Ecotoxicological strategies employing biochemical markers and organisms to monitor the efficacy of malathion photolysis treatment.

The objective of this study was to assess the photolysis-mediated degradation of malathion in standard and commercial formulations, and to determine the toxicity of these degraded formulations. Degradation tests were carried out with 500 µg L-1 of malathion and repeated three times. The initial and residual toxicity was assessed by using Lactuca sativa seeds for phytotoxicity, Stegomyia aegypti larvae for acute toxicity, and Stegomyia aegypti mosquitoes (cultivated from the larval stage until emergence as mosquitoes) to evaluate the biochemical markers of sublethal concentrations. For the standard formulations the photolytic process efficiently reduced the initial concentration of malathion to levels below the regulatory limits however, the formation of byproducts was revealed by chromatography, which allowed for a more complete proposal of photolytic-mediated malathion degradation route. The degraded formulations inhibited the growth of L. sativa seeds, while only the untreated formulations showed larvicidal activity and mortality. Both formulations slightly inhibited acetylcholinesterase activity in S. aegypti mosquitoes, while the standard formulation decreased and the commercial formulation increased glutathione S-transferase activity. However, there were no significant differences for superoxide dismutase, esterase-α, esterase-ß and lipid peroxidation. These findings indicate that in the absence of the target compound, the presence of byproducts can alter the enzymatic activity. In general, photolysis effectively degrade malathion lower than the legislation values; however, longer treatment times must be evaluated for the commercial formulation.

Surveillance of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) as a Method for Prevention of Arbovirus Transmission in Urban and Seaport Areas of the Southern Coast of Brazil.

Entomological surveillance is a traditional method to measure presence, distribution, and seasonal variation of vectors in urban areas, and is essential to targeted control activities to prevent arbovirus transmission. Ovitraps as one of the main components of surveillance programs, enable determination of female oviposition behavior, as well as identification of seasonal variations of the vector. The goals of this study were 1) to detect the mosquitos (Aedes aegypti Linnaeus) and (Aedes albopictus Skuse) (Diptera:Culicidae), in Paranaguá city, 2) to assess ovitrap positive index (OPI), egg density index (EDI), and their relationship with meteorological variables, and 3) to evaluate the vertical transmission of dengue, Zika, and chikungunya in Ae. aegypti and Ae. albopictus. The study was carried out in urban areas of Paranaguá city, an important port region of Brazil, from June 2017 to November 2018. The city was divided into 16 area-clusters. Three-hundred and thirty-one ovitraps were installed monthly, remaining for four days in selected places. Kernel density maps were done to compare the spatiotemporal distribution of collected eggs. Areas which maintained constant oviposition associated with vector activity were identified and were found to overlap the area-clusters with the highest EDI. As viral RNA was not detected, vertical transmission was likely not a maintenance mechanism of arbovirus circulation in Ae. aegypti and Ae. albopictus vectors. This study reiterates the importance, efficiency, and feasibility of ovitraps to monitor the presence and dynamics of Aedes spp. populations.

Genetic diversity and Kdr mutations of natural Aedes (Stegomyia) aegypti (Diptera: Culicidae) populations of Brazil

ABSTRACT Aedes (Stegomyia) aegypti is an important vector of dengue, yellow fever, chikungunya and Zika virus. It is well known that resistance monitoring and genetic diversity data help designing the vector control programs. This study aimed to evaluate resistance to pyrethroids (PYs) through the frequency of kdr mutations Val1016IIe and F1534C, and the genetic variation of the mitochondrial gene ND4 in six natural populations of A. aegypti from Paraná - Brazil. Adults were obtained from eggs collected from Alvorada do Sul, Marilena, Maringá, Nova Londrina, Paranavaí and São Carlos do Ivaí. From these adults, 345 were used to identify the 1016 and 1534 sites, and 120 were used to perform the ND4 gene analysis. The studied populations from Paraná showed PYs resistance, low gene flow and genetic diversity. Additionally, a relationship was observed among the haplotypes of populations from the Amazon and Southeastern Brazil, Peru, Mexico, and North America.

Accumulation of nylon microplastics and polybrominated diphenyl ethers and effects on gut microbial community of Chironomus sancticaroli.

