Abstract The use of botanical insecticides has increased in recent years due to the demand for effective products, particularly against insects resistant to conventional insecticides. Among these is Aedes aegypti, a well-adapted mosquito to urban environments that opportunistically feeds on humans and animals, contributing to the spread of virus. We evaluated the potential of essential oils (EOs) extracted from Eucalyptus citriodora, Eucalyptus staigeriana, Eucalyptus caryophyllus, and Mentha arvensis in terms of their larvicidal activity against Ae. aegypti. EOs’ compounds were determined using gas chromatography-mass spectrometry (GC-MS). Bioassays were performed on third instar larvae of Ae. aegypti to evaluate the larvicidal effects of EO dilutions in dimethyl sulfoxide (DMSO) at different concentrations. Mortality rates were observed over a 72-hour period to determine the efficacy of the treatments. Citronellal (86.64) predominated in E. citriodora, limonene in E. staigeriana (41.68), eugenol in E. caryophyllus (87.76), and menthol in M. arvensis (51.53%). EOs exhibited larvicidal activity from 10 ppm, with notable efficacy at 85 ppm, in which those from E. staigeriana and M. arvensis caused maximum mortality to Ae. aegypti larvae. Results revealed distinct efficacy patterns among EOs, with E. staigeriana displaying high toxicity within 24 h, achieving LC50 and LC95 values of 47.04 ppm and 97.35 ppm, respectively. Larvicidal effects within 1 h were observed for E. citriodora and E. caryophyllus. This study underscores larvicidal efficacy against Ae. aegypti, notably E. staigeriana, which had the lowest LC50 value. The findings indicate that the tested samples have potential for use as bioinsecticides. products welladapted well adapted animals virus (EOs Ae chromatographymass chromatography mass GCMS. GCMS GC MS . (GC-MS) DMSO (DMSO concentrations 72hour hour 72 treatments 86.64 8664 86 64 (86.64 E 41.68, 4168 41.68 , 41 68 (41.68) 87.76, 8776 87.76 87 76 (87.76) M 51.53%. 5153 51.53% 51 53 (51.53%) 8 2 LC LC5 LC9 4704 47 04 47.0 9735 97 35 97.3 respectively value bioinsecticides (GC-MS 7 86.6 866 6 (86.6 416 41.6 4 (41.68 877 87.7 (87.76 515 51.53 5 (51.53% 470 0 47. 973 9 3 97. 86. (86. 41. (41.6 87. (87.7 51.5 (51.53 (86 (41. (87. 51. (51.5 (8 (41 (87 (51. ( (4 (51 (5

Resumo O uso de inseticidas botânicos tem aumentado nos últimos anos devido à demanda por produtos eficazes, especialmente contra insetos resistentes aos inseticidas convencionais. Entre eles está o Aedes aegypti, um mosquito bem adaptado a ambientes urbanos que se alimenta oportunisticamente de humanos e animais, contribuindo para dispersão de vírus. Avaliamos o potencial de óleos essenciais (OEs) extraídos de Eucalyptus citriodora, Eucalyptus staigeriana, Eucalyptus caryophyllus e Mentha arvensis em relação à sua atividade larvicida contra Ae. aegypti. Os compostos dos OEs foram determinados usando cromatografia gasosa acoplada à espectrometria de massa (GC-MS). Os bioensaios foram realizados em larvas de terceiro instar de Ae. aegypti para avaliar os efeitos larvicidas das diluições de OE em dimetil sulfóxido (DMSO) em diferentes concentrações. As taxas de mortalidade foram observadas ao longo de 72 horas para determinar a eficácia dos tratamentos. O componente citronelal (86,64) predominou em E. citriodora, limoneno em E. staigeriana (41,68), eugenol em E. caryophyllus (87,76) e mentol em M. arvensis (50,73%). Os OEs exibiram atividade larvicida a partir de 10 ppm, com grande eficácia a 85 ppm, onde os de E. staigeriana e M. arvensis causaram mortalidade máxima de larvas de Ae. aegypti. Os resultados revelaram padrões de eficácia distintos entre os OEs, com E. staigeriana exibindo maior toxicidade dentro de 24 horas, alcançando valores de CL50 e CL95 de 47,04 ppm e 97,35 ppm, respectivamente. Efeitos larvicidas dentro de 1 hora foram observados para E. citriodora e E. caryophyllus. Este estudo destaca a eficácia larvicida contra Ae. aegypti, especialmente E. staigeriana, que teve o menor valor de CL50. Concluímos que esses OEs são potencialmente larvicidas para o controle eficaz de Ae. aegypti. eficazes convencionais animais vírus (OEs Ae GCMS. GCMS GC MS . (GC-MS) DMSO (DMSO concentrações 7 tratamentos 86,64 8664 86 64 (86,64 E 41,68, 4168 41,68 , 41 68 (41,68) 87,76 8776 87 76 (87,76 M 50,73%. 5073 50,73% 50 73 (50,73%) 8 2 CL CL5 CL9 4704 47 04 47,0 9735 97 35 97,3 respectivamente (GC-MS 86,6 866 6 (86,6 416 41,6 4 (41,68 87,7 877 (87,7 507 50,73 5 (50,73% 470 0 47, 973 9 3 97, 86, (86, 41, (41,6 87, (87, 50,7 (50,73 (86 (41, (87 50, (50,7 (8 (41 (50, ( (4 (50 (5

