Interspecific differences in developmental mode determine early cognitive abilities in teleost fish.

Most studies on developmental variation in cognition have suggested that individuals are born with reduced or absent cognitive abilities, and thereafter, cognitive performance increases with age during early development. However, these studies have been mainly performed in altricial species, such as humans, in which offspring are extremely immature at birth. In this work, we tested the hypothesis that species with other developmental modes might show different patterns of cognitive development. To this end, we analysed inhibitory control performance in two teleost species with different developmental modes, the zebrafish Danio rerio and the guppy Poecilia reticulata, exploiting a simple paradigm based on spontaneous behaviour and therefore applicable to subjects of different ages. Zebrafish hatch as larvae 3 days after fertilisation, and have an immature nervous system, a situation that mirrors extreme altriciality. We found that at the early stages of development, zebrafish displayed no evidence of inhibitory control, which only begun to emerge after one month of life. Conversely, guppies, which are born after approximately one month of gestation as fully developed and independent individuals, solved the inhibitory control task since their first days of life, although performance increased with sexual maturation. Our study suggests that the typical progression described during early ontogeny in humans and other species might not be the only developmental trend for animals' cognition and that a species' developmental mode might determine variation in cognition across subjects of different age.

Zebrafish excel in number discrimination under an operant conditioning paradigm.

Numerical discrimination is widespread in vertebrates, but this capacity varies enormously between the different species examined. The guppy (Poecilia reticulata), the only teleost examined following procedures that allow a comparison with the other vertebrates, outperforms amphibians, reptiles and many warm-blooded vertebrates, but it is unclear whether this is a feature shared with the other teleosts or represents a peculiarity of this species. We trained zebrafish (Danio rerio) to discriminate between numbers differing by one unit, varying task difficulty from 2 versus 3 to 5 versus 6 items. Non-numerical variables that covary with number, such as density or area, did not affect performance. Most fish reached learning criterion on all tasks up to 4 versus 5 discrimination with no sex difference in accuracy. Although no individual reached learning criterion in the 5 versus 6 task, performance was significant at the group level, suggesting that this may represent the discrimination threshold for zebrafish. Numerosity discrimination abilities of zebrafish compare to those of guppy, being higher than in some warm-blooded vertebrates, such as dogs, horses and domestic fowl, though lower than in parrots, corvids and primates. Learning rate was similar in a control group trained to discriminate between different-sized shapes, but zebrafish were slightly more accurate when discriminating areas than numbers and males were more accurate than females. At the end of the experiment, fish trained on numbers and controls trained on areas generalized to the reciprocal set of stimuli, indicating they had used a relational strategy to solve these tasks.

Guppies (Poecilia reticulata) are deceived by visual illusions during obstacle negotiation.

Animals travelling in their natural environment repeatedly encounter obstacles that they can either detour or go through. Gap negotiation requires an accurate estimation of the opening's size to avoid getting stuck or being injured. Research on visual illusions has revealed that in some circumstances, transformation rules used to generate a three-dimensional representation from bidimensional retinal images fail, leading to systematic errors in perception. In Ebbinghaus and Delboeuf illusions, the presence of task-irrelevant elements causes us to misjudge an object's size. Susceptibility to these illusions was observed in other animals, although with large intraspecific differences. In this study, we investigated whether fish can accurately estimate gap size and whether during this process they may be deceived by illusory patterns. Guppies were extremely accurate in gap negotiation, discriminating holes with a 10% diameter difference. When presented with two identical holes surrounded by inducers to produce Ebbinghaus and Delboeuf patterns, guppies misperceived gap size in the predicted direction. So far, researchers have principally considered illusions as useful tools for exploring the cognitive processing underlying vision. Our findings highlight the possibility that they have important ecological implications, affecting the everyday interactions of an animal with its physical world besides its intra- and interspecific relationships.

Environmental enrichment decreases anxiety-like behavior in zebrafish larvae.

