Trinidadian Guppies Research Articles

Do replicates of independent guppy lineages evolve similarly in a predator-free laboratory environment?

The Trinidadian guppy is emblematic of parallel and convergent evolution, with repeated demonstrations that predation regime is a driver of adaptive trait evolution. A classic and foundational experiment in this system was conducted by John Endler 40 years ago, where male guppies placed into low‐predation environments in the laboratory evolved increased color in a few generations. However, Endler's experiment did not employ the now typical design for a parallel/convergent evolution study, which would employ replicates of different ancestral lineages. We therefore implemented an experiment that seeded replicate mesocosms with small founding populations of guppies originating from high‐predation populations of two very different lineages. The different mesocosms were maintained identically, and male guppy color was quantified every four months. After one year, we tested whether male color had increased, whether replicates within a lineage had parallel phenotypic trajectories, and whether the different lineages converged on a common phenotype. Results showed that male guppy color generally increased through time, primarily due to changes in melanic color, whereas the other colors showed inconsistent and highly variable trajectories. Most of the nonparallelism in phenotypic trajectories was among mesocosms containing different lineages. In addition to this mixture of parallelism and nonparallelism, convergence was not evident in that the variance in color among the mesocosms actually increased through time. We suggest that our results reflect the potential importance of high variation in female preference and stochastic processes such as drift and founder effects, both of which could be important in nature.

Competitive asymmetry and local adaptation in Trinidadian guppies.

The outcome of competition between individuals often depends on body size. These competitive asymmetries can drive variation in demographic rates, influencing the ecology and evolution of life histories. The magnitude and direction of such asymmetries differ among taxa, yet little is known empirically about how adaptation to resource limitation alters competitive asymmetries. Here, we investigate the relationship between size-dependent competitive ability and adaptation to resource limitation. We examined size-dependent competition in two ecotypes of Trinidadian guppy, adapted to high or low levels of resource competition. Using aquaria-based competition experiments, we describe how the size and ecotype of competitors influence somatic growth rate, whilst controlling for the confounding effect of niche differentiation. We replicated our study across two independent evolutionary origins of the "competitive" ecotype. The two "competitive" ecotypes differed markedly in size-dependent asymmetry, indicating that adaptation to resource limitation alone is insufficient to explain changes in size-dependent competitive asymmetry. For one origin, the ecotype adapted to resource limitation was a superior competitor over a wide range of size pairings. The equivalence of competitors varied over fivefold, dependent on size and ecotype; in three of four populations, larger individuals had a competitive advantage. Our results demonstrate that competitive asymmetry has strong effects on somatic growth. Because somatic growth contributes to demographic parameters, intraspecific trait variation is likely to play a key role in regulating demographic rates. Our findings imply that the evolution of size-dependent asymmetries under conditions of intense competition is likely to be constrained by niche availability, although further research is needed to verify this.

Habituation and individual variation in the endocrine stress response in the Trinidadian guppy (Poecilia reticulata)

The vertebrate stress response enables individuals to react to and cope with environmental challenges. A crucial aspect of the stress response is the elevation of circulating glucocorticoids. However, continued activation of the stress response under repeated exposure to stressors can be damaging to fitness. Under certain circumstances it may therefore be adaptive to habituate to repeated exposures to a particular stressor by reducing the magnitude of any associated release of glucocorticoids. Here, we investigate whether Trinidadian guppies (Poecilia reticulata) habituate to repeated exposure to a mild stressor, using a waterborne hormone sampling approach that has previously been shown to elicit a stress response in small fish. We also test for individual variation in the extent of habituation to this stressor. Concentrating on freely circulating cortisol, we found that the first exposure to the assay induced high cortisol release rates but that guppies tended to habituate quickly to subsequent exposures. There were consistent differences among individuals in their average cortisol release rate (after accounting for effects of variables such as body size) over repeated exposures. Our analyses did not find evidence of individual differences in habituation rate, although limitations in statistical power could account for this finding. We repeated the analysis for free 11-ketotestosterone, which can also respond to stressors, but found no obvious habituation pattern and no among-individual variation. We also present data on conjugated forms of both hormones, which were repeatable but did not show the expected time-lagged habituation effect. We discuss consistent individual differences around the general pattern of habituation in the flexible stress response, and highlight the potential for individual variation in habituation to facilitate selection against the deleterious effects of chronic stress.

