Evaluación de la eficacia biológica y de la sensibilidad de Aedes aegypti a los insecticidas piretroides deltametrina y ciflutrina durante el brote del virus Zika en Kuna Yala, Panamá / Evaluation of the biological efficacy and susceptibility in Aedes aegypti to the pyrethroid insecticides deltamethrin and cyfluthrin during the Zika virus outbreak in Kuna Yala, Panama

Introducción. El desarrollo de la resistencia a insecticidas de Aedes aegypti representa una gran amenaza para la salud pública. La vigilancia y el monitoreo de la eficacia biológica a los insecticidas y la sensibilidad de las poblaciones de Aedes aegypti es de fundamental importancia para prolongar la vida útil de estas moléculas. Objetivo. Evaluar la eficacia biológica de los insecticidas deltametrina y ciflutrina y la sensibilidad de poblaciones de Aedes aegypti a estos insecticidas durante el brote epidémico de virus del Zika en Kuna Yala, Panamá. Materiales y métodos. Se evaluó la eficacia biológica de la deltametrina y la ciflutrina, y la sensibilidad a estos insecticidas de poblaciones de la cepa Aedes aegypti Ustupo, mediante bioensayos estandarizados por la Organización Mundial de la Salud durante el brote epidémico de virus del Zika en Kuna Yala, Panamá. Resultados. En los bioensayos con Aedes aegypti Ustupo se observó posible resistencia a deltametrina y a ciflutrina con un porcentaje de mortalidad del 95,3 y 94 %, respectivamente. Se registró baja eficacia biológica con la cepa Aedes aegypti Ustupo para la deltametrina y la ciflutrina, con medias de porcentajes de mortalidad de 75 y 31,1 %, en el intradomicilio, mientras que en el peridomicilio fue de 63,7 y 26,1 %, respectivamente. Conclusión. Los resultados de este estudio representan un desafío que debe enfrentar el Programa Nacional de Control de Aedes para lograr cuidar y mantener el efecto tóxico de los insecticidas aplicados contra las poblaciones de Aedes. Es necesario que el Programa Nacional de Control de Aedes establezca unos lineamientos de manejo de la resistencia para caracterizarla y evaluar la distribución geográfica de las poblaciones afectadas. Lo anterior con el propósito de garantizar la sostenibilidad de las intervenciones antivectoriales contra las poblaciones de Aedes.

Introduction. The development of resistance to insecticides in Aedes aegypti represents a major threat to public health. Surveillance and monitoring of the biological efficacy and sensibility of Aedes aegypti populations to insecticides is fundamental to prolong the useful life of insecticide molecules. Objective. To evaluate the biological efficacy of deltamethrin and cyfluthrin and sensibility to insecticides in Aedes aegypti during the zika epidemic outbreak in Kuna Yala, Panama. Materials and methods. We assessed the biological efficacy of deltamethrin and cyfluthrin, and sensibility in the strain Aedes aegypti Ustupo using bioassays standardized by the World Health Organization during the Zika epidemic outbreak in Kuna Yala, Panama. Results. In the bioassays with Aedes aegypti Ustupo, we observed a possible resistance to deltamethrin and cyfluthrin with a mortality rate of 95,3 and 94%, respectively. The obtained results registered low biological efficacy of deltamethrin and cyfluthrin with average percentages of mortality of 75 and 31.1% in the intradomicile, and 63,7 and 26.1% in the peridomicile. Conclusion. The results of this study represent a challenge for the National Aedes Control Program to care for and maintain the toxic effect of insecticides applied against Aedes populations. There is a need for the National Aedes Control Program to establish some guidelines about resistance assessment and resistant populations' geographic distribution to guarantee the sustainability of anti-vector interventions against Aedes populations.