Abstract Around the world, the main problems of livestock are caused by ectoparasites, however, commercial acaracide are toxic to the environment and detrimental to One Health. Therefore, research has increasingly focused on development of natural products as alternatives for tick control. The purpose of this study was to evaluate the larvicidal effect on Rhipicephalus (Boophilus) microplus, through use of essential oils (EOs) extracted from the leaves, flower buds and stems of Tetradenia riparia. The chemical composition of these EOs was determined through gas chromatography coupled to mass spectrometry (GC-MS). They were tested on larvae at concentrations of 100.000 to 40 µg/mL, using the larval packet test and under semi-natural conditions. The main class of compounds in the chemical composition was sesquiterpenes (both oxygenates and hydrocarbons), whereas the predominant compounds in the leaves, flower buds and stems were 14-hydroxy-9-epi-caryophyllene, T-cadinol and 6-7-dehydroroyleanone, respectively. The leaves proved to be the most effective, with highest larvicidal activity (LC99.9 = 83.53 µg/mL). When tested under semi-natural conditions, the oils obtained efficiency above 98% in all compound tests. The results indicated that these EOs were effective against R. (B.) microplus larvae in vitro and ex-situ, proving that this plant has bioactive molecules with significant larvicidal activity. world ectoparasites however Health Therefore control Boophilus (Boophilus (EOs riparia GCMS. GCMS GC MS . (GC-MS) 100000 100 000 100.00 4 µgmL µg mL µg/mL seminatural semi conditions both hydrocarbons, hydrocarbons , hydrocarbons) 14hydroxy9epicaryophyllene, 14hydroxy9epicaryophyllene hydroxyepicaryophyllene 14 hydroxy 9 epi caryophyllene, caryophyllene 14-hydroxy-9-epi-caryophyllene Tcadinol T cadinol 67dehydroroyleanone, 67dehydroroyleanone dehydroroyleanone 6 7 dehydroroyleanone, 6-7-dehydroroyleanone respectively LC99.9 LC999 LC LC99 (LC99. 8353 83 53 83.5 µg/mL. µg/mL) 98 tests R B. B (B. exsitu, exsitu ex situ, situ ex-situ (GC-MS 10000 10 00 100.0 epicaryophyllene 1 LC99. LC9 (LC99 835 8 5 83. (B 1000 0 100. (LC9 (LC

Resumo Os principais problemas para a pecuária estão relacionados às ectoparasitoses, e ao fato dos carrapaticidas apresentarem elevada toxicidade ao meio ambiente e à saúde única. Surgem, então, demandas na busca por inovações e desenvolvimento de produtos naturais, como alternativas para o controle dos carrapatos. O objetivo deste trabalho foi avaliar o potencial da atividade larvicida sobre Rhipicephalus (Boophilus) microplus a partir dos óleos essenciais de Tetradenia riparia (TrOEs) extraídos das folhas, botões florais e caules. A composição química foi determinada por cromatografia gasosa acoplada à espectrometria de massa (GC/MS). Os TrOEs foram testados sobre larvas nas concentrações de 100.000 a 40 µg/mL pelo teste de pacote de larvas e em condições seminaturais. Na composição química, a classe majoritária foi os sesquiterpenos (oxigenados e hidrocarbonetos); já os compostos em destaques foram 14-hidroxy-9-epi-caryophyllene, T-cadinol e 6-7-dehidroroyleanone para folhas, botões florais e caules, respectivamente. As folhas demonstraram ser mais eficientes e com maior poder larvicida (CL99.9 = 83.53 µg/mL). Quando testado em condições seminaturais os óleos obtiveram eficiência acima de 98% em todos os compostos testados. Os resultados indicaram que os TrOEs, foram eficazes sobre as larvas de R. (B.) microplus in vitro e ex-situ, evidenciando que esta planta possui moléculas bioativas com ação larvicidas significativas. ectoparasitoses única Surgem então naturais carrapatos Boophilus (Boophilus (TrOEs caules GC/MS. GCMS GC/MS . GC MS (GC/MS) 100000 100 000 100.00 4 µgmL µg mL oxigenados hidrocarbonetos hidrocarbonetos) 14hidroxy9epicaryophyllene, 14hidroxy9epicaryophyllene hidroxyepicaryophyllene 14 hidroxy 9 epi caryophyllene, caryophyllene 14-hidroxy-9-epi-caryophyllene Tcadinol T cadinol 67dehidroroyleanone dehidroroyleanone 6 7 respectivamente CL99.9 CL999 CL CL99 (CL99. 8353 83 53 83.5 µg/mL. µg/mL) 98 R B. B (B. exsitu, exsitu ex situ, situ ex-situ significativas (GC/MS 10000 10 00 100.0 epicaryophyllene 1 CL99. CL9 (CL99 835 8 5 83. (B 1000 0 100. (CL9 (CL

