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Background The control of housefly, Musca domestica, heavily relies on the application of insecticides. Propoxur, a carbamate, was widely used for vector control. The housefly populations with high propoxur resistance displayed the point mutations and overexpression of the acetylcholinesterase. However, the roles of cytochrome P450 monoxygenases (P450s), as a kind of the important detoxification enzymes, remain poorly understand in the housefly resistant to propoxur. Results P450s were implied to contribute to propoxur resistance based on the synergism of PBO and the increase of P450 enzyme activity in the propoxur resistance near-isogenic line (N-PRS). Five P450 (CYP6G4, CYP6A25, CYP304A1, CYP6D3, and CYP6A1) genes by RNA-seq comparison were significantly up-regulated in the N-PRS strain with >1035-fold resistance to propoxur. A total of thirteen nonsynonymous mutations of three P450 genes (CYP6G4, CYP6D3, and CYP6D8) were found in the N-PRS strain. The amino acid substitutions of CYP6D3 and CYP6D8 were probably not resistance-associated single nucleotide polymorphisms (SNPs) because they were also found in the aabys susceptible strain. However, CYP6G4 variant in the N-PRS strain was not found in the aabys strain. The conjoint analysis of mutations and a series of genetic crosses exhibited that the housefly propoxur resistance was strongly associated with the mutations of CYP6G4 gene. Conclusion Our results suggested that a combination of upregulated transcript levels and mutations of CYP6G4 contributed to propoxur resistance in the housefly. This article is protected by copyright. All rights reserved.

The Ocimum basilicum essential oil (EO) was evaluated for its biological effect on M. domestica. Characterization of O. basilicum EO revealed the presence of methyl chavicol (70.93%), linalool (9.34%), epi-α-cadinol (3.69 %), methyl eugenol (2.48%), γ-cadinene (1.67%), 1,8-cineole (1.30%) and (E)-β-ocimene (1.11%). The basil EO and its constituents methyl chavicol and linalool caused the neuronal response in female adults of M. domestica. Adult female flies showed reduced preference to food source laced with basil EO and methyl chavicol. Substrate treated with EO and methyl chavicol at 0.25% caused an oviposition deterrence of over 80%. The ovicidal effect was high in O. basilicum EO (EC50 9.74mg/dm3) followed by methyl chavicol (EC50 10.67mg/dm3) and linalool (EC50 13.57mg/dm3. On contact toxicity, adults exposed to EO (LD50 10.01 μg/adult) were more susceptible than to methyl chavicol and linalool (LD50 13.62 μg/adult and LD50 43.12 μg/adult respectively). EO and its constituents methyl chavicol and linalool induced the detoxifying enzymes Carboxyl esterase (Car E) and Glutathione S – transferases (GST)

In this study, a mixed-level orthogonal array design was employed for the optimum conditions of breeding housefly larvae by food waste. The results showed that the effects of these factors on the weight of 50 larvae, larvae yield, and crude protein content were the culture substrate ratio > the breeding density > the feeding mode. The optimum conditions for the housefly larvae to convert food waste were as follows: culture substrates ratio 1:3, breeding density 10.0 g/kg, and all substance added on the first day. The optimum food waste mass reduction rate was 79.1-83.6%. The value of the essential amino acids (Eaa)/ the total amino acids (Taa) (45.1%) and E/the nonessential amino acid (Naa) values (0.83%) in the housefly larvae products met the Food and Agricultural Organization (FAO) requirements for feed protein. The crude fat content (30.1 ± 1.18%) was higher than of the housefly larvae after bioconversion of pig manure (22.0%) and the fish meal standard of China. The contents of total nutrients (N+P+K ≥ 5.5%) and heavy metals (Pb ≤ 0.40 mg/kg, Cr ≤ 1.50 mg/kg, Cd ≤ 0.40 mg/kg) in the residues of this study met the Chinese standard for organic fertilizer. Tilapia raised with the dried housefly larvae showed the best growth performance and nutrient concentrations in the experiment groups (p < 0.05). Moreover, the trace elements concentration in tilapia raised with the four kinds of feeds complied with the maximum levels of contaminants in foods in both China and WHO. These findings show that the housefly larvae products that converted food waste are suitable for use in the production of fish feed.

Flonicamid is a chordotonal modulator and novel systemic insecticide that has been used frequently for controlling a broad range of insect pests. The risk of flonicamid resistance was assessed through laboratory selection and determining inheritance pattern and cross-resistance potential to five insecticides in house fly, Musca domestica L. Very low to high flonicamid resistance in M. domestica populations was found compared with the susceptible strain (SS). A flonicamid-selected (Flonica-RS) M. domestica strain developed 57.73-fold resistance to flonicamid screened for 20 generations compared with the SS. Overlapping 95% fiducial limits of LC50 of the F1 and F1ǂ, and dominance values (0.87 for F1 and 0.92 for F1ǂ) revealed an autosomal and incomplete dominant flonicamid resistance. The monogenic model of resistance inheritance suggested a polygenic flonicamid resistance. The Flonica-RS strain displayed negative cross-resistance between flonicamid and sulfoxaflor (0.10-fold) or clothianidin (0.50-fold), and very low cross-resistance between flonicamid and flubendiamide (4.71-fold), spinetoram (4.68-fold), or thiamethoxam (2.02-fold) in comparison with the field population. The estimated realized heritability (h2) value of flonicamid resistance was 0.02. With selection mortality 40–90%, the generations required for a 10-fold increase in LC50 of flonicamid were 94–258 at h2 (0.02) and slope (3.29). Flonicamid resistance was inherited as autosomal, incomplete dominant, and polygenic in the Flonica-RS. Negative or very low cross-resistance between flonicamid and sulfoxaflor, clothianidin, flubendiamide, spinetoram, and thiamethoxam means that these insecticides can be used as alternatives for controlling M. domestica. These data can be useful in devising the management for M. domestica.

Musca domestica is a global insect-pest of human beings and animal agriculture. Pyriproxyfen, a juvenile hormone analog, has shown its potential for effective management of M. domestica. However, lethal and sublethal effects of pyriproxyfen on biological traits and demographic growth parameters of M. domestica are still unknown. The present study investigated the effects of lethal and sublethal concentrations on different biological traits of M. domestica for two generations i.e., exposed parents (F0) and their offspring (F1). Concentration-response bioassays revealed that concentrations of pyriproxyfen that caused 50% (LC50), 25% (LC25), 10% (LC10) and 2% (LC2) mortality of M. domestica were estimated as 0.12, 0.06, 0.03 and 0.01 μg/g, respectively. In the F0 generation, exposure of 3rd instar larvae to these concentrations resulted in a reduced pupation rate, lengthened pupal stage duration, light weight pupae and reduction in adult emergence in a concentration-dependent manner. In the case of F1 generation, similar trend was observed for pupation rate, pupal stage duration, and total developmental period (i.e., egg to adult); however, pupal weight was affected at LC10, LC25, LC50 levels, and adult emergence at only LC25 and LC50 levels. The values of demographic growth parameters, analyzed through age-stage, two-sex life table theory, were significantly decreased at all the levels of pyriproxyfen compared with control. This study highlights that pyriproxyfen has the potential to suppress the population of M. domestica through its lethal and sublethal effects and presents an empirical basis from which to consider management decisions for chemical control in the field.

The Musca domestica salivary gland hypertrophy virus (MdSGHV) substantially enlarges the house fly’s salivary glands and prevents or delays ovarian development in its adult host, but the effect that MdSGHV has on the house fly’s food consumption is currently unknown. Using house flies from a laboratory-reared colony, we evaluated the effect of MdSGHV infection on food consumption over a 7-d period. Both treatment (virus-infected) and control (saline-injected) flies were provided with a choice of 8% sucrose solution and 4% powdered milk solution to determine food preferences. Quantities of each solution consumed were measured every 24 h for each fly to measure food consumptions. Infected house flies were shown to consume less overall of both solutions than house flies injected with saline. The largest consumption discrepancy was seen between female house flies. Healthy female flies with developing ovaries continued to consume a sugar and protein diet, whereas infected female flies fed predominantly on a sugar diet. Additionally, infected male and female flies consumed significantly lower quantities of protein and sucrose than control flies. This suggests that MdSGHV has a negative consumption effect (e.g., hunger, starvation) on its host. Thus, differences in food consumption of infected and control flies probably represent differences in the nutritional requirements of flies resulting from viral infection.

