Allelochemicals-mediated interaction between algae and bacteria: Direct and indirect contact.

Secondary metabolites with bioactivity are allelochemicals. This study adopted direct contact (R0) and indirect contact (separated by 0.45 µm membrane, R1-A for algae, R1-S for sludge) to reveal the role of metabolites especially allelochemicals on interaction of bacteria and algae. Direct contact exhibited better nutrients removal than indirect contact, due to less antibacterial allelochemicals and oxidative stress. Bacterial signaling molecules were not detected. The major algae-derived allelochemicals were 13-Docosenamide, 9-Octadecenamide, n-Hexadecanoic acid, erucic acid, octadecanoic acid, ß-sitosterol, and E,E,Z-1,3,12-Nonadecatriene-5,14-diol. Furthermore, presence of 13-Docosenamide and 9-Octadecenamide was associated with succession of Flavobacterium and suppression of nitrifying bacteria (Nitrosomonas, Ellin6067, and Nitrospira). Direct contact stimulated denitrifying bacteria Saccharimonadales and algae Scenedesmus, whereas indirect contact is friendly to Dechloromonas, Competibacter, nitrifying bacteria, algae Desmodesmus and Dictyosphaerium. This study highlights the essentiality of cell contact of bacteria-algae in establishing synergy, as cell contact mitigates antagonistic effect induced by metabolites.

Differential interference effects of thermal pollution on the induced defense of different body-sized cladocerans.

Climate warming influences the biological activities of aquatic organisms, including feeding, growth, and reproduction, thereby affecting predator-prey interactions. This study explored the variation in thermal sensitivity of anti-predator responses in two cladoceran species with varying body sizes, Daphnia pulex and Ceriodaphnia cornuta. These species were cultured with or without the fish (Rhodeus ocellatus) kairomone at temperatures of 15, 20, 25, and 30 °C for 15 days. Results revealed that cladocerans of different body sizes exhibited varying responses to fish kairomones in aspects such as individual size, first-brood neonate size, total offspring number, average brood size, growth rate, and reproductive effort. Notably, low temperature differently affected defense responses in cladocerans of different body sizes. Both high and low temperatures moderated the intensity of the kairomone-induced response on body size at maturity. Additionally, low temperature reversed the reducing effect of fish kairomone on the total offspring number, average brood size, and reproductive effort in D. pulex. Conversely, it enhanced the increasing effect of fish kairomone on these parameters in C. cornuta. These results suggest that inducible anti-predator responses in cladocerans are modifiable by temperature. The differential effects of fish kairomones on various cladocerans under temperature influence offer crucial insights for predicting changes in predator-prey interactions within freshwater ecosystems under future climate conditions.

Enhancing entomopathogenic nematode efficacy with Pheromones: A field study targeting the pecan weevil.

Consistent efficacy is required for entomopathogenic nematodes to gain wider adoption as biocontrol agents. Recently, we demonstrated that when exposed to nematode pheromone blends, entomopathogenic nematodes showed increased dispersal, infectivity, and efficacy under laboratory and greenhouse conditions. Prior to this study, the impact of entomopathogenic nematode-pheromone combinations on field efficacy had yet to be studied. Steinernema feltiae is a commercially available entomopathogenic nematode that has been shown to increase mortality in insect pests such as the pecan weevil Curculio caryae. In this study, the pecan weevil was used as a model system to evaluate changes in S. feltiae efficacy when treated with a partially purified ascaroside pheromone blend. Following exposure to the pheromone blend, the efficacy of S. feltiae significantly increased as measured with decreased C. caryae survival despite unfavorable environmental conditions. The results of this study highlight a potential new avenue for using entomopathogenic nematodes in field conditions. With increased efficacy, using entomopathogenic nematodes will reduce reliance on conventional management methods in pecan production, translating into more environmentally acceptable practices.

Protocol for monitoring and analyzing pheromone-mediated behavioral response of sea lamprey in a natural system.