RESUMO: O uso contínuo de inseticidas sintéticos para o controle do mosquito vetor de arbovírus, Aedes aegypti, tem levado à seleção natural de populações resistentes a diferentes grupos químicos. Assim, compostos derivados de plantas surgiram como uma alternativa viável para o controle desses vetores. Desse modo, este estudo foi realizado para determinar se o extrato metanólico bruto (CME) das folhas de Clibadium surinamense possui atividade larvicida contra Ae. aegypti. Para isso, Larvas de terceiro e quarto instar de Ae. aegypti foram mantidas em recipientes contendo 99 mL de água e 1 mL de CME diluído em etanol nas concentrações de 250, 500, 750 e 1000 ppm. O grupo controle continha 99 mL de água e 1 mL de etanol. Três ensaios foram realizados em triplicata para cada grupo. Após 24 horas de observação, de acordo com a análise de variância de uma via,os valores de CL50 e CL90 foram de 283 e 430 ppm, respectivamente. Em conclusão, demonstramos pela primeira vez que o CME das folhas de C. surinamense apresenta atividade larvicida contra Ae. aegypti em condições de laboratório.

ABSTRACT: The continuous use of synthetic insecticides for controlling the arboviral vector Aedes aegypti has led to the natural selection of mosquito populations resistant to different chemical groups. Thus, plant-derived compounds have emerged as a viable alternative for vectorcontrol. This study determined whether the crude methanolic extract (CME) from leaves of Clibadium surinamense has larvicidal activity against Ae. aegypti. Third- and fourth-instar Ae. Aegyptilarvae were kept in recipients containing 99 mL of water and 1mL of ethanol-diluted CMEat concentrations of 250, 500, 750, and 1000 ppm. The control group contained 99 mL of water and 1 mL of ethanol. Three trials were performed in triplicate for each group.After 24 hours of treatment, the LC50 and LC90 values were determined to be 283 and 430 ppm, respectively, according to one-way analysis of variance. In conclusion, we have demonstrated for the first time that the CME from leaves of C. surinamense show larvicidal activity against Ae. aegypti under laboratory conditions.

Resumen: La OMS ha señalado que las enfermedades de transmisión vectorial-ETV, engloban más de 17% de las enfermedades infecciosas en todo el mundo, entre ellas el dengue. El presente estudio se encuentra abocado en el Plan de Acción sobre entomología y control de vectores 2018-2023 de la OMS y OPS, respecto a la presencia del Aedes aegypti vector del virus dengue-DEN y su susceptibilidad al control químico, en áreas bajo influencia de asentamientos humanos precarios en el municipio de San Antonio, Central-Paraguay, altamente urbanizado. Un muestreo de campo a través de ovitrampas generó datos de criaderos de Ae. aegypti en áreas bajo influencia de asentamientos humanos precarios con índices de infestación larvaria por Ae aegypti entre 8,6% y 9,5%, superando el valor tolerable < 1% estudiados entre 2018 y 2019. Mediante modelación por DIVAGIS, fueron determinadas las áreas de existencia de Ae aegypti en San Antonio y los distritos vecinos, en relación a las 19 variables bioclimáticas, siendo las variables de mayor influencia el intervalo de temperatura diurna promedio la isotermalidad, temperatura promedio del mes más frio y la precipitación del mes seco, que garantizan la presencia y desarrollo de este vector Aedes aegypti). Los Mapas generados fueron complementados con la presencia de asentamientos humanos e industrias como actividad antrópica insertadas en cuencas hidrográficas y vías fluviales importantes para la instalación del vector en domicilio y peri domicilio. La evaluación de la susceptibilidad/ resistencia de poblaciones silvestres de Ae. aegypti colectado en San Antonio, a larvicida Temefos, mediante bioensayos en condiciones controladas de laboratorio, reveló que las poblaciones circulantes del vector Aedes aegypti, son susceptibles RR<3 al larvicida utilizado por los programas de control vectorial del Ministerio de Salud Pública y Bienestar Social. Todo lo anteriormente expuesto, se encuentra enmarcado en el desarrollo de las metas de los objetivos de desarrollo sostenible, los cuales estipulan que, para el 2030, se debe poner fin a las enfermedades tropicales desatendidas y otras transmisibles por vectores, más específicamente en ODS 3 Salud y Bienestar. Igualmente se circunscribe con el ODS 11, el cual versa sobre ciudades y comunidades sostenibles, debido a que indaga aspectos relativos a la creciente urbanización en la zona de estudio, con varios desafíos, como la expansión urbana no planificada, sin servicios lo que hace que las ciudades sean más vulnerables a las enfermedades. El trabajo desarrollado sienta bases para futuros estudios de empoderamiento ciudadano y desarrollo de una adecuada gobernanza, frente al problema de salud urbana que constituye el DEN.