Musca domestica is one of the major cosmopolitan pests in livestock facilities because it can be both a nuisance and a vector of pathogens to animals. Currently, treatment of animal manure with insect growth regulator (IGR) insecticides is among major practices to control M. domestica throughout the year over wide-ranging environmental temperatures. Fluctuation in daily or seasonal temperature is one of the most established factors impacting toxicity of insecticides against insect pests. In this study, the effect of posttreatment temperature (range, 20-36 °C) on the toxicity of eight IGRs: five chitin synthesis inhibitors (cyromazine, diflubenzuron, lufenuron, novaluron, triflumuron), two juvenile hormone analogs (methoprene, pyriproxyfen), and one ecdysone agonist (methoxyfenozide), was investigated against M. domestica. The toxicity of lufenuron and novaluron increased by 1.78 times over the range of 20-28 °C, and 2.25 and 1.83 times, respectively, over the range of 28-36 °C, with an overall increase by 4.00 and 3.26 times, respectively (i.e., positive temperature coefficient). In contrast, the toxicity of diflubenzuron, pyriproxyfen, and triflumuron decreased by 1.43, 1.89, and 2.10 times, respectively, over the range of 20-28 °C, and 1.70, 2.00, and 1.95 times, respectively, over the range of 28-36 °C, with an overall decrease by 2.43, 3.78, and 4.10 times, respectively. The toxicity of cyromazine, methoprene, and methoxyfenozide did not change significantly. Overall, these data will help stakeholders to choose appropriate insecticides for M. domestica control depending on the prevailing environmental temperature and to avoid misuse of insecticides that ultimately lead to environmental safety.

Ulcerative colitis (UC) is a modern refractory disease with steadily increasing incidence worldwide that urgently requires effective and safe therapies. Therapeutic peptides delivered using nanocarriers have shown promising developments for the treatment of UC. We developed a novel colon-accumulating oral drug delivery nanoplatform consisting of Musca domestica cecropin (MDC) and mesoporous carbon nanoparticles (MCNs) and investigated its effects and mechanism of action for the treatment of UC. Methods: An optimized one-step soft templating method was developed to synthesize MCNs, into which MDC was loaded to fabricate MDC@MCNs. MCNs and MDC@MCNs were characterized by BET, XRD, and TEM. MDC and MDC@MCNs resistance to trypsin degradation was measured through Oxford cup antibacterial experiments using Salmonella typhimurium as the indicator. Uptake of MDC and MDC@MCNs by NCM460 cells was observed by fluorescence microscopy. The biocompatibility of MDC, MCNs, and MDC@MCNs was evaluated in three cell lines (NCM460, L02, and NIH3T3) and C57BL/6 mice. Dextran sulphate sodium was used to establish models of NCM460 cell injury and UC in mice. MTT assay, flow cytometry, and mitochondrial membrane potential assay were applied to determine the effects of MDC@MCNs on NCM460 cells injury. Additionally, a variety of biological methods such as H&E staining, TEM, ELISA, qPCR, Western blotting, and 16s rDNA sequencing were performed to explore the effects and underlying mechanism of MDC@MCN on UC in vivo. Colonic adhesion of MCNs was compared in normal and UC mice. The oral biodistributions of MDC and MDC@MCNs in the gastrointestinal tract of mice were also determined. Results: MDC@MCNs were successfully developed and exhibited excellent ability to resist destruction by trypsin and were taken up by NCM460 cells more readily than MDC. In vitro studies showed that MDC@MCNs better inhibited DSS-induced NCM460 cells damage with lower toxicity to L02 and NIH3T3 cells compared with MDC. In vivo results indicated that MDC@MCNs have good biocompatibility and significantly improved colonic injury in UC mice by effectively inhibiting inflammation and oxidative stress, maintaining colonic tight junctions, and regulating intestinal flora. Moreover, MDC@MCNs were strongly retained in the intestines, which was attributed to intestinal adhesion and aggregation of MCNs, serving as one of the important reasons for its enhanced efficacy after oral administration compared with MDC. Conclusion: MDC@MCNs alleviated DSS-induced UC by ameliorating colonic epithelial cells damage, inhibiting inflammation and oxidative stress, enhancing colonic tight junctions, and regulating intestinal flora. This colon-accumulating oral drug delivery nanoplatform may provide a novel and precise therapeutic strategy for UC.

Background Permethrin has been used frequently in the control of Musca domestica worldwide including Pakistan, with reports of resistance development in different field strains. A near-isogenic line of M. domestica with permethrin resistance (Perm-R) was constructed to elucidate mode of inheritance and cross-resistance analyses. Results Toxicity responses of reciprocal progenies F1 and F1' showed no significant difference in median lethal dose (LD50 ) values, suggesting that permethrin resistance was inherited in an autosomal fashion with neither sex linkage nor maternal effects in the Perm-R strain of M. domestica. Degree of dominance was 0.6 and 0.7 for F1 and F1' progenies, respectively, which suggests that permethrin resistance was expressed as an incompletely dominant trait. Chi-square analyses for self-bred (F2), and backcross progenies (BC1, BC2, BC3 and BC4) revealed significant differences between the observed and expected mortality, indicating the possibility of multiple genes responsible for permethrin resistance. Moreover, the Perm-R strain did not show cross-resistance to propoxur, chlorpyrifos, profenofos or spinetoram. Conclusion Permethrin resistance in the Perm-R strain of M. domestica was inherited as autosomal, incompletely dominant and governed by more than one gene. Lack of cross-resistance between permethrin and propoxur, chlorpyrifos, profenofos or spinetoram provides an opportunity for rotational use of these insecticides in the control of M. domestica. These data could help to manage M. domestica and the problem of permethrin resistance.

The essential oil (EO) of Thymus serpyllum and thymol were evaluated for their insecticidal activity against the housefly (Musca domestica) larvae and pupae. Contact toxicity and fumigation bioassays were used. Chemical composition analysis of T. serpyllum EO by gas chromatographic mass spectrometry (GC-MS) revealed that thymol (41.6%), p-cymene (21.9%), and γ-terpinene (19.2%) were the major components. For larval assays, the LC50 value of T. serpyllum EO was 0.4 μl/cm2 for contact toxicity and 20.9 μl/l for fumigation toxicity. For thymol, the contact toxicity LC50 value was 0.035 μl/cm2 and the fumigation LC50 value was 2.0 μl/l. For the pupal assay, T. serpyllum EO had a percentage inhibition rate (PIR) value of 100% for both contact toxicity (1.0 μl/cm2) and fumigation toxicity assay (25 μl/l), whereas thymol had a PIR of 100% for contact toxicity (0.1 μl/cm2) and fumigation assay (5 μl/l). This study shows that T. serpyllum EO and thymol are toxic to housefly larvae and pupae and have the potential for use in the population control of this species.

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.

