Strains Of House Fly Research Articles

Isolation and characterization of the decreased cuticular penetration mechanism of fluralaner resistance in the house fly, Musca domestica

Decreased cuticular penetration has been documented as a mechanism of resistance in several insects, yet this mechanism remains poorly understood. Levels of resistance conferred, effects of the physicochemical properties on the manifestation of resistance and the effects of different routes of exposure are largely unknown. We recently selected a strain (FlurR) of house fly that was >11,000-fold resistance to fluralaner, and decreased cuticular penetration was one of the mechanisms of resistance (Norris et al., 2023). We sought to isolate the decreased penetration mechanism from FlurR into the background of the susceptible aabys strain, and to characterize the protection it conferred to fluralaner and other insecticides. We successfully isolated the decreased penetration mechanism and found that it conferred 7.1-fold resistance to fluralaner, and 1.4- to 4.9-fold cross-resistance to five other insecticides by topical application. Neither mass, metabolic lability, vapor pressure, nor logP explained the differences in the resistance ratios. The mechanism also conferred cross resistance by residual and feeding exposure, although at reduced levels compared to topical application. Remaining data gaps in our understanding of this mechanism are discussed.

The P450-Monooxygenase Activity and CYP6D1 Expression in the Chlorfenapyr-Resistant Strain of Musca domestica L.

The house fly Musca domestica L. is one of the most common insects of veterinary and medical importance worldwide; its ability to develop resistance to a large number of insecticides is well known. Many studies support the involvement of cytochrome P-450-dependent monooxygenases (P450) in the development of resistance to pyrethroids, neonicotinoids, carbamates, and organophosphates among insects. In this paper, the monooxygenase activity and expression level of CYP6D1 were studied for the first time in a chlorfenapyr-resistant strain of house fly. Our studies demonstrated that P450 activity in adults of the susceptible strain (Lab TY) and chlorfenapyr-resistant strain (ChlA) was 1.56-4.05-fold higher than that in larvae. In females of the Lab TY and ChlA strains, this activity was 1.53- and 1.57-fold higher, respectively (p < 0.05), than that in males, and in contrast, the expression level of CYP6D1 was 21- and 8-fold lower, respectively. The monooxygenase activity did not vary between larvae of the susceptible strain Lab TY and the chlorfenapyr-resistant strain ChlA. Activity in females and males of the ChlA strain exceeded that in the Lab TY strain specimens by 1.54 (p = 0.08) and 1.83 (p < 0.05) times, respectively, with the same level of CYP6D1 expression. PCR-RFLP analysis revealed a previously undescribed mutation in the promoter region of the CYP6D1 gene in adults of the Lab TY and ChlA strains, and it did not affect the gene expression level. The obtained results show that the development of resistance to chlorfenapyr in M. domestica is accompanied by an increase in P450-monooxygenase activity without changes in CYP6D1 expression.

Topical Toxicity and Repellency Profiles of 17 Essential Oil Components against Insecticide-Resistant and Susceptible Strains of Adult Musca domestica (Diptera: Muscidae).

The house fly is a significant pest in agriculture and human health that is increasingly difficult to manage due to multiple limitations including resistance development. To explore alternative pesticides, the topical toxicity and repellency profiles of 17 essential oil components (EOCs) were evaluated against a resistant and a susceptible strain of house fly, Musca domestica L., using topical application and Y-tube olfactometers, respectively. Six of the most toxic EOCs based on the LD50 were further investigated against a susceptible strain of house fly. Thymol, (+)-pulegone, eugenol, and carvacrol were always the top four most toxic chemicals tested against the resistant house fly strain. Little to no resistance was observed to the top six EOCs based on the comparison of the results between resistant and susceptible house fly strains. P-Cymene, citronellic acid, R-(+)-limonene, linalool, γ-terpinene, estragole, and eugenol were repellent to adult house flies at certain concentrations while (-)-carvone and thymol were attractive to adult house flies. This screening of a wide variety of individual EOCs provides a stronger foundation of information for further research. This should encourage further investigation into the topical toxicity and repellency in field studies, which will provide more insight into the performance of biopesticides for house fly management and potential commercialization.

