Use of the Proboscis Extension Response Assay to Evaluate the Mechanism of House Fly Behavioral Resistance to Imidacloprid.

The house fly, Musca domestica L., is a significant human and livestock pest. Experiments used female adult house flies glued onto toothpicks for controlled exposure of their tarsi alone (tarsal assay) or their tarsi and proboscis (proboscis assay) with a sucrose solution containing imidacloprid at either a low (10 µg/mL) or high (4000 µg/mL) concentration. Proboscis extension response (PER) assays were used to characterize the response of imidacloprid-susceptible and behaviorally resistant house fly strains to contact with sucrose solutions containing either a low or high concentration of imidacloprid. In each assay, 150 female flies from each fly strain were individually exposed to sucrose solutions containing either a low or high concentration of imidacloprid by deliberate contact of the fly tarsi to the test solution. The PER for each fly was subsequently recorded at 0, 2, and 10 s following the initial tarsal contact. A significant and rapid reduction in PER was observed only for the behaviorally resistant fly strain and only following contact by the flies' proboscis with the sucrose solution containing the high imidacloprid concentration. The results suggest that chemoreceptors on the fly labellum or internally on the pharyngeal taste organs are involved in the detection of imidacloprid and discrimination of the concentration, resulting in an avoidance behavior (proboscis retraction) only when imidacloprid is at sufficient concentration. Further research is needed to identify the specific receptor(s) responsible for imidacloprid detection.

Highlights of Veterinary Entomology, 2022.

The field of veterinary entomology is dominated by research concerning insects and arthropods that negatively impact the health of domestic animals. A curated selection of peer-reviewed research which was highlighted at the 2022 Joint Meeting between the Entomological Society of America, the Entomological Society of Canada, and the Entomological Society of British Columbia, which prioritized exploring entomology through the lens of art, science, and culture are summarized. Articles are categorized into (i) biting and non-biting flies, (ii) ectoparasites, and (iii) review articles, with the overall goal of showcasing innovative methodologies, addressing overlooked questions in veterinary entomology, and highlighting comprehensive reviews. While not exhaustive, the selected studies represent a myriad of arthropods, methodologies, and perspectives, to inspire future scientists with diverse research avenues and emphasize the continual evolution and importance of entomological studies in today's world.

Behavioral resistance to insecticides: current understanding, challenges, and future directions.

Identifying and understanding behavioral resistance to insecticides is vital for maintaining global food security, public health, and ecological balance. Behavioral resistance has been documented to occur in a multitude of insect taxa dating back to the 1940s, but has not received significant research attention due primarily to the complexities of studying insect behavior and a lack of any clear definition of behavioral resistance. In recent years, a systematic effort to investigate the mechanism(s) of behavioral resistance in pest taxa (e.g. the German cockroach and the house fly) has been undertaken. Here, we practically define behavioral resistance, describe the efforts taken by research groups to elucidate resistance mechanisms, and provide insight on designing appropriate bioassays for investigating behavioral resistance mechanisms in the future.

Evaluation of the inheritance and dominance of behavioral resistance to imidacloprid in the house fly (Musca domestica L.) (Diptera: Muscidae).

The house fly, Musca domestica, is a cosmopolitan species known for its pestiferous nature and potential to mechanically vector numerous human and animal pathogens. Control of adult house flies often relies on insecticides formulated into food baits. However, due to the overuse of these baits, insecticide resistance has developed to all insecticide classes currently registered for use in the United States. Field populations of house flies have developed resistance to imidacloprid, the most widely used neonicotinoid insecticide for fly control, through both physiological and behavioral resistance mechanisms. In the current study, we conducted a comprehensive analysis of the inheritance and dominance of behavioral resistance to imidacloprid in a lab-selected behaviorally resistant house fly strain. Additionally, we conducted feeding preference assays to assess the feeding responses of genetic cross progeny to imidacloprid. Our results confirmed that behavioral resistance to imidacloprid is inherited as a polygenic trait, though it is inherited differently between male and female flies. We also demonstrated that feeding preference assays can be instrumental in future genetic inheritance studies as they provide direct insight into the behavior of different strains under controlled conditions that reveal, interactions between the organism and the insecticide. The findings of this study carry significant implications for pest management and underscore the need for integrated pest control approaches that consider genetic and ecological factors contributing to resistance.

Evaluation of the stability of physiological and behavioral resistance to imidacloprid in the house fly (Musca domestica L.) (Diptera: Muscidae).

