THE GENETICS OF CENTRAL AND MARGINAL POPULATIONS OF DROSOPHILA SERRATA. II. HYBRID BREAKDOWN IN FITNESS COMPONENTS AS A CORRELATED RESPONSE TO SELECTION FOR DESICCATION RESISTANCE

Multiple-peak epistasis is one of the four premises that underlie Wright's shifting-balance theory of evolution. A selection experiment was conducted in an attempt to push different geographic populations to different fitness peaks as a correlated response to selection for an additively controlled character (desiccation resistance). Four populations of Drosophila serrata, sampled from central and marginal areas of its distribution along a 3000-km stretch of Australia's east coast, underwent selection for desiccation resistance for 14 generations. After selection had ceased, control lines from each of the populations were crossed to determine the amount of hybrid breakdown that existed before selection and selected lines were crossed to determine the amount of hybrid breakdown after selection. Hybrid breakdown was measured in three fitness traits: developmental time, viability, and fecundity. When the individual crosses were examined, virtually no evidence was found for hybrid breakdown between these populations. However, the level of hybrid breakdown in development time in the control lines increased as the distance between the populations in the field increased. This relationship was lost in the selected lines. Therefore, selection for desiccation resistance influenced the level of hybrid breakdown in a fitness trait, although selection may need to be maintained for longer than 14 generations if a new relationship between hybrid breakdown and distance is to be formed.     

Large Genetic Change at Small Fitness Cost in Large Populations of Drosophila Melanogaster Selected for Wind Tunnel Flight: Rethinking Fitness Surfaces

The fitness effects of extreme genetic change by selection were studied in large populations subjected to prolonged, intense selection. Two replicate populations of Drosophila melanogaster, with estimated effective sizes 500 <= N(e) <= 1000, were selected for increased performance in a wind tunnel, selecting on average the fastest 4.5% of flies. The mean apparent flying speed of both lines increased from ~2 to 170 cm/sec and continued to respond at diminishing rates, without reaching a plateau, for 100 generations. Competitive fitness tests in generations 50 and 85 showed minimal or no fitness loss in selected lines compared to controls. Sublines relaxed in generations 65 and 85 showed minimal or no regression in apparent flying speed. Hybrid lines, from a cross of selected X control lines in generation 75, responded to reselection saltationally, showing that the chromosomes of the selected lines had been assembled from alleles at many loci, from many different chromosomes in the base population. Thus, major genetic change was achieved, but without the costs usually associated with strong directional selection. Large population size has been interpreted, in opposing models, as either a brake or an accelerator in its effects on long-term change by selection. These results favor the second model, and challenge the concept of rugged fitness surfaces underlying the first model.     

Evidence for a robust sex-specific trade-off between cold resistance and starvation resistance in Drosophila melanogaster

In insects changes in lipid metabolism may underlie a trade-off between cold resistance and starvation resistance. To test this we examined correlated responses in independent sets of Drosophila melanogaster lines selected for increased cold resistance and increased starvation resistance. The starvation lines showed correlated patterns found in other D. melanogaster populations selected for this trait, including higher lipid levels and increased resistance to desiccation, although the selected lines did not show a longer development time as found in some other studies. Consistent with the trade-off hypothesis, selected lines with increased starvation resistance showed decreased resistance to a cold stress as measured by mortality, whereas selected lines with increased cold resistance showed a decrease in starvation resistance. To counter the possibility of inadvertent selection accounting for these patterns, selected and control lines from both selection regimes were crossed to form mass bred populations, which were left for four generations prior to establishing isofemale lines. By scoring starvation and cold resistance in these lines derived from both sets of selection regimes, we confirmed the negative association between resistance to these stresses in females but not in males. Potential implications of this trade-off for surviving cold conditions when food resources are limiting are discussed.     

