Concentration-Dependent Feeding Deterrence to 20-Hydroxyecdysone for Three Subterranean Termite Species (Blattodea: Rhinotermitidae)

Effective active ingredients in toxicant bait formulations must be non-deterrent to insect feeding behavior at lethal concentrations. This study evaluated feeding deterrence for Coptotermes formosanus Shiraki, C. gestroi (Wasmann), and Reticulitermes flavipes (Kollar) when provided access to cellulose impregnated with various concentrations of the insect molting hormone, 20-hydroxyecdysone (20E). Termites were exposed to 20E concentrations of 200, 500, 1000 and 2000 ppm and to noviflumuron at 5000 ppm in a 24 h choice-test, and the mass of substrate consumption from treated and untreated media pads was compared for each treatment. 20E feeding deterrence was detected at 500, 1000 and 2000 ppm for C. gestroi, and at 2000 ppm for C. formosanus. No significant differences in consumption of treated and untreated substrate was detected at any concentration for R. flavipes. Potential methods for reducing deterrence are discussed.

Genetic Manipulation of the Ergot Alkaloid Pathway in Epichloë festucae var. lolii and Its Effect on Black Beetle Feeding Deterrence

Epichloë endophytes are filamentous fungi (family Clavicipitaceae) that live in symbiotic associations with grasses in the sub family Poöideae. In New Zealand, E. festucae var. lolii confers significant resistance to perennial ryegrass (Lolium perenne) against insect and animal herbivory and is an essential component of pastoral agriculture, where ryegrass is a major forage species. The fungus produces in planta a range of bioactive secondary metabolites, including ergovaline, which has demonstrated bioactivity against the important pasture pest black beetle, but can also cause mammalian toxicosis. We genetically modified E. festucae var. lolii strain AR5 to eliminate key enzymatic steps in the ergovaline pathway to determine if intermediate ergot alkaloid compounds can still provide insecticidal benefits in the absence of the toxic end product ergovaline. Four genes (dmaW, easG, cloA, and lpsB) spanning the pathway were deleted and each deletion mutant was inoculated into five different plant genotypes of perennial ryegrass, which were later harvested for a full chemical analysis of the ergot alkaloid compounds produced. These associations were also used in a black beetle feeding deterrence study. Deterrence was seen with just chanoclavine present, but was cumulative as more intermediate compounds in the pathway were made available. Ergovaline was not detected in any of the deletion associations, indicating that bioactivity towards black beetle can be obtained in the absence of this mammalian toxin.

Larvicidal and Antifeedant Activity of Some Plants Extracts Against the Larvae of Helicoverpa Armigera (Hubner)

In the present study, selected some plants, namely, Andrographis paniculata Ness., Cardiospermum halicacabum L., Cassia tora L., Catharanthus roseus L (G) Don., Datura metal L., Eupatorium riparium and Mikania micarantha were screened for their larvicidal and antifeedant activity against the larvae of Helicoverpa armigera (Hubner) under laboratory conditions. The crude extracts of all the selected plants demonstrated a dose dependent increase in bioactivity. However the bioactivity of four plants namely, A.paniculata, Cassia tora L., C.halicacabum L., and Datura metal L. was significantly higher (p ≤ 0.05) than the control and extracts of C.roseus, E. riparium and M.micarantha. Methanol extract of A.paniculata caused highest oral toxicity with larval mortality ranging between 29.00% and 58.22% across the test concentration (0.2%, 0.4% and 1% w/v) while extract of C.tora L., demonstrated the highest feeding deterrence with reduction in larval feeding by 59.92% and 76.61% at 0.2% and 0.4% respectively. Crude extract of C.halicacabum L., leaves demonstrated high oral toxicity and feeding deterrence while extract of D.metel showed moderate level of oral toxicity as well as feeding deterrence at the highest tested concentration. Thus it may be concluded that four out of the selected plants possess insecticidal property and can be further investigated for the development of a potent natural botanical insecticide.

