Chronic low‐dose radiation inhibits cisplatin‐induced formation of tumorous clones in Drosophila melanogaster wts/ + heterozygotes

Ionizing radiation is one of the most extensively studied carcinogens. In contrast to the detrimental effects of high‐dose radiation in carcinogenesis, the biological effects of low‐dose radiation remains poorly understood. In this study, we introduced adult wts/ + heterozygotes of Drosophila melanogaster as transgenic model organisms to determine the tumorigenic activity of low‐dose radiation. The warts (wts) gene is a tumor suppressor gene in mice and humans that is directly involved in cell cycle regulation. Fruit flies at the first larval stage were subjected to ionizing radiation, and then tumorigenic activity was evaluated as the frequency of observed warts tumorous mosaic clones in adult flies. Low‐dose irradiation alone did not cause tumorigenesis in our system. In combined treatment with a chemical carcinogen, chronic irradiation at 0.2 Gy decreased the frequency of tumorous clones induced by cisplatin. These results suggest that low‐dose radiation alone is not deleterious but beneficial in tumorigenesis induced by a chemical carcinogen.     

Selection for divergent emigration rates in laboratory populations ofMusca domestica

High and low emigration rates through a laboratory system were selected for in populations of house flies (Musca domestica L.). Emigration consisted of movement of flies from on plastic box to another by way of a connecting tube. Selection was carried out by first dividing a wild population of flies into two lines and then selecting for movement from the box in one line and against movement from the box in the other line. The selection experiment was performed twice. In both experiments a statistically significant difference between the two strains was obtained in three to four generations of selection. In the second experiment, after 5 genrations the mean number of high emigration rate flies emigrating in 1 hour was 4.37 times the mean number of low emigration rate flies and in 24 hours was 2.81 times the number of low emigration rate flies. The second experiment was terminated after 6 generations, but the first experiment was continued for 35 generations. In this case, divergence ceased in roughly 15 generations. The possible relevance of the findings to laboratory population experiments in which spatial discontinuities are included is discussed.     

Reproductive strategy in Drosophila melanogaster: Significance of a genetic divergence between temperate and tropical populations

Summary Reproductive capacities of tropical and temperate populations of D. melanogaster were compared using three complementary techniques: (1) measure of egg production by females grown in the laboratory under uncrowded conditions and provided as adults with abundant food; (2) study of egg production of flies of unknown ages, collected in nature and then kept in similar conditions; and (3) analysis of ovarian activity of wild females dissected just after their capture.Tropical populations showed a lower fecundity in the laboratory and this was also observed in laboratory reared adults. On the average, flies also appeared to be older in the tropics than in temperate countries. These data, together with ecological observations showing that tropical populations live in a more predictable and stable environment, suggest that temperature populations are r-selected, while tropical ones are K-selected. The study of ovarian activity of wild females failed however to confirm this expectation. Tropical flies, which have a lower genetic fecundity, generally appeared to produce more propagules than did temperate flies. Such a contradiction shows how the ideas of r- and K-selection are difficult to apply to natural populations of Drosophila. Population density and interindividual competition are probably not the main selective forces in nature. Attention must also be paid to the necessity of exploring the environment to find resources, to the role of predation and parasitism, and to the occurrence in temperate countries of seasonal fluctuations with different selective pressures on successive generations.     

Influence of biotic and abiotic factors on the persistence of a Beauveria bassiana biopesticide in laboratory and high-rise poultry house settings