Olfactory-mediated behaviors in fish are often examined in artificial microcosms that enable well-controlled treatments but fail to replicate environmental and social contexts. However, observing these behaviors in nature poses challenges. Here, we describe a protocol for recording sea lamprey (Petromyzon marinus) behaviors in a natural system. We describe steps for administering and verifying accurate odorant concentrations, surveying sea lamprey abundance, and tracking sea lamprey movements. We also detail procedures to analyze treatment effects on pheromone-mediated spawning in a high-density population. For complete details on the use and execution of this protocol, please refer to Scott et al.1.

Spectral sensitivity of click beetles (Coleoptera: Elateridae) and their responses to light stimuli in laboratory and field experiments.

With increasingly fewer insecticides registered to control the larvae of pest click beetles (Coleoptera: Elateridae), integrative beetle management, including pheromone- and light-based trapping of adult beetles, must be explored as an alternative strategy. Here, we analyzed the spectral sensitivity and color preference of 9 elaterids across 6 genera in electrophysiological recordings and in behavioral bioassays. In electroretinogram recordings (ERGs), dark-adapted beetles were exposed to narrow wavebands of light in 10-nm increments from 330 to 650 nm. All beetles proved most sensitive to green (515-538 nm) and ultraviolet (UV) light (~360 nm). In 4-choice bioassay arenas with 3 light emitting diodes (LEDs; green [525 nm], blue [470 nm], red [655 nm]) and a dark control as test stimuli, beetles discriminated between test stimuli, being preferentially attracted to green and blue LEDs. In field experiments, Vernon pitfall traps fitted with a green, blue or white LED captured significantly more male and female Agriotes lineatus and A. obscurus than dark control traps. When traps were baited with green or blue LEDs at light intensities that differed by 10-fold, the traps baited with higher light intensity lures captured numerically more beetles but trap catch data in accordance with light intensity did not differ statistically. Light-based trapping may be a viable tool for monitoring elaterid species known not to have pheromones.

Courtship and mating behavior of Endoclita signifer (Hepialidae: Lepidoptera) and the male sex pheromones in hairbrushes.

Endoclita signifer Walker is the most destructive wood-boring pest of Eucalyptus in China, causing significant economic and ecological damage. As an insect of the primitive Lepidoptera family Hepialidae, E. signifer fly and mat for only 10-20 min at dusk. The courtship and mating behavior of E. signifer adults and whether male moths release sex pheromones are still unknown, especially since transitory flight survival strategies in primitive moths differ from advanced moths like noctuids. In this study, we first observed the courtship and mating behavior of E. signifer by considering the effects of space and then analyzed extracts of male hairbrushes using gas chromatography-electroantennogram detection. Our results indicated that during the courtship period, flying males form courtship fields, lekking, and chase flying females before mating with them; E. signifer were more successful in mating in larger spaces (Length × Width × Height = 9.6 × 7 × 4 m); 5 compounds in the hairbrushes of the male moths which elicited antennal responses of 2 sexes, despite at high concentrations. Combined with it, indicating that communication between male and female may rely on male sex pheromones. These findings can serve as a basis for studying the mechanisms of sex communication in E. signifer and developing sex pheromone-based trapping techniques.

Allelochemicals: A source for developing economically and environmentally friendly plant growth regulators.

Allelochemicals are specific secondary metabolites that can exhibit autotoxicity by inhibiting the growth of the same plant species that produced them. These metabolites have been found to affect various physical processes during plant growth and development, including inhibition of seed germination, photosynthesis, respiration, root growth, and nutrient uptake, with diverse mechanisms involving cell destruction, oxidative homeostasis and photoinhibition. In some cases, allelochemicals can also have positive effects on plant growth and development. In addition to their ecological significance, allelochemicals also possess potential as plant growth regulators (PGRs) due to their extensive physiological effects. However, a comprehensive summary of the development and applications of allelochemicals as PGRs is currently lacking. In this review, we present an overview of the sources and categories of allelochemicals, discuss their effects and the underlying mechanisms on plant growth and development. We showcase numerous instances of key phytohormonal allelochemicals and non-phytohormonal allelochemicals, highlighting their potential as candidates for the development of PGRs. This review aims to provide a theoretical basis for the development of economical, safe and effective PGRs utilizing allelochemicals, and emphasizes the need for further research in this area.

