Impact of SumiLarv® 2MR on Aedes aegypti larvae: a multicenter study in Brazil.

BACKGROUND: Aedes aegypti is associated with dengue, Zika, and chikungunya transmission. These arboviruses are responsible for national outbreaks with severe public health implications. Vector control is one of the tools used to prevent mosquito proliferation, and SumiLarv® 2MR is an alternative commercial product based on pyriproxyfen for larval/pupal control. In this study, the residual effectiveness of SumiLarv® 2MR in different regions of Brazil was evaluated in simulated field conditions. METHODS: We conducted a multicenter study across four Brazilian states-Amapá, Pernambuco, Rio de Janeiro, and São Paulo-given the importance to the country's climatic variances in the north, northeast, and southeast regions and their influence on product efficiency. The populations of Ae. aegypti from each location were held in an insectary. Third-instar larvae (L3) were added every 2 weeks to water containers with SumiLarv® 2MR discs in 250-, 500- and 1000-l containers in Amapá and Rio de Janeiro, and 100-l containers in Pernambuco and São Paulo, using concentrations of 0.04, 0.08, and 0.16 mg/l. RESULTS: Adult emergence inhibition over 420 days was observed in all tests conducted at a concentration of 0.16 mg/l; inhibition for 308-420 days was observed for 0.08 mg/l, and 224-420 days for 0.04 mg/l. CONCLUSIONS: Sumilarv® 2MR residual activity demonstrated in this study suggests that this new pyriproxyfen formulation is a promising alternative for Aedes control, regardless of climatic variations and ideal concentration, since the SumiLarv® 2MR showed adult emergence inhibition of over 80% and residual activity greater than 6 months, a period longer than that recommended by the Ministry of Health of Brazil between product re-application in larval breeding sites.

Regionality in vector control: effect of fluctuating temperature in the susceptibility of Aedes aegypti (Diptera: Culicidae) larvae to Pyriproxyfen.

Using Pyriproxyfen in controlling Aedes aegypti shows great potential considering its high competence in low dosages. As an endocrine disruptor, temperature can interfere with its efficiency, related to a decrease in larval emergence inhibition in hotter environments. However, previous studies have been performed at constant temperatures in the laboratory, which may not precisely reflect the environmental conditions in the field. The aim of this study was to assess the effect of the fluctuating temperatures in Pyriproxyfen efficiency on controlling Aedes aegypti larvae. We selected maximum and minimum temperatures from the Brazilian Meteorological Institute database from September to April for cities grouped by five regions. Five fluctuating temperatures (17-26; 20-28.5; 23-32.5; 23-30.5; 19.5-31 °C) were applied to bioassays assessing Pyriproxyfen efficiency in preventing adult emergence in Aedes aegypti larvae in five concentrations. In the lowest temperatures, the most diluted Pyriproxyfen treatment (0.0025 mg/L) was efficient in preventing the emergence of almost thrice the larvae than in the hottest temperatures (61% and 21%, respectively, p value = 0.00015). The concentration that inhibits the emergence of 50% of the population was lower than that preconized by the World Health Organization (0.01 mg/L) in all treatments, except for the hottest temperatures, for which we estimated 0.010 mg/L. We concluded that fluctuating temperatures in laboratory bioassays can provide a more realistic result to integrate the strategies in vector surveillance. For a country with continental proportions such as Brazil, considering regionalities is crucial to the rational use of insecticides.

Assessment of insecticide resistance of Aedes aegypti (Diptera: Culicidae) populations to insect growth regulator pyriproxyfen, in the northeast region of Brazil.

Vector control has been an essential strategy in Brazil to manage vector-borne diseases, and the use of insecticides plays an important role in this effort. Pyriproxyfen (PPF) has become a common insect growth regulator used to control juvenile stages of mosquitoes by disturbing their growth and development. This study assesses the susceptibility and resistance status of Brazilian Ae. aegypti populations that previously showed low resistance levels to PPF. Eggs of Ae. aegypti were collected from six cities located in the northeast states of Ceará (Quixadá, Icó, and Juazeiro do Norte), and Bahia (Itabuna, Brumado, and Serrinha). We used the Ae. aegypti Rockefeller strain as an experimental control and a strain known to be susceptible to insecticides. Inhibition of emergence rates by 50% of Ae. aegypti populations varied from 0.0098-0.046 µg/L. Mosquitoes from Icó, Serrinha, and Brumado showed low resistance levels [resistance ratio (RR50) = 2.33, 4.52, and 4.83, respectively], whereas moderate levels of resistance were detected in populations from Juazeiro do Norte (RR50=5.83) and Itabuna (RR50=7.88). Aedes aegypti collected from the Quixadá population showed a high resistance level to pyriproxyfen (RR50=11). The evolution of resistance in Brazilian Ae. aegypti populations to PPF can compromise vector control efforts. Continuous monitoring of insecticide resistance in Ae. aegypti is essential for making timely management decisions for effective vector control and management.

