Insecticidal and developmental inhibitory properties of monoterpenes on Culex pipiens L. (Diptera: Culicidae)

Twelve monoterpenes were evaluated for larvicidal and adulticidal activities towards Culex pipiens. Geraniol and cuminaldehyde were the most toxic monoterpenes to larvae, with LC₅₀ values of 38.6 and 38.9 mg/l after 24 h of treatment, respectively, whereas cuminaldehyde was the most potent compound after 48 h of treatment, followed by geraniol and thymol. In fumigant toxicity experiments, (R)-carvone and geraniol were the most toxic monoterpenes against the adults at all three tested concentrations and after both 24 and 48 h. When tested at sublethal concentrations (0.5 LC₅₀), (R)-carvone, (S)-limonene and cuminaldehyde decreased hatchability, pupation and adult emergence and induced high larval mortality. Our results suggest that geraniol, cuminaldehyde and (R)-carvone are promising toxicants against Culex pipiens and could be useful in the search for new natural insecticides.     

Insecticidal activities of monoterpenes and phenylpropenes against <Emphasis Type="Italic">Sitophilus oryzae</Emphasis> and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases

In the present study, six monoterpenes [(?)-citronellal, p-cymene, (?)-menthone, α-pinene, α-terpinene, and (?)-terpinen-4-ol] and two phenylpropenes [trans-cinnamaldehyde and eugenol] were evaluated for their contact and fumigant toxicities against Sitophilus oryzae adults. The effects of these compounds on the mortality of S. oryzae adults in stored wheat and their inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) were examined. The tested compounds showed varying degrees of contact toxicity, with trans-cinnamaldehyde (LC₅₀ = 0.01 mg/cm²) being the most potent compound, followed by (?)-menthone (LC₅₀ = 0.013 mg/cm²) and eugenol (LC₅₀ = 0.015 mg/cm²). In a fumigant toxicity assay, the monoterpenes α-terpinene, p-cymene, and (?)-menthone showed the highest toxicities (LC₅₀ = 50.79, 52.37, and 54.08 μl/L air, respectively). Trans-cinnamaldehyde, (?)-citronellal, and eugenol were the least toxic (LC₅₀ > 100 μl/L air). In general, the oxygenated compounds exhibited high contact toxicities while the hydrocarbon compounds exhibited high fumigant toxicities. When tested for their insecticidal activities against S. oryzae in stored wheat, trans-cinnamaldehyde was found to be the most potent compound, with 73.9% mortality at an application rate of 0.5 g/kg and complete mortality (100%) at 1 and 5 g/kg after 1 week of treatment. All of the tested compounds showed AChE inhibition, although (?)-citronellal and trans-cinnamaldehyde presented the strongest enzyme inhibition, with IC₅₀ values of 18.40 and 18.93 mM, respectively. On the other hand, (?)-terpinene-4-ol exhibited the highest inhibition of ATPases, followed by α-pinene and α-terpinene.     

Studies on the application of a nuclear polyhedrosis virus to control populations of the Egyptian cottonworm, Spodoptera littoralis

Variations in the resistance to nuclear polyhedrosis virus (NPV) were found in three populations of Spodoptera littoralis. The LD₅₀ for the most resistant population was 1.07 × 10⁴ PIB/5th instar larva as compared to 8.4 × 10² and 5.8 × 10² PIB/larva in the other two populations. The effect of NPV persisted in larvae which survived and pupated. Some of the pupae died, and those which survived produced normally shaped adults. While fecundity was sharply reduced in the less resistant populations, the effect on the most tolerant population was less pronounced. A 3-year-old inoculum, stored unprotected from daylight and without cooling, was much less effective even against the most sensitive larval population as compared to a relatively fresh and refrigerated batch. Larvae in their 6th instar proved to be approximately 10-fold more resistant to the NPV than 5th instar ones, while the difference in weight was only about twice. These variations in resistance to NPV are also discussed from the point of view of applying S. littoralis NPV in pest control schemes.     

