Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

In this study, the knockdown and mortality effects of imidacloprid and methomyl were investigated. The residual surface applications were carried out to determine the knockdown effects (KDt₅₀ and KDt₉₅) and mortality (LD₅₀ and LD₉₅) induced by each insecticide. For mortality comparisons, the susceptible house fly (Musca domestica L., Diptera: Muscidae) of a WHO population and three natural field‐collected M. domestica populations from Turkey were used. In conclusion, it was found that the resistance to imidacloprid and methomyl was significantly higher in the field populations when compared to the susceptible population from WHO. The results showed that applicators and pest management decision‐makers should control and conduct an integrated pest management strategy by including biological agents to prevent the development of high levels of resistance in the field populations.     

Mutations in the house fly Vssc1 sodium channel gene associated with super-kdr resistance abolish the pyrethroid sensitivity of Vssc1/tipE sodium channels expressed in Xenopus oocytes

The super-kdr insecticide resistance trait of the house fly confers resistance to pyrethroids and DDT by reducing the sensitivity of the fly nervous system. The super-kdr genetic locus is tightly linked to the Vssc1 gene, which encodes a voltage-sensitive sodium channel alpha subunit that is the principal site of pyrethroid action. DNA sequence analysis of Vssc1 alleles from several independent super-kdr fly strains identified two amino acid substitutions associated with the super-kdr trait: replacement of leucine at position 1014 with phenylalanine (L1014F), which has been shown to cause the kdr resistance trait in this species, and replacement of methionine at position 918 with threonine (M918T). We examined the functional significance of these mutations by expressing house fly sodium channels containing them in Xenopus laevis oocytes and by characterizing the biophysical properties and pyrethroid sensitivities of the expressed channels using two-electrode voltage clamp. House fly sodium channels that were specifically modified by site-directed mutagenesis to contain the M918T/L1014F double mutation gave reduced levels of sodium current expression in oocytes but otherwise exhibited functional properties similar to those of wildtype channels and channels containing the L1014F substitution. However, M918T/L1014F channels were completely insensitive to high concentrations of the pyrethroids cismethrin and cypermethrin. House fly sodium channels specifically modified to contain the M918T single mutation, which is not known to exist in nature except in association with the L1014F mutation, gave very small sodium currents in oocytes. Assays of these currents in the presence of high concentrations of cismethrin suggest that this mutation alone is sufficient to abolish the pyrethroid sensitivity of house fly sodium channels. These results define the functional significance of the Vssc1 mutations associated with the super-kdr trait of the house fly and are consistent with the hypothesis that the super-kdr trait arose by selection of a second-site mutation (M918T) that confers to flies possessing it even greater resistance than the kdr allele containing the L1014F mutation.     

Survey of insect growth regulator (IGR) resistance in house flies (Musca domestica L.) from southwestern Turkey

Insect growth regulators (IGRs) are currently the fastest‐growing class of insecticides, and in Turkey these products represent a new approach to pest control. In recent years, several IGRs were also registered for the control of the house fly, Musca domestica L. (Diptera: Muscidae), in Turkey. A field survey was conducted in the summers of 2006 and 2007 to evaluate resistance to some agriculturally and medically used IGRs on house flies from livestock farms and garbage dumps in the greenhouse production areas (Merkez, Kumluca, Manavgat, and Serik) of Antalya province (Southwestern Turkey). The results of larval feeding assay with technical diflubenzuron, methoprene, novaluron, pyripoxyfen, and triflumuron indicate that low levels (RF<10‐fold) of resistance to the IGRs exist in the house fly populations from Antalya province. Exceptions, however, were two populations, Guzoren and Toptas, from the Kumluca area which showed moderate resistance to diflubenzuron with 11.8‐fold in 2006 and 13.2‐fold in 2007, respectively. We found substantial variation in susceptibility of field‐collected house fly populations from year to year and from product to product. We generally observed an increase in resistance at many localities sampled from 2006 to 2007. The implications of these results to the future use of IGRs for house fly control are discussed. It will be critically important to continue monitoring efforts so that appropriate steps can be taken if resistance levels start to increase.     

