Microsomal metabolism of juvenile hormone analogs in the house fly,Musca domestica L.

Of six juvenile hormone analogs of the alkyl 3,7,11-trimethyl-2,4-dodecadienate type, only the isopropyl ester was strongly morphogenic in the house fly, Musca domestica L. In vitro assays revealed that house fly microsomes contain B-esterases as well as oxidases which metabolize such analogs. However, these esterases did not hydrolyze the isopropyl ester, ZR-515. Enzymes prepared from larvae, pupae, and adults were all active and there was evidence that in the late larval stage the esterase activity was cyclic, showing a minimum in the early third instar and a maximum a few hours later. When microsomes from two susceptible and two resistant house fly strains were compared for metabolic activity against the juvenile hormone analogs, those from the resistant strains were 1.3 to 20 × higher in oxidase activity but there was no difference in esterase activity. The oxidative metabolism of two analogs ZR-515 and 512 was greatly enhanced when the flies were induced with phenobarbital but there was no enhancement in metabolism of three of the remaining analogs and only a slight enhancement of a fourth. It is concluded that the insecticidal action of ZR-515 is largely due to its stability in the presence of the house fly esterases.     

细菌诱导家蝇蛹抗菌物质的分离纯化

家蝇生境中充满病原菌,其有效的防防机制成为研究的热点。通过三氟乙酸缓冲液浸提、热处理、固相萃取以及两次RP-HPLC等方法,从混合细菌液诱导的家蝇蛹中分离纯化出4种具有抗菌活性的物质。结果表明,提取的家蝇蛹粗提液抗菌谱较广,对多种微生物具有不同程度的抗菌作用,且抗菌效果好于天然肽类防腐剂和一些化学防腐保鲜剂。4种抗菌物质中包括3个抗菌肽和1个抗菌化合物。Tricine-SDS-PAGE结果显示3个抗菌肽的分子量分别约为12kDa、8kDa和4kDa。本研究表明细菌诱导家蝇蛹中具有丰富的抗菌物质。     

Genetic studies on glutathione-dependent reactions in resistant strains of the house fly,Musca domestica L

Genetic studies of glutathione-dependent reactions were conducted with a diazinon-resistant house fly strain in which resistance is controlled primarily by genes on chromsome II. The resistant strain was crossed with a susceptible strain which had mutant markers on chromosomes II, III, and V, and the F₁ was backcrossed to the susceptible strain. Glutathione transferase activities of the resultant eight phenotypes were measured using 3,4-dichloronitrobenzene, methyl iodide, and γ-benzene hexachloride as substrates. High levels of all these activities are controlled by gene(s) on chromosome II. Further analysis was made by introducing diazinon resistance into a susceptible strain via genetic crossing-over. Intermediate activity levels for 3,4-dichloronitrobenzene and methyl iodide conjugations were introduced along with intermediate levels of resistance. Assays of individual flies of the synthesized strain revealed they were heterogeneous for glutathione-dependent activities, consisting of individuals with low, intermediate, and high transferase activity. Based on these results, high levels of the glutathione-dependent enzymes are not a major biochemical mechanism responsible for diazinon resistance. It was also demonstrated that glutathione S-aryltransferase and S-alkyltransferase in the house fly, as measured with 3,4-dichloronitrobenzene and methyl iodide, are inseparable genetically and may, therefore, be the same enzyme.     

Genetic and biochemical mechanisms limiting fipronil toxicity in the LPR strain of house fly,Musca domestica

Fipronil is a new insecticide which exerts its toxic action by interacting with the insect GABA-gated chloride channel. Previous studies have shown that cyclodiene-resistant insects have low to moderate levels of cross-resistance to fipronil, while other resistant strains are usually susceptible. In contrast, we recently found a strain (LPR) of house fly (Musca domestica L) with 15-fold cross-resistance to fipronil that was not associated with cyclodiene resistance. Fipronil cross-resistance in LPR was inherited as an intermediately dominant, autosomal, multigenic trait. [¹⁴C]Fipronil was observed to penetrate into LPR flies more slowly than into susceptible flies. S,S,S-tributylphosphorotrithioate and diethyl maleate pretreatment did not reduce the level of fipronil cross-resistance, while piperonyl butoxide resulted in a slight decrease. These results indicate that decreased penetration and monooxygenase-mediated detoxification may be mechanisms contributing to fipronil cross-resistance in the LPR strain. © 1999 Society of Chemical Industry     