Microplastics (MP) are emerging contaminants with the capacity to bind and transport hydrophobic organic compounds of environmental concern, such as polybrominated diphenyl ethers (PBDEs). The aim of this study was to investigate the ingestion of nylon (polyamide) MP alone and when associated with PBDEs and their effects on Chironomus sancticaroli larvae survival and microbiome structure. Survival, PBDE uptake and microbial community composition were measured in fourth instar larvae exposed for 96 h to BDEs- 47, 99, 100 and 153 in the presence and absence of 1% w/w MP in sediment. Microbiome community structures were determined through high throughput sequencing of 16S small subunit ribosomal RNA gene (16S rRNA). Initial experiments showed that larvae ingested MP faster at 0.5% w/w MP, while depuration was more efficient at 1% w/w MP, although retention of MP was seen even after 168 h depuration. No mortality was observed as a result of PBDEs and MP exposure. MP had a negative effect on PBDE concentration within larvae (η2 = 0.94) and a negative effect on sediment concentrations (η2 = 0.48). In all samples, microbial communities were dominated by Alphaproteobacteria, Betaproteobacteria, Actinobacteria and Gammaproteobacteria. Bacterial alpha diversity was not significantly affected by PBDEs or MP exposure. However, the abundance of discrete bacterial taxa was more sensitive to MP (X2 = 45.81, p = 0.02), and PBDE exposure. Our results highlight that C. sancticaroli showed no acute response to MPs and PBDEs, but that MPs influenced bacterial microbiome structure even after only short-term acute exposure.

Impact of polycyclic aromatic hydrocarbons in mangroves from the Colombian pacific coast: Evaluation in sediments and bivalves.

Bivalves and sediments were sampled from mangroves in the Pacific Coast of Colombia to evaluate the concentrations of the 16 polycyclic aromatic hydrocarbons (PAHs) prioritized by U.S.EPA. Mangroves are highly vulnerable to anthropogenic activities, such as oil spills, which affect sediments and the organisms that depend on that ecosystem. Twelve samples of mangrove and non-mangrove (sandy) sediments and 20 samples of Anadara tuberculosa from mangrove were collected in marine and estuarine areas. In sediments and A. tuberculosa, the highest concentration of Æ©16PAHs was found in estuarine mangroves close to the Rosario River mouth, ranging from 171.4 to 564.0 ng g-1 and 31.0 to 169.0 ng g-1, respectively. For the bivalve, the concentrations showed less variability than sediment, with 25% and 20% of bivalve samples exceeding the limits established by the European Regulatory Commission and Ministry of Health and Social Protection of Colombia, respectively, which can cause effects on people's health. The PAHs isomeric ratios determined in sediments indicated that these compounds were originated mainly from petrogenic sources. The PAHs profile reveals the dominance of 3 and 4 rings PAHs in sediments and dominance of 4 rings PAHs in bivalves.

Molecular and biochemical evaluation of effects of malathion, phenanthrene and cadmium on Chironomus sancticaroli (Diptera: Chironomidae) larvae.

In-vitro effects of sub-lethal concentrations of malathion, phenanthrene (Phe) and cadmium (Cd) were tested on Chironomus sancticaroli larvae in acute bioassays by measuring biochemical and molecular parameters. Malathion was evaluated at 0.001, 0.0564 and 0.1006 mg L-1; Phe at 0.0025, 1.25 and 2.44 mg L-1; and Cd at 0.001, 3.2 and 7.4 mg L-1. The recovery test carried out at the highest concentration of each compound showed that survival of larvae exposed to Phe ranged from 4% to 5%, while the effects of malathion and Cd were irreversible, not allowing the emergence of adults. Results showed that malathion and Cd inhibited AChE, EST-α and ES-ß activities at the two highest concentrations. Phe at 0.0025, 1.25 and 2.44 mg L-1; and Cd at 3.2 and 7.4 mg L-1 inhibited glutathione S-transferase activity. Oxidative stress was exclusively induced by the lowest concentration of malathion considering SOD activity once CAT was unaffected by the stressors. Lipid peroxidation was registered exclusively by malathion at the two highest concentrations, and total hemoglobin content was only reduced by Cd at the two highest concentrations. The relationship among biochemical results, examined using the PCA, evidenced that malathion and Cd concentrations were clustered into two groups, while Phe only formed one group. Four hemoglobin genes of C. sancticaroli were tested for the first time in this species, with Hemoglobin-C being upregulated by malathion. The toxicity ranking was malathion > Phe > Cd, while biochemical and molecular results showed the order malathion > Cd > Phe. Our results highlight the importance of combining different markers to understand the effects of the diverse compounds in aquatic organisms.

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.

