Design and elucidation of an insecticide from natural compounds targeting mitochondrial proteins of Aedes aegypti.

Developing new pesticides poses a significant challenge in designing next-generation natural insecticides that selectively target specific pharmacological sites while ensuring environmental friendliness. In this study, we aimed to address this challenge by formulating novel natural pesticides derived from secondary plant metabolites, which exhibited potent insecticide activity. Additionally, we tested their effect on mitochondrial enzyme activity and the proteomic profile of Ae. aegypti, a mosquito species responsible for transmitting diseases. Initially, 110 key compounds from essential oils were selected that have been reported with insecticidal properties; then, to ensure safety for mammals were performed in silico analyses for toxicity properties, identifying non-toxic candidates for further investigation. Subsequently, in vivo tests were conducted using these non-toxic compounds, focusing on the mosquito's larval stage. Based on the lethal concentration (LC), the most promising compounds as insecticidal were identified as S-limonene (LC50 = 6.4 ppm, LC95 = 17.2 ppm), R-limonene (LC50 = 9.86 ppm, LC95 = 27.7 ppm), citronellal (LC50 = 40.5 ppm, LC95 = 68.6 ppm), R-carvone (LC50 = 61.4 ppm, LC95 = 121 ppm), and S-carvone (LC50 = 62.5 ppm, LC95 = 114 ppm). Furthermore, we formulated a mixture of R-limonene, S-carvone, and citronellal with equal proportions of each compound based on their LC50. This mixture specifically targeted mitochondrial proteins and demonstrated a higher effect that showed by each compound separately, enhancing the insecticidal activity of each compound. Besides, the proteomic profile revealed the alteration in proteins involved in proliferation processes and detoxification mechanisms in Ae. aegypti. In summary, our study presents a formulation strategy for developing next-generation natural insecticides using secondary plant metabolites with the potential for reducing the adverse effects on humans and the development of chemical resistance in insects. Our findings also highlight the proteomic alteration induced by the formulated insecticide, showing insight into the mechanisms of action and potential targets for further exploration in vector control strategies.

Epidemiological features of Leishmania infantum in dogs (Canis lupus familiaris) suggest a latent risk of visceral leishmaniasis in the metropolitan area of Bucaramanga, Santander, Eastern Colombia.

Visceral leishmaniasis (VL) is a disease caused by species of the Leishmania donovani complex that is mainly transmitted through the urban cycle involving dogs as the primary reservoir. In Colombia, the incidence of VL is increasing, along with the spread of potential vectors. This study aims to investigate the eco-epidemiological factors associated with Leishmania spp. infection in dogs from the Metropolitan Area of Bucaramanga (MAB), Santander, eastern Colombia, which is a region at risk for VL. We conducted molecular and serological surveillance of Leishmania spp. in 207 dogs from MAB to determine the epidemiological factors associated with infection. Subsequently, we carried out a molecular and serological analysis of phlebotomine and humans, respectively, in areas with a higher prevalence of infection, aiming to describe the main features associated with the transmission cycle. Out of the 207 dogs tested, 37 (17.8%, 95% CI = 12.6-23.1%) were positive for the presence of Leishmania antibodies by the IFAT test, and only 9 (4.3%, 95% CI = 1.55-7.15%) were positive for L. infantum by PCR. Multivariate analyses indicated that canine shelters and dogs with clinical signs commonly associated with canine VL had a higher prevalence of infection (P < 0.05). In the entomological survey, 69 blood-fed female phlebotomine of the genus Lutzomyia were captured in canine shelters, among them, 55% were identified as Lutzomyia camposi, 29% as Lu. ovallesi, 7% as Lu. dubitans, 6% as Lu. torvida, and 3% as Lu. cayennensis. The identified meal sources of the phlebotomine included human, pig, avian, cattle, and porcupine (Coendou quichua) blood. However, no phlebotomine positive for Leishmania spp. were detected by molecular analyses. Finally, 14 humans who had frequent contact with L. infantum-positive dogs were analyzed through rK39 test, but none tested was positive for IgG/IgM antibodies. The molecular and serological analyses indicate the circulation of L. infantum in dogs from MAB, with canine shelters having the highest prevalence of infection. The entomological survey of canine shelters showed a significant diversity of phlebotomine without potential vectors of L. infantum, suggesting the presence of infection in dogs from these areas could take place in other locations or through other transmission routes. The circulation of L. infantum in multiple dogs from MAB suggests a latent risk of zoonotic transmission of VL in these cities.