The development of anxiety disorders is often linked to individuals' negative experience. In many animals, development of anxiety-like behavior is modeled by manipulating individuals' exposure to environmental enrichment. We investigated whether environmental enrichment during early ontogenesis affects anxiety-like behavior in larval zebrafish. Larvae were exposed from hatching to either an environment enriched with 3D-objects of different color and shape or to a barren environment. Behavioral testing was conducted at different intervals during development (7, 14, and 21 days post-fertilization, dpf). In a novel object exploration test, 7 dpf larvae of the two treatments displayed similar avoidance of the visual stimulus. However, at 14 and 21 dpf, larvae of the enriched environment showed less avoidance, indicating lower anxiety response. Likewise, larvae of the two treatments demonstrated comparable avoidance of a novel odor stimulus at 7 dpf, with a progressive reduction of anxiety behavior in the enriched treatment with development. In a control experiment, larvae treated before 7 dpf but tested at 14 dpf showed the effect of enrichment on anxiety, suggesting an early determination of the anxiety phenotype. This study confirms a general alteration of zebrafish anxiety-like behavior due to a short enrichment period in first days of life.

Guppies show sex and individual differences in the ability to inhibit behaviour.

In humans, individual and sex differences have been long reported for several cognitive tasks and are at least in part due to variability in the function that inhibits behaviour (i.e. inhibitory control). Similar evidence of individual and sex differences in inhibitory abilities is also present in other vertebrates, but is scarce outside primates. Experiments on reversal learning, which requires inhibiting behaviours, suggest that this variability may exist in a teleost fish, the guppy, Poecilia reticulata. We tested this hypothesis by observing guppies in an inhibitory task. Guppies were exposed to unreachable prey inside a transparent tube for six trials. Guppies showed a marked reduction in the number of attempts to catch the prey within the first trial and also over repeated trials. We found a striking sex difference in the capacity to inhibit foraging behaviour. Males attempted to attack the prey twice as often as females and showed negligible improvement over trials. Irrespective of sex, individuals remarkably differed in their performance, with some guppies being systematically more skilled than others across the repeated trials. These results confirm that individual and sex differences in the ability to inhibit behaviour are not restricted to humans and other primates, suggesting that they might be widespread among vertebrates. Variability in inhibitory ability provides an explanation for emerging records of variability in other cognitive tasks in fish.

The ontogeny of continuous quantity discrimination in zebrafish larvae (Danio rerio).

Several studies have investigated the ontogeny of the capacity to discriminate between discrete numerical information in human and non-human animals. Contrarily, less attention has been devoted to the development of the capacity to discriminate continuous quantities. Recently, we set up a fast procedure for screening continuous quantity abilities in adult individuals of an animal model in neurodevelopmental research, the zebrafish. Two different sized holes are presented in a wall that divides the home tank in two halves and the spontaneous preference of fish for passing through the larger hole is exploited to measure their discrimination ability. We tested zebrafish larvae in the first, second and third week of life varying the relative size of the smaller circle (0.60, 0.75, 0.86, 0.91 area ratio). We found that the number of passages increased across the age. The capacity to discriminate the larger hole decreased as the ratio between the areas increased. No difference in accuracy was found as a function of age. The accuracy of larval zebrafish almost overlaps that found in adults in a previous study, suggesting a limited role of maturation and experience on the ability to estimate areas in this species.

Honeybees use absolute rather than relative numerosity in number discrimination.

Various vertebrate species use relative numerosity judgements in comparative assessments of quantities for which they use larger/smaller relationships rather than absolute number. The numerical ability of honeybees shares basic properties with that of vertebrates but their use of absolute or relative numerosity has not been explored. We trained free-flying bees to choose variable images containing three dots; one group ('larger') was trained to discriminate 3 from 2, while another group ('smaller') was trained to discriminate 3 from 4. In both cases, numbers were kept constant but stimulus characteristics and position were varied from trial to trial. Bees were then tested with novel stimuli displaying the previously trained numerosity (3) versus a novel numerosity (4 for 'larger' and 2 for 'smaller'). Both groups preferred the three-item stimulus, consistent with absolute numerosity. They also exhibited ratio-dependent discrimination of numbers, a property shared by vertebrates, as performance after 2 versus 3 was better than after 3 versus 4 training. Thus, bees differ from vertebrates in their use of absolute rather than of relative numerosity but they also have some numeric properties in common.

Sex differences in discrimination reversal learning in the guppy.