BACKGROUND: The house fly (Musca domestica L.) is a synanthropic fly species commonly associated with confined animal facilities. House fly control relies heavily on insecticide use. Neonicotinoids are currently the most widely used class of insecticide and have been formulated into granular fly baits since 2002. Physiological resistance to imidacloprid in house flies has been observed to be unstable and decline over time without continual selection pressure, indicating that resistance has a fitness cost to individuals in the absence of exposure to insecticides. The stability of behavioral resistance to imidacloprid in the house fly has not been evaluated. In the current study, we assess the stability of physiological and behavioral resistance in house flies to imidacloprid over time. RESULTS: Physiological susceptibility to imidacloprid varied significantly among three house fly strains examined, with WT-15 exhibiting the greatest susceptibility to imidacloprid with an LC50 and LC95 of 109.29 (95.96-124.49) µg g-1 and 1486.95 (1097.15-2015.23) µg g-1 , respectively. No significant differences in survival were observed across 30 generations of a house fly strain (BRS-1) previously selected for behavioral resistance to imidacloprid with percentage survival ranging from 93.20% at F0 in 2020 to 96.20% survival at F30 in 2022. CONCLUSION: These results have significant implications for the management of house flies exhibiting behavioral resistance in field settings. It appears that standard resistance management tactics deployed to reduce the prevalence of physiological resistance, such as rotating or temporarily discontinuing the use of specific insecticides, may not lead to reduced behavioral resistance to imidacloprid. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Concentration Dependent Feeding on Imidacloprid by Behaviorally Resistant House Flies, Musca domestica L. (Diptera: Muscidae).

The house fly (Musca domestica L.) is a cosmopolitan and synanthropic pest fly commonly associated with confined animal facilities, known to mechanically vector numerous disease-causing pathogens. Control of adult house flies often relies on insecticides formulated into insecticidal baits, though many baits have failed due to insecticide resistance. House fly resistance to imidacloprid, the most widely used neonicotinoid insecticide available for fly control, has evolved through physiological and behavioral mechanisms in field populations. Behavioral resistance to imidacloprid was documented in field populations of flies from southern California dairies. Lab colonies of these flies were established and behavioral resistance to imidacloprid was selected over several generations. The current study examined the ability of these lab-selected flies to feed on varying concentrations of imidacloprid formulated in sucrose, and if these flies would demonstrate a feeding preference for different concentrations of imidacloprid when exposed in bioassays. Behaviorally resistant flies preferred to feed on untreated sucrose as opposed to treated sucrose at concentrations greater than 25 µg/g imidacloprid when provided sucrose treated with and without imidacloprid. When provisioned with only sucrose treated with a low and high imidacloprid concentration, flies fed on the low concentrations (≤100 µg/g) imidacloprid but reduced feeding on either treatment when concentrations were >100 µg/g imidacloprid. The current study extends the body of knowledge on house fly behavioral resistance to imidacloprid, which could provide insights into future failures of granular fly baits.

Genetic evaluation and characterization of behavioral resistance to imidacloprid in the house fly.

Insecticide resistance in pest populations is an increasing problem in both urban and rural settings due to over-application of insecticides and lack of rotation among insecticidal chemical classes. The house fly (Musca domestica L.) is a cosmopolitan pest fly species implicated in the transmission of numerous pathogens. The evolution of insecticide resistance long has been documented in house flies, with resistance reported to all major insecticide classes. House fly resistance to imidacloprid, the most widely used neonicotinoid insecticide available for fly control, has evolved in field populations through both physiological and behavioral mechanisms. Previous studies have characterized and mapped the genetic changes that confer physiological resistance to imidacloprid, but no study have examined the genetics involved in behavioral resistance to imidacloprid to date. In the current study, several approaches were utilized to characterize the genetics and inheritance of behavioral resistance to imidacloprid in the house fly. These include behavioral observation analyses, preference assays, and the use of genetic techniques for the identification of house fly chromosome(s) carrying factors. Behavioral resistance was mapped to autosomes 1 and 4. Inheritance of resistance was shown to be neither fully dominant nor recessive. Factors on autosomes 1 and 4 independently conferred contact-dependent avoidance of imidacloprid and a feeding preference for sugar alone or for sugar with dinotefuran, another neonicotinoid insecticide, over imidacloprid. This study serves as the first linkage analysis of a behavioral trait in the house fly, and provides new avenues for research regarding inherited behavior in the house fly and other animals.

Selection, Reversion, and Characterization of House Fly (Diptera: Muscidae) Behavioral Resistance to the Insecticide Imidacloprid.

Insecticide resistance in pest populations is an increasing problem in both urban and rural settings caused by over-application of insecticides and lack of rotation among chemical classes. The house fly (Musca domestica L.) is a cosmopolitan fly species implicated in the transmission of numerous pathogens, and which can be extremely pestiferous when present in high numbers. The evolution of insecticide resistance has long been documented in house flies, with resistance reported to all major insecticide classes. House fly resistance to imidacloprid, the most widely used neonicotinoid insecticide available for fly control, has been selected for in field populations through both physiological and behavioral resistance mechanisms. In the current study, house flies collected from a southern California dairy were selectively bred for behavioral resistance to imidacloprid, without increasing the physiological resistance profile of the selected flies. Flies were also successfully selected for behavioral susceptibility to imidacloprid. The rapid selection for either behavioral resistance or behavioral susceptibility suggests that inheritable alleles conferring behavioral resistance were already present in the wild-type fly population collected from the dairy site. The methods used for the specific selection of behavioral resistance (or susceptibility) in the fly population will be useful for further studies on the specific mechanisms conferring this resistance. House fly behavioral resistance was further investigated using behavioral observation and feeding preference assays, with resistance determined to be both contact-dependent and specific to the insecticide (imidacloprid) rather than to a non-insecticidal component of a bait matrix as previously documented.