Evaluación de la sensibilidad a organofosforados en poblaciones de Aedes aegypti (L. ) (Diptera: Culicidae) del departamento de La Guajira, Colombia / Evaluation of susceptibility to organophosphates in populations of Aedes aegypti (L. ) (Diptera: Culicidae) in La Guajira, Colombia

Introducción. El dengue es un problema de salud pública para el departamento de La Guajira. El control se ha enfocado en el vector con el uso de insecticidas, entre ellos los organofosforados. Objetivo. Evaluar el estado de la sensibilidad a insecticidas organofosforados de quince poblaciones de Aedes aegypti (L.) en el departamento de La Guajira, Colombia. Materiales y métodos. Se realizaron bioensayos para temefos, malatión y pirimifos- metil en larvas de tercer estadio y mosquitos adultos de Ae. aegypti en los municipios de Albania, Barrancas, Dibulla, Distracción, El Molino, Fonseca, Hatonuevo, La Jagua del Pilar, Maicao, Manaure, Riohacha, San Juan del Cesar, Uribia, Urumita y Villanueva, siguiendo la metodología de la Organización Mundial de la Salud (OMS) y la técnica de botellas usando la guía de los de los Centers for Disease Control and Prevention, respectivamente. Se determinó la sensibilidad por medio de la relación de resistencia a CL50 y CL95 (RRCL50, RRCL95) para temefos y a dosis y tiempo diagnóstico para temefos, malatión y pirimifos-metil en las poblaciones de campo evaluadas, usando como control la cepa sensible Rockefeller. Resultados. Las 15 poblaciones del departamento de La Guajira son sensibles a: temefos (relación de la resistencia a CL50<5,0; relación de resistencia a CL95<5,0; 98 a 100 % de mortalidad); pirimifos-metil (99 a 100 % de mortalidad) y malatión (100 % de mortalidad). Conclusión. Con base en los resultados obtenidos, es factible el uso de temefos, malatión y pirimifos-metil para el control de Ae. aegypti en las poblaciones evaluadas.

Introduction. Dengue is a public health problem in La Guajira region. Control has focused on the vector using insecticides, including organophosphates. Objective. To evaluate the state of susceptibility to organophosphates insecticides in fifteen Aedes aegypti (L.) populations in La Guajira, Colombia. Materials and methods. We collected samples of third-instar larvae and adult mosquitoes of Ae. aegypti in the municipalities of Albania, Barrancas, Dibulla, Distracción, El Molino, Fonseca, Hatonuevo, La Jagua del Pilar, Maicao, Manaure, Riohacha, San Juan del Cesar, Uribia, Urumita, Villanueva. Bioassays for temefos, malathion, and pirimiphos-methyl were carried out following the methodology of the World Health Organization, and the bottle technique using the guidance of the Centers for Disease Control and Prevention. Susceptibility to temefos was determined through the resistance ratio between lethal concentration 50 and lethal concentration 95; for the compounds temefos, malathion and pirimiphos-methyl, susceptibility was calculated using diagnostic dose and diagnostic time in the populations evaluated. Rockefeller susceptible strain was used as a control. Results. All evaluated populations of Ae. aegypti from La Guajira were found to be susceptible to temefos (ratio resistance to CL50<5.0; ratio resistance to CL95<5.0; 98 - 100 % mortality); pirimiphosmethyl (99 - 100 % mortality), and malathion (100 % mortality). Conclusion. Based on the results, the use of temefos, malathion, and pirimiphosmethyl is feasible for the control of Ae. aegypti in the evaluated populations.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Abstract Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Resumo A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-1, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-1 em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-1 como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-1 para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

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 / 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.

Resistência das populações de Aedes (Stegomyia) aegypti (Linnaeus, 1762) (Insecta, Diptera, Culicidae) aos inseticidas utilizados para o controle: estado da arte do conhecimento / Resistance of Aedes (Stegomyia) aegypti (Linnaeus, 1762) (Insecta, Diptera, Culicidae) populations to insecticides used for control: art state of knowledge

