Design, Synthesis, and Bioactivity of Trifluoroethylthio-Substituted Phenylpyrazole Derivatives.

As the first marketed phenylpyrazole insecticide, fipronil exhibited remarkable broad-spectrum insecticidal activity. However, it poses a significant threat to aquatic organisms and bees due to its high toxicity. Herein, 35 phenylpyrazole derivatives containing a trifluoroethylthio group on the 4 position of the pyrazole ring were designed and synthesized. The predicted physicochemical properties of all of the compounds were within a reasonable range. The biological assay results revealed that compound 7 showed 69.7% lethality against Aedes albopictus (A. albopictus) at the concentration of 0.125 mg/L. Compounds 7, 7g, 8d, and 10j showed superior insecticidal activity for the control of Plutella xylostella (P. xylostella). Notably, compound 7 showed similar insecticidal activity against Aphis craccivora (A. craccivora) compared with fipronil. Potential surface calculation and molecular docking suggested that different lipophilicity and binding models to the Musca domestica (M. domestica) gamma-aminobutyric acid receptors may be responsible for the decreased activity of the tested derivatives. Toxicity tests indicated that compound 8d (LC50 = 14.28 mg/L) induced obviously 14-fold lower toxicity than fipronil (LC50 = 1.05 mg/L) on embryonic-juvenile zebrafish development.

House fly larval grazing alters dairy cattle manure microbial communities.

BACKGROUND: House fly larvae (Musca domestica L.) require a live microbial community to successfully develop. Cattle manure is rich in organic matter and microorganisms, comprising a suitable substrate for larvae who feed on both the decomposing manure and the prokaryotic and eukaryotic microbes therein. Microbial communities change as manure ages, and when fly larvae are present changes attributable to larval grazing also occur. Here, we used high throughput sequencing of 16S and 18S rRNA genes to characterize microbial communities in dairy cattle manure and evaluated the changes in those communities over time by comparing the communities in fresh manure to aged manure with or without house fly larvae. RESULTS: Bacteria, archaea and protist community compositions significantly differed across manure types (e.g. fresh, aged, larval-grazed). Irrespective of manure type, microbial communities were dominated by the following phyla: Euryarchaeota (Archaea); Proteobacteria, Firmicutes and Bacteroidetes (Bacteria); Ciliophora, Metamonanda, Ochrophyta, Apicomplexa, Discoba, Lobosa and Cercozoa (Protists). Larval grazing significantly reduced the abundances of Bacteroidetes, Ciliophora, Cercozoa and increased the abundances of Apicomplexa and Discoba. Manure aging alone significantly altered the abundance bacteria (Acinetobacter, Clostridium, Petrimonas, Succinovibro), protists (Buxtonella, Enteromonas) and archaea (Methanosphaera and Methanomassiliicoccus). Larval grazing also altered the abundance of several bacterial genera (Pseudomonas, Bacteroides, Flavobacterium, Taibaiella, Sphingopyxis, Sphingobacterium), protists (Oxytricha, Cercomonas, Colpodella, Parabodo) and archaea (Methanobrevibacter and Methanocorpusculum). Overall, larval grazing significantly reduced bacterial and archaeal diversities but increased protist diversity. Moreover, total carbon (TC) and nitrogen (TN) decreased in larval grazed manure, and both TC and TN were highly correlated with several of bacterial, archaeal and protist communities. CONCLUSIONS: House fly larval grazing altered the abundance and diversity of bacterial, archaeal and protist communities differently than manure aging alone. Fly larvae likely alter community composition by directly feeding on and eliminating microbes and by competing with predatory microbes for available nutrients and microbial prey. Our results lend insight into the role house fly larvae play in shaping manure microbial communities and help identify microbes that house fly larvae utilize as food sources in manure. Information extrapolated from this study can be used to develop manure management strategies to interfere with house fly development and reduce house fly populations.

Enterobacter hormaechei in the intestines of housefly larvae promotes host growth by inhibiting harmful intestinal bacteria.