Recent studies suggest the potential for use of oil formulations of the entomopathogenic fungus, Beauveria bassiana, as residual sprays for control of house flies in poultry production facilities. The current study investigated the influence of biotic and abiotic factors on biopesticide persistence. We found that flies physically removed and deactivated conidia, with higher fly densities and greater cumulative exposure hastening the decline. Nonetheless, very low densities of viable conidia were still able to cause rapid mortality, suggesting the potential for relatively long re-treatment intervals as fly populations are controlled. Considering abiotic factors, we found that fungal spray treatments remained viable for up to 13 weeks under laboratory conditions. Periodic exposure of flies to the spray residue showed high levels of mortality, with very little decline in mortality rate over time. Equivalent treatments placed in a commercial poultry house showed much more rapid decline. One trial at the end of summer showed conidia to remain viable up to seven weeks. However, repeats during the winter months revealed decay in 1–2 weeks, with fly mortality rates influenced accordingly. The exact reasons for the more rapid decay remain unclear but could be linked to high concentrations of ammonia in the basement areas, especially during winter when ventilation is minimal. The combined data suggest the potential for adaptive treatment regimes with weekly spray intervals in conditions with very high fly populations and/or high ammonia levels, and potentially monthly spray intervals when fly populations and ammonia levels are reduced.     

The fly factor phenomenon is mediated by interkingdom signaling between bacterial symbionts and their blow fly hosts

We tested the recent hypothesis that the"fly factor"phenomenon(food cur-rently or previously fed on by flies attracts more flies than the same type of food kept inccessible to flies)is mediated by bacterial symbionts deposited with feees or regur-gitated by feeding flies.We allowed laboratory-reared black blow flies,Phormia regina(Meigen),to feed and de fecate on bacterial Luria-Bertani medium solidified with agar,and isolated seven morphologically distinct bacterial colonies.We identified these us-ing matrix-assisted laser desorption/ionization mass spectrometry and sequencing of the 165 rRNA gene.In two-choice laboratory experiments,traps baited with cultures of Pro-teus mirabilis Hauser,Morganella morganii subsp.sibonii Jensen,or Serratia marcescens Bizio,captured significantly more flies than corresponding control jars baited with tryptic soy agar only.A mixture of seven bacterial strains as a trap bait was more attractive to flies than a single bacterial isolate(M.m.siboni).In a field experiment,traps baited with agar cultures of P:mirabilis and M.m siboni in combination captured significantly more flies than lraps baited with either bacterial isolate alone or the agar control.As evident by gas chromatography-mass spectrometry,the odor profiles of bacterial isolates differ,which may explain the additive effect of bacteria to the attractiveness of bacterial trap baits.As"generalist bacteria,"P mirabilis and M.m.sibonii growing on animal protein(beef liver)or plant protein(tofu)are similarly effective in attracting flies.Bacteria-derived airborne semiochemicals appear to mediate foraging by flies and to inform their feeding and oviposition decisions.     

Bacterial growth properties at low optical densities

A method for accurate quantification of growth rate and yield of bacterial populations at low densities was developed with a modified version of a stepwise linear model for fitting growth curves based on optical density measurements, and adapted to measurements at low optical densities in 96-well microtiter plates. The method can be used for rapid and precise estimates of growth rate and yield, based on optical density measurements of large numbers of cultures of Escherichia coli. E. coli B lines were serially propagated at low glucose concentration during a long-term evolution experiment. Growth rate and yield of populations sampled from each of 12 lines that evolved for 20,000 generations under these conditions and two ancestral clones was measured. Populations were grown at three different glucose concentrations. Consistent with earlier findings, statistical analysis showed that both exponential growth rate and yield per unit of glucose differed significantly between the three glucose concentrations tested. Significant adaptation of the evolved populations to the nutrient conditions in which they evolved for 20,000 generations was observed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The spatial patterns of parasitism of eumenid wasps,Anterhynchium flavomarginatum andOrancistrocerus drewseni by the miltogrammine flyAmobia distorta

1. Spatial patterns of parasitism of eumenid wasps Anterhynchium flavomarginatum and Orancistrocerus drewseni by the miltogrammine fly Amobia distorta were studied in Kyoto, Japan during 1980–1984.
2. In generations of low (<5%) and medium (5–20%) parasitism, percent parasitism per shed (the habitat of the hosts) increased as a function of host density. Conversely, in generations of high (>20%) parasitism, percent parasitism was rather constant over different host densities.
3. The spatial distributions of adult miltogrammine flies among sheds were censused in generations of low and medium parasitism. The frequency of observations of adult miltogrammine flies was higher at sheds of higher host density (aggregative behavioral response), but on the other hand, the adult miltogrammine flies distributed in an underdispersed (or regular) manner in relation to other conspecifics.
4. The spatially density independent relationship between host density and percent parasitism in generations of high parasitism was explained in relation to parasitoid dispersal from patches of high parasitoid density.