Insecticidal activity of essential oils from American native plants against Aedes aegypti (Diptera: Culicidae): an introduction to their possible mechanism of action.

Searching for new bioactive molecules to design insecticides is a complex process since pesticides should be highly selective, active against the vector, and bio-safe for humans. Aiming to find natural compounds for mosquito control, we evaluated the insecticidal activity of essential oils (EOs) from 20 American native plants against Aedes aegypti larvae using bioassay, biochemical, and in silico analyses. The highest larvicide activity was exhibited by EOs from Steiractinia aspera (LC50 = 42.4 µg/mL), Turnera diffusa (LC50 = 70.9 µg/mL), Piper aduncum (LC50 = 55.8 µg/mL), Lippia origanoides (chemotype thymol/carvacrol) (LC50 = 61.9 µg/mL), L. origanoides (chemotype carvacrol/thymol) (LC50 = 59.8 µg/mL), Hyptis dilatata (LC50 = 61.1 µg/mL), Elaphandra quinquenervis (LC50 = 61.1 µg/mL), and Calycolpus moritzianus (LC50 = 73.29 µg/mL) after 24 h. This biological activity may be related to the disruption of the electron transport chain through the mitochondrial protein complexes. We hypothesized that the observed EOs' effect is due to their major components, where computational approaches such as homology modeling and molecular docking may suggest the possible binding pose of secondary metabolites that inhibit the mitochondrial enzymes and acetylcholinesterase activity (AChE). Our results provided insights into the possible mechanism of action of EOs and their major compounds for new insecticide designs targeting the mitochondria and AChE activity in A. aegypti for effective and safe insecticide.

Synthesis of new series of quinoline derivatives with insecticidal effects on larval vectors of malaria and dengue diseases.

Mosquito borne diseases are on the rise because of their fast spread worldwide and the lack of effective treatments. Here we are focusing on the development of a novel anti-malarial and virucidal agent with biocidal effects also on its vectors. We have synthesized a new quinoline (4,7-dichloroquinoline) derivative which showed significant larvicidal and pupicidal properties against a malarial and a dengue vector and a lethal toxicity ranging from 4.408 µM/mL (first instar larvae) to 7.958 µM/mL (pupal populations) for Anopheles stephensi and 5.016 µM/mL (larva 1) to 10.669 µM/mL (pupae) for Aedes aegypti. In-vitro antiplasmodial efficacy of 4,7-dichloroquinoline revealed a significant growth inhibition of both sensitive strains of Plasmodium falciparum with IC50 values of 6.7 nM (CQ-s) and 8.5 nM (CQ-r). Chloroquine IC50 values, as control, were 23 nM (CQ-s), and 27.5 nM (CQ-r). In vivo antiplasmodial studies with P. falciparum infected mice showed an effect of 4,7-dichloroquinoline compared to chloroquine. The quinoline compound showed significant activity against the viral pathogen serotype 2 (DENV-2). In vitro conditions and the purified quinoline exhibited insignificant toxicity on the host system up to 100 µM/mL. Overall, 4,7-dichloroquinoline could provide a good anti-vectorial and anti-malarial agent.

Impact of Cymbopogon flexuosus (Poaceae) essential oil and primary components on the eclosion and larval development of Aedes aegypti.

The current study describes the effects of sub-lethal concentrations and constituent compounds (citral and geranyl acetate) of Cymbopogon flexuosus essential oil (EO) on the development of Aedes aegypti. We treated eggs with 6, 18, or 30 mg L-1 and larvae with 3 or 6 mg L-1 of EO and its major compounds (citral and geranyl acetate). Citral and geranyl acetate were evaluated at 18, 30, and 42 mg L-1 and compared with commercial growth inhibitors (diflubenzuron and methoprene). We measured larval head diameter, siphon length, and larval length. Finally, we examined concentrations of molt hormone (MH) and juvenile hormone III (JH III) using high-performance liquid chromatography coupled to mass spectrometry. All geranyl acetate concentrations decreased egg hatching, while EO altered molting among larval instars and between larvae and pupae, with an increase in the larval length (3 mg L-1: 6 ± 0.0 mm; 6 mg L-1: 6 ± 0.7 mm) and head width (3 mg L-1: 0.8 ± 0 mm; 6 mg L-1: 0.8 ± 0.0 mm) compared with the control group. We did not detect chromatographic signals of MH and JH III in larvae treated with C. flexuosus EO or their major compounds. The sub-lethal concentrations C. flexuosus EO caused a similar effect to diflubenzuron, namely decreased hormone concentrations, an extended larval period, and death.