Fumigant toxicity of Apiaceae essential oils and their constituents against Sitophilus oryzae and their acetylcholinesterase inhibitory activity

We evaluated the insecticidal and acetylcholinesterase (AChE) inhibition activities of the essential oils and their constituents of 10 Apiaceae on the adult rice weevil, Sitophilus oryzae. Of the 10 species tested, dill (Anethum graveolens), caraway (Carum carvi), and cumin (Cuminum cyminum) essential oils showed strong fumigant toxicity against adult S. oryzae. LC₅₀ values of caraway, dill, and cumin essential oils were 2.45, 3.29, and 4.75 mg/L air, respectively. Among the test compounds, (+)-carvone, (?)-carvone, cuminaldehyde, dihydrocarvone, linalool oxide, carveol, trans-anethole, and neral demonstrated strong fumigant toxicity against adult S. oryzae with LC₅₀ values of 0.61, 0.84, 1.12, 2.92, 3.76, 4.29, 5.02, and 6.60 mg/L air, respectively. α-Pinene showed the strongest AChE inhibition activity followed by β-pinene and limonene. The measured toxicity of the artificial blends of the constituents identified in dill and cumin oils indicated that (+)-carvone and cuminaldehyde were major contributors to the fumigant toxicity of the artificial blend.     

Effectiveness of monoterpenes and phenylpropenes on Sitophilus oryzae L. (Coleoptera: Curculionidae) in stored wheat

Six monoterpenes, ((?)-citronellal, p-cymene, (?)-menthone, α-pinene, α-terpinene and (?)-terpinen-4-ol) and two phenylpropenes, (trans-cinnamaldehyde and eugenol) were tested for their insecticidal activity against the rice weevil, Sitophilus oryzae under laboratory conditions. The bioassays were carried out on wheat at the concentrations of 0.5, 1.0 and 5.0?g/kg. Adult mortality was assessed after 14?days of exposure. After this interval, the treated wheat was retained for an additional period of 90?days, in order to evaluate progeny production and wheat loss. At the concentrations of 0.5 and 1.0?g/kg, trans-cinnamaldehyde and eugenol achieved highest adult mortality. At 5.0?g/kg, all compounds except for p-cymene and α-pinene caused complete (100%) adult mortality. Generally, trans-cinnamaldehyde was the most potent compound causing complete inhibition of progeny at the three concentrations. Similarly, no progeny were observed in the wheat treated with (?)-citronellal, eugenol, p-cymene, (?)-menthone, and (?)-terpinen-4-ol at 5.0?g/kg after 45 and 90?days. Similar trends were noted for wheat weight loss and damage as the treatment with monoterpenes and phenylpropenes at the highest rate preserved the wheat intact and free from damage by S. oryzae for 90?days. Our findings suggest the tested compounds except p-cymene and α-pinene could be recommended for use as part of an integrated pest management program for S. oryzae control in stored wheat.     

Toxicity and repellency of origanum essential oil and its components against Tribolium castaneum (Coleoptera: Tenebrionidae) adults

The components of Origanum vulgare L. essential oil showing insecticidal activity and repellency against red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), adults were analysed by GC-MS. All constituents were identified, and the main components were carvacrol (67.2%), p-cymene (16.2%), γ-terpinene (5.5%), thymol (4.9%), and linalool (2.1%). In a vapor phase fumigant assay, the origanum oil was more effective in closed conditions (LD₅₀ = 0.055 mg/cm³) than in open conditions (LD₅₀ > 0.353 mg/cm³). This suggests that toxicity is exerted largely in the vapor phase. Based on 24-h LD₅₀ values, the toxicity of caryophyllene oxide (0.00018 mg/cm³) was comparable with that of dichlorvos (0.00007 mg/cm³). In addition, thymol, camphene, α-pinene, p-cymene, and γ-terpinene showed good insecticidal activity (LD₅₀ = 0.012–0.195 mg/cm³). In repellency tests using 9 constituents of origanum oil, caryophyllene oxide showed complete repellency at 0.03 mg/cm². Hydrogenated monoterpenoids, such as thymol, α-pinene, carvacrol, and myrcene, elicited strong repellency at 0.03 and 0.006 mg/cm². Repellency depended on both time and concentration. These results indicate that origanum oil and its components could be potential candidates as a fumigant and repellent for managing T. castaneum adults.     