Toxicity assessment of two IGR insecticides on eggs and larvae of the ladybird Eriopis connexa.

BACKGROUND: Eriopis connexa is an important predator in the Neotropical region, associated with pests of economic relevance on horticultural crops in Argentina. The use of broad-spectrum insecticides could reduce the biodiversity of these natural enemies in agroecosystems and put at risk its conservation. New, selective "risk reduced" insecticides could be an alternative to conventional chemical control to promote sustainable agriculture. The goal of this work was to assess the lethal and sublethal effects of two insect growth regulator (IGR) insecticides on eggs and two larval instars of E. connexa exposed to insecticides. RESULTS: Pyriproxyfen and cypermethrin significantly affected egg hatching by 28.8% and 70.4%, respectively. Pyriproxyfen reduced the survival of larvae that emerged by ≈52% from Day (D3)3 after hatching and caused the lengthening of developmental time for both larval and pupal stages. By contrast, teflubenzuron did not reduce hatching and survival but shortened the developmental time of the pupae stage. Cypermethrin reduced the survival of 2nd (L2 ) and 4th (L4 ) larval instars by 36.4% and 74.6%, respectively, and lengthened the development time of L2 . Pyriproxyfen lengthened the development time of L4 and reduced the fecundity and fertility of females. Teflubenzuron reduced survival of L2 and L4 larval instars by 46.9% and 28.6%, respectively, and lengthened the total development time for the larval stage. In addition, teflubenzuron reduced the fecundity and fertility of females. CONCLUSIONS: Both eggs and larvae were susceptible to exposure to IGR, showing lethal and sublethal effects. This study highlights, once again, the higher toxicity of cypermethrin to E. connexa. The toxicity of both IGR insecticides could impair the performance of E. connexa as a biological control agent in agroecosystems. © 2022 Society of Chemical Industry.

Monitoring of Insecticides resistance against cotton Jassid (Amrasca biguttutla biguttutla) under laboratory conditions / Monitoramento da resistência de inseticidas contra o algodão Jassid (Amrasca biguttutla biguttutla) em condições laboratoriais

Development of insecticides resistance mainly hinge with managements techniques for the control of Jassid, Amrasca biguttutla biguttutla. Five insecticides were applied against field collected and laboratory rared jassid populations during the years of 2017 to 2019 to profile their resistance level against field population of jassid through leaf dip method. Very low resistance level was found in jassid against confidor whereas high level of resistance was observed by pyriproxyfen against other test insecticides. Gradual resistance was observed against diafenthiuron. It is concluded that for the management of Jassid repetition of same insecticide should be avoided. The use of confidor may be reduced to overcome resistance against Jassid.(AU)

Desenvolvimento da resistência a inseticidas principalmente em dobradiça com técnicas de manejo para o controle de jassid, Amrasca biguttutla biguttutla. Cinco inseticidas foram aplicados contra populações de jassídeos coletados em campo e em laboratório durante os anos de 2017 a 2019 para traçar o perfil de seu nível de resistência contra populações de jassídeos em campo através do método de imersão nas folhas. Nível de resistência muito baixo foi encontrado em jassid contra confidor, enquanto alto nível de resistência foi observado por piriproxifeno contra outros inseticidas de teste. Resistência gradual foi observada contra o diafenthiuron. Conclui-se que, para o manejo do jassid, a repetição do mesmo inseticida deve ser evitada. O uso de confidor pode ser reduzido para superar a resistência contra jassid.(AU)

