On distinguishing between canonical tRNA genes and tRNA gene fragments in prokaryotes.

Automated genome annotation is essential for extracting biological information from sequence data. The identification and annotation of tRNA genes is frequently performed by the software package tRNAscan-SE, the output of which is listed for selected genomes in the Genomic tRNA database (GtRNAdb). Here, we highlight a pervasive error in prokaryotic tRNA gene sets on GtRNAdb: the mis-categorization of partial, non-canonical tRNA genes as standard, canonical tRNA genes. Firstly, we demonstrate the issue using the tRNA gene sets of 20 organisms from the archaeal taxon Thermococcaceae. According to GtRNAdb, these organisms collectively deviate from the expected set of tRNA genes in 15 instances, including the listing of eleven putative canonical tRNA genes. However, after detailed manual annotation, only one of these eleven remains; the others are either partial, non-canonical tRNA genes resulting from the integration of genetic elements or CRISPR-Cas activity (seven instances), or attributable to ambiguities in input sequences (three instances). Secondly, we show that similar examples of the mis-categorization of predicted tRNA sequences occur throughout the prokaryotic sections of GtRNAdb. While both canonical and non-canonical prokaryotic tRNA gene sequences identified by tRNAscan-SE are biologically interesting, the challenge of reliably distinguishing between them remains. We recommend employing a combination of (i) screening input sequences for the genetic elements typically associated with non-canonical tRNA genes, and ambiguities, (ii) activating the tRNAscan-SE automated pseudogene detection function, and (iii) scrutinizing predicted tRNA genes with low isotype scores. These measures greatly reduce manual annotation efforts, and lead to improved prokaryotic tRNA gene set predictions.

Sperm-depleted males of the two-spotted spider mite can replenish sperm in a few hours.

In many animals, males increase their reproductive success by mating with as many females as possible. The number of females a male can fertilize is often limited by male competition for access to females, sperm competition, and the cost of sperm production. Especially, recent studies have shown that sperm production is more costly than previously expected. In the two-spotted spider mite, Tetranychus urticae Koch, the number of females a male can inseminate is limited mainly by male competition for access to females. However, in the absence of rivals, males mate with so many females that they can become sperm-depleted. Mating without sperm transfer does not produce any offspring, although it takes time and energy. Therefore, a question arises as to why males continue to mate even after sperm depletion. In this study, we hypothesized that males continue to mate because sperm is replenished after a short period. To test the hypothesis, we investigated how long it takes for sperm replenishment after sperm depletion. We found that in 3 h, sperm can be replenished enough to inseminate a few females. As 3 h is sufficiently short not to lose the next mating opportunity, the results support the hypothesis. However, copulation duration in the sperm-replenished males was significantly longer than in the sperm-depleted males but shorter than in males before sperm depletion. To explain the differences, further research would be necessary. In addition, anatomical physiology study in males is also required to confirm that sperm is indeed depleted and replenished.

Males mate with females even after sperm depletion in the two-spotted spider mite.

Generally, males increase their reproductive success by mating with as many females as possible, whereas females increase their reproductive success by choosing males who provide more direct and indirect benefits. The difference in reproductive strategy between the sexes creates intense competition among males for access to females, therefore males spend much energy and time for competition with rival males for their reproduction. However, if they do not need to engage themselves into male competition and females are in no short supply, how many females can a male mate with and fertilize? We address this question in the two-spotted spider mite, Tetranychus urticae Koch. In this study, we investigated how many females a young, virgin male mated in 3 h, and checked whether the mated females were fertilized. We found that on average males mated with 12-13 females (range: 5-25). As latency to next mating did not change with the number of matings, the males are predicted to engage in even more matings if the mating trial were continued beyond 3 h. Copulation durations decreased with the number of matings and typically after 11 copulations with females any further copulations did not lead to fertilization, suggesting that males continued to mate with females even after sperm depletion. We discuss why spider mite males continue to display mating and copulation behaviour even after their sperm is depleted.

Mandrills learn two-day time intervals in a naturalistic foraging situation.

