour tous les indices. La mortalité de la colonie de Rockefeller variait plus rapidement lorsqu'il y avait de légères variations dans la concentration de malathion que la population de Foz do Iguaçu.

Mitochondrial affectation, DNA damage and AChE inhibition induced by Salvia officinalis essential oil on Aedes aegypti larvae.

The aim of this research study was to understand the mechanism of action of Salvia officinalis (Lamiaceae) essential oil (EO) on Aedes aegypti larvae. We evaluated the effect on DNA damage, acetylcholinesterase (AChE) inhibition and mitochondrial enzymatic alterations. The major components were analyzed in silico using OSIRIS and Molispiration free software. Aedes aegypti DNA was extracted from mosquito larvae between third (L3) and fourth (L4) instars to determine the DNA fragmentation or degradation at S. officinalis EO lethal concentrations (LC10, LC20, LC50, and LC90). DNA integrity was assessed in both LCs in larvae treated for 24 h and in larvae homogenized with EO; we also assessed purified DNA larvae by a densitometric analysis. The AChE inhibition was quantified in protein larvae L3-L4 following Ellman's method and the enzymatic activities related to the mitochondrial respiratory chain of mitochondrial proteins was estimated by spectrophotometry. In silico analysis of 1,8-cineol and of α-thujone, major EO components, showed that they were highly permeable in biological membranes without mutagenic risks. Alterations in the integrity of DNA were observed in larvae exposed and homogenized with S. officinalis EO. The EO induced an AChE inhibition of 37 ±â€¯2.6% to IC50. On the other hand, mitochondrial bioenergetics suggest that EO inhibits electrons entry to the respiratory chain, via Complex II. AChE activity alteration causes mortality of individuals, by blocking the insect cholinergic functions. These results indicate that EO affects the integrity of DNA, the mitochondrial respiration chain and the AChE activity.

Neotropical Ablabesmyia Johannsen (Diptera: Chironomidae, Tanypodinae)-part I.

This is a series of three manuscripts about the genus Ablabesmyia. In the first new species are described, three species are re-examined and keys to males, pupae and larvae of known neotropical species are provided and an emended generic diagnosis of Ablabesmyia is given. The second manuscript will analyze the molecular data of Neotropical species and the third one will check, through a phylogenetic analysis, the truth of subgenera. In this study, twenty-four new species of Ablabesmyia Johannsen are described and figured: A. arquata, A. cauame, A. commata, A. communiba, A. cordeiroi, A, depaulai, A. diversa, A. ducke, A. fazzari, A. fusariae, A. gessnerae, A. gigas, A. jaquirana, A. laurindoi, A. manauara, A. martha, A. novema, A. parannulata, A. parareissi, A. pinhoi, A. rafaeli, A. separata, A. strixinoae and A. suiamissu. We re-examined A. cinctipes (Johannsen), A. infumata (Edwards) and A. metica Roback. The species were collected in Brazil, in lentic systems. Keys to males, pupae and larvae of known neotropical species of the genus Ablabesmyia are provided and an emended generic diagnosis of Ablabesmyia is given.

First report on susceptibility of wild Aedes aegypti (Diptera: Culicidae) using Carapa guianensis (Meliaceae) and Copaifera sp. (Leguminosae).

Oils of Carapa guianensis and Copaifera spp. are well known in the Amazonian region as natural insect repellent, and studies have reported their efficiency as larvicide against some laboratory mosquito species. However, in wild populations of mosquitoes, these oils have not yet been evaluated. Thus, the objective of this study was to investigate their efficiency as larvicide in wild populations of Aedes aegypti with a history of exposure to organophosphate. The susceptibility of larvae was determined under three different temperatures, 15°C, 20°C, and 30°C. For each test, 1,000 larvae were used (late third instar and early fourth instar-four replicates of 25 larvae per concentration). Statistical tests were used to identify significant differences. The results demonstrated that as the laboratory A. aegypti, the wild populations of A. aegypti were also susceptible to C. guianensis and Copaifera sp. oils. The lethal concentrations for Copaifera sp. ranged from LC(50) 47 to LC(90) 91 (milligrams per liter), and for C. guianensis, they were LC(50) 136 to LC(90) 551 (milligrams per liter). In relation to different temperature, the effectiveness of the oils on larvae mortality was directly related to the increase of temperature, and better results were observed for temperature at 25°C. The results presented here indicate the potential larvicidal activity of C. guianensis and species of Copaifera, in populations of A. aegypti from the wild. Therefore, the results presented here are very important since such populations are primarily responsible for transmitting the dengue virus in the environment.