Sex-biased expression between guppies varying in the presence of ornamental coloration.

Sex-biased gene expression provides a means to achieve sexual dimorphism across a genome largely shared by both sexes. Trinidadian guppies are ideal to examine questions of sex-bias as they exhibit sexual dimorphism in ornamental coloration with male only expression. Here we use RNA-sequencing to quantify whole transcriptome gene expression differences, with a focus on differential expression of color genes between the sexes. We determine whether males express genes positively correlated with coloration at higher levels than females. We find that all the differentially expressed color genes were more highly expressed by males. Males also expressed all known black melanin synthesis genes at higher levels than females, regardless of whether the gene was significantly differentially expressed in the analysis. These differences correlated with the visual color differences between sexes at the stage sampled, as all males had ornamental black coloration apparent. We propose that sexual dimorphism in ornamental coloration is caused by male-biased expression of color genes.

Artificial Light at Night Affects Emergence from a Refuge and Space Use in Guppies

Artificial light at night (ALAN) is a major form of anthropogenic pollution. ALAN is well known to affect different behaviours during nighttime, when changes in light conditions often have immediate consequences for the trade-offs individuals experience. How ALAN affects daytime behaviours, however, has received far less attention. Here we studied how ALAN affected daytime personality traits and learning ability. We exposed Trinidadian guppies, Poecilia reticulata, for 10 weeks to different ALAN levels: bright light (24 hrs bright light, ~5,000 lx), dim light (12 hrs bright light; 12 hrs dim light, ~0.5 lx) and control (12 hrs bright light; 12 hrs dark). Afterwards, we tested how the treatments affected diurnal emergence from a refuge, space use, activity, sociability and the ability to memorize the location of companion fish. Individuals exposed to the light treatments (both dim and bright light) emerged quicker from a refuge and fish from the bright light treatment spent relatively more time in the open area of the arena. ALAN did not affect any of the other behaviours, although memory could not be tested since fish did not learn the companions’ location. Our results demonstrate that ALAN, next to affecting nocturnal behaviours, can also affect key diurnal behavioural processes, associated with risk-taking.

Transmission risk predicts avoidance of infected conspecifics in Trinidadian guppies.

Associating with conspecifics afflicted with infectious diseases increases the risk of becoming infected, but engaging in avoidance behaviour incurs the cost of lost social benefits. Across systems, infected individuals vary in the transmission risk they pose, so natural selection should favour risk-sensitive avoidance behaviour that optimally balances the costs and benefits of sociality. Here, we use the guppy Poecilia reticulata-Gyrodactylus turnbulli host-parasite system to test the prediction that individuals avoid infected conspecifics in proportion to the transmission risk they pose. In dichotomous choice tests, uninfected fish avoided both the chemical and visual cues, presented separately, of infected conspecifics only in the later stages of infection. A transmission experiment indicated that this avoidance behaviour accurately tracked transmission risk (quantified as both the speed at which transmission occurs and the number of parasites transmitting) through the course of infection. Together, these findings reveal that uninfected hosts can use redundant cues across sensory systems to inform dynamic risk-sensitive avoidance behaviour. This correlation between the transmission risk posed by infected individuals and the avoidance response they elicit has implications for the evolutionary ecology of infectious disease, and its explicit inclusion may improve the ability of epidemic models to predict disease spread.

Differences in neural activity, but not behavior, across social contexts in guppies, Poecilia reticulata