Primates display high efficiency in finding food in complex environments. Knowledge that many plant species produce fruit simultaneously, can help primates to anticipate fruit finding at the start of fruiting seasons. Knowledge of elapsed time can help primates decide when to revisit food trees to find ripened fruit and to return before competitors find these fruits. To investigate whether mandrills are able to learn time intervals of recurring food, we recorded the foraging choices of captive mandrills in a group setting. We used a procedure with renewable food rewards that could be searched for: carrots and grapes, hidden underground in specific places with different renewal intervals (2 and 5 days, respectively). We monitored the first choice of location for individuals, if other individuals had not already searched at the same location, to exclude possible effects of individuals following others rather than relying on memory. Throughout the study, the mandrills became increasingly likely to first search at carrot locations on carrot days, while the probability of them searching at carrot locations decreased on days without carrot. Due to model instability, our results were inconclusive about an effect of grape days on the choice of the mandrills. Cues provided by conspecifics indicating the availability of simultaneously emerging food rewards did not affect the choice of the mandrills. We conclude that mandrills can take into account elapsed time in a foraging context. Thereby, this study indicates how mandrills can use temporal cognitive abilities to overcome temporal challenges of food-finding in a group setting.

A life-history perspective on sexual selection in a polygamous species.

BACKGROUND: Ever since Darwin, evolutionary biologists have studied sexual selection driving differences in appearance and behaviour between males and females. An unchallenged paradigm in such studies is that one sex (usually the male) signals its quality as a mate to the other sex (usually the female), who is choosy in accepting a partner. Here, we hypothesize that in polygamous species these roles change dynamically with the mating status of males and females, depending on direct reproductive costs and benefits of multiple matings, and on sperm competition. We test this hypothesis by assessing fitness costs and benefits of multiple matings in both males and females in a polygamous moth species, as in moths not males but females are the signalers and males are the responders. RESULTS: We found that multiple matings confer fitness costs and benefits for both sexes. Specifically, the number of matings did not affect the longevity of males or females, but only 67% of the males and 14% of the females mated successfully in all five nights. In addition, the female's reproductive output increased with multiple matings, although when paired with a new virgin male every night, more than 3 matings decreased her reproductive output, so that the Bateman gradient for females fit a quadratic model better than a linear model. The male's reproductive success was positively affected by the number of matings and a linear regression line best fit the data. Simulations of the effect of sperm competition showed that increasing last-male paternity increases the steepness of the male Bateman gradient and thus the male's relative fitness gain from additional mating. Irrespective of last-male paternity value, the female Bateman gradient is steeper than the male one for up to three matings. CONCLUSION: Our results suggest that choosiness in moths may well change throughout the mating season, with males being more choosy early in the season and females being more choosy after having mated at least three times. This life-history perspective on the costs and benefits of multiple matings for both sexes sheds new light on sexual selection forces acting on sexual signals and responses.

Why Do Phytoplankton Evolve Large Size in Response to Grazing?

Phytoplankton are among the smallest primary producers on Earth, yet they display a wide range of cell sizes. Typically, small phytoplankton species are stronger nutrient competitors than large phytoplankton species, but they are also more easily grazed. In contrast, evolution of large phytoplankton is often explained as a physical defense against grazing. Conceptually, this explanation is problematic, however, because zooplankton can coevolve larger size to counter this size-dependent escape from grazing. Here, we hypothesize that there is another advantage for the evolution of large phytoplankton size not so readily overcome: larger phytoplankton often provide lower nutritional quality for zooplankton. We investigate this hypothesis by analyzing an eco-evolutionary model that combines the ecological stoichiometry of phytoplankton-zooplankton interactions with coevolution of phytoplankton and zooplankton size. In our model, evolution of cell size modifies the nutrient uptake kinetics of phytoplankton according to known allometric relationships, which in turn affect the nutritional quality of phytoplankton. With this size-based mechanism, the model predicts that low grazing pressure or nonselective grazing by zooplankton favors evolution of small phytoplankton cells of high nutritional quality. In contrast, selective grazing for nutritious food favors evolution of large phytoplankton of low nutritional quality, which are preyed on by medium- to large-sized zooplankton. This size-dependent change in food quality may explain the commonly observed shift from dominance by small picophytoplankton in oligotrophic waters with low grazing pressure to large phytoplankton species in nutrient-rich waters with high grazing pressure.

Males cannibalise and females disperse in the predatory mite Phytoseiulus persimilis.