Exploiting a Y chromosome-linked Cas9 for sex selection and gene drive.

CRISPR-based genetic engineering tools aimed to bias sex ratios, or drive effector genes into animal populations, often integrate the transgenes into autosomal chromosomes. However, in species with heterogametic sex chromsomes (e.g. XY, ZW), sex linkage of endonucleases could be beneficial to drive the expression in a sex-specific manner to produce genetic sexing systems, sex ratio distorters, or even sex-specific gene drives, for example. To explore this possibility, here we develop a transgenic line of Drosophila melanogaster expressing Cas9 from the Y chromosome. We functionally characterize the utility of this strain for both sex selection and gene drive finding it to be quite effective. To explore its utility for population control, we built mathematical models illustrating its dynamics as compared to other state-of-the-art systems designed for both population modification and suppression. Taken together, our results contribute to the development of current CRISPR genetic control tools and demonstrate the utility of using sex-linked Cas9 strains for genetic control of animals.

Model to design insecticides against Aedes aegypti using in silico and in vivo analysis of different pharmacological targets.

Compounds having insecticidal activity can be used to control Aedes aegypti mosquitoes, a major worldwide vector, and several plants have a source of such molecules. A principal component analysis (PCA) was carried out to determine the criterion to select larvicidal metabolites. The insecticidal activity of seven selected metabolites by PCA was validated by determining its lethal concentrations 50 (LC50) by probit analysis. Six of the seven evaluated molecules presented LC50 values <100 ppm. The effects of these six molecules on acetylcholinesterase and the respiratory chain complexes of the mitochondria of Ae. aegypti were evaluated. Four metabolites presenting the highest inhibition effects on these targets were mixed in 11 different combinations, and the percentage of mortality of each mixture on Ae. aegypti larvae were determined. Secondary metabolites such as geranyl acetate, α-humulene, ß-caryophyllene, geraniol, nerol, and n-octanol presented LC50 values under 100 ppm (44, 41, 48, 84, 87, and 98 ppm, respectively), whereas 1,8-cineole presented a LC50 value of 183 ppm. We found that, geranyl acetate, α-humulene, ß-caryophyllene, nerol, n-octanol, and geraniol inhibited at least one of the six targets with an efficiency between 25 and 41%. Overall, the evaluation of the different mixtures revealed a synergistic effect between geranyl acetate and geraniol, and an antagonistic effect between α-humulene and ß-caryophyllene compounds.

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

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

Toxicidad y afectación en la locomoción de Triatoma dimidiata (Latreille 1811) (Hemiptera: Reduviidae: Triatominae) tratados con análogos de girgensohnina / Toxicity and affectation in locomotion of Triatoma dimidiata (Latreille 1811) (Hemiptera: Reduviidae: Triatominae) treated with analogues of girgensohnine

Resumen Introducción: Análogos del alcaloide girgensohnina, diseñados y sintetizados para inhibir a la enzima acetilcolinesterasa, han presentado efecto insecticida sobre insectos vectores de enfermedades. Objetivo: Determinar la actividad insecticida de 12 análogos sintéticos de gingersohnina sobre ninfas del primer estadio de Triatoma dimidiata. Materiales y Métodos: Se tomó como referencia el protocolo de la OMS 2005. Ninfas del primer estadio de T. dimidiata fueron expuestas a los análogos por aplicación tópica y exposición a superficies para determinar las dosis letales (DL50 y DL95) y verificar alteración en la locomoción mediante el software Videomex V. Resultados: El análogo 6 presentó la mayor actividad insecticida a las 72h con una mortalidad del 20,8% ± 3,57 para el tratamiento de topicación 25% ± 0,00 para las superficies no porosas. Las dosis letales de la molécula 6 fueron: DL50 1036,8 ± 0,38 ng/insecto y DL95 3991,4 ± 0,50 ng/insecto respectivamente. Con respecto a la locomoción, el análogo indujo un comportamiento similar al insecticida comercial deltametrina.