     

Differentiation of New Metastatic Variants of B16 Melanoma Under Different Culture Conditions

The purpose of this study was to examine the differentiation of variant tumors of the B16 metastatic melanoma when tumors were grown serially under different culture conditions and transplanted into C57BL/6J black mice, lethal yellow A^y/a, albino c/c, and C⁺/c mutant mice. Morphological and biochemical markers of melanogenesis were examined in cells in culture and in the corresponding tumors. Cellular pigmentation was assessed in terms of the levels of DOPA and 5-S-CD and in terms of tyrosinase activity in the various cell lines and tumors. The observed change from high to low metastatic capacity, which was dependent on culture conditions, appeared to be unrelated to melanogenesis even though changes were observed in the biochemical melanotic phenotype. Overall, tumor cells from spontaneous pulmonary metastases appear to differentiate in ways that are unrelated to the instability of experimental metastatic capacity. The melanotic phenotype in albino c/c and C⁺/c mice was dependent on the phenotype of the parental tumors. A marked difference was observed between two pigmentation compartments, one of which was stable in the B16 control, while the other was unstable in YB16 and MB16 variant cells and in the tumors derived from them. It appears, therefore, that the metastatic capacity of B16 metastatic variants is changeable and is independent of the unstable melanogenic behavior. The production of metastases and the differentiation of tumors in the present experiments appeared to be related to the genetic background of the mice and the epigenetic metabolic environment of tumors and cells.     

Experimentally induced host‐shift changes life‐history strategy in a seed beetle

Expansion of the host range in phytophagous insects depends on their ability to form an association with a novel plant through changes in host‐related traits. Phenotypic plasticity has important effects on initial survival of individuals faced with a new plant, as well as on the courses of evolutionary change during long‐term adaptation to novel conditions. Using experimental populations of the seed beetle that evolved on ancestral (common bean) or novel (chickpea) host and applying reciprocal transplant at both larval and adult stage on the alternative host plant, we studied the relationship between the initial (plastic) phases of host‐shift and the subsequent stages of evolutionary divergence in life‐history strategies between populations exposed to the host‐shift process. After 48 generations, populations became well adapted to chickpea by evolving the life‐history strategy with prolonged larval development, increased body mass, earlier reproduction, shorter lifespan and decreased plasticity of all traits compared with ancestral conditions. In chickpea‐adapted beetles, negative fitness consequences of low plasticity of pre‐adult development (revealed as severe decrease in egg‐to‐adult viability on beans) exhibited mismatch with positive effects of low plasticity (i.e. low host sensitivity) in oviposition and fecundity. In contrast, beetles adapted to the ancestral host showed high plasticity of developmental process, which enabled high larval survival on chickpea, whereas elevated plasticity in adult behaviour (i.e. high host sensitivity) resulted in delayed reproduction and decreased fecundity on chickpea. The analysis of population growth parameters revealed significant fluctuation during successive phases of the host‐shift process in A. obtectus.     

Megaselia scalaris reared on Rhipicephalus (Boophilus) microplus laboratory cultures

Different laboratory cultures of the acarine tick Rhipicephalus (Boophilus) microplus (Canestrini, 1888) (Ixodida: Ixodidae) were infested by small Megaselia scalaris (Loew, 1866) (Diptera: Phoridae) flies. Larvae of this species exhibited opportunistic parasitism predominantly on engorged female ticks, causing severe damage to their cuticle through which the flies were able to reach R. microplus internal organs, on which they fed until developing into pupae in the tick's remains. The flies were kept by continuous propagation on fresh ticks over six generations during which the same parasitoid behaviour was observed. Here we report on an ixodid tick laboratory culture used for rearing M. scalaris.     