Introdução: cerca de quatro bilhões de pessoas residem em áreas com risco de dengue, uma arbovirose transmitida pelo mosquito Aedes (Stegomyia) aegypti (Linnaeus, 1762) (Insecta, Diptera, Culicidae). Na tentativa de combater esse vetor e reduzir a disseminação da dengue, o meio de controle vetorial frequentemente utilizado são os inseticidas. Entretanto, o uso indiscriminado destes no controle do mosquito está relacionado aos mecanismos de resistência desse vetor. Objetivo: mapear as evidências científicas relacionadas à resistência do mosquito A. aegypti aos inseticidas utilizados para o controle populacional. Metodologia: revisão de escopo, segundo metodologia Instituto Joana Briggs, em bases de dados indexadas: PubMed, Embase, CINAHL, Web of Science, SCOPUS e Biblioteca Virtual em Saúde. Através do mnemônico PCC (população, conceito e contexto), elaborou-se a estratégia de busca utilizando descritores do Decs e Mesh. Resultados: foram encontrados 1.631 estudos sobre a temática. Após critérios de elegibilidade e seleção, foram incluídos 30 estudos específicos sobre resistência do Aedes aegypti a inseticidas foram incluídos na revisão. A maior parte tratou de forma experimental e 28 (93,3%) estudos trabalharam o controle químico. Em relação à resistência a inseticidas, em 20 estudos (66,6%) constatou-se resistência metabólica e em 9 (30%) a resistência mediada pela alteração da variabilidade genética. Os estudos que observaram mutação não deixam claro se a mutação gênica é especificamente devido à ação mutagênica aos inseticidas. Conclusões:o A. aegypti desenvolve adaptações que lhe conferem resistência aos inseticidas, sendo que esses mecanismos de resistência estão relacionados à variabilidade genética e a adaptações metabólicas, que são transmitidas a seus descendentes ao longo das gerações. Assim, torna-se necessário um avanço nos estudos visando não apenas identificar e explicar os mecanismos de resistência, mas encontrar meios alternativos de manejo que possam controlar o inseto sem ocasionar resistência aos mesmos.

Introduction: about four billion people live in areas at risk of dengue, an arbovirus transmitted by the Aedes (Stegomyia) aegypti (Linnaeus, 1762) (Insecta, Diptera, Culicidae). To combat this vector and reduce the spread of dengue, the means of vector control often used are insecticides. However, their indiscriminate use in mosquito control is related to the resistance mechanisms of this vector. Objective: to map the scientific evidence related to the resistance of the Aedes aegypti mosquito to the insecticides used to control dengue. Methodology: scope review, according to the Joana Briggs Institute methodology, in indexed databases: PubMed, Embase, CINAHL, Web of Science, SCOPUS and Virtual Health Library. Using the mnemonic PCC (population, concept and context), a search strategy was developed using Decs and Mesh descriptors. Results: 1,631 studies on the subject were found. After eligibility and selection criteria, 30 specific studies on Aedes aegypti resistance to insecticide were included in the review . Most treated experimentally and 28 (93.3%) studies worked on chemical control. Regarding the resistance to insecticide, in 20 studies (66.6%) there was found metabolic resistance and in 9 (30%) resistance mediated by modification on the genetic variability. Studies that have looked at mutation do not make it clear whether the gene mutation is specifically due to mutagenic action of insecticides. Conclusions: Aedes aegypti develops adaptations that gives them resistance to insecticides, and these resistance mechanisms are related to genetic variability and to metabolic adaptations, which are transmitted to its descendants over generations. Thus, it is necessary to perform advance in studies aiming not only to identify and explain the resistance mechanisms, but to find alternative means of management that can control the insect without causing resistance to them.

Advances in receptor-mediated resistance mechanisms of Lepidopteran insects to Bacillus thuringiensis toxin / 生物工程学报

Bacillus thuringiensis is widely used as an insecticide which is safe and environmentally friendly to humans and animals. One of the important insecticidal mechanisms is the binding of Bt toxins to specific toxin receptors in insect midgut and forming a toxin perforation which eventually leads to insect death. The resistance of target pests to Bt toxins is an important factor hampering the long-term effective cultivation of Bt crops and the continuous use of Bt toxins. This review summarizes the mechanism of insect resistance to Bt toxins from the perspective of important Bt toxin receptors in midgut cells of Lepidopteran insects, which may facilitate the in-depth study of Bt resistance mechanism and pest control.

