Cuticular extracts induce aggregation in head lice.

The human head lice Pediculus humanus capitis (De Geer) (Phthiraptera: Pediculidae) are strict, obligate human ectoparasites that spends their entire life cycle in the host and cause skin irritation and derived infections. Despite the health-related importance, few studies have evaluated the chemical communication among these insects. Here, we evaluate the response of lice of both sexes to cuticular extracts using two solvents of different polarity (hexane and methanol). Cuticular extracts that elicited an attraction response towards head lice were analysed by gas chromatography-mass spectrometry (GC-MS) to determine the cuticular lipid profile. Both lice sexes were attracted to the hexane extracts but not the methanol extracts, suggesting the non-polarity of the compounds present in the cuticle. Chemical analyses of hexane extracts from males and females showed high similarity in major compounds. This study provides the first evidence that lice respond to cuticle extracts, which may be important to understand aggregation behaviour.

The Response of Susceptible and Pyrethroid-Resistant Blattella germanica (Dyctioptera: Blattellidae) to Shelter-Associated Cues.

In this work, it was studied the role of faeces in the location and permanence in a shelter in susceptible and pyrethroid-resistant individuals of Blattella germanica (Linnaeus 1767). Additionally, the effect of different concentrations of palmitic acid on the modulation of these behaviours was tested. A shelter constituted by a square cardboard structure was offered to susceptible as well as to resistant specimens. The shelter bases were treated with faecal extracts obtained from susceptible or resistant cockroaches, or with solutions of palmitic acid. The behaviour of susceptible as well as resistant specimens was analysed using infrared videography software. Susceptible's faecal extract attracted both specimens since the time spent by cockroaches to locate the treated shelters was lower, whereas the faecal extract from resistant insects did not elicit any effect on both strains. Faecal extracts showed an arrestant effect on both strains, suggested by the time spent inside the shelter that was significantly higher in their presence. On the other hand, treatment with palmitic acid produced an attractant or a repellent effect depending on the concentration and strain. The tested lower concentration was attractant to susceptible insects, but did not produce any effect on resistant ones. In addition, the higher concentrations did not produce any effect on susceptible individuals, but resulted repellent for resistant ones. Palmitic acid did not produce an arrestant effect on the strains as there was not an increase in time spent inside the shelter in the presence of this substance. An increase in the number of visits to the shelter and to the periphery was also observed in shelters treated with the faecal extract and with the lower concentration of palmitic acid. These results show that compounds of the susceptible faeces were attractant to cockroaches of both strains, while faecal extracts from resistant insects were not. Moreover, a dual effect of palmitic acid was observed, being attractant at low concentrations and repellent as concentration increased. Additionally, a difference in the concentration threshold at which the effect of this substance changes was observed between strains.

Feeding Behaviour of a Pyrethroid-Resistant Strain of the German Cockroach Blattella germanica (Linnaeus, 1767).

Many authors report biological differences among insecticide-resistant pest species, mainly associated with parameters of life history, but only a few studies approach differences at the behavioural level. Feeding behaviour in Blattella germanica (L.) is modulated by the detection of chemical volatiles emitted from food sources in order to be located a long-median distance, and also by the physicochemical properties of food. This work aimed to study the differences in the feeding behaviour of a susceptible and a pyrethroid-resistant strain of B. germanica, in the location and exploitation of a food source. Resistant males showed a lower performance in the feeding behaviour compared to susceptible males. Particularly, the time taken to locate the source was significantly higher in the resistant individuals, suggesting a lower capacity in the detection of food odours. In addition, although the Intake rate was negatively related to the stiffness of food for both strains, resistant individuals showed a lower intake rate compared to susceptible ones, when exploiting a food source of the same stiffness. In a control context, the phenotypic characteristics associated to feeding of pyrethroid-resistant individuals could induce a behavioural resistance mechanism caused by a reduction in the amount of toxic gel bait ingested. If that is the case, resistance of this strain would not be associated to physiological changes affecting the toxicity of the active principle of gel baits, but to a lower efficacy of these products against resistant individuals due to less amount of gel consumed, generating control problems of this pest.

Characterization of the pyrethroid resistance mechanisms in a Blattella germanica (Dictyoptera: Blattellidae) strain from Buenos Aires (Argentina).