Repellent and Feeding Deterrent Activities of Butanolides and Lignans Isolated from Cinnamomum camphora against Tribolium castaneum

Three lignans (1–3) and three butanolides (4–6) were isolated from the lipophilic extract of the Cinnamomum camphora stem bark. The six compounds were identified as (-)-sesamin (1), 9α-hydroxysesamin (2), 9β-hydroxysesamin (3), obtusilactone A (4), isoobtusilactone A (IOA, 5), and isomahubanolide (6) from their spectroscopic data. Four (1, 2 and 5, 6) of them were evaluated for their repellent and feeding deterrent activities against Tribolium castaneum. In this work, the three butanolides (4–6) were confirmed to exist in C. camphora for the first time. Results of bioassays indicated that (-)-sesamin (1), IOA (5), and isomahubanolide (6) displayed certain repellent activities against T. castaneum at 78.63, 15.73, and 3.15 μg/cm2 at 2 h after exposure. Among the three compounds, (-)-sesamin (1) and IOA (5) exerted stronger effects and maintained longer duration of repellency. Furthermore, IOA (5) and isomahubanolide (6) showed good feeding deterrent activity against T. castaneum. IOA (5) was still potently active at low concentrations with the feeding deterrence index (FDI) ranging from 42.85% to 50.66% at 15–1500 ppm. This work provides some evidence for explaining antiinsect properties of the nonvolatile fraction of the C. camphora stem bark and helps promote the development and comprehensive utilization of this tree species.

Terpenoid-Induced Feeding Deterrence and Antennal Response of Honey Bees

Multiple interacting stressors negatively affect the survival and productivity of managed honey bee colonies. Pesticides remain a primary concern for beekeepers, as even sublethal exposures can reduce bee immunocompetence, impair navigation, and reduce social communication. Pollinator protection focuses on pesticide application guidelines; however, a more active protection strategy is needed. One possible approach is the use of feeding deterrents that can be delivered as an additive during pesticide application. The goal of this study was to validate a laboratory assay designed to rapidly screen compounds for behavioral changes related to feeding or feeding deterrence. The results of this investigation demonstrated that the synthetic Nasonov pheromone and its terpenoid constituents citral, nerol, and geraniol could alter feeding behavior in a laboratory assay. Additionally, electroantennogram assays revealed that these terpenoids elicited some response in the antennae; however, only a synthetic Nasonov pheromone, citral, and geraniol elicited responses that differed significantly from control and vehicle detections.

Analysis of phototoxin taste closely correlates nucleophilicity to type 1 phototoxicity

Pigments often inflict tissue-damaging and proaging toxicity on light illumination by generating free radicals and reactive oxygen species (ROS). However, the molecular mechanism by which organisms sense phototoxic pigments is unknown. Here, we discover that Transient Receptor Potential Ankyrin 1-A isoform [TRPA1(A)], previously shown to serve as a receptor for free radicals and ROS induced by photochemical reactions, enables Drosophila melanogaster to aphotically sense phototoxic pigments for feeding deterrence. Thus, TRPA1(A) detects both cause (phototoxins) and effect (free radicals and ROS) of photochemical reactions. A group of pigment molecules not only activates TRPA1(A) in darkness but also generates free radicals on light illumination. Such aphotic detection of phototoxins harboring the type 1 (radical-generating) photochemical potential requires the nucleophile-sensing ability of TRPA1. In addition, agTRPA1(A) from malaria-transmitting mosquitoes Anopheles gambiae heterologously produces larger current responses to phototoxins than Drosophila TRPA1(A), similar to their disparate nucleophile responsiveness. Along with TRPA1(A)-stimulating capabilities, type 1 phototoxins exhibit relatively strong photo-absorbance and low energy gaps between the highest occupied molecular orbital and the lowest unoccupied molecular orbital. However, TRPA1(A) activation is more highly concordant to type 1 phototoxicity than are those photochemical parameters. Collectively, nucleophile sensitivity of TRPA1(A) allows flies to taste potential phototoxins for feeding deterrence, preventing postingestive photo-injury. Conversely, pigments need to bear high nucleophilicity (electron-donating propensity) to act as type 1 phototoxins, which is consistent with the fact that transferring photoexcited electrons from phototoxins to other molecules causes free radicals. Thus, identification of a sensory mechanism in Drosophila reveals a property fundamental to type 1 phototoxins.