Abstract Introduction: Analogs of the girgensohnine alkaloid, designed and synthesized to inhibit acetylcholinesterase enzyme, have presented an insecticidal effect on vectors insects of diseases. Objective: To determine the insecticidal activity of 12 synthetic analogs of gingersohnine in first stage nymphs of Triatoma dimidiata. Materials and methods: WHO protocol 2005, using exploratory doses, was used to determine the best molecule according to its mortality. First stage nymphs of T. dimidiata were exposed to different doses of the analogs by topical application and surface exposure. Lethal doses (LD50 and DL95) and alterations in locomotion using Videomex V software were determined. Results: Analog 6 presented the highest insecticidal activity at 72 h with a mortality of 20.8% ± 3.57 for topical treatment and 25%±0,0 for non-porous surfaces evaluation. Analog 6 lethal doses were LD50 1036.8 ± 0.38 ng/insecto and DL95 3991.4 ± 0.508 ng/insecto, respectively. With regard to locomotion, analogue 6 induced a similar behavior to that observed for commercial insecticide deltametrin.

Synthesis of new α-amino nitriles with insecticidal action on Aedes aegypti (Diptera: Culicidae)

Abstract Aedes aegypti is the principal vector of arboviral pathogens that may cause diseases as dengue fever, chikungunya and zika. The harmful environmental effects of commercial pesticides coalesced with the development of insecticide-resistant populations encourage the discovery and generation of new alternative products as a tool to reduce the incidence of vector-borne diseases. In this work, through the classic three component Strecker reaction of commercial benzaldehydes, cyclic secondary amines and KCN, a new series of nine α-amino nitriles, girgensohnine analogs, has been synthetized and screened for larvicide and adulticide properties against A. aegypti, one of the dominant vectors of dengue, chikungunya and zika in tropical and subtropical areas all over the world. Molecules 3 and 4 were identified as potential larvicidal agents with LC50 values of 50.55 and 69.59 ppm, respectively. Molecule 3 showed 100% of mortality after 2 h of treatment when a concentration of 30 ppm in adulticidal assays was evaluated. Additionally, in order to elucidate the mode of action of these molecules, their acetylcholinesterase (AChE) inhibitory properties were evaluated using the Ellman assay. It was found that the molecules possess a weak AChE inhibitory activity with IC50 values between 148.80 and 259.40 µM, indicating that AChE could not be a principal target for insecticide activity.

Evaluation of the insecticidal activity of essential oils and their mixtures against Aedes aegypti (Diptera: Culicidae)

Abstract The search for new insecticides to control dengue fever, chikungunya, and Zika vectors has gained relevance in the past decades. The aim of the present study was to evaluate the larvicidal action of essential oils (EOs) from Thymus vulgaris, Salvia officinalis, Lippia origanoides, Eucalyptus globulus, Cymbopogon nardus, Cymbopogon martinii, Lippia alba, Pelargonium graveolens, Turnera diffusa, and Swinglea glutinosa on Aedes (Stegomyia) aegypti. The EOs were extracted by microwave-assisted hydrodistillation and characterized by gas chromatography/mass spectrometry (GC/MS). The chemical components of the EOs were identified by linear retention indices and mass spectra. Lethal concentrations (LC50 and LC95) were determined by probit analysis using larvae of Ae. aegypti between the third and the fourth instars. All EOs achieved larvicidal activity at LC50 values lower than 115 mg/L. The lowest LC50 value (45.73 mg/L) corresponded to T. vulgaris EO, whereas C. martinii EO showed the highest LC50 (LC50 = 114.65 mg/L). Some EO mixtures showed lower LC50 than oils used individually, such as the mixtures of L. origanoides + S. glutinosa (LC50 = 38.40 mg/L), T. diffusa + S. glutinosa (LC50 = 63.71 mg/L), and L. alba + S. glutinosa (LC50 = 48.87 mg/L). The main compounds of the EOs with highest larvicidal activity were thymol (42%) and p-cymene (26.4%).

[Insecticidal action of synthetic girgensohnine analogues and essential oils on Rhodnius prolixus (Hemiptera: Reduviidae)].