Monitoring of resistance of insecticides in house fly (Musca domestica) populations in hungary

Samples of 24 house fly (Musca domestica L.) populations were collected from animal farms in Hungary in 1990 and kept in the laboratory to determine their susceptibility to different types of insecticide: organochlorines, organophosphates, carbamates, pyrethroids, macrocyclic lactone and insect growth regulators. The adulticides were tested with topical bioassay in all 24 populations, the larvicides were studied with treated larval medium in 16 populations. The data were expressed as LD₅₀ and LC₅₀ values (ng fly ?1 and mg kg ?1 larval medium respectively). The percentages of populations which had resistance ratios > 10 at LD₅₀ or LC₅₀ were: 63% to DDT, 50% to methoxychlor, 13% to lindane, 83% to malathion, 63% to trichlorfon, 4% to propetamphos, 96% to dioxacarb, 46% to propoxur, 4% to methomyl, 13% to pyrethrum, 96% to bioresmethrin, 63% to permethrin, 58% to cypermethrin, 79% to SK-80, 79% to deltamethrin, 38% to invermectin, 0% to diflubenzuron, 0% to cyromazine. Correlation analysis showed a high degree of positive correlation among the adulticides except for ivermectin, bioresmethrin and SK-80. No cross-resistance was found between the larvicides and the conventional adulticides. Differences of insecticide resistance levels among the populations surveyed were studied by principal component and factor analysis. A fairly good relationship between resistance status and control practices used on farms was revealed. The populations originating from those farms where the application of adulticides had been frequent or regular and where high resistance was shown to most chemicals could be separated from the others.     

Insecticide metabolism by multiple glutathione S-transferases in two strains of the house fly,Musca domestica (L)

Glutathione S-transferases from two strains of house fly have been prepared in a high degree of homogeneity by a procedure involving affinity chromatography and isoelectrofocusing. They fall into two groups in each strain. One group, of isoelectric point greater than pH 6.5, catalyzes the glutathione-dependent degradation of lindane, diazinon and methyl parathion. The other group, of low isoelectric point, has conjugating activity with the model substrate CDNB, but very little activity with the insecticide substrates. In the Cornell R strain the three isoenzyme forms in the high pI group appear to be almost identical in their substrate specificities. In the A strain, it is apparent that the enzyme forms falling into this group vary markedly in substrate specificity. The dehydrochlorination of DDT paralleled very closely the conjugation of the other insecticides catalyzed by the three high pI enzymes in the Cornell R strain. In the A strain, DDT dehydrochlorinase was most strongly associated with a glutathione S-transferase isoelectric at pH 7.1. It is tentatively concluded that multiple genes are involved in the production of the glutathione S-transferases involved in pesticide metabolism in the house fly and that DDT dehydrochlorinase may be derived from some, but not all, of these same genes.     

The potential use of allicin as a biopesticide for the control of the house fly,Musca domestica L.

The house fly, Musca domestica L., is a widespread pest of intensively reared livestock, where its presence negatively affects growth and productivity through the irritation their activity causes. Furthermore, adult flies mechanically vector a wide range of animal and human pathogens and, as such, pose a health risk to both livestock and people. The development of resistance in M. domestica populations to most of the insecticides used against them, coupled with diminishing product availability in many regions, means that new fly control methods and materials are constantly required. In the present study, a formulation of allicin, a compound derived from garlic cloves, was evaluated against the eggs, larvae and adults of Musca domestica L.. An in-diet LD₅₀ of 134 ppm against larvae was achieved, whilst marked ovicidal activity was also recorded. Adults were similarly affected when exposed directly and indirectly to surface residues of the compound. The results indicate that allicin exhibits potential to be used against filth flies through incorporation into their developmental substrates and, potentially, for the control of other economically important dipteran pests.     