Animals are continually faced with the challenge of producing context-appropriate social behaviors. In many instances, appropriate behaviors differ by social situation. However, in some instances, the same behaviors are employed across different social contexts, albeit in response to distinct stimuli and with distinct purposes. We took advantage of behavioral similarities across mating and aggression contexts in guppies, Poecilia reticulata, to understand how patterns of neural activity differ across social contexts when behaviors are nonetheless shared. While there is growing interest in understanding behavioral mechanisms in guppies, resources are sparse. As part of this study, we developed a neuroanatomical atlas of the guppy brain as a research community resource. Using this atlas, we found that neural activity in the preoptic area reflected social context, whereas individual differences in behavioral motivation paralleled activity in the posterior tuberculum and ventral telencephalon (teleost homologs of the mammalian ventral tegmental area and lateral septum, respectively). Our findings suggest independent coding of social salience versus behavioral motivation when behavioral repertoires are shared across social contexts. Choosing behaviors appropriate to the current social situation is of central importance to animals. Interactions with different social partners (e.g., mates, competitors, or offspring) generally require distinct behavioral repertories. However, in some cases, similar behaviors are used across social contexts. The neural mechanisms underlying social behavior are particularly intriguing in these situations, where the same behaviors are produced in response to distinct social stimuli and for distinct purposes. We took advantage of behavioral similarities across mating and aggression interactions in Trinidadian guppies to explore how social information is reflected in the brain when fish perform a common set of behaviors across contexts. We found that activity in distinct brain regions reflects social context versus behavioral motivation, suggesting a means by which social inputs and behavioral outputs can be coded independently of one another.

Wild and laboratory exposure to cues of predation risk increases relative brain mass in male guppies

Abstract There is considerable diversity in brain size within and among species, and substantial dispute over the causes, consequences and importance of this variation. Comparative and developmental studies are essential in addressing this controversy. Predation pressure has been proposed as a major force shaping brain, behaviour and life history. The Trinidadian guppy, Poecilia reticulata, shows dramatic variation in predation pressure across populations. We compared the brain mass of guppies from high and low predation populations collected in the wild. Male but not female guppies exposed to high predation possessed heavier brains for their body size compared to fish from low predation populations. The brain is a plastic organ, so it is possible that the population differences we observed were partly due to developmental responses rather than evolved differences. In a follow‐up study, we raised guppies under cues of predation risk or in a control condition. Male guppies exposed to predator cues early in life had heavier brains relative to their body size than control males, while females showed no significant effect of treatment. Collectively our results suggest that male guppies exposed to predation invest more in neural tissue, and that these differences are at least partly driven by plastic responses. A plain language summary is available for this article.

Evolutionary genetics of personality in the Trinidadian guppy II: sexual dimorphism and genotype-by-sex interactions

Sexual dimorphism in behaviour and personality has been identified in a number of species, but few studies have assessed the extent of shared genetic architecture across the sexes. Under sexually antagonistic selection, mechanisms are expected to evolve that reduce evolutionary conflict, resulting in genotype-by-sex (GxS) interactions. Here we assess the extent of sexual dimorphism in four risk-taking behaviour traits in the Trinidadian guppy, Poecilia reticulata, and apply a multivariate approach to test for GxS interactions. We also quantify the among-individual and genetic covariances between personality and size and growth, which are known a priori to differ between the sexes. We found significant sexual dimorphism in three of the four behaviours, although rmf between sex-specific homologous traits was significantly <+1 for only one behaviour. Using multivariate models, we then estimated sex-specific genetic (co)variance matrices (Gm and Gf) and tested for asymmetry of the cross-trait cross-sex genetic covariance structure (submatrix B). While Gm and Gf were not significantly different from each other overall, their respective leading eigenvectors were poorly aligned. Statistical support for asymmetry in B was found, but limited to a single trait pair for which the cross-sex covariances differed (i.e., COVA(m,f) ≠ COVA(f,m)). Thus, while single- and multi-trait perspectives evidence some GxS, the overall picture is one of similarity between the sexes in their genetic (co)variance structures. Our results suggest behavioural traits related to risk-taking may lack the sex-specific genetic architecture for further dimorphism to evolve under what is hypothesised to be antagonistic selection.