Monitoring of Insecticides resistance against cotton Jassid (Amrasca biguttutla biguttutla) under laboratory conditions / Monitoramento da resistência de inseticidas contra o algodão Jassid (Amrasca biguttutla biguttutla) em condições laboratoriais

Abstract Development of insecticides resistance mainly hinge with managements techniques for the control of Jassid, Amrasca biguttutla biguttutla. Five insecticides were applied against field collected and laboratory rared jassid populations during the years of 2017 to 2019 to profile their resistance level against field population of jassid through leaf dip method. Very low resistance level was found in jassid against confidor whereas high level of resistance was observed by pyriproxyfen against other test insecticides. Gradual resistance was observed against diafenthiuron. It is concluded that for the management of Jassid repetition of same insecticide should be avoided. The use of confidor may be reduced to overcome resistance against Jassid.

Resumo Desenvolvimento da resistência a inseticidas principalmente em dobradiça com técnicas de manejo para o controle de jassid, Amrasca biguttutla biguttutla. Cinco inseticidas foram aplicados contra populações de jassídeos coletados em campo e em laboratório durante os anos de 2017 a 2019 para traçar o perfil de seu nível de resistência contra populações de jassídeos em campo através do método de imersão nas folhas. Nível de resistência muito baixo foi encontrado em jassid contra confidor, enquanto alto nível de resistência foi observado por piriproxifeno contra outros inseticidas de teste. Resistência gradual foi observada contra o diafenthiuron. Conclui-se que, para o manejo do jassid, a repetição do mesmo inseticida deve ser evitada. O uso de confidor pode ser reduzido para superar a resistência contra jassid.

Monitoring of Insecticides resistance against cotton Jassid (Amrasca biguttutla biguttutla) under laboratory conditions / Monitoramento da resistência de inseticidas contra o algodão Jassid (Amrasca biguttutla biguttutla) em condições laboratoriais

Development of insecticides resistance mainly hinge with managements techniques for the control of Jassid, Amrasca biguttutla biguttutla. Five insecticides were applied against field collected and laboratory rared jassid populations during the years of 2017 to 2019 to profile their resistance level against field population of jassid through leaf dip method. Very low resistance level was found in jassid against confidor whereas high level of resistance was observed by pyriproxyfen against other test insecticides. Gradual resistance was observed against diafenthiuron. It is concluded that for the management of Jassid repetition of same insecticide should be avoided. The use of confidor may be reduced to overcome resistance against Jassid.

Desenvolvimento da resistência a inseticidas principalmente em dobradiça com técnicas de manejo para o controle de jassid, Amrasca biguttutla biguttutla. Cinco inseticidas foram aplicados contra populações de jassídeos coletados em campo e em laboratório durante os anos de 2017 a 2019 para traçar o perfil de seu nível de resistência contra populações de jassídeos em campo através do método de imersão nas folhas. Nível de resistência muito baixo foi encontrado em jassid contra confidor, enquanto alto nível de resistência foi observado por piriproxifeno contra outros inseticidas de teste. Resistência gradual foi observada contra o diafenthiuron. Conclui-se que, para o manejo do jassid, a repetição do mesmo inseticida deve ser evitada. O uso de confidor pode ser reduzido para superar a resistência contra jassid.

Monitoring of Insecticides resistance against cotton Jassid (Amrasca biguttutla biguttutla) under laboratory conditions

Abstract Development of insecticides resistance mainly hinge with managements techniques for the control of Jassid, Amrasca biguttutla biguttutla. Five insecticides were applied against field collected and laboratory rared jassid populations during the years of 2017 to 2019 to profile their resistance level against field population of jassid through leaf dip method. Very low resistance level was found in jassid against confidor whereas high level of resistance was observed by pyriproxyfen against other test insecticides. Gradual resistance was observed against diafenthiuron. It is concluded that for the management of Jassid repetition of same insecticide should be avoided. The use of confidor may be reduced to overcome resistance against Jassid.

Resumo Desenvolvimento da resistência a inseticidas principalmente em dobradiça com técnicas de manejo para o controle de jassid, Amrasca biguttutla biguttutla. Cinco inseticidas foram aplicados contra populações de jassídeos coletados em campo e em laboratório durante os anos de 2017 a 2019 para traçar o perfil de seu nível de resistência contra populações de jassídeos em campo através do método de imersão nas folhas. Nível de resistência muito baixo foi encontrado em jassid contra confidor, enquanto alto nível de resistência foi observado por piriproxifeno contra outros inseticidas de teste. Resistência gradual foi observada contra o diafenthiuron. Conclui-se que, para o manejo do jassid, a repetição do mesmo inseticida deve ser evitada. O uso de confidor pode ser reduzido para superar a resistência contra jassid.