Toxicity of spinosad to susceptible and resistant strains of house flies,Musca domestica

The toxicity of spinosad, a new insecticide derived from the bacterium Saccharopolyspora spinosa, was evaluated against susceptible and resistant strains of house fly (Musca domestica L.). Spinosad was highly toxic to house flies based on 72-h LD₅₀ values and the symptoms of poisoning were consistent with a neurotoxic mechanism of action. Spinosad was relatively slow acting, with the maximum toxicity noted at 72 h. Piperonyl butoxide and S,S,S,-tribu-tylphosphorotrithioate synergized the toxicity of spinosad by 3·0- and 1·8-fold, respectively, while diethyl maleate had no significant effect. These results suggest that there is a small degree of monooxygenase-mediated spinosad detoxification in house flies, while hydrolases may be only minimally important and glutathione transferases may have no role. There were no substantial levels of cross-resistance detected, except in the LPR strain where a low 4·3-fold cross-resistance was observed. The cyclodiene-resistant OCR strain was 2·7-fold more sensitive to spinosad than the susceptible strain (CS). These results suggest that cross-resistance may not be a limiting factor for the use of spinosad against house flies. © 1998 Society of Chemical Industry     

Behavioural and electrophysiological investigation of 1-(7-ethoxygeranyl)-2-methylbenzimidazole on the house fly Musca domestica

The insecticidal properties of 1-(7-ethoxygeranyl)-2-methylbenzimidazole (EGMB) were investigated on larval and adult house flies. Unsynergised EGMB gave topical LD₅₀ values of 0.53 μg per female fly on NAIDM strain house flies. When flies were pretreated with 5.2 μg piperonyl butoxide, susceptibility was increased (LD₅₀ 0.12 μg per female fly). House fly larvae were less susceptible to EGMB (LD₅₀ 2.2 μg). Poisoning with EGMB resulted in a rapid reduction in locomotor activity of both larval and adult house flies. This reduction in locomotion was progressive and led to complete paralysis. Various parameters of larval nervous system function were investigated in larvae during these early phases of poisoning. As early as 15 min after dosing larvae with LD₉₅ doses of EGMB, sensory nerves were less responsive. Over a somewhat longer time (2–4 h), neurally evoked contractures were adversely affected by EGMB. In some cases, this effect appeared to be due to reduced postsynaptic potential amplitude; in other instances, it appeared to be due to an effect independent of neuromuscular transmission. The close temporal correlation between behavioural and electrophysiological observations suggests that the nervous and muscular systems are important sites of action of EGMB.     

First report of cyromazine resistance in a population of UK house fly (Musca domestica) associated with intensive livestock production

BACKGROUND: House fly control in livestock‐rearing facilities is heavily reliant on the use of the larvicide cyromazine. While extensive use of this compound has led to the development of resistance in several countries, no elevated tolerance has so far been reported from the United Kingdom. RESULTS: Tolerance to cyromazine in larvae derived from a field strain collected at an intensive pig unit was significantly elevated over that of insects taken from a susceptible laboratory strain. Resistance factors (RFs) of 2.9 and 2.4 were returned for assays initiated with eggs and neonate larvae respectively. The RF for field strain larvae exposed from neonate increased significantly to 3.9 and 5.6 following rounds of selection at 1.0 and then 1.5 mg kg^?1 cyromazine. CONCLUSION: Low‐level resistance to cyromazine in UK house flies is reported here for the first time. The geographic extent of this resistance is unknown but, if widespread, may lead to control failures in the future, and indicates that careful stewardship of this compound in the United Kingdom is now required. © Crown copyright 2010. Reproduced with permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.     

Depolarization of motor nerve terminals by pyrethroids in susceptible and kdr-resistant house flies

When applied at concentrations of one nM or higher to a house fly larval neuromuscular preparation, deltamethrin (DM) and fenvalerate (FV) greatly increased miniature excitatory postsynaptic potential (mepsp) rate and blocked neuromuscular transmission. The DM-induced mepsp discharge was abolished by tetrodotoxin (TTX), removal of Ca²⁺ from the saline, or by application of hyperpolarizing stimuli to the nerve, indicating that it was due to depolarization of the presynaptic terminals. Also, in the presence of TTX, K⁺ depolarization increased mepsp rate at the same external K⁺ concentration before and after DM treatment, confirming that DM released transmitter by depolarizing the nerve terminals rather than by altering the voltage dependence of transmitter release. The potassium channel blocker tetraethylammonium (TEA) increased mepsp rate somewhat, while aconitine (20 μM), which keeps sodium channels open, increased mepsp rate consistently. Pretreatment of nerves with a subthreshold dose of TEA greatly increased the mepsp rate-increasing activity of DM and aconitine, while a subthreshold level of aconitine did not synergize DM. These observations suggest that DM, like aconitine, depolarized nerves by modifying the sodium channels. Knockdown resistant (kdr) larvae were resistant to the depolarizing action of DM and aconitine but not to that of TEA, indicating that the kdr gene produced a modified sodium channel which was less sensitive to the action of pyrethroids and aconitine. During sustained transmitter release by DM, evoked release gradually declined, resulting in a condition called early block in which spontaneous release was high and release could be evoked by electrotonic depolarization of the nerve terminals, but not by a nerve action potential. Early block was probably due to conduction block in the nerve terminals. Early block eventually gave way to late block, characterized by the decline of spontaneous release to subnormal levels and complete failure of evoked release. After late block, the calcium ionophore X-537A could not release transmitter, suggesting that late block was due to depletion of available transmitter. DM did not have a direct effect upon extrasynaptic muscle membrane. However, after late block, muscles were left insensitive to the putative transmitters glutamate and aspartate when these were bath or iontophoretically applied. A low rate of mepsps persisted after late block, indicating that the muscles were still sensitive to the natural transmitters.     