Abstract: WHO has reported that vector-borne diseases-VTDs account for more than 17% of infectious diseases worldwide, including dengue. The present study is addressed in the Action Plan on entomology and vector control 2018-2023 of the WHO and PAHO, regarding the presence of Aedes aegypti as vector dengue virus-DEN and its susceptibility to chemical control, in areas under the influence of precarious human settlements in the municipality of San Antonio, Central-Paraguay, highly urbanized. Field sampling through ovitraps generated data from Ae aegypti hatcheries in areas under the influence of precarious human settlements, with larval infestation index between 8.6% and 9.5%, exceeding the tolerable value < 1% between 2018 and 2019. Areas of the presence of Ae aegypti in San Antonio and neighboring districts were determined by DIVAGIS modeling, in relation to the 19 bioclimatic variables. The more determinant variables were the average of daily temperature, the isothermality, the average of temperature of the coldest month and precipitation of the driest month; all of them ensure the presence and development of Ae. aegypti. The resulting maps were complemented with the presence of human settlements and industries, as anthropic activity, inserted in watersheds and important waterways for the installation of the vector at home and peridomicile. The evaluation of the susceptibility/resistance to larvicide Temefos, of wild populations of Ae. aegypti, collected in San Antonio, by bioassays under controlled laboratory conditions, revealed that the circulating populations of this vector, are susceptible- RR<3 -to this larvicide, highly used by vector control programs of the Ministry of Public Health and Social Welfare. All of the above is framed in the sustainable development goals, which stipulate that, by 2030, neglected tropical diseases and other vector-transmissible diseases must be ended, more specifically in SDG 3-Health and Wellness. It is also circumscribed in SDG 11, which deals with sustainable cities and communities, because it investigates aspects of increasing urbanization in the study area, with several challenges, such as unplanned urban expansion, without basic services, making cities more vulnerable to disease. The work developed provides a basis for future studies on citizen empowerment and the development of good governance in the face of the urban health problem that constitutes the CSP.

Resumen La liberación de productos farmacéuticos se ha incrementado en ecosistemas terrestres y acuáticos poniendo en riesgo a la biota, pudiendo generar múltiples impactos en los organismos, desde modificar la expresión enzimática hasta el impacto intergeneracional en los organismos expuestos. Este estudio evaluó la influencia del fenobarbital e ibuprofeno sobre tolerancia al insecticida temefos en dos poblaciones de Aedes aegypti, La Esperanza (LE) y Rockefeller (Rock). Las larvas I fueron expuestas a 17.7 µg/ml de ibuprofeno y 200 µg/ml de fenobarbital hasta alcanzar el estadio III; posteriormente, se determinó la mortalidad larvaria al temefos (0.005, 0.025 y 0.050 µg/ml) y la actividad enzimática de las esterasas. Se encontró que el fenobarbital favorece una mayor tolerancia a 0.025 µg/ml de temefos, a las 24 horas de exposición, en la población LE (44.00 ± 6.93% de mortalidad) a diferencia de la Rock (97.33 ± 2.67% de mortalidad); además, disminuye la actividad enzimática de las alfa esterasas en los especímenes Rock y LE (0.3892 ± 0.0756 y 0.1722 ± 0.0194, densidad óptica, respectivamente). Asimismo, el ibuprofeno reporta una menor DL90 (0.024 µg/ml temefos) que el testigo (DL90 = 0.039 µg/ml de temefos) a las 2 horas de exposición. Se concluye que el fenobarbital aumenta la tolerancia de larvas de Aedes aegypti al temefos, y el ibuprofeno estimula la actividad de las alfa y beta-esterasas.

Abstract The release of pharmaceutical products has increased in terrestrial and aquatic ecosystems, putting the biota at risk, being able to generate multiple impacts on organisms, from modifying enzymatic expression to intergenerational impact on exposed organisms. This study evaluated the influence of phenobarbital and ibuprofen on tolerance to the insecticide temephos in two populations of Aedes aegypti, La Esperanza (LE) and Rockefeller (Rock). Larvae I were exposed to 17.7 µg/ml ibuprofen and 200 µg/ml phenobarbital until reaching stage III; subsequently, larvae mortality to temephos (0.005, 0.025 and 0.050 µg/ml) and the enzymatic activity of esterases were determined. Phenobarbital was found to favors a higher tolerance to 0.025 µg/ml of temephos, at 24 hours of exposure, in the LE population (44.00 ± 6.93% mortality) as opposed to Rock (97.33 ± 2.67% mortality); furthermore, it decreased enzymatic activity of alpha esterases in Rock and LE specimens (0.3892 ± 0.0756 and 0.1722 ± 0.0194, optical density, respectively). Also, ibuprofen reports a lower LD90 (0.024 µg/ml temephos) than the control (DL90 = 0.039 µg/ml temephos) at 2 hours of exposure with temephos. It is concluded that phenobarbital increases the tolerance of Aedes aegypti larvae to temephos, and ibuprofen stimulates the activity of alpha- and beta-esterases.