Evolutionary genetics of personality in the Trinidadian guppy I: maternal and additive genetic effects across ontogeny

Among-individual variation in behaviour is a widespread phenomenon, with several frameworks developed to explain its existence. Maternal effects, which can have significant influence over evolutionary processes, are an understudied source of behavioural variation. Maternal effects are not necessarily static, however, since their importance can change over offspring ontogeny, typically declining with age relative to additive genetic effects. Here, using a quantitative genetics approach, we test the prediction that maternal effects will influence age-specific risk-taking behaviour in Trinidadian guppies, Poecilia reticulata. Individuals were subject to a single open-field trial as juveniles and up to four repeat trials as adults, with five traits indicative of risk-taking behaviour measured in each trial. We then partitioned phenotypic variance into additive genetic (VA) and maternal identity (VM) components, in addition to testing brood size and maternal weight as specific sources of maternal effects. We found that VM had significant influence over juvenile traits, with very low VA estimates. Whereas, in adults, all traits were significantly heritable, with little support for VM. We also found a strong influence of maternal traits on juvenile behaviours as predicted, with significant, albeit smaller, effects found in adults. Maternal weight was heritable and itself subject to maternal effects. Thus, maternal weight is a likely source of maternal genetic effects that are expected to alter response to selection on personality in this system. More generally, our study highlights that while maternal effects can be an important source of personality variation, this varies over ontogeny of offspring.

Shared and Species-Specific Patterns of Nascent Y Chromosome Evolution in Two Guppy Species.

Sex chromosomes form once recombination is halted around the sex-determining locus between a homologous pair of chromosomes, resulting in a male-limited Y chromosome. We recently characterized the nascent sex chromosome system in the Trinidadian guppy (Poecilia reticulata). The guppy Y is one of the youngest animal sex chromosomes yet identified, and therefore offers a unique window into the early evolutionary forces shaping sex chromosome formation, particularly the rate of accumulation of repetitive elements and Y-specific sequence. We used comparisons between male and female genomes in P. reticulata and its sister species, Endler’s guppy (P. wingei), which share an ancestral sex chromosome, to identify male-specific sequences and to characterize the degree of differentiation between the X and Y chromosomes. We identified male-specific sequence shared between P. reticulata and P. wingei consistent with a small ancestral non-recombining region. Our assembly of this Y-specific sequence shows substantial homology to the X chromosome, and appears to be significantly enriched for genes implicated in pigmentation. We also found two plausible candidates that may be involved in sex determination. Furthermore, we found that the P. wingei Y chromosome exhibits a greater signature of repetitive element accumulation than the P. reticulata Y chromosome. This suggests that Y chromosome divergence does not necessarily correlate with the time since recombination suppression. Overall, our results reveal the early stages of Y chromosome divergence in the guppy.

Heterospecific shoaling in an invasive poeciliid: shared history does not affect shoal cohesion

Social behaviour potentially plays an important role in invasion success. New colonists, for example, may glean useful information about predators and food by interacting with native heterospecifics. The extent to which invaders benefit from such social interactions could hinge on their prior exposure to other species. Here we asked how the shoaling decisions of the Trinidadian guppy, Poecilia reticulata, a successful invasive species, are mediated by their shared history with a heterospecific, the phenotypically similar Micropoecilia picta. To do this, we monitored shoal cohesion in single-species treatments and in treatments where M. picta was present. We predicted that shoal cohesion would be greater in single- than in mixed-species shoals. We also hypothesized that mixed-species shoals consisting of fish with a shared history would be more cohesive than those where the two species had hitherto occurred allopatrically. We found that shoal cohesion did not differ between single- and two-species treatments, or in relation to shared history with M. picta. However, while guppies were more often found in mixed-species than single-species shoals, they were more likely to have a conspecific individual as their nearest neighbour within mixed-species shoals. These results show that guppies willingly shoal with heterospecifics, even in the absence of a shared history, but also that the resulting shoals are not randomly assembled. This flexibility in shoaling may confer a crucial advantage in the initial stages of invasion.

An experimental test of alternative population augmentation scenarios.