Control of pyrethroid-resistant populations of Triatoma infestans, the main vector of Trypanosoma cruzi, by treating dogs with fluralaner in the Argentine Chaco

We assessed whether fluralaner administered to outbred healthy dogs reduced or supressed site infestation and abundance of pyrethroid-resistant populations of Triatoma infestans Klug (Heteroptera: Reduviidae). We conducted a placebo-controlled before-and-after efficacy trial in 28 infested sites in Castelli (Argentine Chaco) over 10 months. All 72 dogs initially present received either an oral dose of fluralaner (treated group) or placebo (control group) at month 0 posttreatment (MPT). Preliminary results justified treating all 38 control-house dogs with fluralaner 1 month later, and 71 of 78 existing dogs at 7 MPT. Site-level infestation and triatomine abundance were evaluated using timed manual searches with a dislodging aerosol. In the fluralaner-treated group, infestation dropped significantly from 100% at baseline to 19% over 6-10 MPT whereas mean abundance fell highly significantly from 5.5 to 0.8-0.9 triatomines per unit effort. In the placebo group, site infestation and mean abundance remained stable between 0 and 1 MPT, and strongly declined after fluralaner administration from 13.0-14.7 - triatomines at 0-1 MPT to 4.0-4.2 over 6-10 MPT. Only one of 81 noninfested sites before fluralaner treatment became infested subsequently. Fluralaner significantly reduced the site-level infestation and abundance of pyrethroid-resistant T. infestans and should be tested more widely in Phase III efficacy trials.

Fluctuating asymmetry and exposure to pyrethroid insecticides in Triatoma infestans populations in northeastern Argentina

Exposure to sublethal doses of insecticide may affect biological traits in triatomines. We investigated the effects of toxicological phenotype (pyrethroid resistance status) and exposure to sublethal doses of deltamethrin on two traits of Triatoma infestans Klug (Heteroptera: Reduviidae) using a phenotypic plasticity experimental design. First-instar nymphs from 14 and 10 full-sib families from pyrethroid-susceptible and pyrethroid-resistant populations, respectively, were used. For the susceptible population, we treated first instars topically with acetone (control) or deltamethrin (treatment) once. For the resistant population, instars were treated once, twice and three times as first, third or fifth-instar nymphs, respectively. We measured cuticle thickness, wing size and wing shape of 484 emerging adults, and tested for treatment effects using mixed ANOVA and MANOVA models. Toxicological phenotype, exposure to deltamethrin and full-sib family exerted significant effects on cuticle thickness, wing size and wing shape. Adult triatomines previously treated with deltamethrin developed significantly thicker cuticles than control triatomines only in the resistant population and significantly bigger wings in both populations. Mean cuticle thickness and wing size increased with increasing exposures to deltamethrin. Exposure to sublethal doses of deltamethrin generated morphological modifications that may affect insect survival and flight dispersal, and hence may have evolutionary and epidemiological consequences.

Estruturação genética e distribuição de mutações kdr em populações de Aedes aegypti (Linnaeus 1762) do estado do Amapá / Genetic structuring and distribution of kdr mutations in populations of Aedes aegypti (Linnaeus 1762) in the state of Amapá