Cannibalism is a widespread phenomenon in nature, often occurring when food is scarce, for example among predators that have overexploited a local prey population. Instead of cannibalising, predators can disperse, thereby avoiding being cannibalised or cannibalising related conspecifics, which results in inclusive fitness loss. Theory on prey exploitation in ephemeral predator-prey systems predicts that predators may be selected to display prudent predation by dispersing early, thus saving food for their remaining offspring. This is especially advantageous when average relatedness in the local population is high. Less prudent predators refrain from dispersing until all prey are exterminated. These prey exploitation strategies may also have repercussions for cannibalism, especially when it is driven by food shortage. We therefore investigated to what extent adult females and males cannibalise or disperse after prey have been exterminated locally. We used two lines of the haplodiploid predatory mite Phytoseiulus persimilis that were selected for early and late dispersal, respectively. In wind tunnels, we observed the cannibalistic and dispersal behaviour of individual adult predators of these lines on a rose leaf with only conspecific larvae as food. Both selection lines behaved similarly, indicating that selection on dispersal behaviour did not result in correlated effects on cannibalism behaviour. Male predators stayed significantly longer on the leaf and engaged more often in cannibalism than females. The results suggest that there might be gender-specific differences in cannibalistic tendency in relation to dispersal. Future theoretical studies on the evolution of cannibalism and dispersal should take differences between the genders into account.

An eco-evolutionary feedback loop between population dynamics and fighter expression affects the evolution of alternative reproductive tactics.

Surprisingly, little is known about how eco-evolutionary feedback loops affect trait dynamics within a single population. Polymorphisms of discrete alternative phenotypes present ideal test beds to investigate this, as the alternative phenotypes typically exhibit contrasting demographic rates mediated through frequency or density dependence, and are thus differentially affected by selection. Alternative reproductive tactics (ARTs), like male fighters and sneakers, are an extreme form of discrete phenotype expression and occur across many taxa. Fighters possess weapons for male-male competition over access to mates, whereas sneakers are defenceless but adopt tactics like female-mimicking. Because fighters in some species mortally injure conspecifics, this raises the question whether fighter expression can feed back to affect population size and structure, thereby altering the selection gradient and evolutionary dynamics of ART expression in an eco-evolutionary feedback loop. Here, we investigated how the eco-evolutionary feedback loop between fighter expression and population size and structure affects the evolution and maintenance of ARTs. We introduced intraspecific killing by fighters in a two-sex, two-ART population model parameterized for the male dimorphic bulb mite (Rhizoglyphus robini) that includes life-history differences between the ARTs and a mating-probability matrix analogous to the classic hawk-dove game. Using adaptive dynamics, we found that the intraspecific killing by fighters can extend the range of life-history parameter values under which ARTs evolve, because fighters that kill other fighters decrease fighter fitness. This effect can be nullified when benefits from killing are incorporated, like increased reproduction through increased energy uptake. The eco-evolutionary feedback effects found here for a dimorphic trait likely also occur in other fitness-related traits, such as behavioural syndromes, parental care and niche construction traits. Current theoretical advances to model eco-evolutionary processes, and empirical steps towards unravelling the underlying drivers, pave the way for understanding how selection affects trait evolution in an eco-evolutionary feedback loop.

Eco-Evolutionary Dynamics of Ecological Stoichiometry in Plankton Communities.

Nitrogen (N) and phosphorus (P) limit primary production in many aquatic ecosystems, with major implications for ecological interactions in plankton communities. Yet it remains unclear how evolution may affect the N∶P stoichiometry of phytoplankton-zooplankton interactions. Here, we address this issue by analyzing an eco-evolutionary model of phytoplankton-zooplankton interactions with explicit nitrogen and phosphorus dynamics. In our model, investment of phytoplankton in nitrogen versus phosphorus uptake is an evolving trait, and zooplankton display selectivity for phytoplankton with N∶P ratios matching their nutritional requirements. We use this model to explore implications of the contrasting N∶P requirements of copepods versus cladocerans. The model predicts that selective zooplankton strongly affect the N∶P ratio of phytoplankton, resulting in deviations from their optimum N∶P ratio. Specifically, selective grazing by nitrogen-demanding copepods favors dominance of phytoplankton with low N∶P ratios, whereas phosphorus-demanding cladocerans favor dominance of phytoplankton with high N∶P ratios. Interestingly, selective grazing by nutritionally balanced zooplankton leads to the occurrence of alternative stable states, where phytoplankton may evolve either low, optimum, or high N∶P ratios, depending on the initial conditions. These results offer a new perspective on commonly observed differences in N∶P stoichiometry between plankton of freshwater and those of marine ecosystems and indicate that selective grazing by zooplankton can have a major impact on the stoichiometric composition of phytoplankton.