Two unique housefly strains, PSS and N-PRS (near-isogenic line with the PSS), were used to clarify the mechanisms associated with propoxur resistance in the housefly, Musca domestica. The propoxur-selected resistant (N-PRS) strain exhibited >1035-fold resistance to propoxur and 1.70-, 12.06-, 4.28-, 57.76-, and 57.54-fold cross-resistance to beta-cypermethrin, deltamethrin, bifenthrin, phoxim, and azamethiphos, respectively, compared to the susceptible (PSS) strain. We purified acetylcholinesterase (AChE) from the N-PRS and PSS strains using a procainamide affinity column and characterized the AChE. The sensitivity of AChE to propoxur based on the bimolecular rate constant (Ki) was approximately 100-fold higher in the PSS strain compared to the N-PRS strain. The cDNA encoding Mdace from both the N-PRS strain and the PSS strain were cloned and sequenced using RT-PCR. The cDNA was 2073 nucleotides long and encoded a protein of 691 amino acids. A total of four single nucleotide polymorphisms (SNPs), I162M, V260L, G342A, and F407Y, were present in the region of the active site of AChE from the N-PRS strain. The transcription level and DNA copy number of Mdace were significantly higher in the resistant strain than in the susceptible strain. These results indicated that mutations combined with the up-regulation of Mdace might be essential in the housefly resistance to propoxur.

The knockdown and adulticidal activities of individual Cymbopogon citratus and Eucalyptus globulus essential oils (EOs) and their combinations were evaluated against three medical insect pests (Aedes aegypti, Aedes albopictus, and Musca domestica) using a WHO susceptibility test. The knockdown and adulticidal activities against the three medical insect pests of combinations of C. citratus and E. globulus EOs were higher than those of individual EOs alone. Combinations of 7.5% C. citratus + 7.5% E. globulus EOs and 10% C. citratus + 10% E. globulus EOs exhibited the highest efficacy against females of the three species with 100% knockdown and mortality rates at 1 and 24 h after exposure, respectively. Their adulticidal activities were equivalent to that of 10% w/v cypermethrin. In contrast, 70% v/v ethyl alcohol negative control was not effective at all. The combinations of EOs showed a synergistic effect, i.e., their adulticidal activity was improved by 0.2 to 100%, with increased knockdown and mortality rates, compared to individual EOs. The highest synergistic effect on effective knockdown and adulticidal activities against females of the three species was achieved by a combination of 2.5% C. citratus + 2.5% E. globulus EOs, with 36.6 to 100% knockdown rate increase and 33.5 to 98.9% mortality rate increase. This study demonstrates that all tested combinations of C. citratus and E. globulus EOs were effective adulticidal agents against females of Ae. aegypti, Ae. albopictus, and M. domestica and have a high potential for development into a botanical insecticide for controlling populations of Aedes mosquitoes and houseflies.

BACKGROUND: The house fly, Musca domestica L. (Diptera: Muscidae) is an important public health pest that serves as a carrier for pathogens transmitting various diseases of man and animals. It is well known for rapid resistance development to insecticides applied for its chemical control. Chlorantraniliprole, an anthranilic diamide, a ryanodine receptor agonist, is a promising agent for the integrated pest management of various insect pests. To design a retrospective resistance management strategy, life history traits of the chlorantraniliprole laboratory‐selected (CTPR‐SEL) and unselected counterpart (UNSEL) sub‐populations of a field strain and their reciprocal crosses were studied. RESULTS: After eight generations of consecutive selection with chlorantraniliprole, a 750‐fold resistance level when compared to a susceptible strain and a 124‐fold resistance level when compared to the UNSEL strain had developed in CTPR‐SEL. Very low cross resistance to bifenthrin but no cross resistance to spinosad and fipronil was observed in the CTPR‐SEL strain. Results of the fitness traits suggest that the CTPR‐SEL has a lower relative fitness (0.34), reduced fecundity, a decrease in eggs hatchability, lower biotic potential and net reproductive rate as compared to the UNSEL strain. Interestingly, chlorantraniliprole resistance was unstable in the CTPR‐SEL. CONCLUSIONS: Fitness costs associated with chlorantraniliprole resistance suggest that the efficacy of this insecticide could be preserved for a prolonged duration of time by alternating its use with insecticides having dissimilar modes of action and no cross resistance. When cross‐resistance is absent, a sequence of two insecticides is expected to be more durable than a mixture unless the population's h² of resistance to the mixture is less than half of the mean of the population's h² of resistance to the two individual components of the mixture. Unstable chlorantraniliprole resistance could also help to sustain its efficacy by being withdrawn from usage for some period of time. © 2019 Society of Chemical Industry

A study was conducted to invesstigate the housefly maggot meal (HMM) as an alternative protein source to replace the soybean meal in broiler chick's diet. A total of 720 1-day-old male broiler chicks were divided into three groups and fed diets formulated with HMM to replace soybean meal at the rate of 0%, 4% and 8%. The study lasted for 42 days in two phases. Results showed that HMM addition did not markedly affect body weight, average daily body weight gain and average daily feed intake of the broiler chicks. Feed conversion ratio increased linearly (1-21 days) in starter or quadratically (22-42 days) in the grower phase. HMM non-significantly increased the feed intake and body weight during the grower phase. Slight changes were observed for decrease of blood biochemical indices in the platelets (day 21), and alkaline phosphatase and lysozyme (day 42), and increase for red blood cells, packed cell volume, total protein and uric acid on day 42; however, the fluctuations were within the physiological range. Non-significant effects were observed for carcass composition and meat quality, except that HMM numerically reduced the shear force of breast muscle (linear, p = .058). These results are the strong evidence that HMM can be used as an alternative protein source at 8% in broiler chick's diet without any adverse effect on chick's performance.

Ninety-four putative G protein-coupled receptors (GPCRs) were identified in the Musca domestica genome. They were annotated and compared with their homologues in Drosophila melanogaster. Phylogenetic analyses of the GPCRs from both species revealed that several family members shared a closer relationship based on the domain architecture. The expression profiles of these genes were examined by quantitative real-time PCR amongst three strains of the house fly, a near-isogenic line strain with imidacloprid resistance (N-IRS), the corresponding susceptible strain (CSS) and another strain derived from field populations with imidacloprid resistance (IRS). We found that five GPCR genes were upregulated in the N-IRS and eight GPCR genes were upregulated in the IRS strains compared to the CSS strain. The transgenic lines of D. melanogaster with the GPCR genes (LOC101899380 in the N-IRS strain and LOC101895664 in the IRS strain) exhibited significantly increased tolerance to imidacloprid, and higher expression of cytochrome P450 genes. Bioinformatic analysis of LOC101899380 was carried out based on its full-length nucleic acid sequence and putative amino acid sequence, and it was named Methuselah-like10 (Mthl10) owing to its homology with D. melanogaster Mthl10. A cell-base cell counting kit-8 toxicity assay demonstrated that the expression of the GPCR gene LOC101899380 in Spodoptera frugiperda (Sf9) cells using a baculovirus-mediated expression system can elevate the cell tolerance to imidacloprid, indirectly supporting the hypothesis that the GPCR gene LOC101899380 plays some role in imidacloprid resistance. These results should be useful for furthering understanding of the regulatory pathway by which house flies develop resistance.

Background It is difficult for insecticide spray droplets to adhere to highly hydrophobic pest surfaces, and this is an important reason behind the low utilization of pesticides. Greater understanding of the wetting behaviors of agro-surfactants on pest surfaces is of great importance in pesticidal applications. Results This research investigated the wetting processes of three surfactant solutions [TritonX-100 (TX-100), sodium dodecyl sulfate (SDS) and dodecyl trimethyl ammonium bromide (DTAB)] on housefly surfaces based on surface tension, contact angles and solid-liquid interaction. As the surfactant concentration increased, the wetting abilities of the solutions improved due a decrease in liquid surface tension. The decrease in contact angles followed an inverted 'S' shape until the concentration exceeded the critical micelle concentration, at which point surfactant adsorption was saturated at the interfaces. The nonionic surfactant TX-100 enhanced wettability more than the ionic surfactants SDS and DTAB. The wetting states of the three surfactant solutions on housefly surfaces transformed to the Wenzel state when the surfactant molecules adsorbed at solid-liquid interfaces (ΓSL ) were 3.09-5.36 times higher than those adsorbed at liquid-air interfaces (ΓLV ). More insecticides attached to housefly surfaces and pesticide utilization was significantly improved after adding TX-100 to the pesticide solution. Conclusion Surfactant TX-100 could be a practical and efficient candidate adjuvant for insecticide spraying of houseflies. This research investigated the wetting mechanism of three different types of surfactant solutions on housefly surfaces, and could provide effective guidance for the preparation of insecticide formulations and pesticide adjuvants with better wettability to improve pesticide utilization. © 2019 Society of Chemical Industry.