Human land use is fragmenting habitats worldwide and inhibiting dispersal among previously connected populations of organisms, often leading to inbreeding depression and reduced evolutionary potential in the face of rapid environmental change. To combat this augmentation of isolated populations with immigrants is sometimes used to facilitate demographic and genetic rescue. Augmentation with immigrants that are genetically and adaptively similar to the target population effectively increases population fitness, but if immigrants are very genetically or adaptively divergent, augmentation can lead to outbreeding depression. Despite well-cited guidelines for the best practice selection of immigrant sources, often only highly divergent populations remain, and experimental tests of these riskier augmentation scenarios are essentially nonexistent. We conducted a mesocosm experiment with Trinidadian guppies (Poecilia reticulata) to test the multigenerational demographic and genetic effects of augmenting 2 target populations with 3 types of divergent immigrants. We found no evidence of demographic rescue, but we did observe genetic rescue in one population. Divergent immigrant treatments tended to maintain greater genetic diversity, abundance, and hybrid fitness than controls that received immigrants from the source used to seed the mesocosms. In the second population, divergent immigrants had a slightly negative effect in one treatment, and the benefits of augmentation were less apparent overall, likely because this population started with higher genetic diversity and a lower reproductive rate that limited genetic admixture. Our results add to a growing consensus that gene flow can increase population fitness even when immigrants are more highly divergent and may help reduce uncertainty about the use of augmentation in conservation.

Developmental plasticity of the stress response in female but not in male guppies.

To survive, animals must respond appropriately to stress. Stress responses are costly, so early-life experiences with potential stressors could adaptively tailor adult stress responses to local conditions. However, how multiple stressors influence the development of the stress response remains unclear, as is the role of sex. Trinidadian guppies (Poecilia reticulata) are small fish with extensive life-history differences between the sexes and population variation in predation pressure and social density. We investigated how sex and early-life experience influence hormonal stress responses by manipulating conspecific density and perceived predation risk during development. In adults, we sampled cortisol twice to measure initial release and change over time in response to a recurring stressor. The sexes differed considerably in their physiological stress response. Males released more cortisol for their body mass than females and did not reduce cortisol release over time. By contrast, all females, except those reared at high density together with predation cues, reduced cortisol release over time. Cortisol responses of males were thus less dynamic in response to current circumstances and early-life experiences than females, consistent with life-history differences between the sexes. Our study underscores the importance of early-life experiences, interacting ecological factors and sex differences in the organization of the stress response.

Erratum: Non-adaptive plasticity potentiates rapid adaptive evolution of gene expression in nature.

This corrects the article DOI: 10.1038/nature15256.

Male guppies compensate for lost time when mating in turbid water

Turbidity (a measure of the cloudiness of water) decreases the visual range of organisms, altering interactions within and between species. For species that visually assess mates, turbidity may affect mating interactions and mate choice. A central question, then, is to what degree organisms plastically adjust mating behaviors to cope with visually altered environments. Here, we investigate the effect of turbidity on the mating behavior of guppies (Poecilia reticulata) in Trinidad, where some streams are increasingly turbid due to upstream rock quarrying. We collected fish from two highly turbid streams (with upstream rock quarries) and two pristine streams (no upstream quarries) in the same drainages. We then observed male mating behaviors with females from the same populations in both turbid and clear assays, recording displays and sneak copulation attempts. Males from turbid streams showed greater individual consistency in mating behaviors. But regardless of provenance, male guppies spent less time associating with females in turbid water overall. When males and females did interact, however, males greatly increased the rate of all mating behaviors in turbid as compared to clear water. Thus, even when lacking a long-term evolutionary history with high turbidity, guppies compensate for lost time when mating in a visually altered environment. Given the global nature of increases in turbidity, understanding behavioral responses to this aquatic anthropogenic change is critical. Here, we investigate the degree to which Trinidadian guppy males in affected areas are able to plastically compensate in turbid water by adjusting mating behaviors. We also test the degree to which previous chronic exposure to turbidity alters this plasticity. In contrast to predictions, we did not find population differences in plasticity; across all populations, however, we found significant plasticity in the guppy mating system in response to anthropogenically increased turbidity. While males spent less time associating with females in turbid water overall, males increased mating effort during periods of association, compensating for lost time in turbid water. We discuss this plasticity and the implications for guppy sexual selection and secondary sexual characteristics such as coloration.