The use of chemical insecticides is the main control method for Blattella germanica worldwide. The prolonged and frequent use of insecticides produced the selection of insecticide-resistant individuals. The German cockroach is one of the most widespread urban pests in Argentina. In the last decades, resistance monitoring studies in this country demonstrated that there is a high prevalence of pyrethroid-resistant populations of B. germanica in the field. In this work, we studied the resistance mechanisms of a field-collected strain of B. germanica at toxicological, enzymatic, and molecular levels. A resistance ratio of 100 was obtained for the resistant strain when it was exposed to ß-cypermethrin. The pretreatment with specific synergists (piperonyl butoxide and triphenyl phosphate) led to a significant increase in the toxicity of the pyrethroid, suggesting an involvement of oxidases and esterases in the detoxification of this insecticide. Moreover, esterase and oxidase activities in the resistant strain were 1.5-fold and 2-fold higher respectively, compared to the susceptible individuals. On the other hand, the voltage-gated sodium channel gene of the resistant cockroaches did not show nucleotidic substitutions in the domain II which are associated to knockdown resistance in this species. These results suggest that the main mechanism of resistance of the studied cockroaches' strain is metabolic, mainly due to an increase in the activity of oxidase and esterase enzymes. The results of this work in addition to other reports found in literature show that the extended use of a single active principle for cockroach control promotes the development of resistance leading to control failure in the field. In contrast, integrated pest management strategies include the use of different control tools in addition to chemical insecticides, which delay the appearance of resistance increasing the efficacy of pest control.

Head lice were also affected by COVID-19: a decrease on Pediculosis infestation during lockdown in Buenos Aires.

Pediculosis is a worldwide disease affecting school-aged children produced by the presence of the head louse, Pediculus humanus capitis De Geer, an obligate ectoparasite on the human scalp feeding exclusively on blood. Transmission occurs primarily through direct physical head-to-head contact. In March 2020, the World Health Organization (WHO) declared the COVID-19 outbreak as a pandemic. COVID-19 is caused by SARS-CoV-2, a variant of the coronavirus. Therefore, on March 18, 2020, the Argentinean government established mandatory isolation for an indefinite period. This obligatory isolation interrupted regular classes avoiding direct contact between children, thus affecting the dispersal route of individuals and the evolution of head louse populations. In this study, we evaluated through an online survey how confinement affected the prevalence of lice during lockdown compared to the situation prior to confinement. The survey allowed to discriminate the different control strategies, the number of treatments, and the amount of insects recorded by parents. Data of 1118 children obtained from 627 surveys were analyzed. As the main result, it was observed that prevalence of lice decreased significantly from before (69.6%) to during (43.9%) COVID-19 lockdown. Moreover, head lice infestation was more effectively controlled in households with up to 2 children in comparison to households with 3 or more children. This is the first study that analyzed the prevalence of head lice during COVID-19 pandemic. In addition, this work demonstrated the impact of social distance in the population dynamics of head lice and how it could affect the control strategies in the future.

Spatial Repellency Caused by Volatile Pyrethroids is Olfactory-Mediated in the German Cockroach Blattella germanica (Dictyoptera: Blattellidae).

Pyrethroids are synthetic insecticides that have a repellent action. This effect has been associated with an increase in the locomotor activity, which causes the avoidance of the insecticide-treated area (excito-repellency). In this work, we studied with behavior and electrophysiological recordings the occurrence of olfactory-mediated repellency caused by pyrethroids of different volatility in the German cockroach Blattella germanica (Linnaeus, 1767). Male cockroaches were spatially repelled when they were exposed to D-allethrin vapors and vapothrin vapors in a dose-dependent manner. No repellency was observed when insects were exposed to permethrin, a non-volatile pyrethroid. To confirm the role of olfaction in this phenomenon, we measured the electrical activity of the cockroaches' antennae in response to these insecticides. There was a significant increase in the electrical activity in response to D-allethrin and vapothrin, but no increase was observed in insects exposed to permethrin. Locomotor activity of cockroaches exposed to pyrethroids was measured in order to discard excito-repellency. No changes in locomotor activity were observed for any of the insecticides. Finally, we found that volatile pyrethroids in the vapor phase cause spatial repellency in cockroaches, being the first report of an olfactory-mediated repellency phenomenon caused by pyrethroids in cockroaches.

Head Lice Recognize and Prefer Head Odor Over Foot and Forearms Odors.