Brazilian Populations of Aedes aegypti Resistant to Pyriproxyfen Exhibit Lower Susceptibility to Infection with Zika Virus.

Zika virus (ZIKV) infection has caused devastating consequences in Brazil as infections were associated with neurological complications in neonates. Aedes aegypti is the primary vector of ZIKV, and the evolution of insecticide resistance (IR) in this species can compromise control efforts. Although relative levels of phenotypic IR in mosquitoes can change considerably over time, its influence on vector competence for arboviruses is unclear. Pyriproxyfen (PPF)-resistant populations of Ae. aegypti were collected from five municipalities located in Northeast of Brazil, which demonstrated different resistance levels; low (Serrinha, Brumado), moderate (Juazeiro do Norte, Itabuna), and high (Quixadá). Experimental per os infection using ZIKV were performed with individuals from these populations and with an insecticide susceptible strain (Rockefeller) to determine their relative vector competence for ZIKV. Although all populations were competent to transmit ZIKV, mosquitoes derived from populations with moderate to high levels of IR exhibited similar or lower susceptibility to ZIKV infection than those from populations with low IR or the susceptible strain. These observations suggest an association between IR and arbovirus infection, which may be attributable to genetic hitchhiking. The use of PPF to control Brazilian Ae. aegypti may be associated with an indirect benefit of reduced susceptibility to infection, but no changes in disseminated infection and transmission of ZIKV among PPF-resistant phenotypes.

Behavior of two classes of organic contaminants in the presence of graphene oxide: Ecotoxicity, physicochemical characterization and theoretical calculations.

Graphene oxide (GO) production has increased considerably and therefore its presence in the environment is inevitable. When in aquatic environment GO can interact with co-existing compounds, modifying their toxicities for several organisms. However, the toxic effects of co-exposure of GO and organic compounds are rarely reported in the literature. Herein, we studied the behavior of four organic aquatic contaminants found in surface water such as 2-phenylbenzotriazoles (non-Cl PBTA-9 and PBTA-9) and phenoxyphenyl pesticides, pyriproxyfen (PYR) and lambdacyhalothrin (LCT), in the presence of GO. GO reduced 90% and 83% of the toxicity of non-Cl PBTA-9 and PBTA for Daphnia. When PBTAs were adsorbed onto GO surface their interactions caused GO agglomeration (up to 20 mm) and consequent precipitation, making PBTAs less bioavailable. PYR and LCT's toxicities increased up to 83% for PYR and 47% for LCT in the presence of GO, because their adsorption on GO lead to the stabilization of the suspensions (up to 0.5 µm). Those particles were then easily ingested and retained in the digestive tract of the daphnids, triggering the Trojan horse effect. Based on theoretical calculations we observed that PBTA compounds are planar, electron-poorer and more reactive than the studied pesticides, suggesting a better stability of the GO/PBTA complexes. PYR and LCT are nonplanar, electron-richer and less reactive towards GO than PBTAs, forming less stable GO complexes that could facilitate the desorption of pesticides, increasing toxic effects. Our results suggest that the properties of the organic toxicants can influence the stability of graphene oxide suspensions, playing a fundamental role in the modulation of their toxicity. Further research is needed for a deep understanding of the behavior of nanomaterials in the presence of contaminants and their effect in the toxicity of aquatic organisms.

Restricted double access mesoporous polypyrrole as adsorbent in pipette-tip solid phase extraction for simultaneous determination of progesterone, pyriproxyfen, and deltamethrin in chicken eggs.

In this study, two sample preparation techniques were evaluated in the simultaneous determination of three compounds with different physicochemical properties, progesterone, pyriproxyfen, and deltamethrin that may be present in the chicken egg. In this procedure, firstly the restricted double access mesoporous polypyrrole was synthesized, which was evaluated as adsorbent in pipette-tip solid phase extraction and dispersive solid phase extraction. After optimizing the extraction parameters, it was found that pipette-tip solid phase extraction was more efficient and, therefore, it was used in the validation and application of the method. The analytical method showed good recoveries, acceptable linearity (r > 0.99), limits of quantification, precision and accuracy, robustness and stability within the limits of the literature. Finally, the developed method was successfully applied in simultaneous determination of analytes in different chicken egg samples. Therefore, this work provided a promising strategy for the extraction of different organic compounds from egg products.