In this study, metabolites involved in the free-biomass filtrates for three endophytic actinomycetes of Streptomyces capillispiralis Ca-1, Streptomyces zaomyceticus Oc-5, and Streptomyces pseudogriseolus Acv-11 were used as biocatalysts for green synthesis of silver nanoparticles (Ag-NPs). Characterization of biosynthesized Ag-NPs was accomplished using UV-Vis spectroscopy, X-ray diffraction patterns (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM-EDX), transmission electron microscopy (TEM), and particle size analyzer. The biosynthesized Ag-NPs showed maximum surface plasmon resonance (SPR) at 440 for strain Ca-1 and 450 for both strains of OC-5 and Acv-11. Nanoparticle spherical shape was recorded with size ranging from 23.77 to 63.14 nm, 11.32 to 36.72 nm, and 11.70 to 44.73 nm for Ca-1, Oc-5, and Acv-11, respectively. SEM-EDX analysis exhibited the weight percentages of 17.3, 22.3, and 48.7% for Ag-NPs synthesized by strains Ca-1, Oc-5 and Acv-11, respectively. The activities of biosynthesized Ag-NPs were concentration dependent and the obtained results confirmed the efficacy of Ag-NPs as antimicrobial agents against Gram-positive and Gram-negative bacteria as well unicellular and multicellular fungi. The MIC for Gram-positive bacteria, Gram-negative bacteria (E. coli), and eukaryotic microorganisms was 0.25 mM with clear zone ranging from 10.3 to 14.6 mm, while MIC for Pseudomonas aeruginosa was 1.0 mM for Ag-NPs synthesized by strain Ca-1 and 0.25 mM for those synthesized by strains Oc-5 and Acv-11. Moreover, Ag-NPs exhibited antimicrobial activity against four plant pathogenic fungi represented by Alternaria alternata, Fusarium oxysporum, Pythium ultimum, and Aspergillus niger at 2.0, 1.5, 1.0, and 0.5 mM of Ag-NPs with different degree. In vitro assessment of the antioxidant efficacy of biosynthesized Ag-NPs was achieved by scavenging assay of H2O2, reducing power of Fe3+, or total antioxidant assay. The results showed that antioxidant activities of Ag-NPs were concentration dependent with the highest activity at Ag-NP concentration of 2.0 mM. Furthermore, the biosynthesized NPs have prospective bioinsecticidal activity against Culex pipiens and Musca domestica. Green synthesis of NPs could be quite potential for the development of new bioactive compounds used in different biomedical applications.

Musca domestica larvae have been used clinically to cure children malnutritional stagnation and low molecular weight peptides (LMWP) of Musca domestica larvae showed more useful bioactivities. But there is no report on anti-diarrhea effects and identification of the LWMP. The purposes of this study were clarifying the anti-diarrhea effects by regulating intestinal microecology and identification of LMWP. In anti-diarrhea test, diarrhea mice were administered LMWP by oral gavage. Then rectal stool indicator bacteria were counted also the identification of rectal stool bacteria were determined by PCR-DGGE. In LMWP identification test, GFC and RP-HPLC were used to separate the peptide. Then the single polypeptide was tested by MALDI TOF and N-terminal sequence analysis. The results of anti-diarrhea showed that LMWP was effective in the inhibition diarrhea in mice. And microbial diversity indices showed that LMWP treatment group exhibited a higher number of bands. The identification test showed that LMWP had four main components (10-30KD, S1, S2, S3), and there were 5, 7 and 4 peaks in S1, S2 and S3, respectively. The the molecular weight of S2-5, S3-2 and S3-3 was 877.053D, 877.0631D and 1069.4391D, respectively. And S3-3 was determined as Chain A, Carboxypeptidase G2. So the hypothesis that intestinal microbiological regulation might be one of the potential anti-diarrhea mechanisms of Musca domestica larvae LMWP which had four main components and one of the single polypeptide was identified could be drawn.

Evolutionary pressure has pushed many extant plants and animals to develop micro/nanostructures on their surfaces to keep them clean. These structures have become ideal models for bio-inspired design. Although microstructures on biological surfaces have been widely studied, little attention has been paid to the combined role of microstructures and animal’s active cleaning behaviors in keeping their surfaces clean. In this study, we explored the relationship between these micro/nanostructures and wettability as well as the role of the housefly cleaning behaviors in keeping their wings clean. Hierarchical structures consisting of microscale macrotrichias with nanoscale grooves on the wings were observed under scanning electron microscope. The wings were hydrophobic (CA = 133.3°) but with high adhesion to water (CAH = 87.5°), indicating that they were non-self-cleaning surfaces. Macroscale droplets standing on the wings could be best described as being in a transitional wetting state between Wenzel and Cassie-Baxter states due to the presence of the nanoscale grooves, which increased the resistance to water penetration. The hydrophobicity decreased (CA = 109.9°) when the nanostructures were removed by coating the wings with a thick layer of polydimethylsiloxane (PDMS). The houseflies could highly efficiently remove the microscale droplets atop the macrotrichias, and reduce bacterial contamination on their wings through grooming and flutter activities. These active cleaning behaviors could offset the absence of self-cleaning properties and play a key role in keeping the wings clean. The results indicate that housefly wings could be used as a template for the design of special functional surfaces. The present findings not only improve our understanding of the wettability and cleaning properties of natural surfaces, but also provide important insights into the design of bio-inspired materials.

Insecticide resistance in house fly populations is a major problem faced by livestock producers worldwide. A survey of insecticide resistance levels and pyrethroid resistance allele frequencies in the United States was conducted in 2008–09, but little is known about how resistance levels have changed over the last 10 years. In addition, new target-site pyrethroid resistance alleles that confer high levels of resistance have been recently identified in the voltage-sensitive sodium channel, and their frequencies in field populations are unknown. Our aim in this study was to reassess the resistance status of house flies from select locations in the United States by examining resistance levels against commonly used insecticides and frequencies of known resistance alleles. House flies were collected from animal production facilities in five different states between 2016 and 2018. Resistance levels to three insecticides (permethrin, tetrachlorvinphos, and methomyl), representing three classes of insecticides (pyrethroids, organophosphates and carbamates) varied geographically and were lowest in the population collected from New Mexico, intermediate in the population collected from Utah, and greatest in the population from Kansas. The recently identified 1B pyrethroid resistance allele increased dramatically in frequency compared to previous reports, most notably in populations from Kansas and Maryland, indicating that it may already be widespread around the United States. Based on comparison with historical data, the population collected from Kansas represents one of the most highly permethrin resistant populations ever sampled. If the alleles responsible for this level of resistance spread, pyrethroids may be of limited use for house fly control in the United States in the near future.

The mortality rate of a field population of house fly (Musca domestica L.) was determined for a granular fly bait containing the active ingredient indoxacarb, which was compared to two commercially available granular fly baits containing either dinotefuran or cyantraniliprole. Indoxacarb was applied at three different application rates 0.498, 0.986, and 1.972 g/m2 (low, medium, and high). Time to 50% mortality was fastest for dinotefuran (5.7 h) and slowest for the low application rate of indoxacarb (10.3 h). Time to 90% mortality was fastest for the high application rate of indoxacarb (27.7 h) and slowest for dinotefuran (51.0 h) and cyantraniliprole (45.9 h). Among the three indoxacarb application rates, the high rate reached both 50 and 90% fly mortality significantly faster than the low rate. The medium rate did not significantly differ from either the high or low application rates. Dinotefuran bait produced greater fly mortality than all other treatments at 30-min post-exposure, with mortality for remaining baits exceeding controls by 3- to 6-h post-exposure. All insecticidal baits produced similar fly mortality by 6-h post-exposure and >94% fly mortality by 96-h post-exposure, indicating that each may be effective in a fly management program. Flies consumed a similar amount of the indoxacarb (regardless of application rate) and dinotefuran baits, but consumed less of the cyantraniliprole bait, suggesting a feeding irritancy or toxicity effect manifested during consumption. Nevertheless, flies consumed enough cyantraniliprole bait to cause mortality similar to other baits by 6-h post-exposure.