Immunity-related genes can serve as early diagnostic markers for infection in housefly rearing systems

Abstract Farming of houseflies, as well as of other insects, could become a sustainable alternative for producing high-protein animal feed. Although promising, this comes with particular challenges, including monitoring insect health in large-scale farming facilities. In this study, we tested if the expression patterns of immunity-related genes could serve as early diagnostic markers for disease and help prevent infection spread with detrimental effects. We introduced a commonly used bacterial entomopathogen, Bacillus thuringiensis, in the housefly larval substrate and monitored larval biomass changes, pupation, and gene expression of three immunity-related genes: PGRP, defensin, and cecropin. We performed the infection assay for two strains of different geographic origins at two temperatures and with two concentrations of the bacterial inoculum. We found that gene expression was faster in indicating differences between larvae grown in treated and untreated substrate compared to monitoring larval growth and pupation. Most strikingly, although there were no significant effects on larval biomass for 48 hours after the substrate inoculation, the effects on gene expression in some cases manifested already two hours post substrate inoculation. As the two housefly strains showed different response speeds in gene upregulation, there is a need for more frequent routine testing to define which response levels could be considered as an alarm. We conclude that monitoring immunity-related genes and microbial surveys of insect-rearing substrates can be used to diagnose infection and prevent its spread early in mass-rearing settings.

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.

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.

High-sugar diet leads to loss of beneficial probiotics in housefly larvae guts.

The housefly (Musca domestica) is a common insect species with only a few recurrent bacterial taxa in its gut microbiota, because the numerous microbial acquisition routes in its septic habitats can favor transient microbes. Here, we investigated the role of the diet on the microbiota and the developmental success of a housefly strain reared on three substrates. We used a control wheat bran-based substrate, and added clotted cream and sucrose to make a high-fat, and a high-sugar substrate, respectively. The conducted survey revealed that, in contrast to the high-fat diet, the high-sugar diet caused lower developmental success and less diverse microbiota, in which several lactobacilli were replaced with Weissella bacterial phylotypes. Cultures with sucrose as the sole carbon source confirmed that a Weissela confusa strain, isolated from larvae, could utilize sucrose more efficiently than other tested lactic acid bacteria; a result also supported by gene function prediction analysis. Enhancing the rearing substrate with Limosilactobacillus fermentum and Lactiplantibacillus plantarum strains, which were isolated from control larvae, could not only revert the negative effect of the high-sucrose diet on development, but also increase the gut bacterial diversity. In our study, we show that the microbiota shifts in response to the high-sucrose diet did not benefit the host, that showed lower developmental success. In contrast, high-sucrose favored specific components of the microbiota, that continued to be enriched even after multiple generations, outcompeting beneficial bacteria. Also, microbiome manipulation showed the potential of probiotics to rescue host performance and restore the microbiome.

Genetics, genomics and mechanisms responsible for high levels of pyrethroid resistance in Musca domestica

Insecticide resistance is both economically important and evolutionarily interesting phenomenon. Identification of the mutations responsible for resistance allows for highly sensitive resistance monitoring and allows tools to study the forces (population genetics, fitness costs, etc.) that shape the evolution of resistance. Genes coding for insecticide targets have many well-characterized mutations, but the mutations responsible for enhanced detoxification have proven difficult to identify.We employed multiple strategies to identify the mutations responsible for the extraordinarily high permethrin resistance in the KS17-R strain of house fly (Musca domestica): insecticide synergist assays, linkage analysis, bulk segregant analyses (BSA), transcriptomics and long read DNA (Nanopore) sequencing. The >85,100-fold resistance in KS17-R was partially suppressed by the insecticide synergists piperonyl butoxide and S,S,S-tributylphosphorothionate, but not by diethyl maleate nor by injection. This suggests the involvement of target site insensitivity, CYP-mediated resistance, possibly hydrolase mediated resistance and potentially other unknown factors. Linkage analysis identified chromosomes 1, 2, 3 and 5 as having a role in resistance. BSA mapped resistance loci on chromosomes 3 and 5. The locus on chromosome 3 was centered on the voltage sensitive sodium channel. The locus on chromosome 5 was associated with a duplication of multiple detoxification genes. Transcriptomic analyses and long read DNA sequencing revealed overexpressed CYPs and esterases and identified a complex set of structural variants at the chromosome 5 locus.