Patterns of reproductive isolation in a haplodiploid - strong post-mating, prezygotic barriers among three forms of a social spider mite.

In speciation research, much attention is paid to the evolution of reproductive barriers, preventing diverging groups from hybridizing back into one gene pool. The prevalent view is that reproductive barriers evolve gradually as a by-product of genetic changes accumulated by natural selection and genetic drift in groups that are segregated spatially and/or temporally. Reproductive barriers, however, can also be reinforced by natural selection against maladaptive hybridization. These mutually compatible theories are both empirically supported by studies, analysing relationships between intensity of reproductive isolation and genetic distance in sympatric taxa and allopatric taxa. Here, we present the - to our knowledge - first comparative study in a haplodiploid organism, the social spider mite Stigmaeopsis miscanthi, by measuring premating and post-mating, pre- and post-zygotic components of reproductive isolation, using three recently diverged forms of the mite that partly overlap in home range. We carried out cross-experiments and measured genetic distances (mitochondrial DNA and nuclear DNA) among parapatric and allopatric populations of the three forms. Our results show that the three forms are reproductively isolated, despite the absence of premating barriers, and that the post-mating, prezygotic component contributes most to reproductive isolation. As expected, the strength of post-mating reproductive barriers positively correlated with genetic distance. We did not find a clear pattern of prezygotic barriers evolving faster in parapatry than in allopatry, although one form did show a trend in line with the ecological and behavioural relationships between the forms. Our study advocates the versatility of haplodiploid animals for investigating the evolution of reproductive barriers.

Sooty mangabeys scavenge on nuts cracked by chimpanzees and red river hogs-An investigation of inter-specific interactions around tropical nut trees.

Carrion scavenging is a well-studied phenomenon, but virtually nothing is known about scavenging on plant material, especially on remnants of cracked nuts. Just like meat, the insides of hard-shelled nuts are high in energetic value, and both foods are difficult to acquire. In the Taï forest, chimpanzees (Pan troglodytes) and red river hogs (Potamochoerus porcus) crack nuts by using tools or strong jaws, respectively. In this study, previously collected non-invasive camera trap data were used to investigate scavenging by sooty mangabeys (Cercocebus atys), two species of Guinea fowl (Agelestres meleagrides; Guttera verreauxi), and squirrels (Scrunidae spp.) on the nut remnants cracked by chimpanzees and red river hogs. We investigated how scavengers located nut remnants, by analyzing their visiting behavior in relation to known nut-cracking events. Furthermore, since mangabeys are infrequently preyed upon by chimpanzees, we investigated whether they perceive an increase in predation risk when approaching nut remnants. In total, 190 nut-cracking events were observed in four different areas of Taï National Park, Ivory Coast. We could confirm that mangabeys scavenged on the nuts cracked by chimpanzees and hogs and that this enabled them to access food source that would not be accessible otherwise. We furthermore found that mangabeys, but not the other species, were more likely to visit nut-cracking sites after nut-cracking activities than before, and discuss the potential strategies that the monkeys could have used to locate nut remnants. In addition, mangabeys showed elevated levels of vigilance at the chimpanzee nut-cracking sites compared with other foraging sites, suggesting that they perceived elevated danger at these sites. Scavenging on remnants of cracked nuts is a hitherto understudied type of foraging behavior that could be widespread in nature and increases the complexity of community ecology in tropical rainforests.

The role of web sharing, species recognition and host-plant defence in interspecific competition between two herbivorous mite species.

When competing with indigenous species, invasive species face a problem, because they typically start with a few colonizers. Evidently, some species succeeded, begging an answer to the question how they invade. Here, we investigate how the invasive spider mite Tetranychus evansi interacts with the indigenous species T. urticae when sharing the solanaceous host plant tomato: do they choose to live together or to avoid each other's colonies? Both species spin protective, silken webs on the leaf surfaces, under which they live in groups of con- and possibly heterospecifics. In Spain, T. evansi invaded the non-crop field where native Tetranychus species including T. urticae dominated. Moreover, T. evansi outcompetes T. urticae when released together on a tomato plant. However, molecular plant studies suggest that T. urticae benefits from the local down-regulation of tomato plant defences by T. evansi, whereas T. evansi suffers from the induction of these defences by T. urticae. Therefore, we hypothesize that T. evansi avoids leaves infested with T. urticae whereas T. urticae prefers leaves infested by T. evansi. Using wild-type tomato and a mutant lacking jasmonate-mediated anti-herbivore defences, we tested the hypothesis and found that T. evansi avoided sharing webs with T. urticae in favour of a web with conspecifics, whereas T. urticae more frequently chose to share webs with T. evansi than with conspecifics. Also, T. evansi shows higher aggregation on a tomato plant than T. urticae, irrespective of whether the mites occur on the plant together or not.