Pyriproxyfen induces intracellular calcium overload and alters antioxidant defenses in Danio rerio testis that may influence ongoing spermatogenesis.

We investigated the in vitro effects of pyriproxyfen on ionic balance in the testis of the zebrafish by measuring 45Ca2+ influx. In vivo pyriproxyfen treatment was carried out to study oxidative stress, and conduct morphological analysis of the testis and liver. Whole testes were incubated in vitro with/without pyriproxyfen (10-12, 10-9 or 10-6 M; 30 min) and 45Ca2+ influx determined. To study pyriproxyfen's mechanism of action, inhibitors/activators of ionic channels or pumps/exchangers, protein kinase inhibitors or a calcium chelator were added 15 min before the addition of 45Ca2+ and pyriproxyfen. We evaluated the in vivo effects of 7 day exposure to waterborne pyriproxyfen (10-9 M) on reactive oxygen species (ROS) formation, lipid peroxidation, and reduced glutathione content (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) and γ-glutamyltransferase (GGT) activity. Morphological analyses of the testis and liver were carried out after in vivo exposure of D. rerio to pyriproxyfen. Pyriproxyfen increased 45Ca2+ influx by opening the voltage-dependent T-type channels (T-type VDCC), inhibiting sarco/endoplasmic reticulum 45Ca2+-ATPase (SERCA) and the NCX exchanger (forward mode) and by mobilizing calcium from stores. The involvement of potassium channels and protein kinase C (PKC) was also demonstrated in pyriproxyfen-induced intracellular calcium elevation. In vivo pyriproxyfen treatment of D. rerio increased lipid peroxidation, decreased GSH content and increased GST activity in testes, in addition to increasing the number and size of spermatogonia cysts and inducing hepatocyte basophilia and dilation of blood vessels in the liver. The toxicity of pyriproxyfen is mediated by calcium overload, increased lipid peroxidation, and a diminished antioxidant capacity in the testis, due to GSH depletion, and altered spermatogenesis. The development of high basophilia in the liver suggests that pyriproxyfen may have estrogenic activity, possibly acting as an endocrine-disruptor. These findings indicate that these alterations may contribute to pyriproxyfen toxicity and spermatogenesis disruption.

Mesoporous molecularly imprinted polymer core@shell hybrid silica nanoparticles as adsorbent in microextraction by packed sorbent for multiresidue determination of pesticides in apple juice.

In this work, we report the synthesis of a mesoporous molecularly imprinted polymer on the surface of silica nanoparticles (core@mMIP) to be applied as adsorbent in microextraction by packed sorbent (MEPS) for selective determination of pesticides in apple juice. The core@mMIP was properly characterized, showing good adhesion of the polymer to the silica core. The best extraction conditions were: 200 µL of ultrapure water as washing solvent, 150 µL of acetonitrile as eluent, 100 µL of sample at pH 2.5, five draw-eject cycles and 8 mg of adsorbent. Thereby, recoveries of 96.12 ± 1.05%, 76.88 ± 6.18% and 76.18 ± 5.57% were obtained for pyriproxyfen (PPX), deltamethrin (DTM) and etofenprox (ETF), respectively. After validation, the method presented linearity in the range of 0.02-10 µg mL-1 (r > 0.99), limit of detection of 0.005 µg mL-1, satisfactory selectivity, and proper precision and accuracy. The method was successfully applied real samples of processed and fresh apple juice.

Pyriproxyfen Exposure Impairs Cognitive Parameters and Alters Cortisol Levels in Zebrafish.