Human head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), are hematophagous parasites that infest human heads. They are extreme host specialists suggesting a strict selection behavior towards the human head by the parasites. Despite the public health relevance of P. humanus capitis, the role of chemical clues to select the human head is not well known. In the present study, we attempted to find out whether head lice recognize and select the odor of the head over the odor of other parts of the human body. Our results using a two-choice olfactometer demonstrated that head lice were highly attracted by the volatile compounds of the human odor air, but they did not show preference for the volatiles of head compared to the volatiles of other parts of the body (forearm or foot). Conversely, when head lice were exposed in an experimental arena to the whole human odor of different parts of the body, they showed a preferential response to the whole head odor compared with the whole foot or forearm odors. These results suggest that head lice can at short distance of the host, be oriented towards the head odor of the human host in the selection of the scalp environment.

N-substituted methyl maleamates as larvicidal compounds against Aedes aegypti (Diptera: Culicidae).

Severe human arboviral diseases can be transmitted by the mosquito Aedes aegypti (L.), including dengue, chikungunya, zika, and yellow fever. The use of larvicides in containers that can result as potential breeding places and cannot be eliminated is the main alternative in control programs. However, their continuous and widespread use caused an increase in insecticide-resistant populations of this mosquito. The aim of this study was to evaluate the effect of three N-substituted methyl maleamates as larvicides on Ae. aegypti, the N-propyl methyl maleamate (PMM), N-butyl methyl maleamate (BMM), and N-hexyl methyl maleamate (HMM). These compounds could have a different mode of action from those larvicides known so far. We evaluated the larva mortality after 1 and 24 h of exposure and we found that mortality was fast and occurs within the first 60 min. HMM was slightly more effective with LC50 values of 0.7 and 0.3 ppm for 1 and 24 h of exposure and LC95 of 11 and 3 ppm. Our results demonstrate that N-substituted methyl maleamates have insecticidal properties for the control of Ae. aegypti larvae. These compounds could become useful alternatives to traditional larvicides after studying their insecticidal mechanism as well as their toxicity towards non target organisms.

Arrestant Effect of Human Scalp Components on Head Louse (Phthiraptera: Pediculidae) Behavior.

Relevant evidence has shown that parasites process host-related information using chemical, visual, tactile, or auditory cues. However, the cues that are involved in the host-parasite interaction between Pediculus humanus capitis (De Geer 1767) and humans have not been identified yet. In this work, we studied the effect of human scalp components on the behavior of adult head lice. Filter paper segments were rubbed on volunteers' scalps and then placed in the experimental arena, where adult head lice were individually tested. The movement of the insects was recorded for each arena using the software EthoVision. Average movement parameters were calculated for the treatments in the bioassays such as total distance, velocity, number of times a head louse crossed between zones of the arena, and time in each zone of the arena. We found that scalp components induced head lice to decrease average locomotor activity and to remain arrested on the treated paper. The effect of the ageing of human scalp samples in the response of head lice was not statistically significant (i.e., human scalp samples of 4, 18, 40, and 60 h of ageing did not elicit a significant change in head louse behavior). When we analyzed the effect of the sex in the response of head lice to human scalp samples, males demonstrated significant differences. Our results showed for the first time the effect of host components conditioning head lice behavior. We discuss the role of these components in the dynamic of head lice infestation.

Modulation of the behavioral and electrical responses to the repellent DEET elicited by the pre-exposure to the same compound in Blattella germanica.