O mosquito Aedes aegypti é vetor competente para a transmissão dos vírus dengue, chikungunya, Zika e febre amarela. O controle químico do vetor é amplamente utilizado, mas vem sendo ameaçado pela disseminação da resistência a inseticidas (RI), caracterizando-se pela seleção de alterações no comportamento ou na fisiologia do mosquito, que fazem com que o inseticida não atinja ou não encontre seu alvo no organismo do inseto. A disseminação da RI ocorre usualmente pelo fluxo gênico entre populações, que no caso do Ae. aegypti é facilitado pela dispersão passiva de seus ovos aderidos a mercadorias. A estruturação genética das populações naturais é, portanto, um fator fundamental que facilita ou dificulta a disseminação de alelos de resistência. Entre os principais mecanismos de RI estão as alterações no gene do canal de sódio regulado por voltagem (NaV), que causam resistência ao efeito knockdown de piretroides, sendo conhecidas como mutações kdr. Populações de Ae. aegypti das localidades de Macapá e Oiapoque, no estado do Amapá, eram geneticamente distintas e apresentavam alelos kdr diferenciados, muito provavelmente devido ao baixo fluxo gênico entre essas duas populações, dificultado pelo acesso terrestre entre essas duas localidades, feito por uma única rodovia que cruza a densa Floresta Amazônica. Não se conhecia, no entanto, como seria a estruturação genética entre as populações de localidades ao longo daquela rodovia e, tampouco sobre ocorrência de mutações kdr. Por isso, avaliamos a frequência das principais mutações kdr em três sítios polimórficos (V410L, V1016I e F1534C) no gene NaV de populações de Ae. aegypti de seis localidades do estado do Amapá (Oiapoque, Calçoene, Porto Grande, Ferreira Gomes, Tartarugalzinho e Macapá), através de genotipagem por qPCR em tempo real. Adicionalmente, investigamos a diversidade e a estruturação genética destas populações via genotipagem de 12 loci de microssatélites. Os alelos kdr R1 (mutante no sítio 1534) e R2 (mutante nos três sítios) foram observados nas seis populações, sendo que a de Oiapoque apresentou a maior frequência do kdr R2, enquanto o kdr R1 foi mais frequente na de Macapá. Genótipos kdr raros em demais regiões do Brasil, sugestivos de duplicações gênicas, foram encontrados em Ae. aegypti de Oiapoque e Calçoene. As análises de microssatélites mostraram baixa ou moderada diferenciação genética par-a-par, sendo que Oiapoque foi a população geneticamente mais distinta das demais e formou praticamente um único grupo genético em uma das análises de estruturação genética. Análise de Componentes Principais (DAPC) também sugeriu a população de Calçoene geneticamente diferenciada das demais, enquanto as outras populações mostraram-se mais similares entre si, sugerindo alto fluxo gênico entre elas. Ainda que a população de Oiapoque seja a mais diferenciada e estruturada, nosso estudo indica que há fluxo gênico entre esta e as demais ao longo da estrada BR-156 (AP), colaborando com a disseminação de alelos vindos da população da Guiana Francesa. Idealmente, medidas de controle químico do vetor devem ser pensadas conjuntamente entre as autoridades de ambos os lados da fronteira binacional.(AU)

Locomotor activity in Aedes aegypti with different insecticide resistance profiles

ABSTRACT OBJECTIVE: To evaluate locomotor activity in four field populations of Ae. aegypti with different insecticide resistance profiles from the state of São Paulo for two years. METHODS: This study comprised the susceptible Rockefeller strain and four populations from São Paulo, Brazil: two considered populations with "reduced susceptibility" to pyrethroids (Campinas and Marília), and two "resistant populations" (Santos and Ribeirão Preto). First, 2016 and 2017 eggs from these five populations were hatched in laboratory. Virgin females underwent experiments under laboratory conditions at 25°C, with 12:12h light/dark (LD) photoperiod; 24-hour individual activity was recorded using a locomotor activity monitor (LAM). RESULTS: In females from 2016 field populations, both resistant populations showed significant more locomotor activity than the two reduced susceptibility populations and the Rockefeller strain (p < 0.05). As for females from 2017 field populations, reduced susceptibility populations showed a significant increased locomotor activity than the Rockefeller strain, but no significant difference when compared to Santos resistant population (p > 0.05). CONCLUSIONS: Our results indicate that insecticide-resistant Ae. aegypti populations show increased locomotor activity, which may affect the transmission dynamics of their arboviruses.

Estado de la resistencia a insecticidas en Triatoma infestans de Argentina

Los insectos triatominos son los vectores de la enfermedad de Chagas. El control químico de estos insectos es la principal herramienta utilizada para reducir la incidencia de la enfermedad. Los piretroides son los principales insecticidas utilizados para el control de los triatominos en todos los países endémicos. La evolución de resistencia a insecticidas convierte en inefciente una exitosa estrategia de control químico. En Argentina se han detectado diferentes focos de resistencia en Triatoma infestans, el vector más importante en el Cono Sur de Sudamérica, asociados a defciencias en el control. Los estudios realizados sobre esta especie muestran que la resistencia es un problema complejo, ya que evolucionó en varias zonas de la distribución geográfca de la especie, se identifcaron diferentes mecanismos de resistencia, está asociada a modifcaciones en diversos procesos biológicos, está vinculada a variaciones ambientales y hay escasez de alternativas de control. En este manuscrito presentamos una revisión del conocimiento sobre la resistencia a piretroides en T. infestans en Argentina.