Passive permeability controls synthesis for the allelochemical sorgoleone in sorghum root exudate.

Competition for soil nutrients and water with other plants foster competition within the biosphere for access to these limited resources. The roots for the common grain sorghum produce multiple small molecules that are released via root exudates into the soil to compete with other plants. Sorgoleone is one such compound, which suppresses weed growth near sorghum by acting as a quinone analog and interferes with photosynthesis. Since sorghum also grows photosynthetically, and may be susceptible to sorgoleone action if present in tissues above ground, it is essential to exude sorgoleone efficiently. However, since the P450 enzymes that synthesize sorgoleone are intracellular, the release mechanism for sorgoleone remain unclear. In this study, we conducted an in silico assessment for sorgoleone and its precursors to passively permeate biological membranes. To facilitate accurate simulation, CHARMM parameters were newly optimized for sorgoleone and its precursors. These parameters were used to conduct 1 µs of unbiased molecular dynamics simulations to compare the permeability of sorgoleone with its precursors molecules. We find that interleaflet transfer is maximized for sorgoleone, suggesting that the precursor molecules may remain in the same leaflet for access by biosynthetic P450 enzymes. Since no sorgoleone was extracted during unbiased simulations, we compute a permeability coefficient using the inhomogeneous solubility diffusion model. The requisite free energy and diffusivity profiles for sorgoleone through a sorghum membrane model were determined through Replica Exchange Umbrella Sampling (REUS) simulations. The REUS calculations highlight that any soluble sorgoleone would quickly insert into a lipid bilayer, and would readily transit. When sorgoleone forms aggregates in root exudate as indicated by our equilibrium simulations, aggregate formation would lower the effective concentration in aqueous solution, creating a concentration gradient that would facilitate passive transport. This suggests that sorgoleone synthesis occurs within sorghum root cells and that sorgoleone is exuded by permeating through the cell membrane without the need for a transport protein once the extracellular sorgoleone aggregate is formed.

Allelochemical root-growth inhibitors in low-molecular-weight cress-seed exudate.

BACKGROUND AND AIMS: Cress seeds release allelochemicals that over-stimulate the elongation of hypocotyls of neighbouring (potentially competing) seedlings and inhibit their root growth. The hypocotyl promoter is potassium, but the root inhibitor was unidentified; its nature is investigated here. METHODS: Low-molecular-weight cress-seed exudate (LCSE) from imbibed Lepidium sativum seeds was fractionated by phase partitioning, paper chromatography, high-voltage electrophoresis and gel-permeation chromatography (on Bio-Gel P-2). Fractions, compared with pure potassium salts, were bioassayed for effects on Amaranthus caudatus seedling growth in the dark for 4 days. KEY RESULTS: The LCSE robustly promoted amaranth hypocotyl elongation and inhibited root growth. The hypocotyl inhibitor was non-volatile, hot acid stable, hydrophilic and resistant to incineration, as expected for K+. The root inhibitor(s) had similar properties but were organic (activity lost on incineration). The root inhibitor(s) remained in the aqueous phase (at pH 2.0, 6.5 and 9.0) when partitioned against butan-1-ol or toluene, and were thus hydrophilic. Activity was diminished after electrophoresis, but the remaining root inhibitors were neutral. They became undetectable after paper chromatography; therefore, they probably comprised multiple compounds, which separated from each other, in part, during fractionation. On gel-permeation chromatography, the root inhibitor co-eluted with hexoses. CONCLUSIONS: Cress-seed allelochemicals inhibiting root growth are different from the agent (K+) that over-stimulates hypocotyl elongation and the former probably comprise a mixture of small, non-volatile, hydrophilic, organic substances. Abundant components identified chromatographically and by electrophoresis in cress-seed exudate fitting this description include glucose, fructose, sucrose and galacturonic acid. However, none of these sugars co-chromatographed and co-electrophoresed with the root-inhibitory principle of LCSE, and none of them (in pure form at naturally occurring concentrations) inhibited root growth. We conclude that the root-inhibiting allelochemicals of cress-seed exudate remain unidentified.