BACKGROUND: As a pervasive insect that transmits a variety of pathogens to humans and animals, the housefly has abundant and diverse microbial communities in its intestines. These gut microbes play an important role in the biology of insects and form a symbiotic relationship with the host insect. Alterations in the structure of the gut microbial community would affect larval development. Therefore, it is important to understand the mechanism regulating the influence of specific bacteria on the development of housefly larvae. METHODS: For this study we selected the intestinal symbiotic bacterium Enterobacter hormaechei, which is beneficial to the growth and development of housefly larvae, and used it as a probiotic supplement in larval feed. 16S rRNA gene sequencing technology was used to explore the effect of E. hormaechei on the intestinal flora of housefly larvae, and plate confrontation experiments were performed to study the interaction between E. hormaechei and intestinal microorganisms. RESULTS: The composition of the gut microflora of the larvae changed after the larvae were fed E. hormaechei, with the abundance of Pseudochrobactrum, Enterobacter and Vagococcus increasing and that of Klebsiella and Bacillus decreasing. Analysis of the structure and interaction of larval intestinal flora revealed that E. hormaechei inhibited the growth of harmful bacteria, such as Pseudomonas aeruginosa, Providencia stuartii and Providencia vermicola, and promoted the reproduction of beneficial bacteria. CONCLUSIONS: Our study has explored the influence of specific beneficial bacteria on the intestinal flora of houseflies. The results of this study reveal the important role played by specific beneficial bacteria on the development of housefly larvae and provide insight for the development of sustained biological agents for housefly control through interference of gut microbiota.

Oral Toxicity of Pseudomonas protegens against Muscoid Flies.

The bioinsecticidal action of Pseudomonas protegens has so far been reported against some target insects, and the mode of action remains unclear. In this study, the pathogenicity potential of a recently isolated strain of this bacterial species against fly larvae of medical and veterinary interest was determined. Preliminary experiments were conducted to determine the biocidal action by ingestion against Musca domestica and Lucilia caesar larvae, which highlighted a concentration-dependent effect, with LC50 values of 3.6 and 2.5 × 108 CFU/mL, respectively. Bacterial septicaemia was observed in the body of insects assuming bacterial cells by ingestion. Such rapid bacterial reproduction in the hemolymph supports a toxin-mediated mechanism of action involving the intestinal barrier overcoming. In order to gain more information on the interaction with the host, the relative time-course expression of selected P. protegens genes associated with virulence and pathogenicity, was determined by qPCR at the gut level during the first infection stage. Among target genes, chitinase D was the most expressed, followed by pesticin and the fluorescent insecticidal toxin fitD. According to our observations and to the diversity of metabolites P. protegens produces, the pathogenic interaction this bacterium can establish with different targets appears to be complex and multifactorial.

Resistance to insect growth regulators and age-stage, two-sex life table in Musca domestica from different dairy facilities.

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.

Comparing the efficacy of nutmeg essential oil and a chemical pesticide against Musca domestica and Chrysomya albiceps for selecting a new insecticide agent against synantropic vectors.

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 ß-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.

Posttreatment temperature influences toxicity of insect growth regulators in Musca domestica.

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.

Exposure Timing and Method Affect Beauveria bassiana (Hypocreales: Cordycipitaceae) Efficacy Against House Fly (Diptera: Muscidae) Larvae.

House flies, Musca domestica L., are widely recognized for their ability to develop resistance to chemical insecticides so alternative control strategies are desired. The use of entomopathogenic fungi such as Beauveria bassiana (Balsamo) Vuillemin to manage house fly populations has shown promising results; however, the success of using this fungus against larval house flies varies widely. The overall objective of this study was to examine factors that may influence efficacy of B. bassiana treatments against larvae. When a high concentration (4 × 1011 conidia/ml) was applied to first- and second-instar larvae in rearing medium, there was a significant reduction in pupation and adult emergence rates. Treating third-instar larvae at the same concentration did not result in a significant reduction of pupation or adult emergence. Temperature (22 versus 32°C) and media composition (diets with- and without propionic acid) did not affect the B. bassiana treatment efficacy against house fly larvae. The narrow time window of vulnerability of larvae and the high doses required to infect them indicate that B. bassiana has little potential as an operational biocontrol agent for house fly larvae.

Ultrastructure of the Immature Stages of Musca domestica (Diptera: Muscidae: Muscinae).