Recent trends in global insecticide use for disease vector control and potential implications for resistance management

Insecticides have played a major role in the prevention, control, and elimination of vector-borne diseases, but insecticide resistance threatens the efficacy of available vector control tools. A global survey was conducted to investigate vector control insecticide use from 2010 to 2019. Out of 140 countries selected as sample for the study, 87 countries responded. Also, data on ex-factory deliveries of insecticide-treated nets (ITNs) were analyzed. Insecticide operational use was highest for control of malaria, followed by dengue, leishmaniasis and Chagas disease. Vector control relied on few insecticide classes with pyrethroids the most used overall. Results indicated that IRS programs have been slow to react to detection of pyrethroid resistance, while proactive resistance management using insecticides with unrelated modes of action was generally weak. The intensive use of recently introduced insecticide products raised concern about product stewardship regarding the preservation of insecticide susceptibility in vector populations. Resistance management was weakest for control of dengue, leishmaniasis or Chagas disease. Therefore, it will be vital that vector control programs coordinate on insecticide procurement, planning, implementation, resistance monitoring, and capacity building. Moreover, increased consideration should be given to alternative vector control tools that prevent the development of insecticide resistance.

History of insecticide resistance of Triatominae vectors

In the last 15 years, different types of Triatominae resistance to different insecticides have been reported; thus, resistance may be more widespread than known, requiring better characterization and delimitation, which was the aim of this review. This review was structured on a literature search of all articles from 1970 to 2015 in the PubMed database that contained the keywords Insecticide resistance and Triatominae. Out of 295 articles screened by title, 33 texts were selected for detailed analysis. Insecticide resistance of Triatomines is a complex phenomenon that has been primarily reported in Argentina and Bolivia, and is caused by different factors (associated or isolated). Insecticide resistance of Triatominae is a characteristic inherited in an autosomal and semi-dominant manner, and is polygenic, being present in both domestic and sylvatic populations. The toxicological profi le observed in eggs cannot be transposed to different stages of evolution. Different toxicological profi les exist at macro- and microgeographical levels. The insecticide phenotype has both reproductive and developmental costs. Different physiological mechanisms are involved in resistance. Studies of Triatomine resistance to insecticides highlight three defi ciencies in interpreting the obtained results: I) the vast diversity of methodologies, despite the existence of a single guiding protocol; II) the lack of information on the actual impact of resistance ratios in the fi eld; and III) the concept of the susceptibility reference lineage. Research on the biological and behavioral characteristics of each Triatominae species that has evolved resistance is required in relation to the environmental conditions of each region.

Knockdown of CYP4PR1, a cytochrome P450 gene highly expressed in the integument tissue of Triatoma infestans, increases susceptibility to deltamethrin in pyrethroid-resistant insects / A eliminação do CYP4PR1, um gene do citocromo P450 altamente expresso no tecido tegumentar de Triatoma infestans, aumenta a suscetibilidade à deltametrina em insetos resistentes aos piretróides

Abstract Metabolic resistance to chemical insecticides implies a greater capacity to detoxify insecticides due to an increase in the expression of genes and/or in the activity of enzymes related to detoxification metabolism. The insect integument is known to participate as the cuticular penetration factor of resistance, but recently this tissue was also linked with metabolic resistance due to P450-dependent detoxification in the Chagas disease vector Triatoma infestans. The objectives of this study were i) to name and classify all P450s known to date in T. infestans, ii) to characterise one of them, CYP4PR1, representing the first member of a new cytochrome P450 subfamily described in insects, and iii) to investigate the potential role of CYP4PR1 in metabolic resistance to deltamethrin in T. infestans. We found that CYP4PR1 is expressed almost exclusively in the integument tissue, and its expression was not induced by deltamethrin. Knockdown of CYP4PR1 by RNA interference in pyrethroid-resistant nymphs caused a significant increment in insect mortality after topical application of two different doses of deltamethrin. These results support the role of the integument on metabolic resistance and suggest that CYP4PR1 might contribute to resistance in integument tissue of T. infestans.