Design of Polymer Carriers for Optimized Pheromone Release in Sustainable Insect Control Strategies.

Semiochemicals such as pheromones play a major role in communication between insects, influencing their spatial orientation, aggregation, defense, and mating. The rational chemical design of precision pheromone-releasing materials are increased the efficiency of pheromone-based plant protection agents. Decades of research is begun to unravel the complex communication structures regulated by semiochemicals, from the neuronal perception of specific chemical substances to the behavioral responses in hundreds of species, including many devastating pest insects. This article summarizes the most effective uses of semiochemicals in agriculture, the behavioral responses of selected target species, and controlled-release strategies based on formulations such as novel fibrous polymer carriers. This study helps scientists, decision-makers, farmers, and the public understand the importance of appropriate mating disruption techniques that reduce the need for broad-spectrum insecticides and limit their impact on non-target and beneficial insects.

Methionol as an aggregation-sex pheromone of the desert-dwelling beetle Eustromula valida (Coleoptera: Cerambycidae).

As part of a long-term project on unraveling the use of pheromones in the large beetle family Cerambycidae, field trials were conducted with generic blends of known cerambycid pheromones at a desert site in southern California. In the first year of testing (2022), the species Eustromula valida (LeConte) (subfamily Cerambycinae, tribe Elaphidiini) and Aethecerinus latecinctus (Horn) (Cerambycinae, Trachyderini) were weakly attracted to one of the lure blends. In follow-up trials in 2023, only E. valida were caught, and collection of volatiles from both sexes of E. valida determined that males sex-specifically produced 3-methylthiopropan-1-ol (methionol), a compound that was not in the tested lure blends. Beetles of both sexes were strongly and specifically attracted to this compound in field bioassays, verifying that it is an aggregation-sex pheromone. No sympatric species were attracted to methionol while it was deployed in the field. Several recent studies have identified methionol as a pheromone component for other cerambycid species in both North and South America, suggesting it may represent another common pheromone component within the Cerambycidae.

Review of surveillance systems for tephritid fruit fly threats in Australia, New Zealand, and the United States.

Many countries conduct fruit fly surveillance but, while there are guidelines, practices vary widely. This review of some countries in the Pacific region demonstrates the diversity of fruit fly surveillance practices. All utilize 3 parapheromones-trimedlure, cuelure, and methyl eugenol-to trap adult male fruit flies. Some target species are not attracted to these compounds so other attractants such as food-based lures are used in certain areas or circumstances. Lure loading and replacement cycles depend on the target species and the local climate. Malathion and dichlorvos (DDVP) are commonly used toxicants, but not in all countries, and other toxicants are being developed to replace these older-generation pesticides. Jackson and Lynfield are commonly used trap designs but newer designs such as cone and Biotrap are being adopted. Local factors such as chemical registrations and climate affect the choice of trap, lure, dispenser, toxicant, and bait concentration. These choices affect the efficacy of traps, in turn influencing optimal trap deployment in space and time. Most states now follow similar practices around trap inspection, servicing, and data handling, but these processes will be disrupted by emerging automated trap technologies. Ultimately, different practices can be attributed to the unique fruit fly risk profiles faced by each state, particularly the suite of fruit flies already present and those that threaten from nearby. Despite the diversity of approaches, international trade in fruit continues with the assurance that fruit fly surveillance practices evolve and improve according to each country's risk profile and incursion experience.