INTRODUCTION: The alkaloid girgensohnine has been used as a natural model in the synthesis of new alkaloid-like alpha-aminonitriles with insecticidal effect against disease vectors. OBJECTIVE: To evaluate the biocide activity of girgensohnine analogues and essential oils of Cymbopogon flexuosus, Citrus sinensis and Eucalyptus citriodora in stage I and stage V Rhodnius prolixus nymphs. MATERIALS AND METHODS: We used a topical application model in tergites and sternites, as well as exposure to treated surfaces with different exploratory doses of each of the molecules and essential oils to determine the lethal doses (LD50 and LD95). RESULTS: Analogue 3 showed the highest insecticidal activity with 83.3±16.7% of mortality when applied on tergites, 38.9±4.8% on sternites and 16.7±0% on treated surfaces in stage I nymphs at 72 hours (h) and 500 mg.L-1. In stage V nymphs, the compounds induced mortality only in sternums (11.1±9.6% for analogue 6 and 5.5±4.7% for analogues 3 and 7 at 72 h and 1500 mg.L-1). The lethal doses for molecule 3 on tergites in stage I nymphs were LD50 225.60 mg.L-1 and LD95 955.90 mg.L-1. The insecticidal effect of essential oils was observed only in stage I nymphs, with 11.1±4.8% for C. flexuosus when applied in sternites, while using exposure to surfaces treated it was 5.6±4.8% for C. sinensis applied on tergites and 8.3±0% on sternites at 72 h and 1000 mg.L-1. CONCLUSION: Synthetic girgensohnine analogues, and C. flexuosus and C. sinensis essential oils showed insecticidal activity in R. prolixus. Analogue 3 showed the greatest insecticidal activity among all molecules and oils evaluated under our laboratory conditions.

Insecticidal action of synthetic girgensohnine analogues and essential oils on Rhodnius prolixus (Hemiptera: Reduviidae) / Acción insecticida de análogos sintéticos de girgensohnina y de aceites esenciales sobre Rhodnius prolixus (Hemiptera: Reduviidae)

ABSTRACT Introduction: The alkaloid girgensohnine has been used as a natural model in the synthesis of new alkaloid-like alpha-aminonitriles with insecticidal effect against disease vectors. Objective: To evaluate the biocide activity of girgensohnine analogues and essential oils of Cymbopogon flexuosus, Citrus sinensis and Eucalyptus citriodora in stage I and stage V Rhodnius prolixus nymphs. Materials and methods: We used a topical application model in tergites and sternites, as well as exposure to treated surfaces with different exploratory doses of each of the molecules and essential oils to determine the lethal doses (LD50 and LD95). Results: Analogue 3 showed the highest insecticidal activity with 83.3±16.7% of mortality when applied on tergites, 38.9±4.8% on sternites and 16.7±0% on treated surfaces in stage I nymphs at 72 hours (h) and 500 mg.L-1. In stage V nymphs, the compounds induced mortality only in sternums (11.1±9.6% for analogue 6 and 5.5±4.7% for analogues 3 and 7 at 72 h and 1500 mg.L-1). The lethal doses for molecule 3 on tergites in stage I nymphs were LD50 225.60 mg.L-1 and LD95 955.90 mg.L-1. The insecticidal effect of essential oils was observed only in stage I nymphs, with 11.1±4.8% for C. flexuosus when applied in sternites, while using exposure to surfaces treated it was 5.6±4.8% for C. sinensis applied on tergites and 8.3±0% on sternites at 72 h and 1000 mg.L-1. Conclusion: Synthetic girgensohnine analogues, and C. flexuosus and C. sinensis essential oils showed insecticidal activity in R. prolixus. Analogue 3 showed the greatest insecticidal activity among all molecules and oils evaluated under our laboratory conditions.

