The mosquito Aedes aegypti is a primary vector for major arboviruses, and its control is mainly based on the use of insecticides. Caffeine and spent coffee grounds (CG) are potential agents in controlling Ae. aegypti by reducing survival and blocking larval development. In this study, we analyzed the effects of treatment with common CG (CCG: with caffeine), decaffeinated CG (DCG: with low caffeine), and pure caffeine on the survival, behavior, and morphology of the midgut of Ae. aegypti under laboratory conditions. Third instar larvae (L3) were exposed to different concentrations of CCG, DCG, and caffeine. All compounds significantly affected larval survival, and sublethal concentrations reduced larval locomotor activity, delayed development, and reduced adult life span. Damage to the midgut of treated larvae included changes in epithelial morphology, increased number of peroxidase-positive cells (more abundant in DCG-treated larvae), and caspase 3-positive cells (more abundant in CCG-treated larvae), suggesting that the treatments triggered cell damage, leading to activation of cell death. In addition, the treatments reduced the FMRFamide-positive enteroendocrine cells and dividing cells compared to the control. CG and caffeine have larvicidal effects on Ae. aegypti that warrant field testing for their potential to control mosquitoes.

The present study was undertaken to assess the impact of a candidate mosquito larvicide, spinosad (8, 17 and 33 μg L−1) on a field population of Daphnia magna under natural variations of water temperature and salinity, using Bti (0.16 and 0.50 μL L-1) as the reference larvicide. Microcosms (125 L) were placed in a shallow temporary marsh where D. magna was naturally present. The peak of salinity observed during the 21-day observation period may have been partly responsible for the decrease of daphnid population density in all the microcosms. It is also probably responsible for the absence of recovery in the microcosms treated with spinosad which caused a sharp decrease of D. magna abundance within the first two days following treatment whereas Bti had no effect. These results suggest that it may be difficult for a field population of daphnids to cope simultaneously with natural (water salinity and temperature) and anthropogenic (larvicides) stressors.

In 2014, as an attempt to address the Zika health crisis by controlling the mosquito population, Brazil took the unprecedented action of applying a chemical larvicide, pyriproxyfen, to drinking water sources. The World Health Organization has established an acceptable daily intake of pyriproxyfen to be 100 μg per kg of body weight per day, but studies have demonstrated that at elevated doses (>5000 mg/kg), there are adverse effects in mice, rats and dogs. To better understand the potential developmental toxicity of pyriproxyfen, we utilized the embryonic zebrafish. Our results demonstrate that the concentration resulting in 50% of animals presenting adverse morphological effects (EC50), including craniofacial defects, was 5.2 μM for daily renewal exposure, and above this concentration, adverse behavioral effects were also observed in animals that followed a static exposure regimen. Thus, zebrafish data suggest that the developmental toxicity of pyriproxyfen may not be limited to insects.

Application of larvicidal oil (LO) is the most common practice in Hong Kong to control mosquitos, and hence prevent mosquito-borne diseases and protect human health. Globally, this study represented the first comprehensive assessment of toxicity and risk posed by LO to marine organisms. We found concentrations of LO ranged from 0.08 to 0.66 mg/L in coastal seawaters of Hong Kong. Waterborne exposure to water-accommodated fractions of LO resulted in growth inhibition to two microalgal species (72-h EC₅₀: 1.92–2.90 mg/L) and acute mortality to three marine animals (96-h LC₅₀: 3.41–8.10 mg/L). From these toxicity results, a concentration that considered to be hazardous to 5% of species (HC₅) was predicted at 1.45 mg/L, while the predicted no-effect concentration was determined to be 0.29 mg/L. The hazard quotient of LO exceeded 1 at 9 out of 15 sites, indicating moderate-to-high ecological risk to exposure of LO in the marine environment of Hong Kong.

