Reversed selection responses in small populations of the housefly (Musca domestica L.)

We compared the efficacy of artificial and natural selection processes in purging the genetic load of perpetually small populations. We subjected replicate lines of the housefly (Musca domestica L.), recently derived from the wild, to artificial selection for increased mating propensity (i.e., the proportion of male–female pairs initiating copulation within 30 min) in efforts to cull out the inbreeding depression effects of long-term small population size (as determined by a selection protocol for increased assortative mating). We also maintained parallel non-selection lines for assessing the spontaneous purge of genetic load due to inbreeding alone. We thus evaluated the fitness of artificially and ‘naturally’ purging populations held at census sizes of 40 individuals over the course of 18 generations. We found that the artificially selected lines had significant increases in mating propensity (up to 46% higher from the beginning of the protocol) followed by reversed selection responses back to the initial levels, resulting in non-significant heritabilities. Nevertheless, the ‘naturally’ selected lines had significantly lower fitness overall (a 28% reduction from the beginning of the protocol), although lower effective population sizes could have contributed to this effect. We conclude that artificial selection bolstered fitness, but only in the short-term, because the inadvertent fixation of extant genetic load later resulted in pleiotropic fitness declines. Still, the short-term advantage of the selection protocol likely contributed to the success of the speciation experiment since our recently-derived housefly populations are particularly vulnerable to inbreeding depression effects on mating behavior.     

Superoxide dismutase in the housefly,Musca domestica (L.)

Four superoxide dismutase (SOD) (E.C. 1.15.1.1) isozymes were present in whole tissue homogenates of Musca domestica when examined by polyacrylamide gel electrophoresis. One of the isozymes contained manganese, and the other three contained copper and zinc. All were observed in each of the body tagma (head, abdomen, and thorax) and at each developmental stage (egg to adult). The copper- and zinc-containing isozymes purified from newly emerged, adult M. domestica had a relative molecular weight of 34,800 as determined by gel filtration chromatography but consisted of two equal-size subunits of 16,000 as measured by sodium dodecylsulfate polyacrylamide gel electrophoresis. An isoelectric point between 4.8 and 5.1 was measured. Approximately 2 mol each of copper and zinc were present per dimer. The three copper, zinc isozymes were identified as charge variants. The amino acid composition of the enzyme was similar to that of copper, zinc-containing superoxide dismutases from other sources. Purified housefly copper, zinc superoxide dismutase was neither deactivated nor able to protect lactic dehydrogenase against deactivation in the presence of light and rose bengal, a known generator of singlet oxygen. The role of SOD in the phototoxic reaction involving rose bengal is discussed.     

Characterization of prostaglandin synthesis in the housefly,Musca domestica (L.)

The in vitro formation of prostaglandins (PG) was examined in the housefly Musca domestica. PG synthetase activity was detected in homogenates of whole insects and in head and thorax, abdomen, ovary and male reproductive tissues. Studies to determine the sub-cellular localization of PG synthetase indicated that the microsomal fraction contained the highest activity. Products obtained from radiolabeled arachidonic acid (20:4) and 8,11,14-eicosatrienoic acid (20:3, n-6) were PGE₂ and PGE₁, respectively, with lower amounts of the PGF series also present. In microsomal preparations from whole insects and reproductive tissues from both males and females, 20:3(n-6) was 2–2.5 times more efficiently converted to PG than was 20:4. Non-steroidal anti-inflammatory drugs (NSAID) fed to houseflies did not inhibit PG production from 20:4, whereas when they were included in microsomal preparations at high levels, they inhibited PG synthesis.     