Incomplete premating and postmating reproductive barriers between two parapatric populations of a social spider mite.

Closely related species with overlapping distributions often show premating reproductive barriers to avoid hybridization. Stigmaeopsis miscanthi (Saito) is a social spider mite infesting Chinese silver grass, and the species consists of two parapatric groups with frequent contacts within the contact zone. They differ in male-male aggressiveness, male morphology, female diapause traits, and life history parameters. There is incomplete but strong post-mating reproductive isolation between the two groups, and their DNA sequences are slightly different, suggesting that they diverged recently. In this study, we investigated premating reproductive barriers. We found that females from different groups frequently shared nest webs, indicating no barriers in the phase of nest establishment. However, inside nests, males from either group showed less courtship behaviour to females of the other group and they copulated less frequently with them when compared to females of the same group. However, the premating reproductive barrier was incomplete and asymmetric. Females of one group frequently resisted courtship by males from the other group, but females of the other group did not. We conclude that some gene flow may occur in the contact zone between the two groups.

Personal experience and reputation interact in human decisions to help reciprocally.

There is ample evidence that human cooperative behaviour towards other individuals is often conditioned on information about previous interactions. This information derives both from personal experience (direct reciprocity) and from experience of others (i.e. reputation; indirect reciprocity). Direct and indirect reciprocity have been studied separately, but humans often have access to both types of information. Here, we experimentally investigate information use in a repeated helping game. When acting as donor, subjects can condition their decisions to help recipients with both types of information at a small cost to access such information. We find that information from direct interactions weighs more heavily in decisions to help, and participants tend to react less forgivingly to negative personal experience than to negative reputation. Moreover, effects of personal experience and reputation interact in decisions to help. If a recipient's reputation is positive, the personal experience of the donor has a weak effect on the decision to help, and vice versa. Yet if the two types of information indicate conflicting signatures of helpfulness, most decisions to help follow personal experience. To understand the roles of direct and indirect reciprocity in human cooperation, they should be studied in concert, not in isolation.

Alternative phenotypes of male mating behaviour in the two-spotted spider mite.

Severe intraspecific competition for mates selects for aggressive individuals but may also lead to the evolution of alternative phenotypes that do not act aggressively, yet manage to acquire matings. The two-spotted spider mite, Tetranychus urticae, shows male mate-guarding behaviour and male-male combat for available females. This may provide opportunity for weaker males to avoid fighting by adopting alternative mating behaviour such as sneaker or satellite tactics as observed in other animals. We investigated male precopulatory behaviour in the two-spotted spider mite by means of video-techniques and found three types of male mating behaviour: territorial, sneaker and opportunistic. Territorial and sneaker males associate with female teleiochrysales and spend much time guarding them. Territorial males are easily disturbed by rival males and engage themselves in fights with them. However, sneaker males are not at all disturbed by rival males, never engage in fights and, strikingly, never face attack by territorial males. Opportunistic males wander around in search of females that are in the teleiochrysalis stage but very close to or at emergence. To quickly classify any given mate-guarding male as territorial or sneaker we developed a method based on the instantaneous response of males to disturbance by a live male mounted on top of a brush. We tested this method against the response of the same males to natural disturbance by two or three other males. Because this method proved to be successful, we used it to collect territorial and sneaker males, and subjected them to morphological analysis to assess whether the various behavioural phenotypes are associated with different morphological characters. However, we found no statistical differences between territorial and sneaker males, concerning the length of the first legs, the stylets, the pedipalps or the body.

Towards evolutionary predictions: Current promises and challenges.