MULTIPLE GENETIC MECHANISMS FOR THE EVOLUTION OF SENESCENCE IN DROSOPHILA MELANOGASTER

We present the results of selection experiments designed to distinguish between antagonistic pleiotropy and mutation accumulation, two mechanisms for the evolution of senescence. Reverse selection for early-life fitness was applied to laboratory populations of Drosophila melanogaster that had been previously selected for late-life fitness. These populations also exhibited reduced early-age female fecundity and increased resistance to the stresses of starvation, desiccation, and ethanol, when compared to control populations. Reverse selection was carried out at both uncontrolled, higher larval rearing density and at controlled, lower larval density. In the uncontrolled-density selection lines, early-age female fecundity increased to control-population levels in response to the reintroduction of selection for early-age fitness. Concomitantly, resistance to starvation declined in agreement with previous observations of a negative genetic correlation between these two characters and in accordance with the antagonistic-pleiotropy mechanism. However, resistance to stresses of desiccation and ethanol did not decline in the uncontrolled-density lines during 22 generations of reverse selection for early-life fitness. The latter results provide evidence that mutation accumulation has also played a role in the evolution of senescence in this set of Drosophila populations. No significant response in early-age fecundity or starvation resistance was observed in the controlled-density reverse-selection lines, supporting previous observations that selection on Drosophila life-history characters is critically sensitive to larval rearing density.     

Response to selection for rapid chill-coma recovery in Drosophila melanogaster: physiology and life-history traits

Resistance to low temperatures can vary markedly among invertebrate species and is directly related to their distribution. Despite the ecological importance of cold resistance this trait has rarely been studied genetically, mainly because low and variable fitness of offspring from cold-stressed mothers makes it difficult to undertake selection experiments and compare cold resistance of parents and offspring. One measure of cold resistance that varies geographically in Drosophila melanogaster and that is amenable to genetic analysis is chill-coma recovery. Three replicate lines of D. melanogaster were selected every second generation, for over 30 generations, for decreased recovery time following exposure to 0 degrees C. Correlated responses were scored to characterize underlying physiological traits and to investigate interactions with other traits. Lines responded rapidly to the intermittent selection regime with realized heritabilities varying from 33% to 46%. Selected lines showed decreased recovery time after exposure to a broad range of low temperatures and also had a lower mortality following a more severe cold shock, indicating that a general mechanism underlying cold resistance had been selected. The selection response was independent of plastic changes in cold resistance because the selected lines maintained their ability to harden (i.e. a short-term exposure to cool temperature resulted in decreased recovery time in subsequent chill-coma assays). Changes in cold resistance were not associated with changes in resistance to high temperature exposure, and selected lines showed no changes in wing size, development time or viability. However, there was a decrease in longevity in the selected lines due to an earlier onset of ageing. These results indicate that chill-coma recovery can be rapidly altered by selection, as long as selection is undertaken every second generation to avoid carry-over effects, and suggest that lower thermal limits can be shifted towards increased cold resistance independently of upper thermal limits and without tradeoffs in many life-history traits.     

Adaptation to fluctuating environments in a selection experiment with Drosophila melanogaster

A fundamental question in life‐history evolution is how organisms cope with fluctuating environments, including variation between stressful and benign conditions. For short‐lived organisms, environments commonly vary between generations. Using a novel experimental design, we exposed wild‐derived Drosophila melanogaster to three different selection regimes: one where generations alternated between starvation and benign conditions, and starvation was always preceded by early exposure to cold; another where starvation and benign conditions alternated in the same way, but cold shock sometimes preceded starvation and sometimes benign conditions; and a third where conditions were always benign. Using six replicate populations per selection regime, we found that selected flies increased their starvation resistance, most strongly for the regime where cold and starvation were reliably combined, and this occurred without decreased fecundity or extended developmental time. The selected flies became stress resistant, displayed a pronounced increase in early life food intake and resource storage. In contrast to previous experiments selecting for increased starvation resistance in D. melanogaster, we did not find increased storage of lipids as the main response, but instead that, in particular for females, storage of carbohydrates was more pronounced. We argue that faster mobilization of carbohydrates is advantageous in fluctuating environments and conclude that the phenotype that evolved in our experiment corresponds to a compromise between the requirements of stressful and benign environments.     

LATITUDINAL CLINE IN DROSOPHILA MELANOGASTER FOR KNOCKDOWN RESISTANCE TO ETHANOL FUMES AND FOR RATES OF RESPONSE TO SELECTION FOR FURTHER RESISTANCE