Occurrence and characterization of a nucleopolyhedrovirus from Spodoptera littoralis (Lepidoptera: Noctuidae) isolated in the azores

A nucleopolyhedrovirus (SpliMNPV-Az) was isolated from diseased larvae of Spodoptera littoralis, collected at the Island of S. Miguel in Azores. The virulence of this isolate was tested against S. littoralis larvae in laboratory. LD₅₀ against 2nd and 3rd instars were not significantly different, 1.44 × 10⁴, 3.89 × 10⁴ OBs per larvae, respectively, but both were significantly different from that against 4th instar, which was 61.3 × 10⁴ OBs per larvae. The complete codons sequence of SpliMNPV-Az Polh gene obtained was 750 bp (NCBI GenBank Accession No. AY600451). This sequence was compared to other 38 polyhedrin genes from NPVs and to 6 granulin genes from GVs and resulted to be identical to the sequence of a SpliMNPV previously published, thus indicating that the natural host of SpliMNPV-Az must be S. littoralis. Genetic distances estimated from restriction enzymes profiles showed SpliMNPV-Az is close to the Egyptian SpliMNPV type B, despite some degree of genetic divergence suggested by slight differences observed on PstI profile.     

Circadian and seasonal study of the cinnamate chemotype from Lippia origanoides Kunth

The leaves of Lippia origanoides Kunth are used in culinary as flavoring regional dishes and remedy for gastrointestinal and respiratory diseases in the Amazon region. The circadian and seasonal study of its essential oil was characterized by GC and GC–MS analysis. The oil components were grouped into monoterpenes, sesquiterpenes and phenylpropanoids, during the dry and rainy season. The main constituents were (E)-methyl cinnamate, (E)-nerolidol, p-cymene, 1,8-cineole, carvacrol, α-pinene, (E)-caryophyllene and γ-terpinene, with great variation throughout the year. In this work, we are reporting the occurrence of a new chemotype for L. origanoides, characterized by an essential oil rich in (E)-methyl cinnamate and (E)-nerolidol, with fruity-woody odor, reminiscent of cinnamon, strawberry and wood. The oil yield varied from 1.7% to 4.6%, which is considered a significant value for the production of essential oils on an industrial scale. This new chemotype may have ecological, chemosystematics and taxonomic significance in the management and economic utilization of the species.     

The insecticidal activity of essential oil constituents against pyrethroid-resistant Anopheles funestus (Diptera: Culicidae)

Malaria vector control relies on the use of insecticides for indoor residual spraying and long-lasting bed nets. However, insecticide resistance to pyrethroids among others, has escalated. Anopheles funestus, one of the major African malaria vectors, has attained significant levels of resistance to pyrethroids. Overexpressed P450 monooxygenases have been previously identified in pyrethroid resistant An. funestus. The escalating resistance against conventional insecticides signals an urgent need for identification of novel insecticides. Essential oils have gained recognition as promising sources of alternative natural insecticides. This study investigated six essential oil constituents, farnesol, (−)-α-bisabolol, cis-nerolidol, trans-nerolidol, methyleugenol, santalol (α and β isomers) and essential oil of sandalwood, for the adulticidal effects against pyrethroid-resistant An. funestus strain. The susceptibility against these terpenoids were evaluated on both pyrethroid-susceptible and resistant An. funestus. Furthermore, the presence of overexpressed monooxygenases in resistant An. funestus was confirmed. Results showed that both the pyrethroid-susceptible and resistant An. funestus were susceptible to three EOCs; cis-nerolidol, trans-nerolidol and methyleugenol. On the other hand, the pyrethroid-resistant An. funestus survived exposure to both farnesol and (−)-α-bisabolol. This study however does not show any direct association of the overexpressed Anopheles monooxygenases and the efficacy of farnesol and (−)-α-bisabolol. The enhanced activity of these terpenoids against resistant An. funestus that has been pre-exposed to a synergist, piperonyl butoxide, suggests their potential effectiveness in combination with monooxygenase inhibitors. This study proposes that cis-nerolidol, trans-nerolidol and methyleugenol are potential agents for further investigation as novel bioinsecticides against pyrethroid-resistant An. funestus strain.     