Musca domestica (Linnaeus, 1758) is a muscoid species that is widespread throughout the world and acts as a mechanical vector of different enteropathogens primarily in underdeveloped countries. The adult and its immature forms are associated with decaying organic matter and can be seen visiting human corpses and animal carcass, and the larvae can also cause an infestation on human and animal wounds, feeding on the tissues. These characteristics make them have a forensic significance and, mainly, a great sanitary importance. This study aimed to analyze and describe morphological aspects of their immature stages of M. domestica, including the eggs, the first-, second-, and third-instar larvae, and the puparium using scanning electron microscopy (SEM). The eggs have standard format that all muscoid flies shared. The exochorion has some modified cells that were used for embryonic respiration. The first-instar larvae present two openings on the posterior spiracle as in the second-instar larvae, but with the last one, we can observe the anterior spiracle structure. The third-instar larvae, as in some other Muscidae species, have a posterior spiracular opening with a sinuous form that is located near the edge of the spiracular plate. The puparium morphology is equal to the third instar with a respiratory structure that helps the breathing during the metamorphosis process. This article supports the need for knowledge over the morphological characteristics of the immature forms of the muscoid Diptera (Linnaeus, 1758) at the same time helping with the correct identification of this insect phase.

Housefly (Musca domestica L.) associated microbiota across different life stages.

The housefly (Musca domestica L.) lives in close association with its microbiota and its symbionts are suggested to have pivotal roles in processes such as metabolism and immune response, but it is unclear how the profound physiological changes during ontogeny affect the housefly's associated microbiota and their metabolic capabilities. The present study applies 16S rRNA gene amplicon sequencing to investigate the development of the host-associated microbiota during ontogeny. The potential for microbiota transfer between developmental stages, and the metabolic potential of these microbiota were evaluated. Representatives of Firmicutes were observed as early colonisers during the larval stages, followed by colonisation by organisms affiliating with Proteobacteria and Bacteroidetes as the flies matured into adults. Microbiota observed across all the developmental stages included Lactococcus, Lactobacillus and Enterococcus, while Weissella and Chishuiella were associated with newly hatched larvae and adults, respectively. Predictive metabolic profiling of the identified microorganisms further suggested that the microbiota and their functional profile mature alongside their host and putative host-microbe relationships are established at different stages of development. The predicted metabolic capability of the microbiota developed from primarily simple processes including carbohydrate and nucleotide metabolisms, to more complex metabolic pathways including amino acid metabolisms and processes related to signal transduction.

Evaluation of Eugenol and (E)-Cinnamaldehyde Insecticidal Activity Against Larvae and Pupae of Musca domestica (Diptera: Muscidae).

Musca domestica L., 1758, is an important mechanical vector of several pathogens for humans and livestock, making it essential to study new alternatives of more efficient and safer control for this dipteran. This study evaluated the toxicity of the phenylpropanoids eugenol and (E)-cinnamaldehyde on its life stages. A contact test with 10 repetitions (n = 10) was performed for each substance concentration on each post-embryonic immature life stage. Both substances presented insecticidal activity on the immature life stages of the dipteran, and secondary effects on development caused by sublethal concentrations. Larvicidal activity was shown from the 1.25 mg/ml concentration by eugenol and from 2.5 mg/ml by (E)-cinnamaldehyde, and both substances had a 100% larval treatment efficacy (LTE) from the 5mg/ml concentration. For pupal treatment, (E)-cinnamaldehyde differed from the control from the 10 mg/ml concentration (P < 0.05), and both phenylpropanoids caused malformation in adults from 10 mg/ml. The highest pupal treatment efficacy (PTE) was obtained from the 30 mg/ml concentration, 67.2% for (E)-cinnamaldehyde, and 32% for eugenol. The products tested in this study showed high larvicidal potential, and both presented pupicidal effects and caused malformation in adults from treated pupae.

Midgut fluxes and digestive enzyme recycling in Musca domestica: A molecular approach.