We have introduced a device for selecting Drosophila for increased resistance to very high concentrations of ethanol fumes. This device has enabled us to: 1) select quickly and easily over a thousand flies at a time, and 2) score the knockdown time of every fly in the distribution, while causing very little injury to the flies. A sample of nine west coast populations of Drosophila melanogaster showed a significant trend toward higher knockdown resistance in more northern populations. A population's level of knockdown resistance was virtually uncorrelated with its alcohol dehydrogenase (Adh) allele frequencies. Five of the above nine populations were then subjected to selection for further knockdown resistance. Each population was divided randomly into four groups of 256 flies: two lines to be selected, and two lines to remain unselected as control lines. In every generation each selected line was measured for knockdown resistance, and the last quartile of flies to be knocked down was saved to continue the selection cycle. Population sizes of the selected and unselected lines were all maintained at 256. Realized heritability, based on the responses to selection of the first four generations, was calculated for each selected line. The five populations were significantly heterogeneous for heritability estimates; the average heritability of the five populations pooled was 0.143 ± 0.019. Over the course of twelve generations, the ten selected lines increased their knockdown times by an average factor of 2.40. Before selection, the five populations were heterogeneous for knockdown resistance, and resistance was greatest among the most northern populations. The amount of change of knockdown resistance over the course of selection was also correlated with latitude: the most southern population increased its knockdown time by a factor of 2.23, and the most northern population increased it by a factor of 2.55. After ten generations of selection, the cline of knockdown resistance was about 4.5 times as steep as that before selection. Small phenotypic differences among populations before selection were thus exaggerated by the action of selection. The differences among populations in their rates of response to selection were attributed to genetic differences that existed before selection. The pattern of change of Adh frequencies over the course of selection was very inconsistent, both among and within populations. From this inconsistency of change of Adh alleles with selection, and the lack of correlation between Adh frequencies and knockdown resistance before selection, we concluded that Adh frequency changes could not have had much effect on the responses of the selected lines.     

Evolution of pre- and post-copulatory traits in female Drosophila melanogaster as a correlated response to selection for resistance to cold stress

Exposure to low temperatures reduces gamete viability and fecundity in females of insect species like Drosophila. Hence, adaptation to cold stress can in principle involve modifications in reproductive traits in females. Studies on resistance to cold stress have mostly addressed the evolution of adult survivorship post cold shock. Very few studies have addressed the evolution of reproductive traits in females in response to cold stress. We have successfully selected replicate populations of Drosophila melanogaster for resistance to cold shock. After 50 generations of selection, we investigated pre- and post-copulatory traits i.e. mating latency, copulation duration, mating frequency and progeny production in female flies exposed to cold shock or control conditions. Post cold shock, females from the selected populations were better at recovery in terms of mating latency, mating success, and progeny production relative to females from the control populations. Performance of the two types of females was not different under control conditions. These findings clearly indicate that adaptation to cold stress involves rapid modification of the reproductive traits.     

Thermal adaptation in<Emphasis Type="Italic">Drosophila serrata</Emphasis> under conditions linked to its southern border: Unexpected patterns from laboratory selection suggest limited evolutionary potential

To investigate the ability ofDrosophila serrata to adapt to thermal conditions over winter at the species southern border, replicate lines from three source locations were held as discrete generations over three years at either 19‡C (40 generations) or temperatures fluctuating between 7‡C and 18δC (20 generations). Populations in the fluctuating environment were maintained either with an adult 0‡C cold shock or without a shock. These conditions were expected to result in temperature-specific directional selection for increased viability and productivity under both temperature regimes, and reduced development time under the fluctuating-temperature regime. Selection responses of all lines were tested under both temperature regimes after controlling for carry-over effects by rearing lines in these environments for two generations. When tested in the 19‡C environment, lines evolving at 19‡C showed a faster development time and a lower productivity relative to the other lines, while cold shock reduced development time and productivity of all lines. When tested in the fluctuating environment, productivity of the 7–18‡C lines selected with a cold shock was relatively lower than that of lines selected without a shock, but this pattern was not observed in the other populations. Viability and body size as measured by wing length were not altered by selection or cold shock, although there were consistent effects of source population on wing length. These results provide little evidence for temperature-specific adaptation inD. serrata —although the lines had diverged for some traits, these changes were not consistent with a priori predictions. In particular, there was no evidence for life-history changes reflecting adaptation to winter conditions at the southern border. The potential forD. serrata to adapt to winter conditions may therefore be limited.     