Abstract The threats posed by insecticide resistance to Aedes aegypti in the context of controlling dengue have led to an urgent search for an environmentally safer alternative chemical with more effective larvicidal properties. Among many molecules tested, 2-methylanthraquinone showed the lowest LC50 for A. aegypti in a previous study and the highest LC50 for zebrafish embryos. Embryos were exposed at concentrations of 1.0, 2.19, 4.78, 10.46, 22.87, 50.0 and 100.0 mg/L, and malformations and mortality were significantly observed only at the highest exposures of 50 and 100 mg/L after 96 h. Micronucleus test and comet assay in zebrafish adults were both negative after exposures at 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L for 96 h. Several biochemical biomarkers were analyzed in adults, and 2-methylanthraquinone did not interfere with acetylcholinesterase activity. The lactate dehydrogenase activity was higher at concentrations of 25 and 100 mg/L. Glutathione-S-Transferase (GST) activities were tested in the gill and body (muscle tail). The gill was more sensitive than body for GST activity after exposure to 2-methylanthraquinone, showing the highest activities, and 2-methylanthraquinone showed low toxicity to a non-target organism.

Abstract The use of biocidal larvicides in the control against mosquitoes is important in reducing the adverse effects on non-target organisms and the environment. In this study, it was aimed to determine the most effective concentrations and durations of Bacillus thrungiensis var. israilensis and spinosad larvicides for controlling of the Culex sp. larvae collected from Kahramanmaraş region. For this pupose, Culex sp. larvae were exposed to Bacillus thrungiensis var. israilensis at the concentrations of 0, 0.025, 0.05, 0.1, 0.2, 0.5, 1.0, ve 2.0 µL/200 cm2 and exposed to spinosad at the concentrations of 0, 0.0075, 0.015, 0.03, 0.06, 0.15, 0.3, ve 0.6 µL /200 cm2 along 5 different times (3, 6, 12, 18 and 24 hours). According to the results of the variance and probit analysis, over 90% of mosquito larvae were observed to have died in almost all of the trial periods, but more than 90% of the larvae were found to have died considerably even1/10(For Bti:1 µL/200cm2, For Spinosad:0.3 µL/ 200cm2) concentration after 24 hours which is recommended concentration. The results of the study showed that both bio-larvicides used gave similar results and were an effective controlling tool for mosquitoes. For this reason, it is important to use any of the two larvicides in consideration of local conditions and non-target organisms. For the observation of environmental health and mosquito resistance, making such studies at certain intervals is extremely important in terms of effective control with mosquitoes.

ABSTRACT Background: Aedes aegypti is currently controlled with synthetic larvicides; however, mosquitoes have become highly resistant to these larvicides and difficult to eradicate. Studies have shown that insecticides derived from fungal extracts have various mechanisms of action that reduce the risk of resistance in these mosquitoes. One possible mechanism is uncontrolled production of reactive oxygen species (ROS) in the larvae, which can cause changes at the cellular level. Thus, the crude extract of Xylaria sp. was evaluated to investigate the oxidative effect of this extract in A. aegypti larvae by quantifying the oxidative damage to proteins and lipids. Methods: The larvicidal potential of the crude extract of Xylaria sp. Was evaluated, and the extract was subsequently tested in human lung fibroblasts for cytotoxicity and ROS production. ROS level was quantified in the larvae that were killed following exposure to the extract in the larvicide test. Results: The crude extract of Xylaria sp. Caused cytotoxicity and induced ROS production in human lung fibroblasts and A. aegypti larvae, respectively. In the larvicide trial, the extract showed an LC50 of 264.456 ppm and an LC90 of 364.307 ppm, and was thus considered active. The extract showed greater oxidative damage to lipids and proteins, with LC90 values of 24.7 µmol MDA/L and 14.6278 ×10-3 nmol carbonyl/ mg protein, respectively. Conclusions: Crude extracts of Xylaria sp. induced oxidative stress that may have caused the mortality of A. aegypti larvae.

Abstract Tetradenia riparia (Hochst.) Codd (Lamiaceae) is a species native to the African continent and used as an insect repellent. The objective of the study was to evaluate the larvicidal potential of essential oils (EOs) from the leaves, flower buds, and stem of T. riparia, collected in winter against Aedes aegypti larvae. The EOs were extracted by hydrodistillation (3 h) and identified by GC/MS. The EOs were tested against larvae of A. aegypti at concentrations ranging from 12500 to 1.5 µg/mL for 24 h. The insecticide activity was evaluated by probit analysis, and the anticholinesterase activity was determined by bioautographic method. The results of the class projection indicated sesquiterpenes as the majority class, corresponding to 60.66% (leaves), 64.70% (flower buds) and 83.99% (stem), and the bioassays on A. aegypti larvae indicated LC50 of 1590, 675 and 665 µg/mL, respectively. The anticholinesterase activity indicated that the EO of the leaves inhibited the enzyme at a concentration of 780 µg/mL, and those from the flower buds and stem inhibited up to 1560 µg/mL. The results indicated weak activity of essential oils against A. aegypti larvae.