RESUMEN Introducción. El alcaloide natural girgensohnina se ha usado como modelo en la síntesis de nuevos análogos de alcaloidales alfa-aminonitrílicos con efecto insecticida en vectores de enfermedades. Objetivo. Evaluar la actividad biocida de análogos de girgensohnina y de aceites esenciales de las plantas Cymbopogon flexuosus, Citrus sinensis y Eucalyptus citriodora en ninfas de estadios I y V de Rhodnius prolixus. Materiales y métodos. Se empleó la aplicación tópica en terguitos, esternitos y superficies tratadas con diferentes dosis exploratorias de cada una de las moléculas y aceites esenciales para determinar las dosis letales (LD50 y LD95). Resultados. El análogo 3 tuvo la mayor actividad insecticida, con una mortalidad de 83,3±16,7% en los terguitos, de 38,9±4,8 % en los esternitos y de 16,7±0 % a las 72 horas en ninfas de estadioI expuestas a superficies tratadas y 500 mg.L-1. En las ninfas de estadio V solo se presentó mortalidad en los esternitos (11,1±9,6 % con el análogo 6 y 5,5±4,7 % con los análogos 3 y 7 a las 72 h y 1.500 mg.L-1). Las dosis letales para la molécula 3 en los terguitos de ninfas de estadio I fueron las siguientes: DL50, 225,60 mg.L-1 y DL95, 955,90 mg.L-1. En cuanto a los aceites esenciales, el efecto insecticida solo se presentó con C. flexuosus (11,1±4,8%) en los esternitos de ninfas de estadio I expuestas a superficies tratadas; con C. sinensis (5,6±4,8%) en los terguitos y en los esternitos (8,3±0%) a las 72 horas y 1.000 mg.L-1. Conclusión. Los análogos sintéticos del alcaloide girgensohnina y los aceites esenciales de C. flexuosus y C. sinensis exhibieron actividad insecticida en R. prolixus. El análogo 3 exhibió la mayor actividad insecticida de todas las moléculas evaluadas bajo las condiciones de laboratorio.

Insecticidal and Repellent Activity of Several Plant-Derived Essential Oils Against Aedes aegypti.

We examined the pupicidal, adulticidal, repellent, and oviposition-deterrent activities of essential oils (EOs) from Lippia alba, L. origanoides, Eucalyptus citriodora, Cymbopogon citratus, Cymbopogon flexuosus, Citrus sinensis , Cananga odorata , Swinglea glutinosa, and Tagetes lucida plants against Aedes aegypti under laboratory conditions. Pupicidal and adulticidal activities were assessed at exploratory concentrations of 250, 310, and 390 parts per million (ppm); and 30, 300, and 1,000 ppm, respectively. The greatest pupicidal activity was exhibited at 390 ppm with a 24-h exposure by L. origanoides, and 390 ppm with a 48-h exposure by Citrus sinensis . Lippia origanoides killed all adult mosquitoes at 300 ppm after 120 min of exposure. Only L. origanoides and E. citriodora EOs, applied at 1,000 ppm to human skin, produced the greatest repellency (100%) to host-seeking Ae. aegypti after 2 min of exposure; the repellency decreased between 12% and 10% after 15 min. Complete oviposition deterrence by gravid Ae. aegypti was observed for E. citriodora EOs at 200 ppm with an oviposition activity index of -1.00. These results confirm that the EOs assessed in this study have insecticidal, repellent, and oviposition-deterrent activities against the dengue vector, Ae. aegypti.

Abundância e agregação de ovos de Aedes aegypti L. e Aedes albopictus (Skuse) (Diptera: Culicidae) no norte e noroeste do Paraná / Abundance and aggregation egg of Aedes aegypti L. and Aedes albopictus (Skuse) (Diptera: Culicidae) in the north and northwest of the State of Paraná, Brazil

Avaliou-se a abundância e agregação dos ovos de Aedes aegypti L. e Aedes albopictus Skuse em Cambé, Ibiporã, Jacarezinho, Maringá e Paranavaí, PR, utilizando-se armadilhas de oviposição. De 225 armadilhas instaladas, 100 registraram-se positivas para ovos. Coletaram-se 4140 ovos, demonstrando distribuição altamente agregada. Ambas as espécies foram encontradas em Cambé, Jacarezinho, Maringá e Paranavaí. Ae. albopictus não foi detectado em Ibiporã, porém onde esteve presente foi menos abundante. A relação entre sexos aproximou-se de 1:1, para Ae. aegypti. Na comparação dos números de adultos coletados entre as duas espécies, obteve-se correlação negativa nas amostras de Maringá e Cambé, o que foi atribuído à sazonalidade dessas populações. A coexistência dessas espécies indica que elas estão sobre pressão dos programas de controle, sendo, portanto necessárias avaliações específicas.