Aegle marmelos (L.) Correa belongs to the family Rutaceae is generally known as “bael fruit tree” occuring across the south Asian countries. The current investigation screened the main derivatives from crude ethanolic extracts of the Bael tree leaf and evaluated activity effects on the larvae and adults of Aedes aegypti (L.) Dengue vector mosquito and a non-target aquatic predator. The GC-MS results showed that the peak area was found to be profound in N-methyl-1-adamantaneacetamide (N-M 1a) followed by oleic acid (OA) with 63.08 and 11.43% respectively. The larvicidal activity against the fourth instar larvae and the crude Ex-Am showed prominent mortality rate (93.60%) at the maximum dosage of 100 ppm. The mortality rate of N-M 1a and OA was occurred at 10 ppm (97.73%) and 12 ppm (95.4%). The repellent activity was found to be prominent at crude Ex-Am (50 ppm) as compared to the pure compounds (N-m 1a and OA) with maximum protection time up to 210 min. The non-target screening of Ex-Am, N-M 1a, and OA on mosquito predator Tx. splendens showed that they are scarcely toxic even at the maximum dosage of 1000 ppm (34.13%), 100 ppm (27.3%), and 120 ppm (31.3%) respectively. Thus, the present investigation clearly proved that the crude Ex-Am and their major derivatives Nm 1-a and OA showed their acute larval toxicity as well as potential mosquito repellent against the dengue mosquito and eco-safety against the mosquito predator.

Aedes aegypti is the main mosquito vector of dengue, zika, chikungunya, and yellow fever diseases. The low effectiveness of vector control options is mainly related to the increased insect’s resistance and to the toxicity of products used for non-target organisms. The development of new environmentally friendly and safer products is imperative. Technical cashew nut shell liquid (tCNSL), mostly composed by cardanol (C), is an abundant by-product of the cashew (Anacardium occidentale L.) production chain, available at low cost, and with proven larvicidal activity. However, chemical modifications in both tCNSL and cardanol were required to increase their water solubilities. Our objectives were to synthesise and characterise sustainable, low-cost and easy-to-use multiple function products based on tCNSL, cardanol, and the sulphonates obtained from both; and to evaluate all these products efficacy as surfactants, larvicidal, and antimicrobial agents. None of the sulphonates presented antimicrobial and larvicidal activities. tCNSL and cardanol were successfully emulsified with sodium technical cashew nut shell liquid sulphonate (NatCNSLS, complex mixture of surfactants). The emulsions obtained presented larvicidal activity due to the presence of tCNSL and cardanol in their composition. Our results showed that the tCNSL+NatCNSLS mixture emulsion was an effective larvicide and surfactant multiple function product, with high availability and easy-to-use, which can facilitate its large-scale use in different environments. Graphical abstract

Diflubenzuron (DFB) is a larvicide widely used to control Aedes aegypti populations as an alternative to organophosphates (OPs), with a specific mechanism of action for insects by inhibiting their chitin synthesis. However, DFB is used extensively in urban and rural environments, having the aquatic environment as the major receptor. Thus, the present study aimed to investigate the toxicity of DFB-based formulation and compare it with the toxicity of the OP temephos (TMP)-based formulation, a larvicide still used to control A. aegypti, on freshwater fishes Oreochromis niloticus and Hyphessobrycon eques. Organisms were submitted to acute (48 h) and prolonged (7 days) exposures, in the presence and absence of organic sediment, seeking interactions between chemical and sediment. Histopathological analyses were performed on O. niloticus gills and liver. According to 48-h median lethal concentration (LC₅₀), DFB- and TMP-based formulations were classified as harmful and toxic to fish, respectively, following the Globally Harmonized System of Classification (GHS). After prolonged exposure to sublethal concentrations, DFB-based formulation decreased H. eques body weight at concentrations 272-fold lower than its LC₅₀. Ultrastructural responses of O. niloticus indicated edemas and aneurisms on gills, and hepatocyte hypertrophy and vascular congestion of the liver. TMP-based formulation also induced pyknotic nuclei, which may lead to irreversible necrosis. The addition of organic sediment did not alter the larvicide toxicity, suggesting that larvicides remained available to the organisms. Altogether, these results suggest that as an insect-specific pesticide, DFB still induces mortality and tissue damage in fishes; thus, both larvicides pose risks to fishes.