Improved vector control of Triatoma infestans limited by emerging pyrethroid resistance across an urban-to-rural gradient in the Argentine Chaco

Background: The sustainable elimination of Triatoma infestans in the Gran Chaco region represents an enduring challenge. Following the limited effects of a routine pyrethroid insecticide spraying campaign conducted over 2011­2013 (first period) in Avia Terai, an endemic municipality with approximately 2300 houses, we implemented a rapid-impact intervention package to suppress house infestation across the urban-to-rural gradient over 2015­2019 (second period). Here, we assess their impacts and whether persisting infestations were associated with pyrethroid resistance. Methods: The 2011­2013 campaign achieved a limited detection and spray coverage across settings (< 68%), more so during the surveillance phase. Following community mobilization and school-based interventions, the 2015­2019 program assessed baseline house infestation using a stratified sampling strategy; sprayed all rural houses with suspension concentrate beta-cypermethrin, and selectively sprayed infested and adjacent houses in urban and peri-urban settings; and monitored house infestation and performed selective treatments over the follow-up. Results: Over the first period, house infestation returned to pre-intervention levels within 3­4 years. The adjusted relative odds of house infestation between 2011­2013 and 2015­2016 differed very little (adj. OR: 1.17, 95% CI 0.91­1.51). Over the second period, infestation decreased significantly between 0 and 1 year post-spraying (YPS) (adj. OR: 0.36, 95% CI 0.28­0.46), with heterogeneous effects across the gradient. Mean bug abundance also dropped between 0 and 1 YPS and thereafter remained stable in rural and peri-urban areas. Using multiple regression models, house infestation and bug abundance at 1 YPS were 3­4 times higher if the house had been infested before treatment, or was scored as high-risk or non-participating. No low-risk house was ever infested. Persistent foci over two successive surveys increased from 30.0 to 59.3% across the gradient. Infestation was more concentrated in peridomestic rather than domestic habitats. Discriminating-dose bioassays showed incipient or moderate pyrethroid resistance in 7% of 28 triatomine populations collected over 2015­2016 and in 83% of 52 post-spraying populations. Conclusions: The intervention package was substantially more effective than the routine insecticide spraying campaign, though the effects were lower than predicted due to unexpected incipient or moderate pyrethroid resistance. Increased awareness and diagnosis of vector control failures in the Gran Chaco, including appropriate remedial actions, are greatly needed.

Insecticidal efficacy of fluralaner (Bravecto®) against Triatoma brasiliensis, a major vector of Trypanosoma cruzi in Brazil

Background: Triatomines are responsible for the vector transmission of the protozoan parasite Trypanosoma cruzi, which causes Chagas disease. Triatoma brasiliensis is the main vector of the parasite in Brazil, and dogs are an important reservoir of the parasite. The aim of this study was to evaluate the insecticidal effect of fluralaner (Bravecto®) on T. brasiliensis after a blood meal in treated dogs. Methods: Healthy mongrel dogs (n = 8) were recruited from the Zoonoses Control Center (ZCC) in the city of Natal, Rio Grande do Norte, Brazil, and randomized into two groups, a fluralaner (Bravecto®)-treated group (n = 4) and a control group (n = 4). Colony-reared third-, fourth- and fifth-instar nymphs of T. brasiliensis nymphs (n = 10) were allowed to feed on dogs from both groups for 30-40 min, once monthly, for up to 12 months. Bug mortality was observed up to 5 days after each blood meal. Results: Mortality in triatomines which had a blood meal on fluralaner (Bravecto®)-treated dogs was 100% for up to 7 months after treatment, with mortality decreasing to 66.4% after 8 months, 57% after 9 months, 35% after 10 months, 10% after 11 months and 0% after 12 months. The mortality of triatomines that fed on non-treated control dogs was always ≤ 2.5%. Conclusions: Our results suggest that fluralaner (Bravecto®) treatment of dogs induces long-term mortality of T. brasiliensis after the blood meal. This is a potential approach to be used to control vector transmission of T. cruzi, the etiological agent of Chagas disease, especially in endemic areas.