Joint action of l-leucine and sodium phosphate buffer solutions as feeding stimulants for protein-deprived females of the house fly Musca domestica

ABSTRACT. Potentiation in joint action was demonstrated between solutions of L-leucine and sodium phosphate buffer (pH 6.3) as feeding stimulants for protein-deprived females of the house fly, Musca domestica L. Both components alone elicited feeding. In two-choice feeding tests, mixtures consisting of equi-stimulating concentrations of the two components were taken in greater quantities than either component alone at twice the concentration in the mixture.
The presence of 1×10^-1 M phosphate buffer markedly lowered the threshold for detection of L-leucine. The presence of phosphate buffer strengthened the preferences shown by flies given choices of concentrations of L-leucine differing by a factor of 2 and enabled them to display preferences at lower concentrations.
The presence of 1×10^-3 M L-leucine increased, somewhat, the ability of flies to detect low concentrations of phosphate buffer. Its presence had relatively little effect on the strength of preference shown between two-fold differences in concentration of phosphate buffer when the higher concentration was 6.3×10^-3 M or less, but markedly strengthened the preferences when the higher concentration was 2.5×10^-2M or greater. Leucine increased the optimal concentration of phosphate buffer by a factor of more than 2 and converted 2×10^-1 M phosphate buffer from a mild feeding deterrent to a powerful feeding stimulant.     

Innate control of local search behaviour in the house fly, Musca domestica

Abstract. The genetic basis of variation in local searching behaviour in the house fly, Musca domestica L., was examined by estimating the heritability of locomotory parameters using parent—offspring regressions. Pathways of adult flies were videotaped for 3 min after ingestion of a 1 μl drop of 0.25 M sucrose in a 75 cm diameter circular arena with minimal external cues. Locomotory variables computed from digitized walking pathways, including locomotory rate, turning rates per unit of distance and time, turn bias, number of stops, and stop duration, were significantly heritable, but heritabilities (additive genetic variance ÷ total phenotypic variance) was generally low (<0.25). Phenotypic variance in search parameters increased with time since ingestion of the sucrose drop, but heritabilities were highest when only the first 30 s of the searching bout were examined, and declined as longer periods were considered. Significant genetic correlations among search parameters suggest that many of these characters are influenced pleiotropically by the same genes or closely linked genes. However, some aspects of local search were inherited independently (e.g. stop duration and locomotory rate). The independence of controlling mechanisms was supported by observed differences between genetic and environmental correlations. This genetic independence could enhance the likelihood of appropriate population responses to temporally or spatially changing selective forces.     

Host age and pathogen exposure level as factors in the susceptibility of the house fly,Musca domestica (Diptera: Muscidae) to Beauveria bassiana

Adult house flies, Musca domestica L., of four ages, <1, 3, 7, and 14 day post-eclosion, were exposed to three strains of Beauveria bassiana (P89, L90 and 447). Flies were exposed to moistened filter paper treated with either a low (1.57×10⁴ conidia/cm²) or high (1.57×10⁵ conidia/cm²) concentration of each fungal strain for 6 h. Strain 447 was superior to the two house fly-derived B. bassiana strains in inducing host infection and mortality. Significant spikes in infection and mortality occurred as early as 5 days post-exposure with higher concentration exposures acting more quickly. Few differences were observed in either infection or mortality among the four fly age classes. On Day 10 post-exposure, 77% of the high-concentration, 447-exposed flies were infected, compared with only 24% of the flies from the P89 low-concentration exposure. Potential applications of these results in integrated house fly management programs are discussed.     

Three mutations identified in the voltage-sensitive sodium channel alpha-subunit gene of permethrin-resistant human head lice reduce the permethrin sensitivity of house fly Vssc1 sodium channels expressed in Xenopus oocytes