Determining the source of house flies (Musca domestica) using stable isotope analysis

BACKGROUND: Intensive livestock units frequently produce flies in large numbers that, on migration, cause nuisance to the occupants of neighbouring dwellings. The resolution of such problems is often reliant on the unequivocal identification of the origin of the flies, particularly when several potential sources exist. This study evaluated stable isotope analysis as a method for differentiating adult houseflies (Musca domestica) on the basis of their dietary history so as to determine their likely source. RESULTS: Flies were reared in the laboratory on several substrates, including chicken and cattle manure, laboratory diet and household vegetable waste. Different fly parts (wings, heads and legs) and whole flies were analysed immediately after eclosion and after 10 days. The δ¹³C and δ¹⁵N values for adults that had developed on each diet type were highly distinct. Both isotopic ratios altered markedly after maintaining the flies for 10 days on a diet of cane sugar solution. CONCLUSIONS: Stable isotope analysis readily differentiated flies that had developed on a range of substrates. The technique, therefore, shows potential to be employed to determine the likely source of various nuisance insects, and to contribute to the abatement of such problems. Copyright © 2011 Society of Chemical Industry     

麦麸饲料中细菌对家蝇幼虫生长发育的影响

从饲料用麦麸中分离出的12种细菌均能支持无菌家蝇Musca domesticaL.幼虫在胰化酪蛋白大豆卵黄琼脂(Trypticase Soy Egg Yolk Agar,TSEYA)培养基中完成整个生长发育过程。1)幼虫在接种香味类香味菌Myroides odoratimimus的TSEYA培养基中生长时间最短,仅需97.61±1.14h;2)幼虫在接种醋酸钙不动杆菌Acinetobacter calcoaceticus的TSEYA培养基中的化蛹率可达到86.81%;3)从接种嗜水汽单胞菌Aeromonas hydrophila的TSEYA培养基中得到的蝇蛹重量最高,达到20.15±0.23mg/个;4)除铜绿假单胞菌Pseudomonas aeruginosa饲养的家蝇羽化率较低(60.87%)外,其余各种细菌饲养的羽化率在84.33%～97.47%之间。此外,枯草芽孢杆菌Bacillussubtilis、香味类香味菌、聚团肠杆菌Enterobacter agglomerans以及成团肠杆菌Pantoea agglomerans可作为单一营养来源支持幼虫完成整个生长发育过程。对枯草芽孢杆菌、金黄色葡萄球菌S...     

Susceptibility to thiamethoxam of Musca domestica from Danish livestock farms

BACKGROUND: Neonicotinoid baits are currently replacing anticholinesterase baits for control of adult houseflies (Musca domestica L.). Introduction of new insecticides includes evaluation of their cross-resistance potential, which was assessed for thiamethoxam in field populations from Denmark. RESULTS: In feeding bioassay with a susceptible strain, thiamethoxam LC(50) at 72 h was 1.7 microg thiamethoxam g(-1) sugar, making it 19-fold, 11-fold and threefold more toxic to houseflies than azamethiphos, methomyl and spinosad respectively. The field populations were 6-76-fold resistant to thiamethoxam. There was no correlation between the toxicities of thiamethoxam and spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The toxicity in feeding bioassay at 72 h of imidacloprid in a susceptible strain was 32 microg imidacloprid g(-1) sugar at LC(50), making it 19-fold less toxic to houseflies than thiamethoxam. There was a strong significant correlation between the toxicities of thiamethoxam and imidacloprid in field populations.CONCLUSION: Neonicotinoid-resistant houseflies were present at a detectable and noticeable level before thiamethoxam and imidacloprid were introduced for housefly control in Denmark. The toxicity of thiamethoxam is explained by other parameters than the toxicities of spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The cross-resistance between thiamethoxam and imidacloprid indicates a coincidence of mechanism of the toxicity and resistance in the field populations.     