Morphometric variations in the housefly,Musca domestica (L.) with latitude

Morphologic analysis of geographic strains of Musca domestica carried out on natural and laboratory experiments starting with 400 and 800 eggs showed phenotypic variations related with latitude. Females of the natural populations showed clines for several morphological traits of the wing, whereas male flies showed a reduction in the dispersion measures (s ² and CV) of wing width and length. The same reduction was obtained for males, females and total number of flies of the natural populations in dispersion of the number of bristles on the fourth abdominal sternite. A significant negative correlation was observed for the head width of females and for the total number of flies emerged in the laboratory experiments started with 400 eggs. All flies produced by the experiments starting with 800 eggs showed a reduction in variability of dispersion of the bristles on the fourth abdominal sternite in the strains obtained from locations south of the area analyzed. Evolutionary aspects of these correlation coefficients between morphometric traits and latitude are discussed.     

Ordered membrane particles in rhabdomeric microvilli of the housefly (Musca domestica L.)

Rhabdomeric microvilli of the housefly were freeze-fractured (FF) and thin sectioned (TS) for ultrastructural examination. Ordered files of closely packed membrane particles (82 Å wide, 250 Å long) were seen (FF) on the microvillar membrane (usually E face). The long axis of each particle was canted about 45° to that of the microvillus. Occasionally particles in this array appeared on the P face. It is hypothesized that ordered particles may represent either a photopigment precursor stock, a second photolabile pigment, or the newly discovered sensitizing, UV-absorbing, photostable visual pigment. In the underlying membrane leaflet (P face) were found spherical (85 Å diameter) unoriented particles in a concentration of about 6,000/μm². The size, shape and density of these structures are compatible with those of rhodopsin particles. These particles also covered the basal area of each microvillus. The findings from TS material were difficult to correlate with those from FF replicas. At high magnification the former showed that the plasma membrane of the transected microvillus is composed of spherical, hollow subunits (averaging 43 Å diameter), sometimes fused to form double, 86 Å units. These substructures were closely packed and continuous around the microvillus. This beaded plasma membrane, in rare cases, was doubled around the microvillus. In other instances the plasma membranes were continuous between neighboring microvilli. The physiological implications of these ultrastructural features are discussed.     

Occurrence and metabolism of prostaglandins in the housefly,Musca domestica (L.)

Prostaglandins (PGs) of the E and F series were quantified in the housefly by radioimmunoassay (RIA). Whole insects and reproductive tissues from both sexes contained PGE₍₁₊₂₎ and PDG_2α which increased in amount with age. PGF_2α levels were higher than PGE series in extracts of whole male and female insects and in ovaries. Male reproductive tissues contained higher amounts of PGE₍₁₊₂₎ than PGF_2α. Gas chromatographic-mass spectrometric (GC-MS) analyses of the products formed after injection of arachidonic acid (20:4) and eicosatrienoic acid (20:3, n-6) into male and femal insects demonstrated the conversion of 20:4 to PGF_2α and 20:3 to PGF_1α. Radiolabeled 20:4 injected into houseflies was rapidly converted to PGE₂ and PGF_2α. The catabolism of PGE₂ was more rapid than PGF_2α in males, whereas in females, PGE₂ and PGF_2α were converted to more polar products at similar rates. Radiolabeled 20:4 injected in the hemolymph was incorporated into the reproductive tracts of male insects. About 2.1% of the total radioactivity from [³H]20:4 injected into males just prior to mating was transferred to females during mating. Thus, PG are formed from 20:4 in male and female houseflies. During mating, 20:4 is transferred from males to females where it can be metabolized to PGF_2α.     

Insecticide-insensitive acetylcholinesterase from a laboratory selected and a field strain of housefly (Musca domestica) (L.)

1. Acetylcholinesterase from the heads of a strain of houseflies selected for resistance to the carbamate insecticide methomyl, and from a methomyl-resistant field strain was found to be less sensitive to inhibition by methomyl than that from a susceptible strain. 2. The enzyme from resistant insects was also more tolerant to malaoxon, dichlorvos and bomyl but not to azamethiphos. 3. The decrease in sensitivity to inhibition appeared to be due to an increase in affinity for substrate.     

Testing alternative methods for purging genetic load using the housefly (Musca domestica L.)