THE GENETICS OF CENTRAL AND MARGINAL POPULATIONS OF DROSOPHILA SERRATA. I. GENETIC VARIATION FOR STRESS RESISTANCE AND SPECIES BORDERS

A selection experiment was used to determine if levels of genetic variance in an ecologically important trait, desiccation resistance, were different in central and marginal populations. Four populations of Drosophila serrata were sampled from central and marginal areas of its distribution, along a 3000-km stretch of Australia's east coast. Rainfall patterns along this stretch of coastline change from a tropical cycle in the north to a temperate cycle in the south. Replicate lines from the four populations underwent selection for desiccation resistance for 14 generations. Realized heritabilities calculated after 10 and 14 generations of selection indicated that the four populations differed significantly in the level of genetic variation for desiccation resistance available to selection. Populations from the more southern marginal areas had lower realized heritabilities than more northern central populations. However, a corresponding increase in mean desiccation resistance toward the margin was not found. A mechanism by which D. serrata seemed to have responded to selection was a reduction in the extent that metabolic rate was increased when flies were exposed to low humidity. This response indicates genetic variation for the control of metabolic rate. In contrast, increased desiccation resistance was not associated with lipid or glycogen levels. Increased resistance to desiccation was accompanied by increased starvation resistance, but radiation resistance was not affected. Selection did not affect the degree that replicate lines or populations had diverged.     

Long-term selection for a quantitative character in large replicate populations of Drosophila melanogaster

Summary Six replicate lines of Drosophila melanogaster, which had been selected for increased abdominal bristle number for more than 85 generations, were assayed by hierarchical analysis of variance and offspring on parent regression immediately after selection ceased, and by single-generation realised heritability after more than 25 generations of subsequent relaxed selection.Half-sib estimates of heritability in 5 lines were as high as in the base population and much higher than observed genetic gains would suggest, excluding lack of sufficient additive genetic variance as a cause of ineffective selection in these lines. Also, there was considerable diversity among the six lines in composition of phenotypic variability: in addition to differences in the additive genetic component, one or more of the components due to dominance, epistasis, sex-linkage or genotype-environment interaction appeared to be important in different lines.Even after relaxed selection, single-generation realised heritabilities in four lines were as high as in the base population. As a large proportion of total genetic gain must have been made by fixation of favourable alleles, the compensatory increase of genetic variability has been sought in a genetic model involving genes at low initial frequencies, enhancement of gene effects during selection and/or new mutations.     

Costs of resistance in the Drosophila-macrocheles system: a negative genetic correlation between ectoparasite resistance and reproduction

Genetic variation for parasite resistance occurs in most host populations. Costs of resistance, manifested as reduced fitness of resistant genotypes in the absence of parasitism, can be an important factor contributing to the maintenance of this variation. One powerful tool for detecting costs of resistance is the study of correlated responses to artificial selection. Provided that experimental lines are recently derived from large outbreeding populations, and that inbreeding is minimized during the experiment, correlated responses to selection are expected to be strong indicators of pleiotropy. We artificially selected for elevated behavioral resistance against an ectoparasitic mite (Macrocheles subbadius) in replicate populations of the fly Drosophila nigrospiracula. Resistance was modeled as a threshold trait, and the realized heritability of resistance was estimated to be 12.3% (1.4% SE) across three replicate lines recently derived from nature. We contrasted the longevity and fecundity of resistant and control (unselected) flies under a variable thermal environment. We report that reduced fecundity is a correlated response to artificial selection for increased resistance, and that the strength of this effect increases from 25 degrees to 29 degrees C. In contrast, longevity differences were not detected between resistant and control lines at either temperature. These findings are robust as they were confirmed with an independent set of experimental lines. Thus, our results identify a negative genetic correlation between ectoparasite resistance and an important life-history trait. That a correlated response was only detected for fecundity, and not longevity, suggests that the genetic correlation is attributable to pleiotropic effects with narrower effects than reallocation of a general resource pool within the organism, although other interpretations are discussed. Combined with fluctuating parasite-mediated selection and temperature, the presence of this trade-off may contribute to the maintenance of genetic variation for resistance in natural populations.     

A Drosophila laboratory evolution experiment points to low evolutionary potential under increased temperatures likely to be experienced in the future

The ability to respond evolutionarily to increasing temperatures is important for survival of ectotherms in a changing climate. Recent studies suggest that upper thermal limits may be evolutionary constrained. We address this hypothesis in a laboratory evolution experiment, encompassing ecologically relevant thermal regimes. To examine the potential for species to respond to climate change, we exposed replicate populations of Drosophila melanogaster to increasing temperatures (0.3 °C every generation) for 20 generations, whereas corresponding replicate control populations were held at benign thermal conditions throughout the experiment. We hypothesized that replicate populations exposed to increasing temperatures would show increased resistance to warm and dry environments compared with replicate control populations. Contrasting replicate populations held at the two thermal regimes showed (i) an increase in desiccation resistance and a decline in heat knock‐down resistance in replicate populations exposed to increasing temperatures, (ii) similar egg‐to‐adult viability and fecundity in replicate populations from the two thermal regimes, when assessed at high stressful temperatures and (iii) no difference in nucleotide diversity between thermal regimes. The limited scope for adaptive evolutionary responses shown in this study highlights the challenges faced by ectotherms under climate change.     