Pyriproxyfen is one of the most used larvicides and insecticides; it acts as an analog of juvenile insect hormone (a growth regulator). It is highly toxic during all stages of mosquito development, suppresses metamorphosis, and interferes in insect reproduction and proliferation. Pyriproxyfen and its main metabolite have been shown to affect brain development in rodents. This compound is employed mainly to eliminate outbreaks of the genus Aedes, even in potable water. Despite the increasing number of toxicological studies about larvicides and insecticides-with an indication of continuous use-there have been few studies about the effects of pyriproxyfen in non-target species such as fish. This study evaluated the effects of pyriproxyfen on behavioral, cognitive, and endocrine parameters in zebrafish. We exposed adult zebrafish to different pyriproxyfen (Pestanal®) concentrations (0.125, 0.675, and 1.75 mg/l) for 96 h. We analyzed behavioral parameters, memory, cortisol levels, and gene expression of glucocorticoid receptor (gr) and corticotrophin-releasing factor (crf) after pyriproxyfen exposure. This exposure did not alter locomotion (distance or mean speed), anxiety-like behavior (latency to enter to the top zone of the tank or time in the top zone of the tank), and social or aggressive behavior. However, there was impaired inhibitory avoidance memory at all tested pyriproxyfen concentrations. Cortisol levels were reduced in exposed groups when compared to control or vehicle. However, gr and crf gene expression in pyriproxyfen-treated animals were unaltered when compared to control or vehicle groups. Taken together, these findings indicate that pyriproxyfen may induce cognitive impairment and altered cortisol levels in zebrafish, a non-target species.

Impact of pyriproxyfen on virus behavior: implications for pesticide-induced virulence and mechanism of transmission.

BACKGROUND: More than 3 years since the last Zika virus (ZIKV) outbreak in Brazil, researchers are still deciphering the molecular mechanisms of neurovirulence and vertical transmission, as well as the best way to control spread of ZIKV, a flavivirus. The use of pesticides was the main strategy of mosquito control during the last ZIKV outbreak. METHODS: We used vesicular stomatitis virus (VSV) tagged with green fluorescent protein (GFP) as our prototypical virus to study the impact of insecticide pyriproxyfen (PPF). VZV-GFP infected and uninfected Jurkat, HeLa and trophoblast cells were treated with PPF and compared to untreated cells (control). Cell viability was determined by the MTT assay. Cell morphology, presence of extracellular vesicles (EVs), virus infection/GFP expression as well as active mitochondrial levels/localization were examined by confocal microscopy. RESULTS: PPF, which was used to control mosquito populations in Brazil prior to the ZIKV outbreak, enhances VSV replication and has cell membrane-altering properties in the presence of virus. PPF causes enhanced viral replication and formation of large EVs, loaded with virus as well as mitochondria. Treatment of trophoblasts or HeLa cells with increasing concentrations of PPF does not alter cell viability, however, it proportionately increases Jurkat cell viability. Increasing concentrations of PPF followed by VSV infection does not interfere with HeLa cell viability. Both Jurkats and trophoblasts show proportionately increased cell death with increased concentrations of PPF in the presence of virus. CONCLUSIONS: We hypothesize that PPF disrupts the lipid microenvironment of mammalian cells, thereby interfering with pathways of viral replication. PPF lowers viability of trophoblasts and Jurkats in the presence of VSV, implying that the combination renders immune system impairment in infected individuals as well as enhanced vulnerability of fetuses towards viral vertical transmission. We hypothesize that similar viruses such as ZIKV may be vertically transmitted via EV-to-cell contact when exposed to PPF, thereby bypassing immune detection. The impact of pesticides on viral replication must be fully investigated before large scale use in future outbreaks of mosquito borne viruses.

Measuring mosquito control: adult-mosquito catches vs egg-trap data as endpoints of a cluster-randomized controlled trial of mosquito-disseminated pyriproxyfen.