The toxicity of Origanum vulgare essential oil to the housefly Musca domestica L. was evaluated. The major constituents of the O. vulgare essential oil by gas chromatographic mass spectrometry (GC-MS) analysis were carvacrol (58.13%), p-cymene (17.85%), thymol (8.15%), γ-terpinene (4.96%), and linalool (3.69%). Toxicity of O. vulgare essential oil against larvae and pupae was evaluated using fumigation and contact assays. The contact toxicity (LC50) of O. vulgare essential oil and carvacrol for larvae was 0.23 and 0.03 μL/cm2, respectively. The fumigation toxicity (LC50) of O. vulgare essential oil and carvacrol for larvae was 9.52 and 2.78 μL/L, respectively. Pupal toxicity was evaluated by percentage inhibition rate (PIR). PIR of O. vulgare essential oil at 0.25 μL/cm2 was 90.9% for the contact assay and 100% at 20 μL/L for the fumigation assay. PIR of carvacrol was 29.5% (0.025 μL/cm2) and 81.8% (1.25 μL/L) for the contact toxicity and fumigation assay, respectively. O. vulgare essential oil and carvacrol have significant toxicity to the housefly and are potential insecticides for housefly control.

Background Permethrin (a pyrethroid) has been recommended for the management of a number of insect pests, including Musca domestica. Recently, resistance to permethrin has been reported from the Punjab province, Pakistan, which necessitates the need to manage the resistance problem. For this reason, a field strain of M. domestica was selected with permethrin for ten consecutive generations (G1-G10) to determine the risk assessment, the rapidity with which permethrin resistance can develop, and the presence or absence of the cross-resistance phenomenon. Results The results revealed that permethrin resistance increased very quickly in response to selection experiments. The resistance ratio (RR) value increased by 16.04- to 605.92-fold and the LD50 value increased by 100.22 to 3787.08 ng fly-1 after selection experiments. Moreover, the increase in LD50 value remained stable when the selected strain (Perm-SEL) was cultured in the absence of permethrin for the next five generations (G11-G15). The results revealed the possibility of minor cross-resistance in the Perm-SEL strain with β-cyfluthrin and deltamethrin, and lack of cross-resistance with imidacloprid and spinosad. The realized heritability (h2 ) value for the Perm-SEL strain was 0.23. Assuming a mean slope value of 1.27 and an h2 value of 0.23, then ten to four generations would be estimated for a 10-fold increase in the LD50 values at 50-90% selection intensity, respectively. Conclusion The results of present study revealed high risk for permethrin resistance development under continuous selection pressure. Permethrin resistance developed very quickly under laboratory conditions, which might reflect the likelihood of resistance evolution in field conditions if permethrin selection pressure holds for a long time. © 2019 Society of Chemical Industry.

Based on the "cluster effect" and the structure characters of acetylcholinesterase (AChE; EC 3.1.1.7), a new series of 1,2,4-triazolin-3-one and phthalimide heterodimers were designed, synthesized, and evaluated as potent dual acetylcholinesterase inhibitors (AChEIs). Most of the synthesized compounds showed good in vitro inhibitory activities towards both Drosophila melanogaster acetylcholinesterase (DmAChE) and Musca domestica acetylcholinesterase (MdAChE). Among them, 5g was found to be the most potent anti-AChE derivate (5g, IC50 = 8.07 µM to DmAChE, IC50 = 32.24 µM to MdAChE). It was 2.31- and 1.35-fold more active than the positive control ethion (CP, IC50 = 18.62 µM to DmAChE, IC50 = 43.56 µM to MdAChE). The docking model study revealed that 5g possessed the fitted spatial structure and bound to the central pocket and peripheral site of DmAChE. Moreover, most compounds demonstrated high insecticidal activity to Lipaphis erysimi and Tetranychus cinnabarinus at the concentration of 300 mg/L.

Housefly (Musca domestica) Larvae powder (HL) is rich in antioxidants. As oxidative stress is considered as one of the main pathogenesis in Alzheimer's Disease (AD), this study was designed to explore the protective effects of HL as an antioxidant on APP/PS1 mice. 2-Month-old APP/PS1 mice were divided into a model control (MC) group, a Donepezil group and a HL group, and C57BL/6 mice were used as the normal control (NC) group. After 180 days of treatment, the memory ability was measured by Morris Water Maze (MWM). The presence of Aβ and the expression of Uncoupling Protein 4 (UCP4) and CyclinD1 were detected by immunohistochemistry. The expressions of Superoxide Dismutase 1 (SOD1), Catalase (CAT) and Mitogen-activated Protein Kinase (MAPK) signal pathways were measured by western blotting. Compared with untreated APP/PS1 mice, the memory abilities of the HL-treated mice were significantly improved. Furthermore, the HL treatment not only down-regulated the deposition of Aβ and the expression of CylinD1, but also increased both the mRNA and protein levels of SOD, CAT, and UCP4, and enhanced the phosphorylation of JNK and P38 MAPK activation. In conclusion, these results suggest that HL may have a protective effect against memory impairment and prevent oxidative stress-induced injury via the regulation of UCP4 and CyclinD1 and the modulation of JNK and P38 MAPK signaling in AD.

Filth flies, including house flies, Musca domestica L., develop in animal manure. Adult house flies often are controlled with pesticides such as imidacloprid. How imidacloprid disseminates and persists after it contaminates manure was measured at a dairy farm. A week after application of imidacloprid via fly bait to cattle manure, a mean of approximately 4 ppm of imidacloprid, and as high as 15 ppm, was quantifiable up to 12 cm from the application site, but not farther. Laboratory experiments addressed the impact of 15 ppm of imidacloprid in manure on egg-to-adult development of house flies and on the biological control ability of a house fly pupal parasitoid, Spalangia endius Walker. In uncontaminated manure, 93% of eggs developed to adults, versus 7% in contaminated manure. In the parasitoid experiment, fly pupae were placed in contaminated or uncontaminated manure with or without S. endius. In the absence of S. endius, nearly 100% of flies emerged, with or without imidacloprid. In the presence of S. endius, only 11% of flies emerged from uncontaminated manure, versus 36% from contaminated manure; and parasitoids emerged from 82% of hosts in uncontaminated manure versus 53% in contaminated manure. These results suggest that realistic concentrations of imidacloprid in filth fly breeding habitat may interfere with house flies developing to the pupal stage, but also with parasitoids locating and utilizing house flies. However, after 1 wk, the effects on parasitoids will be low 12 cm beyond where bait was applied.

Most dietary lipids are triacylglycerols (TAGs) and phosphatides that are digested by TAG lipases and phospholipases (PLIPs), respectively, originating fatty acids (FA). The genome of Musca domestica has genes coding for phospholipases A1 (1PLIP), A2 (2PLIP), B (BPLIP), and acid lipases (ALIP), as for proteins involved in activation, binding, and metabolism of FA, which expression in the larval midgut was evaluated by RNA-seq. Some of the codified proteins were identified in midgut microvillar-enriched membrane by proteomics. 1PLIPs are the most expressed PLIPs, mainly in anterior midgut whereas 2PLIPs, and BPLIP in middle and posterior midgut, and ALIPs between middle and posterior regions. Absorption of FAs is putatively accomplished by proteins involved in FA activation (acyl-CoA synthetases) found in microvillar-enriched membrane preparations. Furthermore, FA uptake could be enhanced by proteins that bind FAs (FA-binding proteins) and its activated form (acyl-CoA binding proteins) mainly expressed in posterior midgut. Activated FAs could have different fates: synthesis of diacylglycerol (DAG) and TAG through monoacylglycerol and glycerol-3-phosphate pathways; synthesis of phosphatides; energy source by β-oxidation. Most genes coding for enzymes of those routes is expressed mainly at the end of posterior midgut. Data suggest that phosphatides are digested in anterior midgut by Md1PLIPs, releasing lysophosphatides that emulsify fats to be digested by MdALIPs in the middle and posterior midgut. Most resulting FAs is absorbed in the posterior midgut, where they follow the synthesis of DAG, TAG, and phosphatides or are oxidized along the midgut, mainly in highly metabolic middle and posterior midgut regions.