Insights sobre a evolução e dispersão da resistência aos piretróides entre Triatoma infestans andinos silvestres da Bolívia

Abstract: Sylvatic populations of Triatoma infestans represent a challenge to Chagas disease control as they are not targeted by vector control activities and may play a key role in post-spraying house reinfestation. Understanding sylvatic foci distribution and gene flow between sylvatic and domestic populations is crucial to optimize vector control interventions and elucidate the development and spread of insecticide resistance. Herein, the genetic profiles of five Andean T. infestans populations from Bolivia with distinct insecticide susceptibility profiles were compared. Multilocus genotypes based on eight microsatellites and the DNA sequence of a fragment of the cytochrome B (cytB) gene were obtained for 92 individuals. CytB haplotypes were analyzed with previously reported Bolivian T. infestans haplotypes to evaluate putative historical gene flow among populations. Each specimen was also screened for two nucleotide mutations in the sodium channel gene (kdr), related to pyrethroid resistance (L1014 and L9251). Significant genetic differentiation was observed among all populations, although individuals of admixed origin were detected in four of them. Notably, the genetic profiles of adjacent domestic and sylvatic populations of Mataral, characterized by higher levels of insecticide resistance, support their common ancestry. Only one sylvatic individual from Mataral carried the kdr mutation L1014, suggesting that this mechanism is unlikely to cause the altered insecticide susceptibility observed in these populations. However, as the resistance mutation is present in the area, it has the potential to be selected under insecticidal pressure. Genetic comparisons of these populations suggest that insecticide resistance is likely conferred by ancient trait(s) in T. infestans sylvatic populations, which are capable of invading domiciles. These results emphasize the need for stronger entomological surveillance in the region, including early detection of house invasion, particularly post-spraying, monitoring for resistance to pyrethroids and the design of integrative control actions that consider sylvatic foci around domestic settings and their dispersion dynamics.

Evaluation of the insecticide susceptibility profile in Cimex lectularius (Hemiptera: Cimicidae) in Belo Horizonte (Brazil)

Abstract INTRODUCTION: Since 2013, major Cimex lectularius infestations have been detected in public shelters in Belo Horizonte (Brazil). Due to this, insecticide resistance has been investigated as one of the possible causes for the failure to control bedbugs. METHODS: Cimicids were subjected to bioassays according to the World Health Organization recommendations using deltamethrin and all commercial insecticides available for control of Cimex in Brazil. RESULTS: Cimicids were deltamethrin resistant and presented indicative of resistance to other insecticides, except for propoxur 1%. CONCLUSIONS: The commercial insecticides have a limited effect on bedbug populations, which may justify the unsatisfactory control observed in the shelters studied.

Molecular variation of Plasmodium vivax dehydrofolate reductase in Mexico and Nicaragua contrasts with that occurring in South America / Variación molecular de la dehidrofolato reductasa de Plasmodium vivax en México y Nicaragua contrasta con la que ocurre en Sudamérica

Abstract: Objective: To research mutations associated to pyrimethamine resistance in dihydrofolate reductase (pvdhfr) of Plasmodium vivax from Mexico and Nicaragua and compare it to that reported in the rest of America. Materials and methods: Genomic DNA was obtained from P. vivax-infected blood samples. A pvdhfr gene fragment was amplified and sequenced. The identified gene variations were compared to those observed in other affected sites of America. Results: No mutations in pvdhfr were detected in P. vivax from Mexico and Nicaragua. One synonymous change and variation in the repeat domain was detected in Nicaraguan parasites. In South America, a high frequency of variant residues 58R and 117N associated to pyrimethamine resistance was reported. Conclusions: The lack of polymorphisms associated with pyrimethamine resistance suggests that drug-resistant P. vivax has not penetrated Mesoamerica, nor have local parasites been under selective pressure. These data contribute to establish the basis for the epidemiological surveillance of drug resistance.

Resumen: Objetivo: Determinar mutaciones en la dihydrofolato reductasa deP. vivax (Pvdhfr) en parásitos de México y Nicaragua, y comparar con lo reportado en América. Material y métodos: Del ADN de sangres infectadas con P. vivax de pacientes, el gen pvdhfr se amplifico y secuenció, y se contrastócon lo observado en América. Resultados: No se detectaron mutaciones asociadas con la resistencia debida a pirimetamina. Los parásitos de Nicaragua tuvieron una mutación sinónima y variación en la región repetida. Se reportaron frecuentes mutaciones asociadas con la resistencia a la pirimetamina en Sudamérica. Conclusiones: La ausencia de polimorfismos en Pvdhfr sugiere que no se han seleccionado ni introducido parásitos resistentes en la zona de estudio, lo que resulta muy útil para la vigilancia epidemiológica.