Density-dependent natural selection does not increase efficiency

Summary Populations ofDrosophila melanogaster kept at high population density (K-selected) for 125 generations have higher larval viability than populations kept at low densities (r-selected) when both are raised under crowded conditions. In additionK-selected adults that emerge from crowded cultures are larger than theirr-selected counterparts. These differences cannot be explained by differences in efficiency of food use. The minimum food required for successful pupation is actually greater in theK-selected populations. I conjecture that there may be a trade-off between minimum food requirements and competitive ability, which has changed substantially in theK-selected populations. The possibility thatK-selected larvae can dig more more deeply and gain access to unused food is examined and rejected as a possible explanation of the viability differences. Evidence is provided supporting the hypothesis that the differences in viability may be due to an increased tendency of theK-selected larvae to pupate off the surface of the medium.     

Spontaneous and Ethyl Methanesulfonate-Induced Mutations Controlling Viability in DROSOPHILA MELANOGASTER. II. Homozygous Effect of Polygenic Mutations

Polygenic mutations affecting viability were accumulated on the second chromosome of Drosophila melanogaster by treating flies with EMS in successive generations. The treated chromosomes were later made homozygous and tested for their effects on viability by comparison of the frequency of such homozygotes with that of other genotypes in the same culture. The treated wild-type chromosomes were kept heterozygous in Pm/+ males by mating individual males in successive generations to Cy/Pm females. The number of generations of accumulation was 1 to 30 generations, depending on the concentration of EMS. A similar experiment for spontaneous polygenic mutations was also conducted by accumulating mutations for 40 generations. The lower limit of the spontaneous mutation rate of viability polygenes is estimated to be 0.06 per second chromosome per generation, which is about 12 times as high as the spontaneous recessive lethal mutation rate, 0.005. EMS-induced polygenic mutations increase linearly with the number of treated generations and with the concentration of EMS. The minimum mutation rate of viability polygenes is about 0.017 per 10(-4)m, which is only slightly larger than the lethal rate of 0.013 per 10(-4) m. The maximum estimate of the viability reduction of a single mutant is about 6 to 10 percent of the normal viability. The data are consistent with a constant average effect per mutant at all concentrations, but this is about three times as high as that for spontaneous mutants. It is obvious that one can obtain only a lower limit for the mutation rate, since some mutants may have effects so near to zero that they cannot be detected. The possibility of measuring something other than the lower limit is discussed. The ratio of the load due to detrimental mutants to that caused by lethals, the D/L ratio, is about 0.2 to 0.3 for EMS-induced mutants, as compared to about 0.5 for spontaneous mutants. This is to be expected if EMS treatment produces a large fraction of small deletions and other chromosome rearrangements which are more likely to be lethal.     

Clinal and seasonal variations in initial retention capacity of virginDrosophila melanogaster females as a strategy for fitness

Summary The study of the first laboratory generation of isofemale lines ofDrosophila melanogaster, founded by wild flies collected in populations of various geographical origins, has shown that virgin females differ in their capacity to control egg deposition. This capacity to delay the ovulation process when no sexual partner is available is genetically determined. The present study shows that the frequency distribution of the various phenotypes varies progressively along a latitudinal cline from Afrotropical to European regions, or from neotropical to nearctic areas, while a few oriental populations appear off the cline. This variation in initial retention capacity, added to a similar variation in number of ovarioles conditioning a potential increase in fecundity for mated flies, induces variation in reactivity to insemination which may be an adaptation to a variable environment. Moreover, similar study of F1 virgin females from isofemale lines founded by wild flies collected in Spring, Summer and Autumn in two French populations over ten successive years revealed that seasonal genetic variations occur in this capacity to postpone egg-laying in case of non-insemination. The frequency of the different phenotypes follows cyclical seasonal variations. Long retention phenotypes are more frequent in Spring and Autumn samples, whereas short retention phenotypes, closer to the ancestral African type, are more common in Summer generations. This balancing selection temporarily modifies the genetic equilibrium of the population, enhancing its competitive ability. Temperature appears to be the primordial selective factor both in geographical and in seasonal variations.     