The Tibellus oblongus spider is an active predator that does not spin webs and remains poorly investigated in terms of venom composition. Here, we present a new toxin, named Tbo-IT2, predicted by cDNA analysis of venom glands transcriptome. The presence of Tbo-IT2 in the venom was confirmed by proteomic analyses using the LC-MS and MS/MS techniques. The distinctive features of Tbo-IT2 are the low similarity of primary structure with known animal toxins and the unusual motif of 10 cysteine residues distribution. Recombinant Tbo-IT2 (rTbo-IT2), produced in E. coli using the thioredoxin fusion protein strategy, was structurally and functionally studied. rTbo-IT2 showed insecticidal activity on larvae of the housefly Musca domestica (LD100 200 &mu, g/g) and no activity on the panel of expressed neuronal receptors and ion channels. The spatial structure of the peptide was determined in a water solution by NMR spectroscopy. The Tbo-IT2 structure is a new example of evolutionary adaptation of a well-known inhibitor cystine knot (ICK) fold to 5 disulfide bonds configuration, which determines additional conformational stability and gives opportunities for insectotoxicity and probably some other interesting features.

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.

Housefly, Musca domestica L. is a pest of public health importance and is responsible for spreading diseases like typhoid, diarrhoea, plague etc. Indiscriminate reliance on synthetic insecticides has led to development of insecticide resistance and ill effect to humans and nontarget animals. This demands an alternative and safer pest control option. This study evaluates the biological effect of Piper betle L essential oil and its constituent eugenol, eugenol acetate, and β - caryophyllene on the housefly. The major components present in P. betel EO were safrole (44.25%), eugenol (5.16%), β -caryophyllene (5.98%), β -selinene (5.93%), α-selinene (5.27%) and eugenol acetate (9.77%). Eugenol caused 4.5fold higher ovicidal activity (EC50 86.99 μg/ml) than P. betle EO (EC50 390.37 μg/ml). Eugenol caused fumigant toxicity to adults (LC50 88.38 mg/dm3). On contact toxicity by topical application, eugenol acetate, eugenol and β-caryophyllene caused higher mortality to larval and adult stages than EO. FESEM (Field Emission Scanning Electron Microscope) images reveal that exposure to P. betle EO causes the shrinkage of the larval cuticle. Both EO and eugenol induced the detoxifying enzymes Carboxyl esterase (Car E) and Glutathione S – transferases (GST) in larvae and adults. EO and eugenol at 0.2% caused effective repellence and oviposition deterrence to M. domestica adults and this merits their use as alternative strategy to manage M. domestica.

Veterinary and medical entomologists who are involved in research on pest control often need to perform dose–response bioassays and analyze the results. This article is meant as a beginner’s guide for doing this and includes instructions for using the free program R for the analyses. The bioassays and analyses are described using previously unpublished data from bioassays on house flies, Musca domestica Linnaeus (Diptera: Muscidae), but can be used on a wide range of pest species. Flies were exposed topically to beta-cyfluthrin, a pyrethroid, or exposed to spinosad or spinetoram in sugar to encourage consumption. LD50 values for beta-cyfluthrin in a susceptible strain were similar regardless of whether mortality was assessed at 24 or 48 h, consistent with it being a relatively quick-acting insecticide. Based on LC50 values, spinetoram was about twice as toxic as spinosad in a susceptible strain, suggesting a benefit to formulating spinetoram for house fly control, although spinetoram was no more toxic than spinosad for a pyrethroid-resistant strain. Results were consistent with previous reports of spinosad exhibiting little cross-resistance. For both spinosad and spinetoram, LC50 values were not greatly different between the pyrethroid-resistant strain and the susceptible strain.

Among the vectorial insect pests, the domestic house fly (Musca domestica L., Diptera: Muscidae) is a ubiquitous livestock pest with the ability to develop resistance and adapt to diverse climates. Successful management of the house fly in various locations requires information about its resistance development and life table features. The status of insect growth regulators resistance and life table features on the basis of age, stage, and two sexes of the house fly from five different geographical locations of Riyadh, Saudi Arabia: Dirab, Al-Masanie, Al-Washlah, Al-Uraija and Al-Muzahmiya were therefore investigated. The range of resistance levels were 3.77–8.03-fold for methoxyfenozide, 5.50–29.75 for pyriproxyfen, 0.59–2.91-fold for cyromazine, 9.33–28.67-fold for diflubenzuron, and 1.63–8.25-fold for triflumuron in five populations of house fly compared with the susceptible strain. Analysis of life history parameters—such as survival rate, larval duration, pupal duration, pre-female duration, pre-male duration, adult and total pre-oviposition periods, longevity of male, oviposition period, female ratio, and fecundity female-1—revealed significant variations among the field populations. Additionally, demographic features—including the generation time, the finite and intrinsic rates of increase, doubling time, and net reproductive rate—varied significantly among the field populations. These results will be helpful in planning the management of the house fly in geographically isolated dairies in Saudi Arabia.

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.

Peritrophic matrix/membrane (PM) critically prevents the midgut of insects from external invasion by microbes. The proteins in the peritrophic membrane are its major structural components. Additionally, they determine the formation and function of this membrane. However, the role of PM proteins in immune regulation is unclear. Herein, we isolated a novel PM protein (MdPM-17) from Musca domestica larvae. Further, the function of MdPM-17 in regulating host innate immunity was identified. Results showed that the cDNA of MdPM-17 full is 635 bp in length. Moreover, it consists of a 477-bp open reading frame encoding 158 amino acid residues. These amino acid residues are composed of two Chitin-binding type-2 domain (ChtBD2) and 19 amino acids as a signal peptide. Moreover, tissue distribution analysis indicates that MdPM-17 was enriched expressed in midgut, and moderate levels in the fat body, foregut, and malpighian tubule. Notably, MdPM-17 recombinant protein showed high chitin-binding capacity, thus belongs to the Class III PM protein group. MdPM-17 protein silencing via RNA interference resulted in the expression of antimicrobial peptide (defensin, cecropins, and diptericin) genes, and this occurred after oral inoculation with exogenous microbes Escherichia coli (Enterobacteriales:Enterobacteriaceae), Staphylococcus aureus (Bacillales:Staphylococcaceae), and Candida albicans (Endomycetales:Saccharomycetaceae)). Therefore, all the antimicrobial peptide (AMP) gene expression levels are high in MdPM-17-depleted larvae during microbial infection compared to controls. Consequently, these findings indicate that MdPM-17 protein is associated with the antibacterial response from the housefly.

The insecticidal activity of Myristica fragrans (Houtt) essential seed oil, (Nutmeg) was evaluated against Musca domestica (Linnaeus) and Chrysomya albiceps (Wiedemann); both important infectious pathogenic disease vectors. The oil was extracted by distillation, and 21 components were identified during chemical analysis; principally β-pinene (26%), α-pinene (10.5%), Sabinene (9.1%) and γ-terpinen (8.5%). Insecticidal properties were identified through larvicide and adulticide tests. Using the immersion method, the oil at 5% was found to be very effective (90 ± 1%) against M. domestica larvae. The results for adulticide activity varied by fly species, dosage, time, and method of exposure. Topical application (on the insect thorax) was more toxic to C. albiceps, where the lethal concentration at 50% (LC50) was 2.02 ± 0.56, and 8.57 ± 2.41 for the common flies. When the insects were exposed to oil impregnated paper, the results were similar for M. domestica and C. albiceps adults with respective LC50 values of 2.74 ± 0.24, and 3.65 ± 0.48. Thus, the results demonstrated that M. fragrans oil presents insecticidal activity and can be used for control of Musca domestica and Chrysomya albiceps.