When a population faces long-term inbreeding, artificial selection, in principle, can enhance natural selection processes for purging the exposed genetic load. However, strong purge pressures might actually decrease fitness through the inadvertent fixation of deleterious alleles and allelic combinations. We tested lines of the housefly (Musca domestica L.) for the effectiveness of artificial selection to promote the adaptation to small population size. Specifically, replicate populations were held at average census sizes of 54 for nine generations or 30 for 14 generations while being subjected to artificial selection pressure for increased fitness in overall mating propensity (i.e., the proportion of virgin male–female pairs initiating copulation within 30 min), while also undergoing selection to create differences among lines in multivariate components of courtship performance. In the 14-generation experiment, a subset of the lines were derived from a founder-flush population (i.e., derived from three male–female pairs). In both experiments, we also maintained parallel non-selection lines to assess the potential for natural purging through serial inbreeding alone. Sub-populations derived from a stock newly derived from the wild responded to artificial selection for increased mating propensity, but only in the short-term, with eventual rebounds back to the original levels. Serial inbreeding in these lines simply reduced mating propensity. In sub-populations derived from the same base population, but 36 generations later, both artificial selection and serial inbreeding increased mating propensity, but mainly to restore the level found upon establishment in the laboratory. Founder-flush lines responded as well as the non-bottlenecked controls, so we base our major conclusions on the comparisons between fresh-caught and long-term laboratory stocks. We suggest that the effectiveness of the alternative purge protocols depended upon the amount of genetic load already exposed, such that prolonged periods of relaxed or altered selection pressures of the laboratory rendered a population more responsive to purging protocols.     

Mitochondrial diversity of Musca domestica housefly populations in the Asian and western Pacific biogeographical regions

Houseflies (Musca domestica L., Diptera: Muscidae) are cosmopolitan, colonizing, and eusynanthropic. Their distribution in the Malaysian archipelago provides an opportunity to study successive waves of colonization and extinction during the Pleistocene and Recent epochs. We scored single-strand conformation polymorphisms (SSCPs) at 16S2 and COII mitochondrial loci in 47 housefly samples from the Australian, Austro-Malayan, Indo-Malayan, Manchurian and Indo-Chinese subregions of Wallace's zoogeographical classification. We discuss the results in light of the Pleistocene vs. post-Pleistocene dispersal and faunal exchange in the Asia-Pacific area. Fourteen haplotypes were detected, of which 10 were confined to a single subregion. No haplotype was ubiquitous and only one was found in four subregions. Population diversity, HS, was greatest in the Indo-Malayan (0.36) and heterogeneous among subregions. The mean subregional diversity was 0.21 +/- 0.03, representing the probability that two randomly chosen flies, from any subregion, had different haplotypes. The hierarchical partition of diversity indicated restricted maternal gene flow among subregions (GRT = 0.60, Nm approximately 0.32). These results suggest long-standing genetic isolation of houseflies in the Malaysian archipelago and support the hypothesis that they dispersed widely during the Pleistocene. Haplotypes common among mainland populations but shared with island groups in low frequencies (<1%) indicate surprisingly little recent gene flow.     

The genetic basis of malathion resistance in housefly (Musca domestica L.) strains from Turkey

Organophosphate insecticide (parathion/diazinon) resistance in housefly (Musca domestica L.) is associated with the change in carboxylesterase activity. The product of MdalphaE7 gene is probably playing a role in detoxification of xenebiotic esters. In our research, we have isolated, cloned and sequenced the MdalphaE7 gene from 5 different Turkish housefly strains. High doses of malathion (600 microg/fly) were applied in a laboratory environment for one year to Ceyhan1, Ceyhan2, Adana and Ankara strains while no insecticide treatment was performed in the laboratory to Kirazli strain. Trp251 --> Ser substitution was found in the product of MdalphaE7 gene in all malathion resistant and Kirazli stocks. In addition, we checked the malathion carboxylesterase (MCE), percent remaining activities in acetylcholinesterase (AChE), glutathion-S-transferase (GST), and general esterase activities in all 5 strains used in this study. In comparing with universal standard sensitive control WHO, a high level of MCE and GST activities were observed while lower level of general esterase activities was detected in the tested strains. In addition, a higher percent remaining activities in AChE than WHO susceptible strain were observed in all malathion resistant strains.     