Evolution has traditionally been a historical and descriptive science, and predicting future evolutionary processes has long been considered impossible. However, evolutionary predictions are increasingly being developed and used in medicine, agriculture, biotechnology and conservation biology. Evolutionary predictions may be used for different purposes, such as to prepare for the future, to try and change the course of evolution or to determine how well we understand evolutionary processes. Similarly, the exact aspect of the evolved population that we want to predict may also differ. For example, we could try to predict which genotype will dominate, the fitness of the population or the extinction probability of a population. In addition, there are many uses of evolutionary predictions that may not always be recognized as such. The main goal of this review is to increase awareness of methods and data in different research fields by showing the breadth of situations in which evolutionary predictions are made. We describe how diverse evolutionary predictions share a common structure described by the predictive scope, time scale and precision. Then, by using examples ranging from SARS-CoV2 and influenza to CRISPR-based gene drives and sustainable product formation in biotechnology, we discuss the methods for predicting evolution, the factors that affect predictability and how predictions can be used to prevent evolution in undesirable directions or to promote beneficial evolution (i.e. evolutionary control). We hope that this review will stimulate collaboration between fields by establishing a common language for evolutionary predictions.

Group selection and inclusive fitness are not equivalent; the Price equation vs. models and statistics.

It is often suggested that any group selection model can be recast in terms of inclusive fitness. A standard reference to support that claim is "'Quantitative genetics, inclusive fitness, and group selection" by Queller (1992) in the American Naturalist 139 (3), 540-558. In that paper the Price equation is used for the derivation of this claim. Instead of a general derivation, we try out a simple model. For this simple example, we find that the result does not hold. The non-equivalence of group selection and kin selection is therefore not only an important finding in itself, but also a case where the use of the Price equation leads to a claim that is not correct. If results that are arrived at with the Price equation are not correct, they can typically be repaired by adding extra assumptions, or explicitly stating implicit ones. We give examples with relatively mild and with less mild extra assumptions. We also discuss why the Price equation is often referred to as dynamically insufficient, and we try to find out what Price's theorem could be.

Artificial selection for timing of dispersal in predatory mites yields lines that differ in prey exploitation strategies.

Dispersal is the main determinant of the dynamics and persistence of predator-prey metapopulations. When defining dispersal as a predator exploitation strategy, theory predicts the existence of a continuum of strategies: from some dispersal throughout the predator-prey interaction (the Milker strategy) to dispersal only after the prey had been exterminated (the Killer strategy). These dispersal strategies relate to differences in prey exploitation at the population level, with more dispersal leading to longer predator-prey interaction times and higher cumulative numbers of dispersing predators. In the predatory mite Phytoseiulus persimilis, empirical studies have shown genetic variation for prey exploitation as well as for the timing of aerial dispersal in the presence of prey. Here, we test whether artificial selection for lines that differ in timing of dispersal also results in these lines differing in prey exploitation. Six rounds of selection for early or late dispersal resulted in predator lines displaying earlier or later dispersal. Moreover, it resulted-at the population level-in predicted differences in the local predator-prey interaction time and in the cumulative numbers of dispersers in a population dynamics experiment. We pose that timing of dispersal is a heritable trait that can be selected in P. persimilis, which results in lines that show quantitative differences in local predator-prey dynamics. This opens ways to experimentally investigate the evolution of alternative prey exploitation strategies and to select for predator strains with prey exploitation strategies resulting in better biological control.

Population bottleneck has only marginal effect on fitness evolution and its repeatability in dioecious Caenorhabditis elegans.

The predictability of evolution is expected to depend on the relative contribution of deterministic and stochastic processes. This ratio is modulated by effective population size. Smaller effective populations harbor less genetic diversity and stochastic processes are generally expected to play a larger role, leading to less repeatable evolutionary trajectories. Empirical insight into the relationship between effective population size and repeatability is limited and focused mostly on asexual organisms. Here, we tested whether fitness evolution was less repeatable after a population bottleneck in obligately outcrossing populations of Caenorhabditis elegans. Replicated populations founded by 500, 50, or five individuals (no/moderate/strong bottleneck) were exposed to a novel environment with a different bacterial prey. As a proxy for fitness, population size was measured after one week of growth before and after 15 weeks of evolution. Surprisingly, we found no significant differences among treatments in their fitness evolution. Even though the strong bottleneck reduced the relative contribution of selection to fitness variation, this did not translate to a significant reduction in the repeatability of fitness evolution. Thus, although a bottleneck reduced the contribution of deterministic processes, we conclude that the predictability of evolution may not universally depend on effective population size, especially in sexual organisms.