Aedes aegypti is the main vector of yellow fever, chikungunya, Zika, and dengue worldwide and is managed by using chemical insecticides. Though effective, their indiscriminate use brings in associated problems on safety to non-target and the environment. This supports the use of plant-based essential oil (EO) formulations as they are safe to use with limited effect on non-target organisms. Quick volatility and degradation of EO are a hurdle in its use; the present study attempts to develop nanoemulsions (NE) of Trachyspermum ammi EO and its constituent thymol using Tween 80 as surfactant by ultrasonication method. The NE of EO had droplet size ranging from 65 ± 0.7 to 83 ± 0.09 nm and a poly dispersity index (PDI) value of 0.18 ± 0.003 to 0.20 ± 0.07 from 1 to 60 days of storage. The NE of thymol showed a droplet size ranging from 167 ± 1 to 230 ± 1 nm and PDI value of 0.30 ± 0.03 to 0.40 ± 0.008 from 1 to 60 days of storage. The droplet shape of both NEs appeared spherical under a transmission electron microscope (TEM). The larvicidal effect of NEs of EO and thymol was better than BEs (Bulk emulsion) of EO and thymol against Ae. aegypti. Among the NEs, thymol (LC₅₀ 34.89 ppm) had better larvicidal action than EO (LC₅₀ 46.73 ppm). Exposure to NEs of EO and thymol causes the shrinkage of the larval cuticle and inhibited the acetylcholinesterase (AChE) activity in Ae. aegypti. Our findings show the enhanced effect of NEs over BEs which facilitate its use as an alternative control measure for Ae. aegypti.

Populations of Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) have shown resistance to insecticides of the carbamate and organophosphate classes. The objective of this study was to assess the susceptibility of C. quinquefasciatus larvae to essential oils from leaves of Eugenia uniflora L., Melaleuca armillaris (Sol. ex Gaertn.) Sm., and Schinus molle L and C. quinquefasciatus larvae’s biochemical responses after their exposure to these leaves. The essential oils were chemically analyzed by GC and GC/MS. First, the lethal concentration for 50% (LC₅₀) values was estimated using different concentrations of essential oils and probit analysis. The larvae were exposed for 1 h at the LC₅₀ estimated for each essential oil. The susceptibility of the larvae to essential oils was evaluated using the following biochemical parameters: concentrations of total protein and reduced glutathione; levels of production of hydrogen peroxide and lipid peroxidation; and the activity of the enzyme acetylcholinesterase (AChE). The main chemical constituents in E. uniflora were E-β-ocimene, curzerene, germacrene B, and germacrone; in M. armillaris were 1,8-cineole and terpinolene; and in S. molle were sabinene, myrcene, and sylvestrene. The essential oils had LC₅₀ values between 31.52 and 60.08 mg/L, all of which were considered effective. All of them also promoted changes in biochemical parameters when compared to the control treatment. The essential oils of S. molle and E. uniflora inhibited the activity of the AChE enzyme, and the essential oil of M. armillaris increased it. All essential oils had larvicidal activity against C. quinquefasciatus, but the essential oil of E. uniflora was the most efficient. Thus, the findings of the present study suggest that the essential oil of E. uniflora can be considered promising for the development of botanical larvicides.

The control of storage insect pests is largely based on synthetic pesticides. However, due to fast growing resistance in the targeted insects, negative impact on humans and non-target organisms as well as the environment, there is an urgent need to search some safer alternatives of these xenobiotics. Many essential oils (EOs) and their bioactive compounds have received particular attention for application as botanical pesticides, since they exhibited high insecticidal efficacy, diverse mode of action, and favourable safety profiles on mammalian system as well as to the non-target organisms. Data collected from scientific articles show that these EOs and their bioactive compounds exhibited insecticidal activity via fumigant, contact, repellent, antifeedant, ovicidal, oviposition deterrent and larvicidal activity, and by inhibiting/altering important neurotransmitters such as acetylcholine esterase (AChE) and octopamine or neurotransmitter inhibitor γ-amino butyric acid (GABA), as well as by altering the enzymatic [superoxide dismutase (SOD), catalase (CAT), peroxidases (POx), glutathione-S-transferase (GST) and glutathione reductase (GR)] and non-enzymatic [glutathione (GSH)] antioxidant defence systems. However, in spite of promising pesticidal efficacy against storage pests, the practical application of EOs and their bioactive compounds in real food systems remain rather limited because of their high volatility, poor water solubility and susceptibility towards degradation. Nanoencapsulation/nanoemulsion of EOs is currently considered as a promising tool that improved water solubility, enhanced bio-efficacy, stability and controlled release, thereby expanding their applicability.