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

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. 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. 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.

Multiple-P450 Gene Co-Up-Regulation in the Development of Permethrin Resistance in the House Fly, Musca domestica

This paper reports a study conducted at the whole transcriptome level to characterize the P450 genes involved in the development of pyrethroid resistance, utilizing expression profile analyses of 86 cytochrome P450 genes in house fly strains with different levels of resistance to pyrethroids/permethrin. Interactions among the up-regulated P450 genes and possible regulatory factors in different autosomes were examined in house fly lines with different combinations of autosomes from a resistant house fly strain, ALHF. Eleven P450 genes that were significantly up-regulated, with levels > 2-fold those in the resistant ALHF house flies, were in CYP families 4 and 6 and located on autosomes 1, 3 and 5. The expression of these P450 genes was regulated by trans- and/or cis-acting factors, especially on autosomes 1 and 2. An in vivo functional study indicated that the up-regulated P450 genes also conferred permethrin resistance in Drosophila melanogaster transgenic lines. An in vitro functional study confirmed that the up-regulated P450 genes are able to metabolize not only cis- and trans-permethrin, but also two metabolites of permethrin, PBalc and PBald. In silico homology modeling and the molecular docking methodology further support the metabolic capacity of these P450s for permethrin and substrates. Taken together, the findings of this study highlight the important function of multi-up-regulated P450 genes in the development of insecticide resistance in house flies.

Lipid quantity and composition vary between three European house fly strains

AbstractLipids are an important class of nutrients for insects, as they are the main energy source during metamorphosis and crucial for survival and reproduction. Lipid reserves in insects are mostly accumulated during larval development and can be influenced by environmental factors such as diet and temperature. Genetic factors can also affect larval lipid content and composition. Larvae of the house fly, Musca domestica L. (Diptera: Muscidae), are a valuable emerging resource for the insect feed sector, fueling a growing interest in the factors regulating larval lipid content. We investigated whether strains of three geographical origins vary in ontogeny, quantity, and composition of stored lipids in response to temperature. Italian, Spanish, and Dutch house fly larvae were reared on a similar diet at 25 and 35 °C. Larval dry weight and total lipid content were determined in early‐, mid‐, and late‐third instars by extraction of soluble lipids. The fatty acid composition was analyzed in late‐third instars using an improved extraction and derivatization protocol. We found that strain, larval stage, and temperature affected larval dry weight and absolute and relative lipid content, with significant interaction effects between these three factors. All flies were reared in a common environment, indicating a genetic component to lipid storage. Analysis of lipid composition identified 11 fatty acids, including some rarely reported in the literature. Five fatty acids accounted for 80 and 81% of the total fatty acid methyl esters detected at 25 and 35 °C, respectively. An effect of temperature but not strain was evident on the composition of the fatty acids. The observed differences in lipid content among strains and temperature conditions could be of interest for commercial rearing of the house fly.

Selection for, and characterization of, fluralaner resistance in the house fly, Musca domestica