BACKGROUND: Aedes aegypti and Culex quinquefasciatus are the main urban vectors of arthropod-borne viruses causing human disease, including dengue, Zika, or West Nile. Although key to disease prevention, urban-mosquito control has met only limited success. Alternative vector-control tactics are therefore being developed and tested, often using entomological endpoints to measure impact. Here, we test one promising alternative and assess how three such endpoints perform at measuring its effects. METHODS: We conducted a 16-month, two-arm, cluster-randomized controlled trial (CRCT) of mosquito-disseminated pyriproxyfen (MD-PPF) in central-western Brazil. We used three entomological endpoints: adult-mosquito density as directly measured by active aspiration of adult mosquitoes, and egg-trap-based indices of female Aedes presence (proportion of positive egg-traps) and possibly abundance (number of eggs per egg-trap). Using generalized linear mixed models, we estimated MD-PPF effects on these endpoints while accounting for the non-independence of repeated observations and for intervention-unrelated sources of spatial-temporal variation. RESULTS: On average, MD-PPF reduced adult-mosquito density by 66.3% (95% confidence interval, 95% CI: 47.3-78.4%); Cx. quinquefasciatus density fell by 55.5% (95% CI: 21.1-74.8%), and Ae. aegypti density by 60.0% (95% CI: 28.7-77.5%). In contrast, MD-PPF had no measurable effect on either Aedes egg counts or egg-trap positivity, both of which decreased somewhat in the intervention cluster but also in the control cluster. Egg-trap data, therefore, failed to reflect the 60.0% mean reduction of adult Aedes density associated with MD-PPF deployment. CONCLUSIONS: Our results suggest that the widely used egg-trap-based monitoring may poorly measure the impact of Aedes control; even if more costly, direct monitoring of the adult mosquito population is likely to provide a much more realistic and informative picture of intervention effects. In our CRCT, MD-PPF reduced adult-mosquito density by 66.3% in a medium-sized, spatially non-isolated, tropical urban neighborhood. Broader-scale trials will be necessary to measure MD-PPF impact on arboviral-disease transmission.

Avaliação da toxicidade pré-natal: estudo de teratogenicidade do inseticida piriproxifeno em ratos Wistar / Prenatal toxicity assessment: teratogenicity study of the insecticide pyriproxyfen in Wistar rats

Este estudo investigou a toxicidade pré-natal do inseticida piriproxifeno em ratos Wistar, de forma a detectar possíveis alterações no desenvolvimento fetal da progênie exposta durante o período organogênico. Três doses de piriproxifeno (100, 300 e 500mg.kg-1) foram administradas por via oral às progenitoras, do sexto ao 15º dia de gestação. Os fetos foram submetidos à técnica de diafanização modificada descrita por Taylor e Van Dyke, para avaliação de malformações e alterações esqueléticas. Os resultados não demonstraram a ocorrência de toxicidade materna sistêmica ou alterações nos índices reprodutivos avaliados. Malformações ou alterações teratogênicas não foram detectadas, no entanto alterações esqueléticas sugestivas de retardo no desenvolvimento foram observadas especialmente nas doses mais altas testadas (300mg.kg-1 e 500mg.kg-1). Considerando-se a situação complexa de risco para a saúde humana, mostra-se importante a execução de investigações adicionais, de modo a contribuir para a adequada avaliação de risco do piriproxifeno em água potável.(AU)

This study investigated the prenatal toxicity of the insecticide pyriproxyfen in Wistar rats to detect the possible changes in the fetal development of the progeny exposed during the organogenic period. Three doses of pyriproxyfen (100, 300, and 500mg.kg-1) were administered orally to the progenitors, from day 6 to 15 of gestation. The fetuses were processed using the Taylor and Van Dyke modified diaphanization technique to evaluate malformations and skeletal changes. The results did not demonstrate the occurrence of systemic maternal toxicity or changes in the reproductive indexes evaluated. Malformations or teratogenic changes were not detected, however, skeletal changes suggestive of developmental delay were observed, especially in the highest doses tested (300 mg.kg-1 and 500 mg.kg-1). Owing to the potentially complex situation regarding its risk to human health, it is important that further studies be performed to contribute to the risk assessment of the addition of pyriproxyfen in drinking water.(AU)

Avaliação da toxicidade pré-natal: estudo de teratogenicidade do inseticida piriproxifeno em ratos Wistar / Prenatal toxicity assessment: teratogenicity study of the insecticide pyriproxyfen in Wistar rats