Egeria densa organic extracts: an eco-friendly approach to suppress Microcystis aeruginosa growth through allelopathy.

Macrophytes play a significant role in shaping plankton communities by shading, controlling water turbulence, and nutrient availability, while also producing allelochemicals causing varying effects on different organisms. Many researchers have shown that when live macrophytes are present, they inhibit cyanobacteria. However, their widespread use is often limited due to numerous concerns, including invasive characteristics. This study focused on the applicability of Egeria densa extracts to suppress Microcystis aeruginosa. We employed pure water and dimethyl sulfoxide, to obtain compounds from E. densa. The results revealed that E. densa aqueous extracts stimulated M. aeruginosa growth, whereas organic extracts exhibited suppression. Specifically, at low concentrations of organics extracts (0.5 and 1 g/L), after day 4, the growth inhibition was confirmed by significantly higher (p < 0.05) stress levels shown in cells treated with low concentrations. The highest inhibition rate of 32% was observed at 0.5 g/L. However, high concentrations of organic extracts (3 and 6 g/L), showed increased growth compared with control. These results suggest that high concentrations of organic extracts from E. densa potentially suppress allelochemical-induced M. aeruginosa inhibition due to high nutrient availability. In comparison with an aqueous solvent, the use of organic solvent seems to be more effective in efficiently extracting allelochemicals from E. densa.

Diet-derived male sex pheromone compounds affect female choice in a noctuid moth.

Sexual signals often function in species recognition and may also guide mate choice within a species. In noctuid moths, both males and females may exercise mate choice. Females of the tobacco budworm Chloridea virescens prefer to mate with larger males, but the signal(s) underlying female choice remain unknown. Male hairpencil volatiles are emitted during close range courtship displays. However, previously identified male hairpencil volatiles, namely acetate esters, aldehydes, alcohols, and fatty acids, are not associated with female choice. Recently, two new hairpencil compounds were identified that elicit strong electrophysiological responses in female antennae: methyl salicylate (MeSA) and δ-decalactone. In this study, we investigated the effect of larval diet and adult feeding on MeSA and δ-decalactone content in hairpencils and determined whether these compounds are involved in female choice. We found that larval diet affected MeSA content in hairpencils, but not δ-decalactone. Conversely, adult feeding affected the level of δ-decalactone, but not MeSA: sugar-water feeding increased δ-decalactone content compared to plain water. In two-choice assays, females mated more with males that had higher amounts of δ-decalactone, and less with males with higher amounts of MeSA.

Dosage impact of submerged plants extracts on Microcystis aeruginosa growth: From hormesis to inhibition.

Allelopathy has been demonstrated to be an environmentally friendly way to control harmful algal blooms. Allelochemicals of submerged plants have attracted extensive research due to their bioavailability. The dose-response of submerged plant extracts on algae growth is worth further study to improve the efficiency of bioremediation. In this study, the ultrasonic-enzymatic assistance method was utilized to extract allelochemicals from Ceratophyllum, Myriophyllum spicatum, and Vallisneria. The effects of low-dosage and high-dosage extracts on the growth of Microcystis aeruginosa were compared based on cell biomass and morphology, photosynthetic parameters, reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) levels. The results showed that the three submerged plant extracts exhibited hormetic effects at low dosages and inhibitory effects at high dosages on algal growth. Within 48 h of cultivation, the enzymatic activities of Microcystis aeruginosa fluctuated, suggesting that the extracts of the three submerged plants induced different oxidative reactions. After 120 h of cultivation with high-dosage extracts, the physiological and biochemical reactions of Microcystis aeruginosa significantly decreased, indicating the effectiveness of the allelopathy of Ceratophyllum, Myriophyllum spicatum, and Vallisneria extracts in controlling algal blooms. The phenomenon of hormesis and inhibition effect confirmed a significant dose-response relationship between the allelochemicals of submerged plant extracts and Microcystis aeruginosa, which could be attributed to the composition and content of allelochemicals. These findings highlight the importance of the relative concentration of the biological algaecide and will benefit other researchers in determining the safe dosage of plant allelochemicals when used in water.