Expression of the heat-shock protein HSP70 in Drosophila buzzatii lines selected for thermal resistance

The level of HSP70 expression induced by a non-lethal high temperature was examined in lines selected for increased thermal resistance and in corresponding control lines of Drosophila buzzatii, in order to test if selection for high temperature resistance leads to an increased level of HSP70 expression. The lines used were selected for up to 64 generations either as adults or through all larval stages. In adult selection lines, hard selection was implemented every second generation after mild heat hardening. In larval selection lines, larvae were exposed each generation to laboratory "natural" selection. Generally lines selected as adults showed a higher HSP70 expression than did controls, both in third instar larvae and in adults. A strong negative response to selection of HSP70 expression was found in all lines that were selected at cycling temperatures during larval development. The results suggests that a trade off between heat resistance in form of HSP70 expression and fecundity/fertility are responsible for the level of HSP70 expression. The effect of the different methods of selection on HSP70 expression suggests that heat resistance constitutes more than one trait.     

Does increased heat resistance result in higher susceptibility to predation? A test using Drosophila melanogaster selection and hardening

Heat resistance of ectotherms can be increased both by plasticity and evolution, but these effects may have trade‐offs resulting from biotic interactions. Here, we test for predation costs in Drosophila melanogaster populations with altered heat resistance produced by adult hardening and directional selection for increased heat resistance. In addition, we also tested for genetic trade‐offs by testing heat resistance in lines that have evolved under increased predation risk. We show that while 35/37 °C hardening increases heat resistance as expected, it does not increase predation risk from jumping spiders or mantids; in fact, there was an indication that survival may have increased under predation following a triple 37 °C compared to a single 35 °C hardening treatment. Flies that survived a 39 °C selection cycle showed lower survival under predation, suggesting a predation cost of exposure to a more severe heat stress. There was, however, no correlated response to selection because survival did not differ between control and selected lines after selection was relaxed for one or two generations. In addition, lines selected for increased predation risk did not differ in heat resistance. Our findings suggest independent evolutionary responses to predation and heat as measured in laboratory assays, and no costs of heat hardening on susceptibility to predation.     

PARALLEL CHANGES IN HOST RESISTANCE TO VIRAL INFECTION DURING 45,000 GENERATIONS OF RELAXED SELECTION

The dynamics of host susceptibility to parasites are often influenced by trade‐offs between the costs and benefits of resistance. We assayed changes in the resistance to three viruses in six lines of Escherichia coli that had been evolving for almost 45,000 generations in their absence. The common ancestor of these lines was completely resistant to T6, partially resistant to T6* (a mutant of T6 with altered host range), and sensitive to λ. None of the populations changed with respect to resistance to T6, whereas all six evolved increased susceptibility to T6*, probably ameliorating a cost of resistance. More surprisingly, however, the majority of lines evolved complete resistance to λ, despite not encountering that virus during this period. By coupling our results with previous work, we infer that resistance to λ evolved as a pleiotropic effect of a beneficial mutation that downregulated an unused metabolic pathway. The strong parallelism between the lines implies that selection had almost deterministic effects on the evolution of these patterns of host resistance. The opposite outcomes for resistance to T6* and λ demonstrate that the evolution of host resistance under relaxed selection cannot be fully predicted by simple trade‐off models.     