Cell damage induced by polybrominated diphenyl ethers on Chironomus sancticaroli (Diptera: Chironomidae)

Abstract Histological effects of polybrominated diphenyl ethers (PBDEs) were observed in Chironomus sancticaroli larvae which underwent acute exposure. 2,2′,4-triBDE (BDE-17), 2,2′,4,4′-tetraBDE (BDE-47) and 2,2′,4,4′,5-pentaBDE (BDE-99) were evaluated at 0.5, 2.0 and 20 μg L-1. Cytoplasm vacuolisation of oenocytes was observed in the larvae exposed to BDE-17 and BDE-47. Cuénot cells were disrupted at the brush border as an effect of the three evaluated congeners highlighting BDE-47 at 2.0 μg L-1; 60% of larvae displayed this disruption. The midgut showed changes in the morphology of apex cells located next to the lumen of region I exposed to BDE-17 and BDE-47, while BDE-99 induced a narrowing of the lumen diameter. Significant cytoplasm vacuolisation of the larvae exposed to BDE-47 and BDE-99 was observed in region II of the midgut. Salivary glands showed acidophilic granules in the cytoplasm exposed to BDE-17 and BDE-47. The results showed that the tissues of C. sancticaroli were sensitive to flame retardants; these histopathologies can compromise the health and physiology of this organism, highlighting the concern with the presence of PBDEs in freshwater sediments.

Behavior of detoxifying enzymes of Aedes aegypti exposed to girgensohnine alkaloid analog and Cymbopogon flexuosus essential oil.

Because mosquito control depend on the use of commercial insecticides and resistance has been described in some of them, there is a need to explore new molecules no resistant. In vivo effects of girgensohnine analog 2-(3,4-dimethoxyphenyl)-2-(piperidin-1-yl)acetonitrile DPPA and Cymbopogon flexuosus essential oil CFEO, on the detoxifying enzymes acetylcholinesterase (AChE), glutathione-S-transferase (GST), nonspecific esterases (α- and ß-), mixed function oxidases (MFO) and p-NPA esterases were evaluated on a Rockefeller (Rock) and wild Aedes aegypti population from Santander, Colombia (WSant). The action was tested after 24h of exposure at concentrations of 20.10, 35.18 and 70.35mgL-1 of DPPA and 18.45, 30.75 and 61.50mgL-1 of CFEO, respectively. It was found that AChE activity of Rock and WSant was not influenced by the evaluated concentration of DPPA and CFEO (p>0.05), while MFO activity was significantly affected by all CFEO concentrations in WSant (p<0.05). GST, α- and ß-esterase activities were affected in Rock exposed at the highest CFEO concentration, this concentration also modified ß-esterases activity of WSant. DPPA and CFEO sublethal doses induced inhibition of AChE activity on untreated larvae homogenate from 12 to 20% and 18 to 26%, respectively. For untreated adult homogenate, the inhibition activity raised up to 14 to 27% for DPPA and 26 to 34% for CFEO. Elevated levels of detoxifying enzymes, found when CFEO was evaluated, showed a larval sensitivity not observed by the pure compound suggesting that DPPA, contrary to CFEO, was not recognized, transformed or eliminated by the evaluated detoxifying enzymes.

Polybrominated diphenyl ethers (PBDEs) effects on Chironomus sancticaroli larvae after short-term exposure.

In-vivo effects of polybrominated diphenyl ethers (PBDEs) containing 3, 4 and 5 bromine atoms were tested on fourth-instar larvae of Chironomus sancticaroli (Diptera: Chironomidae) after 48h of exposure, by measuring the activity of the acetyl cholinesterase, alpha and beta esterases and glutathione S-transferase. The PBDE congeners 2,2',4-triBDE (BDE-17), 2,2',4,4'-tetraBDE (BDE-47) and 2,2',4,4',5-pentaBDE (BDE-99) were evaluated at 0.5, 1.0, 2.0 and 3.0ngmL-1. Acetyl cholinesterase activity decreased significantly (p≤0.05) at all evaluated concentrations of the three PBDE congeners, except for larvae exposed to BDE-17 at 1.0 and 2.0ngmL-1. The significant inhibition of acetyl cholinesterase activity ranged from 18% (BDE-47 at 0.5ngmL-1) to 72% (BDE-47 at 2.0ngmL-1). The enzymes alpha and beta esterase were also affected by the three congeners, reducing their activity from 14% (BDE-99 at 1.0ngmL-1) to 52% (BDE-47 at 2.0ngmL-1) and from 7% (BDE-99 at 2.0ngmL-1) to 34% (BDE-47 at 3.0ngmL-1) respectively. Substantial increments in glutathione S-transferase activity were similarly observed, varying from 138% (BDE-99 2.0 at ng mL-1) to 346% (BDE-17 at 1.0ngmL-1). DNA strand breaks were detected exclusively in larvae exposed to BDE-99 at 2.0 and 3.0ngmL-1 (H=11.7, p=0.019). These results showed that C. sancticaroli larvae were sensitive to the PBDEs treatments under the experimental conditions.