The abundance and aggregation of eggs of Aedes aegypti L. and Aedes albopictus Skuse was evaluated in the municipalities of Cambé, Ibiporã, Jacarezinho, Maringá and Paranavaí, in the State of Paraná, Brazil by means of oviposition traps. Of the 225 installed traps, 100 were registered as positive for eggs; 4140 eggs were collected, thus demonstrating an highly aggregate distribution. Both species were registered in Cambé, Jacarezinho, Maringá and Paranavaí. Ae. albopictus was generally less abundant and was not present in Ibiporã nor in the oviposition traps of a second collection of Maringá. The relation between sexes for Ae. aegypti was approximately 1:1. In the comparison of the number of adults collected between the two species, a negative correlation was obtained in the samples of Maringá and Cambé, what was attributed the seasonality of these populations. The coexistence of these species indicates that both are under pressure by the control programs, therefore specific evaluations are necessary.

[Abundance and aggregation egg of Aedes aegypti L. and Aedes albopictus (Skuse) (Diptera: Culicidae) in the north and northwest of the State of Paraná, Brazil]. / Abundância e agregação de ovos de Aedes aegypti L. e Aedes albopictus (Skuse) (Diptera: Culicidae) no norte e noroeste do Paraná

The abundance and aggregation of eggs of Aedes aegypti L. and Aedes albopictus Skuse was evaluated in the municipalities of Cambé, Ibiporã, Jacarezinho, Maringá and Paranavaí, in the State of Paraná, Brazil by means of oviposition traps. Of the 225 installed traps, 100 were registered as positive for eggs; 4140 eggs were collected, thus demonstrating an highly aggregate distribution. Both species were registered in Cambé, Jacarezinho, Maringá and Paranavaí. Ae. albopictus was generally less abundant and was not present in Ibiporã nor in the oviposition traps of a second collection of Maringá. The relation between sexes for Ae. aegypti was approximately 1:1. In the comparison of the number of adults collected between the two species, a negative correlation was obtained in the samples of Maringá and Cambé, what was attributed the seasonality of these populations. The coexistence of these species indicates that both are under pressure by the control programs, therefore specific evaluations are necessary.

Larvicidal action of ethanolic extracts from fruit endocarps of Melia azedarach and Azadirachta indica against the dengue mosquito Aedes aegypti.

Ethanolic extracts from the kernels of ripe fruits from the Indian Lilac Melia azedarach and from the well-known Neem tree, Azadirachta indica were assayed against larvae of Aedes aegypti, the mosquito vector of dengue fever. The lethality bioassays were carried out according to the recommendations of the World Health Organization. Extracts were tested at doses ranging from 0.0033 to 0.05 g% in an aqueous medium for 24 and 48 h, at 25 or 30 degrees C, with or without feeding of the larvae. LC50, LC95 and LC99 were determined. Both seed extracts proved lethal for third to fourth instar larvae. Non-fed A. aegypti larvae were more susceptible to Azadirachta extracts at both temperatures. Under a more realistic environmental situation, namely with fed larvae at 25 degrees C, the death rates caused by the Melia extract were higher, although at 30 degrees C the extract of Azadirachta had an even higher lethality. Inter allia, the LC50 values for the crude extracts of these two members of the Meliaceae ranged from 0.017 to 0.034 g% while the LC99 values ranged from 0.133 to 0.189 g%. Since no downstream processing was undertaken to purify the active agents in the extracts, our findings seem very promising, suggesting that it may be possible to increase the larvicidal activity further by improving the extraction and the fractionation of the crude limonoids, for instance removing the co-extracted natural fats.

[A simulation model for the control of the Aedes aegypti, the mosquito vector of dengue and yellow fever, by the crustacean Mesocyclops spp]. / Modelo de simulación para el control del mosquito Aedes aegypti, transmisor del dengue y la fiebre amarilla, por el crustáceo Mesocyclops spp.

OBJECTIVE: A simulation model is presented to show the predation dynamics of Mesocyclops spp. over Aedes aegypti. METHODS: The system is represented through four differential equations. H'(t), quantity of eggs; L'(t), quantity of larvae; A'(t), quantity of adults and C'(t), quantity of copepods. Initially the equations are of the classic predator-prey type, according to Lotka (1924) and Volterra (1926). Then it is modified into a system with functional response for invertebrates, according to Holling. RESULTS: The first system effectively controls and stabilizes the mosquito population, while the second suggests that copepods may be ineffective as mosquito controllers. CONCLUSIONS: The need to study predator-prey systems (copepodos-mosquitos) with projects that integrate laboratory and of field tests is recognized. Only then will it be possible to establish the validity of predators as effective biological controllers of mosquitoes.