Infectivity of monoaxenic subcultures of the arbuscular mycorrhizal fungus Glomus versiforme associated with Ri-T-DNA-transformed carrot root

  Glomus versiforme was associated in vitro with Ri-T-DNA-transformed carrot root and after 4 months of cultivation, numerous axenic arbuscular mycorrhizal (AM) propagules were obtained. Three successive generations of spores and mycorrhizal root pieces were obtained by reassociating a 4-month-old root piece with a new carrot root. A biological test was conducted to assess the infectivity of the three generations of inoculum. Both for spores and mycorrhizal root pieces, a significant decrease in infectivity was observed among the successive generations. Mycorrhizal root pieces however, exhibited, higher infectivity than spores. These results show the possibility of maintaining AM fungi cultures in axenic conditions for a long time and raise questions about the loss of infectivity among the generations. The necessity of maintaining the same strains in vitro and in vivo in two separate collections is discussed. Received: 15 January 1996 / Received revision: 28 May 1996 / Accepted: 16 June 1996     

The contribution of ancestry,chance, and past and ongoing selection to adaptive evolution

The relative contributions of ancestry, chance, and past and ongoing election to variation in one adaptive (larval feeding rate) and one seemingly nonadaptive (pupation height) trait were determined in populations ofDrosophila melanogaster adapting to either low or high larval densities in the laboratory. Larval feeding rates increased rapidly in response to high density, and the effects of ancestry, past selection and chance were ameliorated by ongoing selection within 15–20 generations. Similarly, in populations previously kept at high larval density, and then switched to low larval density, the decline of larval feeding rate to ancestral levels was rapid (15-20 generations) and complete, providing support for a previously stated hypothesis regarding the costs of faster feeding inDrosophila larvae. Variation among individuals was the major contributor to variation in pupation height, a trait that would superficially appear to be nonadaptive in the environmental context of the populations used in this study because it did not diverge between sets of populations kept at low versus high larval density for many generations. However, the degree of divergence among populations (FST) for pupation height was significantly less than expected for a selectively neutral trait, and we integrate results from previous studies to suggest that the variation for pupation height among populations is constrained by stabilizing selection, with a flat, plateau-like fitness function that, consequently, allows for substantial phenotypic variation within populations. Our results support the view that the genetic imprints of history (ancestry and past selection) in outbreeding sexual populations are typically likely to be transient in the face of ongoing selection and recombination. The results also illustrate the heuristic point that different forms of selection-for example directional versus stabilizing selection—acting on a trait in different populations may often not be due to differently shaped fitness functions, but rather due to differences in how the fitness function maps onto the actual distribution of phenotypes in a given population. We discuss these results in the light of previous work on reverse evolution, and the role of ancestry, chance, and past and ongoing selection in adaptive evolution.     

Effects on ADH activity and distribution,following selection for tolerance to ethanol in Drosophila melanogaster

Strains of Drosophila melanogaster homozygous for either the Adh ^F or the Adh ^S allele were kept on food supplemented with ethanol for 20 generations. These strains (FE and SE) were tested for tolerance to ethanol and compared with control strains (FN and SN). The E strains showed increased tolerance to ethanol both in the adult and in the juvenile life stages. In adults the increase in tolerance was not accompanied by an increase in overall ADH activity. However, there were changes in the distribution of ADH over the body parts. Flies of the FE strain possessed significantly more ADH in the abdomen, compared with FN. Another set of FN and SN populations were started both on standard food and on ethanol food with reduced yeast concentrations. After 9 months ADH activities were determined in flies from these populations which had been placed on three different media: the food the populations had been kept on, regular food and regular food supplemented with ethanol. The phenotypic effects of yeast reduction on ADH activity were considerably, but longterm genetic effects were limited.     