Abstract Plutella xylostella (L.) is responsible for considerable vegetable crop losses in the metropolitan region of Manaus, Brazil. In recent decades, essential oils have been investigated as an alternative to synthetic insecticides. The genusPiperis widely distributed in Amazonia and essential oils from these plants have insecticidal properties. This study describes the chemical composition of the essential oils fromPiper capiterianumandPiper krukoffiias well as the lethal and sublethal effects onP. xylostella. The phytotoxicity of the oils on the host plant was also evaluated. Globulol was the major constituent of theP. krukoffiioil ando-cymene was the major constituent of theP. capitarianumoil. The oil fromP. capiterianumexhibited greater toxicity to larvae and eggs. This oil also presented greater repellant action, feeding deterrence and mild phytotoxicity to the host plant (Brassicae oleraceae). The findings suggest that this oil can be used in the preparation of a formulated insecticide for the management ofP. xylostellain different development phases. However, further studies are needed to evaluate the effect of this oil on crops under field conditions as well as non-target organisms and determine the cost-benefit ratio of a product formulated withP. capitarianumoil.

ABSTRACT Objective To evaluate the larvicidal effect in petroleum ether of the extracts of Allium sativum (garlic) and Annona muricata (soursop) on larvae in stage IV of Aedes aegypti under laboratory conditions. Methods Different bioassays (treatments) were performed in 6 factors for Annona muricata and 7 concentrations of Allium sativum, with four replications and one control. Mortality readings were taken at 2, 12, 24, 36 and 48 hours. The data obtained statistically (Abbott correction and ANOVA analysis) were validated, in addition, the concentrations and lethal times for both extracts were determined with a Probit analysis. Results It was obtained that, in a 48 hour period, the treatment of 500 ppm of the extract of Annona muricata resulted in a mortality of 97%, while the treatment of 2000 ppm with Allium sativum reached a mortality of 85%. The lethal time 50 (50% mortality) for Annona muricata, was obtained in the treatment of 200 ppm within 24 hours, in the case of Allium sativum it was in the treatment of 1200 ppm before 48 hours. For the lethal time 90 for Annona muricata, obtain the treatment of 400 ppm before 40 hours, for the case of Allium sativum, the lethal time 90 (90% mortality) could not be obtained experimentally, it was determined by means of a linear mathematical model, resulting in 51 hours. Conclusion Both species affected larvicidal effect. However, the most efficient and effective extract as a larvicide is that of Annona muricata, which allows giving a natural, viable, economical and biodegradable alternative for the control of larvae of this species.

RESUMEN Objetivo Evaluar el efecto larvicida en éter de petróleo de los extractos de Allium sativum (ajo) y Annona muricata (guanábana) sobre larvas en IV estadio de Aedes aegypti en condiciones de laboratorio. Métodos Se realizaron diferentes bioensayos (tratamientos) en 6 concentraciones para Annona muricata y 7 concentraciones de Allium sativum, con cuatro repeticiones y un control. Se tuvo lecturas de mortalidad a las 2, 12, 24, 36 y 48 horas. Se validaron los datos obtenidos estadísticamente (corrección de Abbott y Análisis ANOVA). Además, se determinaron las concentraciones y tiempos letales para ambos extractos con un análisis Probit. Resultados Se obtuvo que, en un periodo de 48 horas, el tratamiento de 500 ppm del extracto de Annona muricata logró una mortalidad del 97%, mientras que el tratamiento de 2000 ppm con Allium sativum logró alcanzar una mortalidad del 85%. El tiempo letal 50 (50% de mortalidad) para Annona muricata, se obtuvo en el tratamiento de 200 ppm antes de 24 horas, para el caso de Allium sativum fue en el tratamiento de 1200 ppm antes de 48 horas. Para el tiempo letal 90 (90% de mortalidad) para Annona muricata, se obtuvo en el tratamiento de 400 ppm antes de 40 horas. Para el caso de Allium sativum, el tiempo letal 90 no fue posible obtenerlo experimentalmente. Se determinó por medio de un modelo matemático lineal, que dio como resultado 51 horas. Conclusión Ambas especies poseen efecto larvicida. Sin embargo, el extracto más eficiente y efectivo como larvicida es el de Annona muricata, lo que permite dar una alternativa natural, viable, económica y biodegradable para el control de larvas de esta especie.

RESUMEN Aedes aegypti L. es el mosquito que transmite el virus de la fiebre amarilla, el dengue, el zika y el chikungunya. La prevención y el control de tales enfermedades generalmente dependen del uso de productos químicos, que causan daños al hombre y al medio ambiente. Por ello, los aceites vegetales con acción larvicida se utilizan como alternativa para controlar este insecto. El objetivo de este trabajo fue evaluar el potencial larvicida de aceites vegetales de Caryocar coriaceum, Mauritia flexuosa, Carapa guianensis, Copaifera langsdorffii, Ricinus communis y Cocos nucifera contra A. aegypti. El experimento se dividió en dos bioensayos. En el primero se realizó un diseño completamente al azar con siete tratamientos (seis aceites vegetales a 500 ppm y un control y cuatro repeticiones). Se evaluó el número de larvas muertas a las 24, 48, 72, 96 y 120 h de exposición. En el segundo bioensayo, se utilizaron los aceites vegetales más eficientes (C. coriaceum y M. flexuosa) en concentraciones 0, 500, 1000, 1500, 2000 y 2500 ppm, con cuatro repeticiones. El número de larvas muertas se evaluó según el primer bioensayo. Todos los aceites utilizados tienen efecto larvicida sobre larvas de A. aegypti de tercer estadio, con mayor eficacia a las 120 h de exposición. Los aceites de C. coriaceum y M. flexuosa mostraron mejor eficacia en el control de larvas, siendo la dosis de 2500 ppm la más recomendada. El LD10, LD50 y LD90 del aceite de M. flexuosa recomendado para controlar larvas son 234, 648 y 1794 ppm, respectivamente.