House flies, Musca domestica (L), are the mechanical vector of >100 human and animal pathogens, including those that are antibiotic-resistant. Given that house flies are associated closely with human and livestock activity, they present medical and veterinary health concerns. Although there are numerous strategies for control of house fly populations, chemical control has been favored in many facilities. Products with pyrethroid active ingredients have been used predominantly for >35 years in space sprays. As a result, strong selection for pyrethroid resistance has led to reduced control of many populations. Reliance on a limited number of insecticides for decades has created fly control problems necessitating the discovery and formulation of new control chemistries.Fluralaner is a relatively new insecticide and acaricide (first reported in 2010), belonging to the isoxazoline class. These insecticides target the glutamate- and gamma-aminobutyric acid-gated (GABA) chloride channels, which is a different mode of action from other insecticides used against house flies. Although is it not currently registered for house fly control in the United States, previous work has shown that fluralaner is highly toxic to house flies and that there was limited cross-resistance found in laboratory strains having high levels of resistance to other insecticides.Herein, we characterized the time and age dependency of fluralaner toxicity, detected cross-resistance in populations from across the United States, and selected a highly resistant (>11,000-fold) house fly strain. We found that the fluralaner LD50 of 18–24 h old flies was 2-fold higher than for 5–6 d old flies. This appears to be due to more rapid penetration of fluralaner into the 5–6 d old flies. Fluralaner resistance was inherited as an intermediate to incompletely dominant trait and was mapped to chromosomes 5 and 3. Resistance could be suppressed to 7-fold with piperonyl butoxide, suggesting that cytochrome P450 (CYP)-mediated detoxification was a major mechanism of resistance. Decreased penetration was also demonstrated as a mechanism of resistance. The utility of fluralaner for house fly control is discussed.

Resistance to insecticides of housefly Musca domestica in the center of the European part of Russia

Under laboratory conditions, resistance to insecticides from various chemical classes of several strains of the housefly, Musca domestica, obtained from populations collected at facilities in the Moscow and Kaluga regions (KSK-1 in a cowshed and KSK-2 in the stable of an equestrian sports club, Krasnogorsk – at a food facility, Kaluga – in the compost of a garden non-profit partnership). It was found that when insecticides were applied topically, houseflies of all the studied strains hadthe highest resistance to cypermethrin (RF=75–900x) and to neonicotinoid compounds thiamethoxam and clothianidin (RF=95–330x). High resistance to fipronil was found in two strains of fly (RF= 46–75x). Strain KSK-1 was weakly tolerant to chlorpyrifos (RF=3.7x), while the other strains were susceptible. All the studied house fly strains were more susceptible to indoxacarb and chlorfenapyr than the laboratory strain S-NIID. The data indicate that studied housefly strains are multiresistant to almost all commonly used insecticides. Possible mechanisms of insect resistance to insecticides are discussed. Frequent use of insecticides of various chemical classes led to resistance in the target pest. Widespread use of pyrethroids in animal facilities has led to high resistance to them in the house fly because places where larvae hatch are constantly polluted with these insecticides. High resistance to neonicotinoids is also due to their frequent use as poisoned baits in livestock facilities. To the contrary, the studied housefly strains remained susceptible to the tested oxadiazines and pyrroles, chemical classes that were recently introduced to Russia.

Houseflies harbor less diverse microbiota under laboratory conditions but maintain a consistent set of host-associated bacteria

The housefly (Musca domestica) is a wide-ranging insect, often associated with decaying matter from livestock and humans. The septic environments in which houseflies live are believed to be a rich source for microbial acquisition. Although the housefly can harbor a wide range of microorganisms, it is not yet well known which microbes are always recurrent, which are dispensable and which environmentally dependent. In the present study, we aim at identifying which microbes are recurrently associated with the housefly gut throughout the species’ life cycle and whether their acquisition relies on the fly’s living environment. We surveyed three housefly strains—two of them kept under standard laboratory conditions for a long time and one wild-caught. To track any shifts happening throughout the lifecycle of the housefly and to test the consistency of the revealed microbial communities, we sampled houseflies at five developmental stages over the course of four consecutive generations. Both the bacterial and fungal microbiota of five developmental stages were studied for all samples, using amplicon sequencing for the 16S and ITS1 rRNA gene, respectively. Results revealed diverse microbial communities yet consistent for each of the two distinct sampling environments. The wild-caught population showed a more diverse and more distinct gut microbiota than the two laboratory strains, even though the strain was phylogenetically similar and shared geographic origin with one of them. Two bacterial genera, Myroides and Providencia, and two yeasts, Trichosporon and Candida tropicalis, were present in all sampled larvae and pupae, regardless of the strain. Analysis of the provided diet revealed that the flies acquired the yeasts through feeding. Our main findings show that houseflies might lose microbial diversity when reared in controlled environments, however they can maintain a consistent set of bacteria. We conclude that although the environment can facilitate certain microbial transmission routes for the housefly, and despite the fungal microbiota being largely acquired through diet, the larval bacterial gut microbiome remains relatively consistent within the same developmental stage.