Insects under different stimuli from the environment modify behavioural responses due to changes in the sensitivity of neurons at the peripheral and/or at the central level of the nervous system. This phenomenon is called neuronal plasticity, and sensory adaptation is an example of it. An insect repellent is a chemical that produces oriented movements of the insects away from its source. In this work we studied the modulation of the behavioural and electrical response to the repellent N, N-diethyl-3-methylbenzamide (DEET) in males of the German cockroach B. germanica produced by previous exposure to the same repellent. Methods. We determined repellency using a circular arena, one half of which was treated with DEET. The time spent by insects in each half of the arena was measured, and a repellency coefficient (RC) was calculated. The RCs of pre-exposed and non-pre-exposed insects were compared. To determine a possible role of nitric oxide in the modulation of the response to DEET after pre-exposure, the nitric oxide donor S-nitroso-acetyl-cysteine (SNAC) was applied on cockroaches' antennae. The electrical activity of the cockroaches' antennae in response to DEET was recorded using electroantennogram (EAG) technique. The response to DEET was recorded also after a long stimulation with the same repellent, and after topical application of SNAC and dbcGMP (a cGMP analogue) on the antennae. Results. We found that previous exposure of B. germanica males to the repellent DEET produced an increase of the repellency at the behavioural level, measured as RC. A possible role of nitric oxide (NO) in the transduction pathway of this phenomenon is suggested, since treatment of the cockroaches with the NO donor SNAC also produced an increase of the repellency elicited by DEET. On the other hand, the response of the cockroaches' antennae exposed to DEET was determined electrophysiologically. The electrical activity in response to DEET decreased when the insects' antennae were stimulated with a long pulse of the repellent. The activity of the antennae was restored after 10 min. Treatment of the antennae either with SNAC or dbGMPc also produced a decrease in the response of the antennae to the repellent. Discussion.The previous exposure to a chemical stimulus can modify the behaviour associated to the same stimulus, increasing or decreasing the behavioural response. In the case of DEET we found that pre-exposure increased DEET repellency in male cockroaches. We also found NO involvement in a similar phenomenon. On the other hand, the test showed that DEET is perceived by insects' antennae as an odour. A long exposure of the antennae to DEET caused a transient decrease in the response of the antennae to the same compound. The same effect was achieved by treating the antennae with SNAC or dbcGMP, suggesting the involvement of the NO/cGMP system in the transduction pathway of the sensory adaptation phenomenon elicited by an odour in this species.

The use of Aedes aegypti larvae attractants to enhance the effectiveness of larvicides.

Aedes aegypti (L.) is an important dengue, chikungunya, and yellow fever vector. Immature stages of this species inhabit human-made containers placed in residential landscapes, and the application of larvicides inside containers that cannot be eliminated is still considered a priority in control programs. Larvicidal efficacy is influenced by several factors, including the formulation used, the water quality, and the susceptibility of larvae, among others. If an attractant can be incorporated into a slow-release larvicide formulation, it will be feasible to direct the larvae into the source of insecticide and thereby improving its efficacy. We studied the influence of 1-octen-3ol and 3-methylphenol on the rate of Ae. aegypti larvae mortality using the larvicides Bacillus thuringiensis var. israelensis (Bti), temephos, and spinosad. These chemicals were combined with the larvicides mixed with agar during the bioassays. Mortality was registered every 10 min, and a lethal time 50 (LT50) was calculated. The inclusion of the Ae. aegypti larvae attractants with the larvicides into a solid agar matrix improved their efficiency obtaining a strong and marked reduction in the LT50 compared with the use of larvicides alone.

Oviposition Behavior in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Response to the Presence of Heterospecific and Conspecific Larvae.

In mosquitoes, location of suitable sites for oviposition requires a set of visual, tactile, and olfactory cues that influences females before laying their eggs. The ability of gravid females to distinguish among potential oviposition sites that will or will not support the growth, development, and survival of their progeny is critical. Aedes aegypti (L.) and Aedes albopictus (Skuse) share ecological niches, being highly competitive in larval stage. We studied the oviposition behavior of both species in the presence of larvae of one or the other species (heterospecific or conspecific larvae). The number of eggs laid by gravid females on oviposition sites (water with different or the same species of Aedes larvae) were compared. The presence and density of heterospecific or conspecific larvae had a positive or negative effect on the ovipositional responses, measured as an oviposition activity index. For both species, the oviposition was not affected by heterospecific larvae with densities between 10 and 100 larvae in water, but a strong attractant behavior was observed for a density of 500 larvae in water. For Ae. albopictus in the presence of larvae of the same species (conspecific oviposition), we observed an attractant effect for larvae density of 10 but not for 100 or 500 larvae in water. Instead, for Ae. aegypti, we observed attraction only for 100 larvae, not for 10 or 500 larvae. Results presented here provide an additional insight about oviposition behavior responses of gravid females in the presence of conspecific and heterospecific larvae in breeding sites.

Behavioral Response of Aedes aegypti (Diptera: Culicidae) Larvae to Synthetic and Natural Attractants and Repellents.