Point mutations in the para-orthologous sodium channel alpha-subunit of the head louse (M815I, T917I, and L920F) are associated with permethrin resistance and DDT resistance. These mutations were inserted in all combinations using site-directed mutagenesis at the corresponding amino acid sequence positions (M827I, T929I, and L932F) of the house fly para-orthologous voltage-sensitive sodium channel alpha-subunit (Vssc1(WT)) gene and heterologously co-expressed with the sodium channel auxiliary subunit of house fly (Vsscbeta) in Xenopus oocytes. The double mutant possessing M827I and T929I (Vssc1(MITI)/Vsscbeta) caused a approximately 4.0mV hyperpolarizing shift and the triple mutant, Vssc1(MITILF)/Vsscbeta, caused a approximately 3.2mV depolarizing shift in the voltage dependence of activation curves. Vssc1(MITI)/Vsscbeta, Vssc1(TILF)/Vsscbeta, and Vssc1(MITILF)/Vsscbeta caused depolarizing shifts ( approximately 6.6, approximately 7.6, and approximately 8.8mV, respectively) in the voltage dependence of steady-state inactivation curves. The M827I and L932F mutations reduced permethrin sensitivity when expressed alone but the T929I mutation, either alone or in combination, virtually abolished permethrin sensitivity. Thus, the T929I mutation is the principal cause of permethrin resistance in head lice. Comparison of the expression rates of channels containing single, double and triple mutations with that of Vssc1(WT)/Vsscbeta channels indicates that the M827I mutation may play a role in rescuing the decreased expression of channels containing T929I.     

Role of prestomal teeth in feeding of the house fly, Musca domestica (Diptera; Muscidae)

Ultrastructural studies of the mouthparts of Musca domestica L. show that the fly's prestomal teeth are more damaging to host tissues than considered previously. When fed on tissue culture or pig cornea, SEM revealed that M. domestica were able to tear and suck up cells. This process occurs so rapidly, and so few cells are involved, that the damage is not perceptible to the naked eye. Except for the prestomal teeth, the mouthparts have few other structures which can inflict the damage observed.     

Mechanisms of regulation of epithelial sodium channel by SGK1 in A6 cells

The serum and glucocorticoid induced kinase 1 (SGK1) participates in the regulation of sodium reabsorption in the distal segment of the renal tubule, where it may modify the function of the epithelial sodium channel (ENaC). The molecular mechanism underlying SGK1 regulation of ENaC in renal epithelial cells remains controversial. We have addressed this issue in an A6 renal epithelial cell line that expresses SGK1 under the control of a tetracycline-inducible system. Expression of a constitutively active mutant of SGK1 (SGK1T(S425D)) induced a sixfold increase in amiloride-sensitive short-circuit current (Isc). Using noise analysis we demonstrate that SGK1 effect on Isc is due to a fourfold increase in the number of functional ENaCs in the membrane and a 43% increase in channel open probability. Impedance analysis indicated that SGK1T(S425D) increased the absolute value of cell equivalent capacitance by an average of 13.7%. SGK1T(S425D) also produced a 1.6-1.9-fold increase in total and plasma membrane subunit abundance, without changing the half-life of channels in the membrane. We conclude that in contrast to aldosterone, where stimulation of transport can be explained simply by an increase in channel synthesis, SGK1 effects are more complex and involve at least three actions: (1) increase of ENaC open probability; (2) increase of subunit abundance within apical membranes and intracellular compartments; and (3) activation of one or more pools of preexistent channels within the apical membranes and/or intracellular compartments.     

The role of intracellular sodium in the regulation of NMDA-receptor-mediated channel activity and toxicity

Sodium (Na⁺) is the major cation in extracellular space and, with its entry into cells, may act as a critical intracellular second messenger that regulates many cellular functions. Through our investigations of mechanisms underlying the activity-dependent regulation of N-methyl-d-aspartate (NMDA) receptors, we recently characterized intracellular Na⁺ as a possible signaling factor common to processes underlying the upregulation of NMDA receptors by non-NMDA glutamate channels, voltage-gated Na⁺ channels, and remote NMDA receptors. Furthermore, although Ca²⁺ influx during the activation of NMDA receptors acts as a negative feedback mechanism that downregulates NMDA receptor activity, Na⁺ influx provides an essential positive feedback mechanism to overcome Ca²⁺-induced inhibition, thereby potentiating both NMDA receptor activity and inward Ca²⁺ flow. NMDA receptors may be recruited to cause excitoxicity through a Na⁺-dependent mechanism. Therefore, the further characterization of mechanisms underlying the regulation of NMDA receptors by intracellular Na⁺ is essential to understanding activity-dependent neuroplasticity in the nervous system.     