Susceptibility of Heliothis armiger to a commercial nuclear polyhedrosis virus

The susceptibility of Heliothis armiger larvae of different ages to a commercial nuclear polyhedrosis virus (NPV), Elcar, was determined by bioassay. The median lethal dosage (LD₅₀) increased 150-fold during the first week of larval life at 25°C, i.e., during development to early fourth instar, but daily feeding rate and thus potential virus acquisition also increased. A linear relationship was determined between log LD₅₀ and larval length, indicating that larval length constitutes a useful index for estimating the susceptibility of larval populations. Median lethal times (LT₅₀s) were similar for larvae tested at ages of 0 to 7 days and ranged from 3.6 to 8.0 days at 30°C. The amount of virus produced in a single, infected neonate was equivalent to 1.4 × 10⁶ LD₅₀s for neonates, a 900,000-fold increase on the dose supplied. The data support the practice of directing the NPV against neonates, but, on the basis of larval susceptibility alone, the age of larvae at treatment may not always be critical.     

Soil contact toxicity of insecticides to the european earwigForficula auricularia [Dermaptera]

The soil contact effects of several commonly used agricultural insecticides on 2nd instar and adultForficula auricularia (L.) were studied in the laboratory. The earwigs were individually confined on soil surfaces sprayed under a track sprayer. Pirimicarb had no effect on either adult or 2nd instar earwigs. The LD_50,s of 2nd instar earwigs for γ HCH, fenitrothion, DDT and dimethoate were all lower than their normal field rates. Second instars were more susceptible to dimethoate and cypermethrin than adults. The pyrethroids cypermethrin and deltamethrin had LD_50,s above normal field rates. However, high levels of knockdown were induced by both insecticides at concentrations below normal application rates. The effects of this in the field remain unknown.     

Polyphenolic extracts from pomegranate and watermelon wastes as substrate to fabricate sustainable silver nanoparticles with larvicidal effect against Spodoptera littoralis

The agricultural wastes adversely affect the environment; however, they are rich in polyphenols; therefore, this study aimed to employ polyphenol-enriched waste extracts for silver nanoparticles synthesis, and study the larvicidal activity of silver nanoparticles fabricated by pomegranate and watermelon peels extracts (PPAgNPs and WPAgNPs) against all larval instars of Spodoptera littoralis. The polyphenol profile of pomegranate and watermelon peel extracts (PP and WP) and silver nanoparticles was detected by HPLC. The antioxidant activity was estimated by DPPH, and FARP assays and the antimicrobial activity was evaluated by disc assay. The Larvicidal activity of AgNPs against Egyptian leaf worm was performed by dipping technique. The obtained AgNPs were spherical with size ranged 15–85 nm and capped with proteins and polyphenols. The phenolic compounds in silver nanoparticles increased about extracts; therefore, they have the best performance in antioxidant/reducing activity, and inhibit the growth of tested bacteria and yeast. The PPAgNPs were the most effective against the first instar larvae instar (LC₅₀ = 68.32 µg/ml), followed by pomegranate extract with (LC₅₀ = 2852 µg/ml). The results indicated that obvious increase in polyphenols content in silver nanoparticles enhance their larvicidal effect and increasing mortality of 1^st larval of S. littoralis Egyptian leafworms causing additive effect and synergism. We recommend recycling phenolic enriched agricultural wastes in producing green silver nanoprticles to control cotton leafworm that causes economic loses to crops.     

Castor and camphor essential oils alter hemocyte populations and induce biochemical changes in larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