Este estudo investigou a toxicidade pré-natal do inseticida piriproxifeno em ratos Wistar, de forma a detectar possíveis alterações no desenvolvimento fetal da progênie exposta durante o período organogênico. Três doses de piriproxifeno (100, 300 e 500mg.kg-1) foram administradas por via oral às progenitoras, do sexto ao 15º dia de gestação. Os fetos foram submetidos à técnica de diafanização modificada descrita por Taylor e Van Dyke, para avaliação de malformações e alterações esqueléticas. Os resultados não demonstraram a ocorrência de toxicidade materna sistêmica ou alterações nos índices reprodutivos avaliados. Malformações ou alterações teratogênicas não foram detectadas, no entanto alterações esqueléticas sugestivas de retardo no desenvolvimento foram observadas especialmente nas doses mais altas testadas (300mg.kg-1 e 500mg.kg-1). Considerando-se a situação complexa de risco para a saúde humana, mostra-se importante a execução de investigações adicionais, de modo a contribuir para a adequada avaliação de risco do piriproxifeno em água potável.(AU)

This study investigated the prenatal toxicity of the insecticide pyriproxyfen in Wistar rats to detect the possible changes in the fetal development of the progeny exposed during the organogenic period. Three doses of pyriproxyfen (100, 300, and 500mg.kg-1) were administered orally to the progenitors, from day 6 to 15 of gestation. The fetuses were processed using the Taylor and Van Dyke modified diaphanization technique to evaluate malformations and skeletal changes. The results did not demonstrate the occurrence of systemic maternal toxicity or changes in the reproductive indexes evaluated. Malformations or teratogenic changes were not detected, however, skeletal changes suggestive of developmental delay were observed, especially in the highest doses tested (300 mg.kg-1 and 500 mg.kg-1). Owing to the potentially complex situation regarding its risk to human health, it is important that further studies be performed to contribute to the risk assessment of the addition of pyriproxyfen in drinking water.(AU)

Assessing the efficacy of two new formulations of larvicide pyriproxyfen for the control of Aedes aegypti using dissemination stations in two sites of Rio de Janeiro city

BACKGROUND Aedes aegypti is the primary transmitter of several arbovirus with great impact in human health. Controlling vector mosquitoes is an essential and complex task. One promising control method is to use mosquitoes as a vehicle to disseminate tiny particles of juvenile-killing insecticides, such as pyriproxyfen (PPF), to breeding sites. OBJECTIVES We aimed to investigate the capacity of Ae. aegypti to disseminate two new formulations of PPF in two sites of Rio de Janeiro city for assessment of the efficacy of these products. METHODS Dissemination stations impregnated with powder and liquid new formulations of PPF were installed in two test sites. Ovitraps were used in the test sites and in a control site for monitoring the presence of Ae. aegypti throughout eggs collection. FINDINGS Entomological indices indicated that the new formulations of PPF were efficient in reducing eggs abundance. Liquid formulation performed better than powder formulation. Ready-to-use formulations of PPF can be quickly applied in the field and can be replaced after a few months. MAIN CONCLUSIONS New formulations of PPF associated with mosquito dissemination approach make a valuable vector control strategy, managing to cover places of difficult access for whatever reason. New formulations application requires less labour, being economically attractive.

Susceptibility of Aedes aegypti populations to pyriproxyfen in the Federal District of Brazil

Abstract INTRODUCTION: In Brasilia, pyriproxyfen (PPF; 0.01 mg/L) has been used for the larval control of Aedes aegypti mosquitoes since 2016. Information on the susceptibility of Ae. aegypti to PPF, and the development of resistance in populations from the Federal District of Brazil (FD) is limited. It is essential to monitor the susceptibility of Ae. aegypti to insecticides in order to improve vector control strategies. This study aimed to evaluate the susceptibility of Ae. aegypti populations from five areas of Brasilia to PPF. METHODS: We performed dose-response tests to estimate the emergence inhibition and resistance ratio of each field population, including the Rockefeller reference population. We also analyzed egg positivity, and the density and mortality of larvae and pupae. RESULTS: Populations from Vila Planalto (RR50=1.7), Regiment Guards Cavalry (RR50=2.5), and Sub-secretary of Justice Complex (RR50=3.7) presented high susceptibility to PPF, while the RR values of populations from Lago Norte (RR50=7.7) and Varjão (RR50=5.9) were moderately high, suggesting the emergence of insipient resistance to PPF in Brasilia. At 30 ng/mL, the highest larvae mortality rate was 2.7% for the population from Lago Norte, while that of pupae was 92.1% for Varjão and Vila Planalto. CONCLUSIONS: The five populations of Ae. aegypti from the FD are susceptible to PPF and there is a need to monitor the susceptibility of Ae. aegypti in new areas of the FD.