Metabolism of permethrin isomers in American cockroach adults,house fly adults,and cabbage looper larvae

Forty-two insect metabolites of [1RS,trans]-and [1RS,cis]-permethrin are tentatively identified in studies with Periplaneta americana adults, Musca domestica adults, and Trichoplusia ni larvae involving administration of ¹⁴C preparations labeled in either the alcohol or acid moieties. The less-insecticidal trans isomer is generally metabolized more rapidly than the more-insecticidal cis isomer, particularly in cabbage looper larvae, and metabolites retaining the ester linkage appear in larger amount with cis-permethrin. Although the dichlorovinyl group effectively blocks oxidation in the acid side chain, the permethrin isomers are metabolized by hydrolysis and hydroxylation at the geminal-dimethyl group (either trans- or cis-methyl substituent) and the phenoxybenzyl group (predominantly at the 4′-position in all species but also at the 6-position in house flies). The alcoholic and phenolic metabolites are excreted as glucosides, and the carboxylic acids are excreted as glucosides and amino conjugates (glycine, glutamic acid, glutamine, and serine) with considerable species variation in the preferred conjugating moiety.     

Genetics of resistance of pyrethroid-selected houseflies,Musca domestica L.

Four resistance factors were isolated genetically from the NPR strain of houseflies (Musca domestica L.), which resists natural pyrethrins, and were characterised toxicologically. The four factors were : pen, which reduces the rate of penetration of insecticides through the cuticle; kdr-NPR, a general pyrethroid resistance mechanism unaffected by the synergist sesamex; py-ses, a mechanism of resistance to natural pyrethrins that can be suppressed by sesamex; and py-ex, a factor that gives strong resistance to synergised natural pyrethrins and to the new synthetic esters, e.g. resmethrin, but little or none to natural pyrethrins alone.     

Carbofuran triggers flight motor output in pyrethroid-blocked reflex pathways of the house fly

The initiation of flight from loss of tarsal contact (flight reflex responses) and the tergotrochanteral muscle (TTM) responses evoked by brain stimulation were analyzed during carbofuran, permethrin, deltamethrin, and DDT poisoning in the house fly. Blockage of the flight reflex by LD₅₀ doses of permethrin or deltamethrin was rapid, but the effects of DDT on the flight reflex took hours to develop. In addition, carbofuran treatment induced spontaneous flight in blocked preparations by an action in the central nervous system. This result suggests that pyrethroid blockage of the flight reflex was due to an action on sensory nerves, since the central flight program and its associated efferent systems were functionally intact. The relevance of this finding in terms of pyrethroid knockdown is discussed. The TTM response was unaffected by permethrin or deltamethrin both early and late in the poisoning process, possibly because the evoked TTM response does not involve peripheral sensory nerves, which seem to be important sites of pyrethroid action early in poisoning. Carbofuran induced repetitive firing and blockage of the TTM response within 1 hr, but normal responses were observed late in poisoning, which is consistent with the reversibility of carbamate inhibition of acetylcholinesterase. DDT caused no change in the evoked TTM response until bursts were recorded about 15 hr after treatment; this was another example of a slowly developing DDT effect. The protracted development of various DDT actions was concordant with a hypothesis of reduced efficacy at a proposed target site, viz., the sodium channels of nerve membranes.     

Linkage group analysis of nerve insensitivity in a pyrethroid-resistant strain of house fly

Nerve insensitivity was a major factor of mechanism of resistance to pyrethroids in the 228e2b strain of house fly. Reciprocal crosses between the resistant and susceptible SRS strains showed that resistance to permethrin was recessive in nerve sensitivity. Linkage group analysis by the F₁ male backcross method using multichromosomal marker strains was investigated electrophysiologically, following 10⁻⁵ M permethrin application to the exposed thoracic ganglia of the backcross progenies. Results of the experiment showed that the recessive genetic factor responsible for nerve insensitivity to permethrin in the resistant strain is located only on the third chromosome.