Two plant essential oils; camphor and castor were tested for insecticidal and antifeedant activity against the 4th instar larvae of Spodoptera littoralis, a serious pest on cotton in Egypt. Also the impact of LC₁₀ of both oils on some physiological parameters in larvae was studied by using leaf dipping technique. Analysis of both oils using GC–MS revealed several insecticidal and antifeedant compounds. Our results showed higher insecticidal activity and antifeedant index of camphor oil against S. littoralis. The LC₅₀ and the antifeedant indices were 163.1, 246.8?mg/ml and 12.69, 6.62% for camphor and castor bean oil, respectively. The total hemocyte count (THC) and differential hemocyte count (DHC) were reduced significantly after 48?h of treatment compared to controls. Both oils reduced all types of hemocytes except plasmatocytes which were reduced only by castor oil. Camphor oil decreased total proteins and carbohydrates while castor oil targeted only carbohydrate content. Both oils didn't affect the amount of total lipids. Lipase, α-amylase and glucose-6-phosphate dehydrogenase (G6PD) enzyme activities were increased significantly in larvae treated with camphor oil than other treatments. These results clearly indicate that castor and camphor oils can affect the nutritional status in S. littoralis larvae, thereby changing the internal metabolic processes in the larvae which make them as potential control agents in IPM programs against S. littoralis.     

Insecticidal and feeding deterrent activities of essential oils in the cabbage looper,Trichoplusia ni (Lepidoptera: Noctuidae)

Ten essential oils were tested against the cabbage looper, Trichoplusia ni larvae for contact, residual and fumigant toxicities and feeding deterrent effects. Against third instar T. ni, Syzygium aromaticum (LD₅₀ = 47.8 μg/larva), Thymus vulgaris (LD₅₀ = 52.0 μg/larva) (the two positive controls) and Cinnamomum glanduliferum (LD₅₀ = 76.0 μg/larva) were the most toxic via topical application. Litsea pungens (LD₅₀ = 87.1 μg/larva), Ilex purpurea (LD₅₀ = 94.0 μg/larva), Cinnamomum cassia (LD₅₀ = 101.5 μg/larva) and Litsea cubeba (LD₅₀ = 112.4 μg/larva) oils were equitoxic. Thymus vulgaris (LC₅₀ = 4.8 mg/ml) and S. aromaticum (LC₅₀ = 6.0 mg/ml) oils were the most toxic in residual bioassays. Cymbopogon citratus (LC₅₀ = 7.7 mg/ml) and C. cassia (LC₅₀ = 8.5 mg/ml) oils were equitoxic followed by Cymbopogon nardus (LC₅₀ = 10.1 mg/ml) in this bioassay. The remaining five oils showed little or no residual effects. In a fumigation bioassay, L. cubeba (LC₅₀ = 16.5 μl/l) and I. purpurea (LC₅₀ = 22.2 μl/l) oils were the most toxic. Cinnamomum glanduliferum (LC₅₀ = 29.7 μl/l) and Sabina vulgaris (LC₅₀ = 31.2 μl/l) oils were equitoxic. Interestingly, S. aromaticum did not exhibit any fumigant toxicity. Cymbopogon citratus, C. nardus and C. cassia strongly deterred feeding by third instar T. ni (DC₅₀s = 26.9, 33.8 and 39.6 μg/cm², respectively) in a leaf disc choice bioassay. The different responses of T. ni larvae to the oils in different bioassays suggest that these essential oils exhibit different modes of action. Based on their comparable efficacy with essential oils already used as active ingredients in many commercial insecticides (i.e. clove oil and thyme oil), some of these essential oils may have potential as botanical insecticides against T. ni.     

Pharmacological interactions of essential oil constituents on the in vitro growth of Plasmodium falciparum

The pharmacological properties of essential oils have been widely studied. However the interaction and contribution of the individual constituents assayed singularly and in combination remains relatively unexplored. To investigate these possible interactions, various essential oil constituents (EOC's) were combined and the interactions on their antimalarial and toxicity properties determined using the tritiated hypoxanthine incorporation and the tetrazolium-based cellular viability assays, respectively. In combination, two inactive EOC's (IC50 values greater than 1 mM), ρ-cymene and carvacrol interacted in a synergistic manner (∑ FIC = 0.02) against a chloroquine-resistant strain of Plasmodium falciparum. This interaction was comparable to that between the potent E- and Z-(±)-nerolidol (IC50 value: 0.9 ± 0.3 μM) and the standard antimalarial, quinine (IC50 value: 0.13 ± 0.04 μM) (∑ FIC = 0.01). However, this latter combination also potentiated the toxicity (∑ FIC = 0.001) of each molecule. A similar increase in toxicity was noted between ρ-cymene and γ-terpinene, as well as E- and Z-(±)-nerolidol and (−)-pulegone. These results show that the pharmacological activities of individual EOC's can vary greatly when used in combination, and show potential as adjuvants in the elimination of malaria infections.     