Aedes aegypti (L.) (Diptera: Culicidae) is the key vector of three important arboviral diseases: dengue, yellow fever, and chikungunya. Immature stages of this species inhabit human-made containers placed in residential landscapes. In this study, we evaluated a few compounds in a sensitive behavioral assay with Ae. aegypti larvae. The orientation of larvae to different compounds was surveyed using a performance index (PI). The PI represents the response to each odorant, where a value of +1 is indicative of full attraction and -1 represents complete repulsion. The widely used insect repellent N, N-diethyl-m-toluamide elicited a significantly negative PI, as did acetophenone and indole. A yeast extract, a known food source, elicited a significantly positive PI, as did 2-methylphenol, 1-octen-3-ol, 3-methylphenol, and fish food. On the other hand, no response was observed for the essential oil of Eucalyptus grandis x Eucalyptus camaldulensis at the concentration evaluated. Pretreatment of larvae with N-ethylmaleimide and ablation of the antennae resulted in a suppression of behavioral responses. The overall mobility of ablated larvae was indistinguishable from unablated controls, and absence of any visible locomotor dysfunction was observed. This work is a contribution to the study of the chemical ecology of disease vectors with the aim of developing more efficient tools for surveillance and control.Natural and synthetic compounds attractive to Ae. aegypti larvae should be incorporated into integrated pest management programs through the use of baited traps or by improving the efficacy of larvicides commonly used in control campaigns.

Effect of release rate and enantiomeric composition on response to pheromones of Megaplatypus mutatus (Chapuis) in poplar plantations of Argentina and Italy.

Megaplatypus mutatus (=Platypus sulcatus Chapuis) is an Ambrosia beetle native to South America, which was recently introduced in Italy and its presence there is causing severe damage to the local poplar plantations. The male M. mutatus pheromone is composed of (S)-(+)-6-methyl-5-hepten-2-ol [(+)-sulcatol], 6-methyl-5-hepten-2-one (sulcatone) and 3-pentanol. A series of field trials testing dose, blend and enantiomer composition performed in Argentina and Italy evaluated attraction and found that the optimal release rate of pheromone components as baits in cross vane baited traps (CIPEIN-CV) was 6, 6 and 30 mg day−1 of sulcatone, (+)-sulcatol and 3-pentanol, respectively. It was also determined that racemic sulcatol is as effective as the pure (+)-isomer for the purpose of beetle catch, due to the inert nature of the (−)-isomer allowing the usage of low cost racemic sulcatol instead of highly expensive (+)-sulcatol. The results of our work contribute to the development of pheromone-based local technologies with low environmental impact and low cost for control or monitoring of an important pest.

Comparative toxicity of oxygenated monoterpenoids in experimental hydroalcoholic lotions to permethrin-resistant adult head lice.

The use of botanical compounds such as essential oils has recently become the subject of great interest as a natural means of pest control because of their ovicidal, larvicidal, or adulticidal activity against various insect species including head lice. We tested and compared the efficacy of pure oxygenated monoterpenoids that are main ingredients of essential oils of good biological activity. We used pulegone and citral, components of Aloysia citrodora, and geraniol, citronellol, and linalool, components of Geranium sp. oil. We found that citronellol and geraniol showed the highest knockdown and mortality effect (>60%) on adults of both sexes (50:50%) and third-stage nymphs. Pulegone, linalool, and citral showed knockdown percentages between 42 and 55%, and mortality percentages between 47 and 53%. A simple linear regression analysis showed statistically significant relationships between the studied toxic effects and viscosity of the monoterpenoids (p < 0.05), but not with their partition coefficient (log P).

Comparison of three types of traps baited with sexual pheromones for ambrosia beetle Megaplatypus mutatus (Coleoptera: Platypodinae) in poplar plantations.

Megaplatypus mutatus (Coleoptera: Platypodinae) is an ambrosia beetle native to South America that only attacks standing live trees and is a serious problem for commercial poplar (Populus L.; Salicaceae) plantations in Argentina. The development of traps baited with synthetic pheromones that can be used for monitoring M. mutatus in infested poplar plantations is an important goal in preventive programs. Pioneer male M. mutatus emit a pheromone composed mainly by (+)-sulcatol and sulcatone. In the current study, we tested their release rates from several polymeric reservoir systems, to develop and manufacture a pheromone-releasing device. The efficacy of three different types of traps was evaluated in the field. Single funnel traps equipped with cross-vanes (CIPEIN-CV) captured significantly more insects than multiple funnel traps (LINDGREN) and simple funnel traps (CIPEIN-F).

Sensitivity of Aedes aegypti adults (Diptera: Culicidae) to the vapors of Eucalyptus essential oils.