Перспективы применения приманок на основе современных действующих веществ для борьбы с комнатными мухами

Topical application revealed the high resistance of field populations of houseflies to pyrethroids (500–900?), neonicotinoids (345–500?), phenylpyrazoles (46–75?), susceptibility to FOS (0.6–1.4?) and oxadiazines (0.2?). With intestinal exposure, resistance rates are somewhat lower, but resistance remains at a high level to fipronil (33–76?), thiamethoxam (80–105?) and clothianidin (80–98?). Susceptibility to indoxacarb (0.2–1.4?) was revealed. The comparative efficacy of two modern baits based on indoxacarb and dinotefuran for resistant and susceptible strains of house flies was studied. Both insecticides are highly toxic to house flies in the form of commercially available baits. Indoxacarb and dinotefuran can be recommended for introduction into rotation schemes of insecticides in the form of food baits to control adult flies. Keywords: dinotefuran, indoxacarb, house flies, insecticide resistance, baits.

Resistance status and the rate of resistance development in Musca domestica Linnaeus (Diptera: Muscidae) to permethrin and imidacloprid in Indonesia

A previous report has indicated that in many regions of Indonesia, populations of Musca domestica have shown very high resistance to permethrin and low resistance to imidacloprid. In this study, the resistance status to permethrin and imidacloprid was updated using a topical application and feeding bioassay. Six housefly strains originated from six highly populated cities in Indonesia, namely Serang (SRG), Jakarta (JKT), Bandung (BDG), Semarang (SMG), Yogyakarta (JOG), and Surabaya (SBY). A seventh strain (Danish Pest Infestation Laboratory) served as the control. Each strain was tested for resistance to the two insecticides. The rate of development of resistance to the two insecticides was measured over ten generations. The results indicated that all field strains showed very high resistance to permethrin. The highest resistance level recorded was in the SRG strain (RR50 = 2880), and the lowest was in the JKT strain (RR50 = 520). Repeated application of permethrin over ten generations increased the resistance level by about 2.7–32.73-fold as compared to the level of their respective parental populations. On the other hand, most strains showed low to moderate resistance to imidacloprid, in which the SRG strain had the highest resistance level (RR50 = 15.5) and the SBY strain had the lowest (RR50 = 2.0). Repeated application of imidacloprid over ten generations increased the resistance level by about 3.25–17.41-fold. The findings, which is the second report of housefly resistance in Indonesia since 2016, provide a crucial foundation for developing appropriate housefly integrated pest management strategies in highly populated areas in Indonesia.

Evaluation of triflumuron and pyriproxyfen as alternative candidates to control house fly, Musca domestica L. (Diptera: Muscidae), in Riyadh city, Saudi Arabia.

This study was conducted to determine the susceptibility and resistance of some house fly strains of Musca domestica L. to the insect growth regulator insecticides triflumuron and pyriproxyfen in some locations in Riyadh city. Field-collected strains of M. domestica L. from five sites in Riyadh city that represented five slaughterhouse sites where flies spread significantly were tested against triflumuron and pyriproxyfen. Triflumuron LC50 values for the five collected strains ranged from 2.6 to 5.5 ppm, and the resistance factors (RFs) ranged from 13-fold to 27-fold that of the susceptible laboratory strain. Pyriproxyfen LC50 values for the field strains ranged from 0.9 to 1.8 ppm with RFs of 3-fold to 5-fold. These results indicate that pyriproxyfen is an effective insecticide to control house flies and should be used in rotation with other insecticides in the control programs applied by Riyadh municipality.