Insects are reported to have water midgut countercurrents fluxes powering enzyme recovery before excretion, usually known as enzyme recycling. Up to now there is a single, and very incomplete, attempt to relate transporters and channels with countercurrent fluxes. In this work, M. domestica midgut water fluxes were inferred from the concentration of ingested and non absorbable dye along the midgut, which anterior midgut was divided in two sections (A1, A2), the middle in one (M) and the posterior midgut in four (P1, P2, P3, and P4), which led to the finding of additional sites of secretion and absorption. Water is secreted in A1 and A2 and absorbed at the middle midgut (M), whereas in posterior midgut, water is absorbed at P2 and secreted in the other sections, mainly at P4. Thus, a countercurrent flux is formed from P4 to P2. To disclose the involvement of the known water transporters Na+:K+:2Cl- (NKCC) and K+:Cl- (KCC), as well as the water channels aquaporins in water fluxes, their expression was evaluated by RNA-seq analyses from triplicate samples of seven sections along the midgut. MdNKCC1 was expressed in A1, MdNKCC2 was expressed in M1 and P2 and MdKCC in middle and in the most posterior region, thus apparently involved in secretion, absorption and both, respectively. MdNKCC2, MdKCC and aquaporins MdDRIP1 and 2 were confirmed as being apical by proteomics of purified microvillar membranes. The role of NKCC and KCC on midgut water fluxes was tested observing the effect of the inhibitor furosemide. The change of trypsin distribution along the posterior midgut and the increase of trypsin excretion in the presence of furosemide lend support to the proposal that countercurrent fluxes power enzyme recycling and that the fluxes are caused by NKCC and KCC transporters helped by aquaporins.

Involvement of TRAF6 in regulating immune defense and ovarian development in Musca domestica.

The tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) is a key cytoplasm signaling adaptor that mediates signals activated by TNFR superfamily and the interleukin-1/Toll-like receptor (IL-1/TLR) superfamily. In the present research, a housefly Musca domestica TRAF6 (MdTRAF6) gene is identified and characterized, with a 51.7-kDa protein possessing a RING domain and a conserved C-terminal TRAF homology MATH domain encoded. MdTRAF6 is widely expressed in diverse tissues with high expression levels in gut and fat body, which is of the highest levels in adult in all growth stages. The expression of MdTRAF6 could be remarkably induced by bacterial challenge, and the silencing MdTRAF6 could alter the expressions of NF-κB-like genes (relish and dorsal) and antimicrobial peptide genes (cecropin, diptericin, attacin, muscin), thus leading elevated mortalities of larvae followed by bacterial infection. Inspiringly. MdTRAF6-depleted adult flies display higher mortality, lower fertility and reduced survival of offspring than the controls. Further investigation reveals that knockdown of MdTRAF6 disturbs the ovarian development and impaires the expressions of vitellogenin and vitellogenin receptor genes in the adult females. All these phenotypes show crucial roles of MdTRAF6 in innate immunity via positive regulation of the Toll pathway and negative regulation of the Imd pathway, and in reproduction by maintaining ovarian development.

Heat Adaptation of the House Fly (Diptera: Muscidae) and Its Associated Parasitoids in Israel.

Insects are ectothermic organisms; hence, all aspects of their biology are strongly influenced by ambient temperatures. Different insect species respond differently with phenotypic plasticity and/or genetic adaptation to changing temperatures. Here, we tested the thermal adaptation of the house fly and three of its parasitoids species by comparing life-history parameters in populations from a hot climate region (Jordan Valley) and from a moderate-climate region (Galilee). No significant differences were found between the two house fly populations, both under hot and moderate experimental conditions. Life-history parameters of the parasitoids (Muscidifurax raptor Girault & Sanders, Spalangia endius Walker, and Spalangia cameroni Perkins [Hymenoptera: Pteromalidae]) varied markedly between origins, species, sexes, and experimental conditions. Of the three species tested, only M. raptor collected in the Jordan Valley proved better adapted to experimental heat conditions, compared to its counterpart population that was collected in the Galilee. Additionally, we tested the effect of elevating temperatures on a house fly lab population for 17 consecutive generations and found no evidence for heat adaptation. We discuss our results in the context of house fly control and global warming.

Insecticidal and mosquito repellent efficacy of the essential oils from stem bark and wood of Hazomalania voyronii.

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.

Amorphous calcium phosphate in the pupal cuticle of Bactrocera dorsalis Hendel (Diptera: Tephritidae): A new discovery for reconsidering the mineralization of the insect cuticle.