Some effects of CGA 72662 on larval development in the housefly,Musca domestica (L.)

Summary

Houseflies reared on artificial diet supplemented with a range of concentrations of CGA 72662 suffered severe developmental disruption at the larval stage. Death was preceded by the appearance of a range of cuticular lesions and abnormalities. These cuticular abnormalities have been observed under the light and electron microscopes.     

Polymorphism of M factors in populations of the housefly, Musca domestica L., in Turkey

M factors, which determine maleness in Musca domestica, were found on the second, third, fourth and fifth linkage groups in housefly populations of Turkey. As in European populations, the male-determining factor was more frequently located on linkage group III (MIII). Some males homozygous or double heterozygous for M factors were identified. Deviations from a 1:1 sex ratio in favour of males, as well as mosaics for somatic marker mutations and sexual mosaics (gynandromorphs), were also observed. The results reveal an extensive polymorphism in the sex-determining system.     

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 domestica L．幼虫在胰化酪蛋白大豆卵黄琼脂（TrypticaseSoy EggYolkAgar,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Myroidesodoratimimus的TSEYA培养基中生长时间最短,仅需97．61&#177;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&#177;0．23mg／个;4）除铜绿假单胞菌Pseudomonasaeruginosa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Bacillus subtilis、香味类香味菌、聚团肠杆菌Enterobacter agglomerans以及成团肠杆菌Pantoeaagglomerans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Staphylococcusaurous、成团肠杆菌、大肠杆菌Escherichiacoli、奇异变形杆菌Proteusmirabilis以及香味类香味菌中各种营养成分进行分析,结果发现,6种细菌均能提供大量的维生素如（0．105～1．08g／kg）。在氨基酸方面,香味类香味菌和枯草芽孢杆菌中的10种昆虫必需氨基酸含量与总氨基酸含量之比（Essential amino acid／Totalaminoacid,EAA／TAA）最高,而金黄色葡萄球菌最低。这个比例与蛹重呈正相关（p=0．031）。在脂肪酸相对含量方面,金黄色葡萄球菌具有最高的饱和脂肪酸含量（76．38％）,香味类香味菌含有56．79％的不饱和脂肪酸,而枯草芽孢杆菌则具有最高的支链脂肪酸含量（42．16％）。     

The male-determining activity on the Y chromosome of the housefly (Musca domestica L.) consists of separable elements.

In the common housefly, the presence or absence of a male-determining factor, M, is responsible for sex determination. In different strains, M has been found on the Y, on the X, or on any of the five autosomes. By analyzing a Y-autosomal translocation and a ring-shaped, truncated Y chromosome, we could show that M on the Y consists of at least two regions with M activity: One of them can be assigned to the short arm of the Y chromosome (MYS), which is largely C-banding negative, the other region lies on the C-banding positive long arm of the Y, including the centromeric part (MYL). Each region alone behaves as a hypomorphic M factor, causing many carriers to develop as intersexes of the mosaic type instead of as males. When introduced into the female germ line by transplantation of progenitor germ cells (pole cells), the MYS shows an almost complete maternal effect that predetermines 96% of the genotypic female (NoM) animals to develop as males. In contrast, the MYL has largely lost its maternal effect, and most of the NoM animals develop as females. Increasing the amount of product made by either of the two hypomorphic M factors (by combining the MYS and MYL or two MYS) leads to complete male development in almost every case. We thus assume that the Y chromosome carries at least two copies of M, and that these are functionally equivalent.