ABSTRACT Aedes aegyptiL. is the mosquito vector of yellow fever, dengue, zika, and chikungunya viruses. The prevention and control of such diseases usually rely on the use of chemicals, that can cause harm to human health and the environment. Vegetable oils with larvicidal activity are used as an alternative tool to control this insect. This study aimed to evaluate the larvicidal activity of vegetable oils fromCaryocar coriaceum,Mauritia fIexuosa,Carapa guianensis, Copaifera langsdorffii, Ricinus communisandCocos nuciferaagainstA. aegypti larvae. The experiment was divided into two bioassays. In the first, a completely randomized design was used with seven treatments (six vegetable oils at 500 ppm and one control with four replications). The number of dead larvae was evaluated 24, 48, 72, 96, and 120 h after exposure. In the second bioassay, the most efficient vegetable oils from the first bioassay (C. coriaceumandM. flexuosa) were used at the concentrations of 0, 500, 1000, 1500, 2000, and 2500 ppm, with four replications. The number of dead larvae was evaluated according to the first bioassay. All oils used had larvicidal activity on third-instar stage larvae ofA. aegypti, with greater efficiency 120 h after exposure. The oils ofC. coriaceumandM. flexuosaat 2500 ppm had the best efficacy in the larvae control. The LD10, LD50, and LD90 ofM. flexuosaoil recommended for controlling larvae are 234, 648, and 1794 ppm, respectively.

Resumen Introducción: Los fitoinsecticidas son alternativas para el control de insectos en diferentes etapas, Gallesia integrifolia (Spreng.) Harms, familia Phytolacaceae, conocida popularmente como pau d’alho, árbol de ajo y guararema en Brasil, es conocida por su fuerte olor aliáceo debido a la presencia de moléculas de azufre en la planta. Esta especie presenta actividad biológica y potencial efecto insecticida que aún no está explotado. Objetivo: El objetivo de este estudio fue evaluar la actividad biológica del extracto crudo etanólico de las hojas, flores y frutos de G. integrifolia en el control de las larvas y pupas de la tercera etapa de Aedes aegypti. Métodos: El material botánico fue recolectado en la ciudad de Umuarama, Paraná, Brasil (23º46’16” S & 53º19’38” W), a una altitud de 442 m y los frutos de G. integrifolia fueron recolectados en mayo de 2017 y las hojas y flores en diciembre del mismo año. Los extractos crudos de hojas, flores y frutos de G. integrifolia se prepararon por técnica de maceración dinámica. La composición química de los extractos se determinó por cromatografía de gases acoplada a espectrometría de masas. La actividad insecticida de los extractos crudos de G. integrifolia fue evaluada en larvas y pupas de A. aegypti en concentraciones entre 0.001 a 25 000 mg/mL, y las concentraciones letales que matan 50 % (CL50) y 99.9 % (CL99.9) de larvas fueron determinados por análisis de probit. La actividad anticolinesterasa fue evaluada por método bioautográfico en concentraciones de 0.000095 a 50 mg/mL. Resultados: El rendimiento de los extractos crudos de G. integrifolia fue de 8.2, 9.1 y 17.3 % para flores, frutos y hojas, respectivamente. La composición química de los extractos de G. integrifolia se caracterizó por la presencia de ésteres de ácidos grasos, fitosteroles, vitaminas, diterpenos oxigenados y compuestos organosulfurados. El extracto de las flores presentó alta cantidad de compuestos de azufre (20.2 %) como disulfuro, bis(2-sulfhidril etilo) (11.9 %), 2,3,5-tritiahexano (6.2 %), 1,2,4-tritiolano (1.1 %) y 2,4-ditiapentano (1.1 %). En relación con la actividad insecticida, el extracto de las flores mostró una gran actividad con CL99.9 de 0.032 mg/mL y CL99.9 de 0.969 mg/mL en larvas y pupas de A. aegypti, respectivamente, y la inhibición más alta de la enzima acetilcolinesterasa (0.00019 mg/mL) ex situ. El extracto de las flores presentó actividad anticolinesterasa y larvicida, 12.8 y 35.6 %, respectivamente, mayor que el control temephos. Conclusiones: Este estudio abre nuevas perspectivas sobre el uso de extractos de G. integrifolia como alternativa bioinsecticida para el control de larvas y pupas de A. aegypti.