Efficacy of the Feliway® Classic Diffuser in reducing undesirable scratching in cats: A randomised, triple-blind, placebo-controlled study.

Scratching the environment is a natural behaviour that cats use for communication and physical maintenance purposes, however when it is carried out on household furniture it is considered unacceptable for some owners and even grounds for relinquishment of cats. The objective of this study was to investigate the efficacy of FELIWAY® Classic Diffuser in reducing undesirable scratching (scratching vertical surfaces indoors other than the scratching post) in cats. A 28 day, randomised, triple-blind, placebo-controlled study with a total of 1060 caregiver-cat dyads was conducted. The study contained two groups: the Pheromone Group consisted of caregivers who were given a pheromone diffuser (n = 546) and the Placebo Group consisted of caregivers who were given a placebo diffuser (n = 514). A questionnaire with three subsections was distributed online. The first section, completed by the respondents at day 0, inquired about the cats' daily routines, social and physical environments, behaviour, temperament, and emotional states. The second section filled on day 0, 7, 14, and 28, assessed the Frequency and the Intensity of the undesirable scratching problem and the effectiveness of the product. The last section, filled on the 28th day of the product application, related to the caregivers' opinions about the product and overall outcome. After 28 days the scratching Frequency reduced for 83.5% of the cats in the Pheromone Group and 68.5% for the Placebo Group (p<0.0001). The Intensity was significantly different between treatment groups at D7 (p = 0.0170), at D14 (p = 0.0189) and at D28 (p<0.001). The reduction of the Global Index Score, which was calculated by multiplying the Intensity with the Frequency, was significantly higher for the Pheromone Group (p<0.001). This study provides direct evidence that the use of FELIWAY® Classic diffuser significantly reduces the Frequency, Intensity and the Global Index Score of undesirable scratching.

Dose-responses of seven species of cerambycid beetles to synthesized pheromones in field experiments.

Adult male cerambycid beetles of the subfamilies Cerambycinae and Lamiinae emit aggregation-sex pheromones that attract both sexes, and these chemicals can be utilized for quarantine surveillance for related exotic species which produce the same or similar pheromones. Here, we assess how attraction of 7 cerambycid species to pheromone-baited traps was influenced by the release rates of synthesized pheromones from polyethylene sachet emitters. Compounds tested included racemic 3-hydroxyhexan-2-one, the (R)-enantiomer of which is the sole or major pheromone component of numerous cerambycine species, and 2 compounds that are pheromone components of many lamiine species: (E)-6,10-dimethyl-5,9-undecadien-2-yl acetate (fuscumol acetate) and 6-methylhept-5-en-2-ol (sulcatol). We confirmed release rates of these compounds could be manipulated by varying the doses loaded into emitters. Various doses and concentrations, ranging from very dilute to the neat compounds, were then tested in field trials. Several species were most strongly attracted to the highest dose of a tested compound, one species to an intermediate dose, while another species was equally attracted regardless of dose. Our results demonstrate the importance of characterizing dose-response relationships for cerambycid species targeted by pheromone-based trapping.

Enantiospecific response of Ips avulsus (Coleoptera: Curculionidae: Scolytinae) to ipsdienol depends on semiochemical context.