It is now widely accepted that Hexapoda emerged from Crustacea. Compared to the ubiquitous calcified exoskeleton in crustaceans, a mineralized cuticle in insects is extremely rare. Catecholamine-driven protein cross-links play a leading role in the sclerotization of insect cuticle. In this study, mineralization was discovered in the pupal cuticle of Bactrocera dorsalis (Diptera: Tephritidae), a common pest of fruit farms. We mainly profiled the features of mineralized pupal cuticles from B. dorsalis and its white mutant B. dorsaliswh and unmineralized cuticle from Musca domestica using high-resolution field emission scanning electron microscopy (SEM) and transmission electron microscopy (TEM) combined with structural analysis involving infrared (IR) spectroscopy, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and synchrotron X-ray absorption near-edge structure (XANES) spectroscopy. We also compared the thermodynamic and mechanical properties of different pupal cuticles. The results showed that the pupal cuticles of B. dorsalis contain a phase of stable amorphous calcium phosphate (ACP) with a high level of magnesium, which is mainly distributed in the exocuticle and assists in the formation of a graded, stiffened cuticle structure. Unexpectedly, this ACP possesses a very low Ca/P ratio and has a composition similar to that of CaHPO4·2H2O. The degree of mineralization in the pupal cuticle of B. dorsaliswh (approximately 22 wt%) is significantly greater than that of wild-type B. dorsalis (approximately 12 wt%), which indicates that there may be a connection between the biomineralization and tyrosine-mediated tanning pathways. These findings provide new evidence for the mineralization of the insect cuticle, which may shed new light on the evolutionary mechanism underlying the divergence of cuticle sclerotization between insects and crustaceans.

Geographical Variations in Life Histories of House Flies, Musca domestica (Diptera: Muscidae), in Punjab, Pakistan.

Musca domestica Linnaeus is an important public health pest with the ability to adapt to diverse climates. Assessment of variations in biology and life-history traits of insects along geographical gradients is important for a successful management plan in different regions. We investigated life-history traits and life table parameters of M. domestica from six different geographical regions of Punjab, Pakistan: Rahim Yar Khan (RYK), Bahawalpur (BWP), Multan (MTN), Lahore (LHR), Gujrat (GJT), and Murree (MRE). Overall, M. domestica from localities of lower latitude and elevations with higher mean temperatures completed their development faster than those from localities of higher latitude and elevations with lower mean temperatures. The immature developmental time was the longest for the MRE population that was collected from higher latitude and elevation with cooler climate, whereas the shortest for the RYK population from lower latitude with warmer climate. Pupal weights were heavier for the RYK, BWP, and MTN populations, all were from the lowest latitude and elevations with warmer climate, compared with rest of the field populations. Similarly, rate of adult eclosion, fecundity, egg hatching, longevity, and life table parameters such as intrinsic rate of population increase, mean relative growth rate, net reproductive rate, and biotic potential were significantly higher for the RYK, BWP, and MTN populations compared with the GJT, LHR, and MRE populations of M. domestica. The current results will probably be of importance when planning management of M. domestica in different geographical regions of Pakistan.

Spatiotemporally different expression of alternatively spliced GABA receptor subunit transcripts in the housefly Musca domestica.

Insect γ-aminobutyric acid (GABA) receptors are important as major inhibitory neurotransmitter receptors and targets for insecticides. The housefly GABA receptor subunit gene MdRdl is alternatively spliced at exons 3 (a or b) and 6 (c or d) to yield the variants of ac, ad, bc, and bd combinations. In the present study, the expression of the MdRdl transcript in the body parts and in the developmental stages of the housefly Musca domestica was examined by quantitative polymerase chain reaction using specific primers that amplify the combinations of alternative exons. The results indicated that the transcripts of MdRdl, including four combinations, were highly expressed in the adult stage. MdRdlbd was the most abundant in the adult head. The expression pattern did not change in the adult stage over 7 days after eclosion. The expression level of the MdRdl bd transcript in the female head was similar to that of the male head. In contrast, MdRdl bc was the predominant transcript in the pupal head and the adult leg. Because the homomeric Rdl bc GABA receptor has a high affinity for GABA, our results provide grounds for designing agonist or competitive-antagonist insecticides that target the orthosteric site of the GABA receptor containing this Rdl variant.

Housefly (Musca domestica) larvae powder, preventing oxidative stress injury via regulation of UCP4 and CyclinD1 and modulation of JNK and P38 signaling in APP/PS1 mice.

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 Aß 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 Aß 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.

Molecular machinery of starch digestion and glucose absorption along the midgut of Musca domestica.

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.