Abstract Introduction: Phytoinsecticides are alternatives to control insects in different stages, Gallesia integrifolia (Spreng.) Harms, Phytolacaceae family, popularly known as pau d’alho, garlic tree, and guararema in Brazil, is known due to its strong alliaceous odor because of the presence of sulfur molecules in the plant. This species presents biological activity and potential insecticide effect that is still unexploited. Objective: This study aimed to evaluate the biological activity of the ethanolic crude extract from G. integrifolia leaves, flowers, and fruits on the control of Aedes aegypti third-stage larvae and pupae. Methods: The botanical material was collected in city Umuarama, Paraná, Brazil at the coordinates (23º46’16” S & 53º19’38” WO), and altitude of 442 m, the fruits of G. integrifolia were collected in May and the leaves and flowers in December 2017. The crude extracts of G. integrifolia leaves, flowers, and fruits were prepared by dynamic maceration technique. The chemical composition of the extracts was determined by gas chromatography coupled to a mass spectrometry. The insecticidal activity of the crude extracts of G. integrifolia were carried out on larvae and pupae of A. aegypti in concentrations between 0.001 to 25 000 mg/mL, and afterwards the lethal concentrations that kill 50 % (LC50) and 99.9 % (LC99.9) were determined by probit analysis. Anticholinesterase activity was determined by bioautographic method at concentrations from 0.000095 to 50 mg/mL. Results: The yield of G. integrifolia crude extracts were 8.2, 9.1, and 17.3 % for flowers, fruits, and leaves, respectively. The chemical composition of G. integrifolia extracts was characterized by presence of fatty acid esters, phytosterols, vitamins, oxygenated diterpenes and organosulfur compounds. The flower extract presented the high amount of sulfur compounds (20.2 %) such as disulfide, bis (2-sulfhydryl ethyl) (11.9 %), 2,3,5-trithiahexane (6.2 %), 1,2,4-trithiolane (1.1 %), and 2,4-dithiapentane (1.1 %). Regarding the insecticidal activity, flower extract showed highly active with LC99.9 of 0.032 mg/mL and LC99.9 of 0.969 mg/mL on A. aegypti larvae and pupae, respectively, and the highest inhibition of acetylcholinesterase enzyme (0.00019 mg/mL) ex situ. The flower extract presented anticholinesterase and larvicide activity, respectively, 12.8 % and 35.6 % greater than the control temephos. Conclusions: This study opens new perspectives on the use of extracts from G. integrifolia as a bioinsecticide alternative for the control of A. aegypti larvae and pupae.

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

RESUMEN Aedes aegypti es un mosquito portador del virus del dengue, la fiebre amarilla y otras enfermedades transmitidas a los humanos. Los larvicidas organofosforados se utilizan para controlar la proliferación de este mosquito, el cual ha generado un alto grado de resistencia, por lo que las nuevas alternativas de biolarvicidas formulados con extractos de plantas son de gran interés. Los objetivos de este estudio fueron evaluar el extracto etanólico de hojas de Azadirachta indica como larvicida contra Aedes aegypti e identificar los principales compuestos químicos GC-MS. En el ensayo, se utilizaron tres concentraciones de extracto de etanol (10 mg L-1, 20 mg L-1 y 50 mg L-1), este se realizó por triplicado contra un control positivo (larvicida comercial: Esporas y cristales endotóxicos de Bacillus thuringiensis var. israelensis Serotipo H-14) y un control negativo (agua). Después de 72 horas de incubación, se observó una mayor mortalidad larval (93%) en el extracto etanólico a una concentración de 50 mg L-1, y los extractos a 10 mg L-1 y 20 mg L-1 mostraron una mortalidad larval del 47% y 70% respectivamente. El compuesto con mayor abundancia determinado por el análisis CG-MS fue el fitol (área del 14,4%). Los resultados obtenidos en este estudio demostraron el potencial larvicida del extracto de etanol de A. indica contra las larvas de A. aegypti.

ABSTRACT Aedes aegypti is a mosquito that carries dengue virus, yellow fever and other diseases transmitted to humans. Organophosphorus larvicides are used to control the proliferation of this mosquito, which has generated a high degree of resistance; hence, new alternatives such as bio-larvicides formulated with plant extracts are of great interest. The aims of this study were to evaluate the ethanolic extract of Azadirachta indica leaves as a larvicide against Aedes aegypti and to determine the main compounds present in it by GC-MS. In the assay, three concentrations of ethanolic extract were used (10 mg L-1, 20 mg L-1, and 50 mg L-1). This was performed thrice against a positive control (commercial larvicide: spores and endotoxic crystals of Bacillus thuringiensis var. israelensis Serotype H-14) and negative control (water). After 72 h of incubation, it was observed higher larval mortality (93%) in the ethanolic extract at a concentration of 50 mg L-1; the extracts at 10 mg L-1 and 20 mg L-1 shown larval mortality of 47% and 70%, respectively. The majority compound determined by the GC-MS analysis was phytol (14.4% area). The results obtained in this study demonstrated the larvicidal potential of the ethanolic extract of A. indica against larvae of A. aegypti.