Protection of potato crop against Phthorimaea operculella (Zeller) infestation using frass extract of two noctuid insect pests under laboratory and storage simulation conditions

The oviposition deterrent effect of water extract of Spodoptera littoralis and Agrotis ipsilon larval frass on Phthorimaea operculella adult females was studied using two types of larval food “Natural host and Semi-artificial diet” under laboratory and storage simulation (semi-field) conditions. Extracted frass of fed larvae on semi-artificial diet showed complete oviposition deterrent effect at treatments with 4th, 5th and 6th instars of S. littoralis, also at treatments with 1st–3rd and 6th instars of A. ipsilon, while the same effect was observed when the larvae fed on castor oil leaves as a natural host only at treatment with frass extract of A. ipsilon 6th instar larvae. Presence of low amounts of phenols and flavonoids in water extract of A. ipsilon larval frass resulted in relatively more effect as oviposition deterrent to fertile adult females on treated oviposition sites, while the opposite effect was obtained in S. littoralis larval frass experiments. At semi-field experiments, the percentage reduction of laid eggs reached 100% after two?days at treatments with frass extracts of 4th and 5th S. littoralis larval instars and A. ipsilon 6th instar larvae fed on semi-artificial diet and/or castor oil leaves. Percentage reduction of laid eggs for untreated sacks reached 93.24 and 48.95% after 2 and 30?days, respectively, when placed between treated sacks, in comparison with the mean number of laid eggs for isolated control.     

Efficacy of various synthetic pyrethroid-impregnated encasement materials against house dust mite under laboratory conditions

The acaricidal activity of synthetic pyrethroid and benzyl benzoate against Dermatophagoides pteronyssinus was examined in the laboratory, using a specially designed test set up. On the basis of median lethal dose (LD₅₀) values, the compound found to be most toxic to D. pteronyssinus was benzyl benzoate (LD₅₀ = 50 mg/m²), followed by permethrin (LD₅₀ = 76.7 mg/m²), deltamethrin (LD₅₀ = 146.7 mg/m²), esbioallenthrin (LD₅₀ = 186.6 mg/m²) and lamdacyhalothrin (LD₅₀ = 756.6 mg/m²). Very low toxicity was observed with bifenthrin (LD₅₀ = 5157.8 mg/m²). A laboratory control trial was also carried out to compare the acaricidal activity (residual effect) of four pyrethroids impregnated on woven and non-woven encasement materials against house dust mites during a 4-month period. Of the pyrethroids used in this study, esbioallenthrin demonstrated the highest acaricidal activity, and of the pyrethroid impregnated materials, the non-woven encasement material was more effective than the woven encasement material.     

Silver and zinc oxide nanoparticles induce developmental and physiological changes in the larval and pupal stages of Spodoptera littoralis (Lepidoptera: Noctuidae)

The rapid growth of nanotechnology application in various fields of science led to need of understanding their possible effects on development and physiology of insects before using them as control agents. Instead of feeding on a toxic dose, the impact of silver nanoparticles (AgNPs, 50–60?nm) and zinc oxide nanoparticles (ZnONPs, 10–30?nm) was studied at a nonlethal concentration on the larval stages of Spodoptera littoralis. Late second instar larvae of S. littoralis were treated with water, 10?mg/mL AgNPs and ZnONPs dipped castor leaves for 6?days. Both nanoparticles treated leaves caused reduction in both larval weight gain and pupal weight than water dipped leaves while ZnONPs only caused extended larval period. ZnONPs increased total hemocyte, granular cell and plasmatocyte counts while AgNPs increased plasmatocytes only. ZnONPs decreased the levels of protein, lipids and carbohydrates than control and AgNPs treated larvae. On the contrary, ZnONPs induced significant increase in the activities of amylase, glucose 6 phosphate dehydrogenase (G6PD), lipase as well as two antioxidative enzymes, the catalase and superoxide dismutase. These results clearly show that ZnONPs ingestion interfere with the digestive and immunological physiology as well as the development of S. littoralis.