Bacteria: A novel source for potent mosquito feeding-deterrents

Antibiotic and insecticidal bioactivities of the extracellular secondary metabolites produced by entomopathogenic bacteria belonging to genusXenorhabdushave been identified; however, their novel applications such as mosquito feeding-deterrence have not been reported. Here, we show that a mixture of compounds isolated fromXenorhabdus budapestensisin vitro cultures exhibits potent feeding-deterrent activity against three deadly mosquito vectors:Aedes aegypti,Anopheles gambiae, andCulex pipiens. We demonstrate that the deterrent active fraction isolated from replicate bacterial cultures is highly enriched in two compounds consistent with the previously described fabclavines, strongly suggesting that these are the molecular species responsible for feeding-deterrence. The mosquito feeding-deterrent activity in the putative fabclavine-rich fraction is comparable to or better than that ofN,N-diethyl-3-methylbenzamide (also known as DEET) or picaridin in side-by-side assays. These findings lay the groundwork for research into biologically derived, peptide-based, low–molecular weight compounds isolated from bacteria for exploitation as mosquito repellents and feeding-deterrents.

Bacteria: A novel source for potent mosquito feeding – deterrents

Antibiotic and insecticidal bioactivities of the extracellular secondary metabolites produced by entomopathogenic bacteria belonging to genus Xenorhabdus have been identified; however, their novel applications such as mosquito feeding-deterrence have not been reported. Here, we show that a mixture of compounds isolated from Xenorhabdus budapestensis in vitro cultures exhibits potent feeding-deterrent activity against three deadly mosquito vectors: Aedes aegypti, Anopheles gambiae and Culex pipiens. We further demonstrate that the deterrent-active fraction isolated from replicate bacterial cultures is consistently highly enriched in two modified peptides identical to the previously described fabclavines, strongly suggesting that these are molecular species responsible for feeding-deterrence. The mosquito feeding-deterrent activity in the fabclavines-rich fraction is comparable to or better than that of N, N-diethyl-3-methylbenzamide (also known as Deet) or picaridin in side-by-side assays. Our unique discovery lays the groundwork for research into biologically derived, peptide-based low molecular weight compounds isolated from bacteria for exploitation as mosquito repellents and feeding-deterrents.

DEET as a feeding deterrent

The insect repellent N,N-diethyl-3-methylbenzamide (DEET), is a multimodal compound that acts as a spatial repellent as well as an irritant (contact repellent), thus being perceived by the insect’s olfactory and gustatory systems as an odorant and a tastant, respectively. Soon after DEET was developed, almost 6 decades ago, it was reported that it reduced mosquito feeding on blood mixed with this repellent. It is now known that the mosquito proboscis senses contact repellents with the tips (labella) of the labium, which remain in direct contact with the outer layers of the skin, while the stylets, including the feeding deterrent sensor (labrum), penetrate the skin. We designed a behavioral assay that allowed us to tease apart contact repellency from feeding deterrence. First, we demonstrate here that when DEET was mixed with blood and covered by Parafilm® layers, it did not leak to the outer surface. In our assays, the mean number of landings and duration of contacts with surfaces covering blood mixed with DEET or blood plus solvent (dimethyl sulfoxide) did not differ significantly. The feeding times, however, were significantly different. When blood was mixed either with 0.1 or 1% DEET, female southern house mosquitoes spent significantly less time feeding than the time spent feeding on blood mixed only with the solvent. By contrast, there were no significant differences in the mean times of feeding on blood containing 1% picaridin and blood plus solvent. Like DEET, the contact repellent and insecticide, permethrin, caused a significant reduction in feeding time. We, therefore, concluded, that in this context, DEET and permethrin act as feeding deterrents.