Housefly, Musca (M) domestica L. (Diptera: Muscidae) is a pervasive insect that transmits a variety of pathogens to humans and livestock. Although numerous synthetic pesticides are available to combat houseflies, their ecological and toxicological concerns have led to the exploration of natural products as safer alternatives. The present work was designed to develop an essential oil based controlled-release evaporating tablet (EO-CRT) and investigate its repellency against M. domestica. This study assesses the toxicological impacts of the EO-CRT following its sub-chronic inhalation exposure. Briefly, repellent activity of fourteen essential oils viz. lemon grass, bergamot, mentha, basil, camphor, lavender, clove, patchouli, rosemary, cinnamon, eucalyptus, citronella, jasmine and wild turmeric against M. domestica were screened using the 'Y'-tube olfactometer. The synergistic activity of the best four oils, under preliminary screening, were further evaluated by double and triple blending. The best combination of three oils were finalized for optimization with 17-run, 3-factor, 3-level Box-Behnken design. This was then employed to construct polynomial models and predict the best optimized formulation EO-CRT. EO-CRT was characterized by Differential Scanning Calorimetry (DSC) and Gas Chromatography-Mass Spectroscopy (GC-MS). The efficacy of the EO-CRT against M. domestica was assessed by attraction and repellent assay. Chest X-ray, histopathology and scanning electron microscopy of the exposed lung was performed to study EO-CRT's sub-chronic toxicity on Wistar rats. The EO-CRT showed slow release up to a period of 10 days at room temperature, exhibited 100% repellency (%Error=1.237) against M. domestica and was found to possess all the characteristics of an ideal formulation. Sub-chronic toxicity study further revealed the non-toxic nature of the EO-CRT. Thus, our study provides an assurance that the formulated EO-CRT could be effective not only in repelling the nuisance pest, M. domestica, in human dwellings, but also in minimizing the mechanical transmission of pathogens by it.

Ethnopharmacological relevance The use of Hazomalania voyronii, popularly known as hazomalana, to repel mosquitoes and resist against insect attacks is handed down from generation to generation in Madagascar. In the present study, we investigated the ability of the essential oils (EOs) obtained from the stem wood, fresh and dry bark of H. voyronii to keep important mosquito vectors (Aedes aegypti and Culex quinquefasciatus) away, as well as their toxicity on three insect species of agricultural and public health importance (Cx. quinquefasciatus, Musca domestica and Spodoptera littoralis). Materials and methods Hydrodistillation was used to obtain EOs from stem wood, fresh and dry bark. The chemical compositions were achieved by gas chromatography-mass spectrometry (GC-MS). Toxicity assays using stem wood and bark EOs were performed on larvae of Cx. quinquefasciatus and S. littoralis, and adults of M. domestica by WHO and topical application methods, respectively. Mosquito repellent activity of the most effective EO, i.e. the bark one, was determined on human volunteers by arm-in-cage tests, and results were compared with that of the commercial repellent N,N-ddiethyl-m-toluamide (DEET). Results The H. voyronii EOs were characterized by oxygenated monoterpenes with perilla aldehyde (30.9–47.9%) and 1,8-cineole (19.7–33.2%) as the main constituents. The fresh and dry bark EOs were the most active on Cx. quinquefasciatus and S. littoralis larvae, respectively, with LC50/LD50 of 65.5 mg L−1, and 50.5 μg larva−1; the EOs from wood and fresh bark displayed the highest toxicity on M. domestica (LD50 values 60.8 and 65.8 μg adult−1, respectively). Repellence assay revealed an almost complete protection (>80%) from both mosquito species for 30 min when pure fresh bark EO was applied on the volunteers’ arm, while DEET 10% repelled >80% of the mosquitoes up to 120 min from application. Conclusion The traditional use of the bark EO to repel insects has been demonstrated although an extended-release formulation based on H. voyronii EOs is needed to increase the repellent effect over time. A wide spectrum of insecticidal activity has been provided as well, suggesting a possible use of H. voyronii EOs in the fabrication of green repellents and insecticides useful to control mosquito vectors and agricultural pests.

Marsh rosemary (Ledum palustre, Ericaceae) has been widely used in the traditional medicine of various regions worldwide, and as insect repellent. Little is known on its essential oil insecticidal potential. This study explored the insecticidal effects of the essential oil obtained from L. palustre growing in Poland on selected insect pests and vectors. GC-MS analysis evidenced an uncommon chemotype characterized by ascaridole (35.3% as sum of cis-ascaridole and isoascaridole) and p-cymene (25.5%). The essential oil was effective against Culex quinquefasciatus, Spodoptera littoralis and Musca domestica, showing LC50/LD50 of 66.6 mg L−1, 117.2 μg larva−1 and 61.4 μg adult−1, respectively. It was not toxic to non-target Eisenia fetida earthworms and moderately toxic to Daphnia magna microcrustaceans, over the positive control α-cypermethrin. The essential oil cytotoxicity on human keratinocytes and fibroblasts showed high IC50 values (71.3 and 84.4 μg mL−1, respectively). Comet assay data highlighted no DNA damages. Based on our findings, this essential oil, characterized by the ascaridole/p-cymene chemotype, could be a candidate for the formulation of botanical insecticides; large-scale production of green insecticides by this rare species may be assured by ex situ cultivation and biotechnological techniques.

In this study, two antioxidant peptides were identified and characterized from the alcalase-hydrolysate of housefly (Musca domestica L.) pupae guided by ABTS cation radical scavenging activity. Peptides sequences were identified as DFTPVCTTELGR (DR12, 1338.48 Da) and ARFEELCSDLFR (AR12, 1485.66 Da) using nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both DR12 and AR12 exert strong ABTS cation radical scavenging ability with EC50 values of 0.39 and 0.35 mM, respectively. Moreover, AR12 can effectively protect PC12 cells from oxidative damage induced by hydrogen peroxide (H2O2) by decreasing intracellular reactive oxygen species (ROS) and malonaldehyde (MDA), recovering cellular mitochondrial membrane potential (MMP), and increasing the activity of intracellular superoxide dismutase (SOD). Stability tests suggest that AR12 is competent for the challenge of heating, acid, alkali or simulated gastrointestinal (GI) digestion and exhibits great activity to remove ABTS cation radical. DR12 shows a great stability against heating, but its antioxidative ability declines after being treated with acid, alkali or simulated GI digestion. In general, both DR12 and AR12 identified from housefly pupae hydrolysate stand a chance of being potential antioxidants or precursors to antioxidants and AR12 might be applied in the field of neuroprotection.

Until now there is no molecular model of starch digestion and absorption of the resulting glucose molecules along the larval midgut of Musca domestica. For addressing to this, we used RNA-seq analyses from seven sections of the midgut and carcass to evaluate the expression level of the genes coding for amylases, maltases and sugar transporters (SP). An amylase related protein (Amyrel) and two amylase sequences, one soluble and one with a predicted GPI-anchor, were identified. Three highly expressed maltase genes were correlated with biochemically characterized maltases: one soluble, other glycocalyx-associated, and another membrane-bound. SPs were checked as being apical or basal by proteomics of microvillar preparations and those up-regulated by starch were identified by real time PCR. From the 9 SP sequences with high expression in midgut, two are putative sugar sensors (MdSP4 and MdSP5), one is probably a trehalose transporter (MdSP8), whereas MdSP1-3, MdSP6, and MdSP9 are supposed to transport glucose into cells, and MdSP7 from cells to hemolymph. MdSP1, MdSP7, and MdSP9 are up-regulated by starch. Based on the data, starch is at first digested by amylase and maltases at anterior midgut, with the resulting glucose units absorbed at middle midgut. At this region, low pH, lysozyme, and cathepsin D open the ingested bacteria and fungi cells, freeing sugars and glycogen. This and the remaining dietary starch are digested by amylase and maltases at the end of middle midgut and up to the middle part of the posterior midgut, with resulting sugars being absorbed along the posterior midgut.