Colonization of hosts by bark beetles is generally mediated by aggregation pheromones. Species competing for the same resource can limit interspecific interactions and maintain reproductive isolation by using different pheromones. In the southern United States, 3 sympatric species of Ips breed in pine hosts, each with a different pheromone blend. Ips avulsus (Eichhoff) uses ipsdienol and lanierone; Ips calligraphus (Germar) uses ipsdienol, cis-verbenol, and trans-verbenol; and Ips grandicollis (Eichhoff) uses ipsenol. Different species can also minimize cross-attraction by using different enantiomeric ratios of the same pheromones. Studies on the enantiomeric ratio of ipsdienol used by I. avulsus have come to contradictory conclusions in part because of geographic and seasonal variation. There is growing evidence that semiochemical context, in the form of different co-baits used in trapping experiments, may also play a role in the responses of I. avulsus to enantiomeric ratios of ipsdienol. We conducted a trapping study at 2 locations with traps baited with (+)-ipsdienol or racemic ipsdienol and co-baited with ipsenol, lanierone, or both ipsenol and lanierone. We found context-dependent effects of both lanierone and ipsenol on the response of I. avulsus to ipsdienol. We suggest that responses to different bait and co-bait combinations may have been shaped by different types of interactions such as the absence of conspecifics or a related species, or the presence of beneficial or antagonistic interspecific interactions.

Semiochemicals Produced by Microbes in Mealybug Honeydew Attract Fire Ants.

Semiochemicals produced by microbes in hemipteran honeydew play an important role in mediating the ant-hemipteran interaction. However, the specific volatile compounds and their origins remain unclear. Here, we found that honeydew produced by Phenacoccus solenopsis exhibited strong attractiveness to fire ant workers, but sterilized honeydew was much less attractive. Four compounds were identified from the honeydew volatiles, including 1-octen-3-ol, limonene, phenylethyl alcohol, and 2,4-ditert-butylphenol. All the identified compounds triggered strong electroantennography response of fire ants and showed repellence at relatively high concentrations while attractiveness at low concentrations to fire ant workers. Furthermore, six bacterial isolates were identified based on 16S rRNA sequencing, namely, Bacillus, Brachybacterium, Kocuria, Microbacterium, Pseudomonas, and Staphylococcus. Fire ants exhibited a strong preference for Brachybacterium, Kocuria, and Microbacterium, suggesting that these bacterial isolates are involved in the attracting effect of honeydew. Both limonene and phenylethyl alcohol were detected from Brachybacterium, while limonene was only detected from Kocuria, and phenylethyl alcohol was exclusively detected from Microbacterium. Reinoculation of these bacteria restored the attractiveness of honeydew to fire ants, and the active compounds, limonene and phenylethyl alcohol, were detectable in bacteria-reinoculated honeydew. Collectively, our results reveal the active compounds in hemipteran honeydew and their association with honeydew bacteria. The findings will contribute to the development of novel attractants for efficient monitoring of fire ants.

The behavioral response to the putative necromones from dead Tribolium castaneum (Coleoptera: Tenebrionidae) in traps by conspecifics as a function of density and time since capture.

Long-term trapping programs of stored product pests provide information for timely and accurate pest management. Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) is a highly successful external-infesting grain pest and is frequently monitored using a commercial pitfall trap that combines pheromonal and kairomonal stimuli. However, an often overlooked component of lure-based traps is the potential for the volatile plume to change over time as individuals are captured. These now-dead insects may then release necromones altering the captures of conspecifics. In this study, we evaluated changes in (i) the behavior of T. castaneum and (ii) the relative change in volatiles over time since dead insects were added and among different densities of dead conspecifics in a commercially available kairomone oil. We used multiple behavior assays, including wind tunnel, release-recapture, and 2-way olfactometer, and performed chemical analyses via headspace collection and gas chromatography coupled with mass spectrometry. Tribolium castaneum response to the kairomone lure was not consistent among assays of density of conspecifics between 4 and 40 adults after 24 or 96 h, or time of seeding over 1-96 h or 8-11 days prior. Tested strains collected in 2012 and 2019 ruled out strain-specific differences. Oil batch effects were also ruled out as a factor contributing to the response of T. castaneum. The relative volatile composition was generally stable among the treatments despite using different seeding densities and seeding times. Given that attraction and relative volatile composition were generally unaffected by prior captures, long-term monitoring programs may be robust in their interpretability over time.