Vapors of essential oils extracted from various species of Eucalyptus (E. gunnii, E. tereticornis, E. grandis, E. camaldulensis, E. dunnii, E. cinerea, E. saligna, E. sideroxylon, E. globulus ssp. globulus, E. globulus ssp. maidenii, E. viminalis and the hybrids E. grandisxE. tereticornis and E. grandisxE. camaldulensis) and their major components were found to be toxic to Aedes aegypti adults, the yellow fever mosquito. An aliquot of each oil was placed in a cylindrical test chamber and the number of knocked-down mosquitoes was recorded as function of time. Knockdown time 50% was then calculated. Results showed that E. viminalis had the fastest knockdown time at of 4.2 min, on the same order as dichlorvos, a standard knockdown agent. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect. The correlation between KT(50) values and calculated vapor pressures of the essential oil components showed that the fumigant activity of simple organic compounds in insects is correlated with their volatility.

Epidemiology of Pediculosis capitis in elementary schools of Buenos Aires, Argentina.

The infestation with the human obligate ectoparasite Pediculus humanus capitis De Geer is a common public health problem affecting mainly schoolchildren worldwide. The aim of the present study was to investigate the infestation levels of head lice in elementary schools from Buenos Aires with resistant levels to permethrin >100. A total of 1,856 children aged 3-13 years old from eight selected elementary schools were examined for head lice. Pediculosis was observed in all the studied schools. The overall infestation rate was 29.7%. Girls were statistically significant more infested than boys, with infestation rate values of 36.1% and 26.7%, respectively (P < 0.0001). Only 42 of the infested girls (12%) and 23 of the infested boys (11.4%) had >10 lice on their hair. The proportions of infested children-both girls and boys-in each age group were not found to differ significantly from one another. The infestation rate among schools varied from 19.12% to 42.74%. This indicated that pediculosis is relatively common in elementary schools from Buenos Aires, and those levels are of epidemic importance. The differences of pediculosis among the studied schools could be explained by the different control strategies applied by parents or advisors to eradicate head lice.

3-pentanol: a new attractant present in volatile emissions from the ambrosia beetle, Megaplatypus mutatus.

Megaplatypus mutatus (=Platypus mutatus) (Coleoptera: Platypodidae) is an ambrosia beetle that is native to South America. It attacks only standing live trees and causes severe stem breakage and death in commercial poplar (Populus) plantations. Previous work showed that male M. mutatus emits a sex pheromone composed mainly of (+)-sulcatol and sulcatone. We collected male volatile emissions during the hours of maximum emergence by using a specific polar microextraction phase; analyzed the extract by GC-MS; and tested the biological activity of selected compounds in the extract with a walking behavioral assay. Female M. mutatus emerged primarily between 7 and 11 h. In the chemical analyses of volatiles, a third compound, 3-pentanol, was identified in a small percentage of samples. Walking behavioral bioassays with video image analysis showed that at the doses tested, 3-pentanol elicited an attractive response from females.

Larvicidal effect of Eucalyptus grandis essential oil and turpentine and their major components on Aedes aegypti larvae.

In the search for new alternatives for the control of Aedes aegypti the larvicidal activity of Eucalyptus grandis essential oil and pine resin essential oil (turpentine) and their major components (alpha- and beta-pinene and 1,8-cineole) was determined. Gas chromatography-mass spectroscopy analysis of E. grandis essential oil revealed that its major components are alpha-pinene and 1,8-cineole. Similar analysis of turpentine obtained by distillation of the resin pitch of conifers showed that alpha- and beta-pinene are the only major components. Third and early 4th instars of the CIPEIN-susceptible strain of Ae. aegypti were exposed to acetonic solutions of E. grandis essential oil, turpentine, and their major components for 24 h. Turpentine, with an LC50 of 14.7 ppm, was more active than the essential oil of E. grandis (LC50: 32.4 ppm). Larvicidal activity of the essential oil components showed that alpha- and beta-pinene present low LC50 values (15.4 and 12.1 ppm, respectively), whereas pure 1,8-cineole showed an LC50 of 57.2 ppm. These results suggest that alpha-pinene in E. grandis and alpha- and beta-pinene in turpentine serve as the principal larvicidal components of both oils. Results obtained on larvicidal effects of essential oil of Eucalyptus grandis and turpentine could be considered a contribution to the search for new biodegradable larvicides of natural origin.