A model for blood meal digestion and fat metabolism in male tsetse flies (Glossinidae)

Abstract. Fat and haematin levels of mature male Glossina morsitans morsitans Westwood were estimated at different times after feeding at temperatures between 15 and 30°C. Flies were kept (largely inactive) in 7.5 × 2.5 cm tubes, or in actograph cages, where flight activity increased with time after feeding. Haematin excretion was modelled as a series of three first order reactions, all with the same rate parameter. The model accounted for > 98% of the variance in mean haematin in each of seven experiments; the rate parameter increased linearly with temperature and activity level. A similar approach was adopted for modelling fat metabolism. The rate coefficients of lipogenesis increased with temperature, and that for lipolysis with temperature, activity level and their interaction. All experiments were analysed simultaneously to provide equations predicting haematin or fat levels for all times, for active or inactive flies, and for temperatures between 15 and 30°C. Haematin exhibited large variations between individuals, but for active flies the expected haematin content at a given time varied little between flies kept at 25 and at 30°C. In inactive flies kept at 25°C, lipogenesis peaked at ≈ 24 h and lipolysis at ≈ 48 h. For active flies the times were 12 and 24 h, respectively; both rates were about twice as high as in inactive flies. Active flies produced (up to 1 mg) more fat out of a given size of blood meal than inactive flies. Curves of fat content against logarithm of haematin content differed little with temperature, and can therefore be useful for comparative studies of field populations of tsetse.     

Single and multigenerational responses of body mass to atmospheric oxygen concentrations in Drosophila melanogaster : evidence for roles of plasticity and evolution

Greater oxygen availability has been hypothesized to be important in allowing the evolution of larger invertebrates during the Earth’s history, and across aquatic environments. We tested for evolutionary and developmental responses of adult body size of Drosophila melanogaster to hypoxia and hyperoxia. Individually reared flies were smaller in hypoxia, but hyperoxia had no effect. In each of three oxygen treatments (hypoxia, normoxia or hyperoxia) we reared three replicate lines of flies for seven generations, followed by four generations in normoxia. In hypoxia, responses were due primarily to developmental plasticity, as average body size fell in one generation and returned to control values after one to two generations of normoxia. In hyperoxia, flies evolved larger body sizes. Maximal fly mass was reached during the first generation of return from hyperoxia to normoxia. Our results suggest that higher oxygen levels could cause invertebrate species to evolve larger average sizes, rather than simply permitting evolution of giant species.     

Genetic and environmental control of melanotic tumors indrosophila

These investigations are concerned with the expression of the genemt which is responsible for the development of pseudotumors depending on different internal and external conditions.Chromosomal rearrangements such as inversions are powerful modifiers ofmt. The Y chromosome has a strongly suppressing effect which is evidently due to its heterochromatin content and particularly to its nucleolus organizer.The view is presented that inversions inDrosophila populations can be considered as master mutations which by themselves change the expression of a large number of genes affecting quantitative characters of ecological importance.High temperature decreases, low temperature increases the percentage of tumorous flies. There is also a profound effect of nutrition on the percentage of tumorous individuals, some substances, especially certain amino acids, furthering the development of tumors, other amino acids suppressing the development of tumors markedly.There are parental effects on the expression ofmt produced by genetic as well as environmental conditions; they may be more strongly paternal than maternal.There are also grandparental effects due to genetic as well as environmental influences. It was not tested if the extrachromosomal transmission of the changedmt expression lasts beyond the second generation.In a discussion of the results it is argued that a slight change in the entire system of message transmission, such as the RNA content of the cell, is more important in terms of differentiation and evolution than the profound change in an individual message due to the mutation of a single gene or DNA particle.This recearch was partly supported by NIH Grant GM 701-05.     

Activity rhythm of Drosophila kept in complete darkness for 1300 generations

Locomotor activity rhythms of dark stock flies of Drosophila melanogaster kept in complete darkness for 700 to 1340 generations were examined. The stock was established by the late Prof. S. Mori in November 1954 to investigate long-term effects of darkness on organisms. The activity of flies was recorded under three types of light conditions: DD after LD12:12, and DD after exposure to a 3.5 h (P3) or 7.5 h (P7) light pulse. In all of these conditions, the experimental dark flies exhibited clear circadian rhythms similar to those of control light flies. We compare our results with those of various studies on troglobites.