Genetics and mechanism of resistance to chlorantraniliprole in Musca domestica L. (Diptera: Muscidae).

The house fly, Musca domestica L. is an important mechanical vector of different pathogens of medical and veterinary importance. It is an organism well-known for its ability to develop insecticide resistance. In the current study, we investigated the genetic basis and mechanism of chlorantraniliprole resistance in a field strain of house fly by selecting it artificially in the laboratory with a commercial formulation of chlorantraniliprole (CTPR-SEL). After seven generations of consecutive selection with chlorantraniliprole, CTPR-SEL strain developed a 644-fold resistance compared with the Susceptible strain and a 3-fold resistance compared with the field strain. Reciprocal crossing between the CTPR-SEL and Susceptible homozygous strains revealed an autosomal and incomplete dominant mode of resistance to chlorantraniliprole. A direct test using a monogenic inheritance model based on chi-square analysis revealed that the resistance was governed by more than one gene. Bioassays with synergists indicated that esterases might be involved in the resistance of house fly to chlorantraniliprole. These findings may be helpful to the development of an improved strategy for chlorantraniliprole resistance management in house fly.

Inheritance, stability, cross-resistance, and life history parameters of a clothianidin-selected strain of house fly, Musca domestica Linnaeus.

The house fly, Musca domestica L., is a cosmopolitan insect pest of public and animal health importance that serves as a mechanical vector of pathogens. Aimed at prospective resistance management to reduce environmental pollution, we characterized the inheritance pattern, realized heritability, fitness cost, cross resistance, stability and mechanism of clothianidin resistance in M. domestica that were collected from the poultry farm. By continuous selection with clothianidin for 11 generations, the clothianidin selected M. domestica strain (Clotha-SEL) developed a 3827-fold resistance compared to a susceptible strain. However, resistance to clothianidin was proved to be unstable when selection with clothianidin was removed for five generations (G7 to G12). Inheritance pattern analysis at G8 of Clotha-SEL (RR=897) revealed that resistance to clothianidin was polygenic, autosomal and incompletely dominant. Realized heritability (h2) for resistance value was 0.38 (at G11) in the tested strain. Synergist bioassays showed that microsomal oxidases and esterases might not contribute significantly in resistance evolution. Fitness costs of clothianidin resistance were present, for example, reduction in growth potential of the Clotha-SEL strain in comparison to the untreated counterpart strain (UNSEL) was observed. No cross resistance to bifenthrin and fipronil and a very low cross-resistance to spinosad were observed. These insecticides could be alternated with clothianidin as an insecticide resistance management tool to sustain its efficacy for a longer time period. These results shall be utilized to devise a proactive resistance management strategy for use of clothianidin against M. domestica that will be helpful to alleviate the allied threats to environmental and human health.

Toxicity of fluralaner, a companion animal insecticide, relative to industry-leading agricultural insecticides against resistant and susceptible strains of filth flies

Filth flies cause billions of dollars of losses annually to the animal production industry. Fluralaner is a relatively new pesticide currently sold for control of fleas, ticks, and mites on companion animals and poultry. We examined the efficacy of fluralaner against three species of filth flies. Insecticide-susceptible horn flies and stable flies were tested topically. Fluralaner outperformed permethrin by > 2-fold for the horn flies but underperformed permethrin by > 45-fold for stable flies at 24 h. House flies were tested topically with fluralaner in comparison to permethrin at 48 h and orally with fluralaner in comparison to imidacloprid at 24 h. Topical fluralaner was 6- to 28-fold as toxic as permethrin in four pyrethroid-resistant strains and not significantly less toxic than permethrin in a susceptible strain and a mildly pyrethroid-resistant strain. There was slight cross-resistance between topically applied fluralaner and permethrin in all five insecticide-resistant strains tested. Oral fluralaner was more toxic than imidacloprid in all four house fly strains tested, 9- to 118-fold as toxic. Oral cross-resistance between imidacloprid and fluralaner was not detected, but imidacloprid resistance was not high in any of the tested strains. Fluralaner shows promise for control of horn flies and house flies.