Among the various glutathione transferase (GST) isozymes in insects, the delta- and epsilon-class GSTs fulfill critical functions during the detoxification of insecticides. We crystalized MdGSTD1, the major delta-class GST isozyme in the housefly (Musca domestica), in complex with glutathione (GSH) and solved its structure at a resolution of 1.4 Å. The overall folding of MdGSTD1 resembled other known delta-class GSTs. Its substrate binding pocket was exposed to solvent and considerably more open than in the epsilon-class GST from M. domestica (MdGSTE2). However, their C-terminal structures differed the most because of the different lengths of the C-terminal regions. Although this region does not seem to directly interact with substrates, its deletion reduced the enzymatic activity by more than 70%, indicating a function in maintaining the proper conformation of the binding pocket. Binding of GSH to the GSH-binding region of MdGSTD1 results in a rigid conformation of this region. Although MdGSTD1 has a higher affinity for GSH than the epsilon class enzymes, the thiol group of the GSH molecule was not close enough to serine residue 9 to form a hydrogen-bond with this residue, which is predicted to act as the catalytic center for thiol group deprotonation in GSH.

Owing to thioether diverse physicochemical properties by non-covalent interactions with bio-macromolecules, thioether derivatives containing heterocyclic moiety are known for their interesting insecticidal bioactivities and attracting considerable attention as neuroactive insecticides. Here we synthesis a series of novel thioether bridged N-phenylpyrazole derivatives incorporating various (hetero)aromatic substituents into 4-position of the pyrazole ring. Structure-activity relationship (SAR) studies resulted in compounds 6d and 7d with the most potent insecticidal activity among the series containing various substituted benzene substituents (LC50 = 13.70–25.47 μg/g). Further optimization to increase the lipophilicity and charge density of aromatic substituents of compounds 6d and 7d resulted in compounds 12d, 14d and 16d with sulfur-containing heterocycle substituents possessing good insecticidal activity against Musca domestica L. among the series (LC50 = 0.67–1.30 μg/g). The thioether bridge N-phenylpyrazole derivatives, which exhibit different length of the spacer arm introduced between N-phenylpyrazole moiety and the (hetero)aromatic substituents, were also prepared and evaluated. By contrast, the insecticidal activities of compounds containing the short thioether bridge, 1,2-bis((hetero)aromatic thio) ethane, are higher than that containing the long thioether bridge, 1,3-bis((hetero)aromatic thio) propane. The results of molecular docking and pharmacophore analyses indicated A299, T303, and L306 of a subunit were essential to form non-covalent interactions contacts with the ligands. Specially, the sulfur-containing heterocycle substituent derivatives 12d and 14d as the sterically favored areas could form the important hydrophobic interactions with the deeper residue P295.

Essential oils are considered an alternative for replacement of conventional insecticides that have provoked an increasing number of resistant species and damages to health. The objective of this work was to investigate the insecticidal activity of Lavandula dentata L. (Lamiales: Lamiaceae) oil against the species Musca domestica L. (Diptera: Muscidae) and Chrysomya albiceps Wiedemann (Diptera: Calliphoridae). Extraction of the essential oil from the aerial parts of the plant was carried out using hydro-distillation. Its principal compounds (1,8-cineol, camphor, and linalool oxide) were identified. Insecticidal activity was determined by evaluating adulticidal effect using topical application methods and exposure to impregnated paper; larvicidal effect was determined using immersion method. The essential oil presented toxicity in M. domestica and C. albiceps adults; the lethal concentration values (LC50) in the superficial application method were respectively 3.13 ± 0.64 and 1.39 ± 0.19% live weight (l/v). Toxicity was also found in the oil impregnated paper exposure test; the LC50 results for M. domestica and C. albiceps were respectively 4.15 ± 0.64 and 5.14 ± 0.81%. Larvicidal effect was observed on third stage M. domestica larvae when exposed to an oil concentration of 2.5% (m/v). In addition, the oil was evaluated for cytotoxicity, mutagenicity, and genotoxicity in human cells, and the in vitro safety of this oil in human cells was verified. Thus, L. dentata oil presented insecticidal activity in M. domestica and C. albiceps and can be used as an alternative for control of these dipterans.

Work was undertaken to investigate the potential use of housefly (Musca domestica) larvae reared on broiler manure as a source of nutrition for poultry production in the United Kingdom. Nutritional analysis showed that larvae have a high (>45% dry wt.) protein content and a favorable amino acid profile that is rich in key amino acids, such as lysine and methionine. A broiler digestibility trial was carried out to determine the apparent ileal digestibility coefficients (AIDC) and true ileal digestibility coefficients (TIDC) of amino acids (AA) from insect larval meal (ILM) from M. domestica and fishmeal (FM) in broiler chickens. This was calculated using multiple linear regression technique based upon 3 inclusions of each protein source in a semisynthetic diet. One-hundred-forty-four day-old male (Ross 308) broilers were fed from hatch on a commercial starter diet for 20 days. Experimental diets were fed from d 21 to 28, and feed intakes were measured daily. On d 28, the trial was terminated, ileal digesta were collected for the determination of AIDC and TIDC of AA, and inflammatory responses (gizzard erosion and eye discharge) were measured. No significant differences were observed in digestibilities between protein sources for any AA. Furthermore, ILM feeding did not induce gizzard erosion or eye discharge at any inclusion. These results provide strong evidence to suggest that ILM of the common housefly can provide a successful alternative protein source to FM in broiler diets.

In this study, starvation-, thermal- and heavy metal-associated expression of four small Musca domestica HSPs (abbreviated as MdomHSPs and they are MdomHSP10, MdomHSP27, MdomHSP27.1 and MdomHSP27.2) were determined. The following results were found: All MdomHSPs were significantly higher expressed during the active larval and adult stages than the egg and pupal stages; All MdomHSPs were expressed at relatively equal levels in the head, thorax and abdomen of adults; The expression of MdomHSP27 was significantly down-regulated in 4-day-old larvae that were starved for 6h, while the other 3 MdomHSPs were not significantly affected; Thermal treatment altered the expression of MdomHSPs in 4-day-old larvae: MdomHSP10 was significantly down-regulated in 4-day-old larvae that were maintained at 4°C and 37°C than in those that were maintained at 25°C; Lead, cadmium and chromium exposure influenced larval expression of MdomHSPs to varying degrees. The expression dynamic profile of MdomHSPs would contribute to the understanding of their physiological role in M. domestica.

In this study, a novel heterozygous antimicrobial peptide MLH was synthesized, expressed, purified, and characterized. The peptide Md-cec-LL-37_Hp (MLH) was selected through bioinformatic analysis using musca domestica antimicrobial peptide (Cec-Med), human antimicrobial peptide LL-37, and helicobacter pylori antimicrobial peptide (Hp) as parent peptides. The target gene was synthesized by overlap extension PCR (SOE-PCR) and connected to the expression vector pET-32a (+), and the recombinant plasmid pET-32a-MLH was transformed to Escherichia coli for constructing pET-32a-MLH/BL21 (DE3). Isopropyl β-D-thiogalactoside (IPTG) was used to induce protein expression, and SDS-PAGE and western blot were adopted to test the target protein. And fermentation condition was optimized to get the mass expression of the fusion protein. The Ni2+ affinity chromatographic column was used to purify. Active heterozygous peptide was obtained after renaturation. Finally, the activity of the heterozygous antimicrobial peptide was identified. The fusion peptide showed significant antimicrobial effect on both E. coli and Staphylococcus aureus.