Evaluation of Beauveria bassiana applications against adult house fly, Musca domestica, in commercial caged-layer poultry facilities in New York state

Applications of a commercially produced Beauveria bassiana product, balEnce, were compared with pyrethrin treatments for the control of adult house flies in New York high-rise, caged-layer poultry facilities. An integrated fly management program, which included the release of house fly pupal hymenopteran parasitoids, was used at all facilities. Adult house fly populations were lower in B. bassiana-treated facilities during the spray and post-spray periods, as recorded on spot cards. Concurrently, the numbers of house fly larvae recovered in B. bassiana-treated facilities were less than one-half that of the pyrethrin-treated facilities. House fly pupal parasitism levels were low, but similar under both treatment regimes. The numbers of adult and larval Carcinops pumilio, a predatory beetle, recovered from B. bassiana-treated facilities were 43 and 66% greater than from the pyrethrin-treated facilities, respectively.     

Development and physiology of follicular atresia during ovarian growth in the house fly, Musca domestica

Atretic follicles regularly occur in the ovary of the house fly, Musca domestica. The frequency of ovarian follicular atresia and the proportion of atretic follicles per ovary are related to the stage of oögenesis and to the age of the females. Only vitellogenic follicles may become atretic. The atresia may occur at any stage of vitellogenesis, though most follicles become atretic in mid-vitellogenesis. Atretic follicles are completely resorbed within 24–36 hr. The follicle cells may play a synthesizing role during growth and disintegrating one during follicle resorption. The induction of glycogen synthesis by the cessation of RNA and protein synthesis and by vitellogenesis in normal follicles is discussed. The same processes occur prematurely in the atretic follicle which can be thus distinguished by a high content of glycogen.     

Effects of disassembly of actin microfllaments on the AVP-induced regulation of sodium channel densities in frog skin epithelium

Vasopressin (AVP) stimulates the absorption of Na⁺ across frog skin epithelium by increasing the number of open apical channels (N_o) mainly through a large expansion of the total number of channels (N_T) at the membrane. This study investigates with blocker-induced noise analysis the potential role of actin in the regulation of AVP-induced changes in channel densities. Particularly we examined the idea that actin regulates the insertion of channels into apical cell membranes, consistent with the model for its hydrosmotic action. Treating cells with cytochalasin B (CB) for 2 h to disassemble the prominent subapical actin network did not significantly alter the stimulation of the I_sc by AVP. Importantly, the treatment had no significant effect on the AVP-induced activation of Na⁺ channels, nor on the increases in N_T indicating that an intact actin network is not required for the natriferic actions on the channels. This result is disparate from well known effects of CB on the AVP-induced hydrosmotic response and we assume that these distinct responses are produced by different mechanisms. Other mechanisms need to be considered to explain the mechanism whereby new Na⁺ channels are recruited to the apical membrane. In particular, mechanisms of intracellular trafficking involved in the redistribution of epithelial Na⁺ channels remain unresolved and need to be refined.     

Proliferation of embryonic cardiomyocytes in zebrafish requires the sodium channel scn5Lab

In mice, homozygous deletion of the cardiac sodium channel Scn5a results in defects in cardiac morphology and embryonic death before robust sodium current can be detected. In zebrafish, morpholino knockdown of cardiac sodium channel orthologs scn5Laa and scn5Lab perturbs specification of precardiac mesoderm and inhibits growth of the embryonic heart. It is not known which developmental processes are perturbed by sodium channel knockdown and whether reduced cell number is from impaired migration of cardiac progenitors into the heart, impaired myocyte proliferation, or both. We found that embryos deficient in scn5Lab displayed defects in primary cardiogenesis specific to loss of nkx2.5, but not nkx2.7. We generated kaede reporter fish and demonstrated that embryos treated with anti‐scn5Lab morpholino showed normal secondary differentiation of cardiomyocytes at the arterial pole between 30 and 48 h post‐fertilization. However, while proliferating myocytes were readily detected at 48 hpf in wild type embryos, there were no BrdU‐positive cardiomyocytes in embryos subjected to anti‐scn5Lab treatment. Proliferating myocytes were present in embryos injected with anti‐tnnt2 morpholino to phenocopy the silent heart mutation, and absent in embryos injected with anti‐tnnt2 and anti‐scn5Lab morpholinos, indicating cardiac contraction is not required for the loss of proliferation. These data demonstrate that the role of scn5Lab in later heart growth does not involve contribution of the secondary heart field, but rather proliferation of cardiomyocytes, and appears unrelated to the role of the channel in cardiac electrogenesis. genesis 51:562–574. © 2013 Wiley Periodicals, Inc.