No apparent cost of evolved immune response in Drosophila melanogaster

Maintenance and deployment of the immune system are costly and are hence predicted to trade‐off with other resource‐demanding traits, such as reproduction. We subjected this longstanding idea to test using laboratory experimental evolution approach. In the present study, replicate populations of Drosophila melanogaster were subjected to three selection regimes—I (Infection with Pseudomonas entomophila), S (Sham‐infection with MgSO₄), and U (Unhandled Control). After 30 generations of selection flies from the I regime had evolved better survivorship upon infection with P. entomophila compared to flies from U and S regimes. However, contrary to expectations and previous reports, we did not find any evidence of trade‐offs between immunity and other life history related traits, such as longevity, fecundity, egg hatchability, or development time. After 45 generations of selection, the selection was relaxed for a set of populations. Even after 15 generations, the postinfection survivorship of populations under relaxed selection regime did not decline. We speculate that either there is a negligible cost to the evolved immune response or that trade‐offs occur on traits such as reproductive behavior or other immune mechanisms that we have not investigated in this study. Our research suggests that at least under certain conditions, life‐history trade‐offs might play little role in maintaining variation in immunity.     

Evolvability of Hsp70 expression under artificial election for inducible thermotolerance in independent populations of Drosophila melanogaster

To test whether expression of the inducible heat-shock protein Hsp70 increases under selection for inducible thermotolerance in Drosophila melanogaster, we performed artificial selection on replicate sets of Drosophila lines founded from two independent populations. Selection entailed pretreatment at 36 degrees C to induce thermotolerance and Hsp70 expression, followed by a more severe heat shock, whose temperature varied between sexes and among generations to achieve 50% mortality. Inducible thermotolerance increased slowly and continuously in selected lines and was 37%-50% greater than in controls after 10-11 generations. Lines founded from the two populations differed in their coevolution of Hsp70 expression. In lines founded from Evolution Canyon, Israel, Hsp70 level initially increased and thereafter was unchanged; replicate lines exhibited two temporal patterns of response to selection. In lines founded from Australia, Hsp70 levels increased throughout selection. In both cases, however, the increase in Hsp70 level averaged only 15%, suggesting that pleiotropy in Hsp70 function constrains evolutionary increase in its expression.     

INBREEDING: ITS EFFECT ON RESPONSE TO SELECTION FOR PUPAL WEIGHT AND THE HERITABLE VARIANCE IN FITNESS IN THE FLOUR BEETLE,TRIBOLIUM CASTANEUM

We report our studies of the effect of inbreeding on the response to selection for increased pupal weight in the flour beetle, Tribolium castaneum. We also report the effects of inbreeding and selection for pupal weight on the heritable variation in fitness and fitness components. We created replicate and independent inbred lines with F-values of 0.00, 0.375, and 0.672, by 0, 2, and 5 generations, respectively, of brother-sister mating of adult beetles from an outbred stock population. Subsequently, we imposed artificial within-family selection for increased pupal weight in each of 15 inbred lines for eight generations; each line had its own paired, unselected control. We compared the response to selection across the three levels of inbreeding with theoretical expectation, and investigated the effects of inbreeding and selection on fitness variation among families within all 30 selected and control lines. Among-line variation in pupal weight increased with increased inbreeding prior to selection but diminished with directional selection. Inbreeding reduced the realized heritability of pupal weight concordant with quantitative predictions of additive theory. Mean fitness, measured in several ways, declined with inbreeding and declined further with selection. In contrast, the genetic variation for fitness in the inbred and selected lines lines equalled or exceeded that of the outbred controls. Our results suggest that inbreeding and selection may affect traits in different ways depending on the relative amounts of additive and nonadditive genetic variation.     

Effect of rate of inbreeding on inbreeding depression in Drosophila melanogaster

Summary This experiment was designed to study the relationship between rate of inbreeding and observed inbreeding depression of larval viability, adult fecundity and cold shock mortality in Drosophila melanogaster. Rates of inbreeding used were full-sib mating and closed lines of N=4 and N=20. Eight generations of mating in the N=20 lines, three generations in the N=4 lines and one generation of full-sib mating were synchronised to simultaneously produce individuals with an expected level of inbreeding coefficient (F) of approximately 0.25. Inbreeding depression for the three traits was significant at F=0.25. N=20 lines showed significantly less inbreeding depression than full-sib mated lines for larval viability at approximately the same level of F. A similar trend was observed for fecundity. No effect of rate of inbreeding depression was found for cold shock mortality, but this trait was measured with less precision than the other two. Natural selection acting on loci influencing larval viability and fecundity during the process of inbreeding could explain these results. Selection is expected to be more effective with slow rates of inbreeding because there are more generations and greater opportunity for selection to act before F=0.25 is reached. Selection intensities seem to have been different in the three traits measured. Selection was most intense for larval viability, less intense for fecundity and, perhaps, negligible at loci influencing cold shock mortality.