Female assessment of male functional fertility during mate choice in a promiscuous fish

AbstractSexual selection should favour females that can assess the functional fertility of available sexual partners and avoid mating with recently mated, sperm‐depleted males. Our current understanding of the sensory mechanism(s) underlying female assessment of males based on their functional fertility and avoidance of sperm‐depleted males is incomplete. Female Trinidadian guppies (Poecilia reticulata) are known to avoid mating with males that they had previously observed mating with other females. Here, we investigated experimentally the proximate sensory cues that they use to distinguish between paired size‐ and colour‐matched mated and unmated males in the absence of visual public information on their prior mating histories. When only water‐borne chemical cues from the males were available, females avoided the previously mated male and preferred the unmated one, but they chose randomly when only male visual cues (and no chemical cues) were available. They also preferred unmated over mated males when freely swimming with them in a more sensorially complex environment with multiple male cues (i.e., visual, chemical and mechanical cues) concurrently available. Females exhibited no preference for either stimulus males when both were unmated, irrespectively of the sensory environment. These novel results suggest that, in the absence of prior visual public information on the recent mating histories of males, female guppies use olfactory cues putatively emitted by mated males to avoid mating with them. The source and nature of the implicated olfactory cues and the fitness benefits gained by female guppies in sexually preferring males that have not recently mated remain unknown and warrant further research.

How pre- and postcopulatory sexual selection influence male mating decisions in a promiscuous species

When females mate multiply, male reproductive success depends on both pre- and postcopulatory processes, including female choice and sperm competition. However, these processes can favour different mating tactics in males. Here we used the Trinidadian guppy, Poecilia reticulata, system to understand how this conflict is resolved. We asked whether knowledge of recent female mating history leads males to adjust their mating effort with respect to the time devoted to mating activity, and the frequency and the sequence of mating tactics employed. To do this we quantified male mating behaviour in three competitive scenarios: (1) Single, when a focal male arrives near a single female and remains alone with her; (2) First, when a focal male is joined by a rival male; and (3) Second, when a focal male arrives after a rival male. We hypothesized that males adjust their behaviour based on arrival order. If female sequential mate choice is the main process shaping male mating behaviours (favouring First males in guppies), males should avoid competition and invest most when Single. Alternatively, if last-male sperm precedence is the major driver of decision making, males should invest more in mating attempts in the Second scenario. Greatest investment when First implies an intermediate strategy. We found that order of arrival influenced mating decisions with most mating activity during the First rather than the Single and Second scenarios. This result suggests that both pre- and postcopulatory processes influence mating investment, and that individual males make contingent decisions to maximize both mating and fertilization success.

Authentic Science with Live Organisms Can Improve Evolution Education

Evolution education in the United States remains controversial and challenging. This is in part due to the difficulty educators face when trying to overcome students' preexisting beliefs about evolution, which can bias assimilation of information and inhibit learning. We propose that the most effective way to overcome such belief persistence is through an engaging, hands-on inquiry approach that mimics the scientific process used to study evolution. Although this teaching approach, known as authentic science, has gained recognition for its effectiveness in the classroom, it has not been widely applied to teach evolution. We describe how an authentic science approach can be used to teach evolution by natural selection, and provide a formula for the development of such programs. Following this blueprint, we developed a program using Trinidadian guppies and implemented it in 7th grade classrooms in Colorado. Pre- and post-program assessments revealed significant increases in both the understanding and acceptance of evolution among participants. Authentic science experiments using locally adapted populations of live organisms may be able to overcome belief persistence and improve student understanding of evolution.

Small Fish, Big Questions: Inquiry Kits for Teaching Evolution

Evolution education poses unique challenges because students can have preconceptions that bias their learning. Hands-on, inquiry approaches can help overcome preset beliefs held by students, but few such programs exist and teachers typically lack access to these resources. Experiential learning in the form of self-guided kits can allow evolution education programs to maximize their reach while still maintaining a high-quality resource. We created an inquiry-based kit that uses live Trinidadian guppies to teach evolution by natural selection using the VIST (Variation, Inheritance, Selection, Time) framework. Our collaborative team included evolutionary biologists and education specialists, and we were able to combine expertise in evolution research and inquiry-based kit design in the development of this program. By constructing the kits with grant funds slated for broader impacts and maintaining them at our university's Education and Outreach Center, we made these kits freely available to local schools over the long term. Students and teachers have praised how clearly the kits teach evolution by natural selection, and we are excited to share this resource with readers of The American Biology Teacher.