In several mammalian and avian species, females show a higher performance than males in tasks requiring cognitive flexibility such as the discrimination reversal learning. A recent study showed that female guppies are twice as efficient as males in a reversal learning task involving yellow-red discrimination, suggesting a higher cognitive flexibility in female guppies. However, the possibility exists that the superior performance exhibited by females does not reflect a general sex difference in cognitive abilities, but instead, is confined to colour discrimination tasks. To address this issue, we compared male and female guppies in two different discrimination reversal learning tasks and we performed a meta-analysis of these experiments and the previous one involving colour discrimination. In the first experiment of this study, guppies were tested in a task requiring them to learn to select the correct arm of a T-maze in order to rejoin a group of conspecifics. In experiment 2, guppies were observed in a numerical task requiring them to discriminate between 5 and 10 dots in order to obtain a food reward. Although females outperformed males in one condition of the T-maze, we did not find any clear evidence of females' greater reversal learning performance in either experiment. However, the meta-analysis of the three experiments supported the hypothesis of females' greater reversal learning ability. Our data do not completely exclude the idea that female guppies have a generally higher cognitive flexibility than males; however, they suggest that the size of this sex difference might depend on the task.

Do domestic dogs (Canis lupus familiaris) perceive the Delboeuf illusion?

In the last decade, visual illusions have been repeatedly used as a tool to compare visual perception among species. Several studies have investigated whether non-human primates perceive visual illusions in a human-like fashion, but little attention has been paid to other mammals, and sensitivity to visual illusions has been never investigated in the dog. Here, we studied whether domestic dogs perceive the Delboeuf illusion. In human and non-human primates, this illusion creates a misperception of item size as a function of its surrounding context. To examine this effect in dogs, we adapted the spontaneous preference paradigm recently used with chimpanzees. Subjects were presented with two plates containing food. In control trials, two different amounts of food were presented in two identical plates. In this circumstance, dogs were expected to select the larger amount. In test trials, equal food portion sizes were presented in two plates differing in size: if dogs perceived the illusion as primates do, they were expected to select the amount of food presented in the smaller plate. Dogs significantly discriminated the two alternatives in control trials, whereas their performance did not differ from chance in test trials with the illusory pattern. The fact that dogs do not seem to be susceptible to the Delboeuf illusion suggests a potential discontinuity in the perceptual biases affecting size judgments between primates and dogs.

Development and testing of a rapid method for measuring shoal size discrimination.

The shoal-choice test is a popular method to investigate quantity discrimination in social fish based on their spontaneous preference for the larger of two shoals. The shoal-choice test usually requires a long observation time (20-30 min), mainly because fish switch between the two shoals with low frequency, thus reducing the possibilities of comparison. This duration limits the use of the shoal-choice test for large-scale screenings. Here, we developed a new version of the shoal-choice test in which the subject was confined in a large transparent cylinder in the middle of the tank throughout the experiment to bound the minimum distance from the stimulus shoals and favour switching. We tested the new method by observing guppies (Poecilia reticulata) in a 4 versus 6 fish discrimination (experiment 1). The new method allowed for a faster assessment of the preference for the larger shoal (<5 min), resulting in potential application for large population screenings. Guppies switched five times more frequently between the two shoals and remained close to the first chosen shoal ten times less compared to experiments with the old method. In experiment 2, we found that with the new method guppies were able to discriminate up to 5 versus 6 fish, a discrimination that was not achieved with the classical method. This last result indicates that minor methodological modifications can lead to very different findings in the same species and suggests that caution should be exercised when interpreting inter-specific differences in quantitative abilities.

Experimental setting affects the performance of guppies in a numerical discrimination task.

A recent study found that guppies (Poecilia reticulata) can be trained to discriminate 4 versus 5 objects, a numerical discrimination typically achieved only by some mammals and birds. In that study, guppies were required to discriminate between two patches of small objects on the bottom of the tank that they could remove to find a food reward. It is not clear whether this species possesses exceptional numerical accuracy compared with the other ectothermic vertebrates or whether its remarkable performance was due to a specific predisposition to discriminate between differences in the quality of patches while foraging. To disentangle these possibilities, we trained guppies to the same numerical discriminations with a more conventional two-choice discrimination task. Stimuli were sets of dots presented on a computer screen, and the subjects received a food reward upon approaching the set with the larger numerosity. Though the cognitive problem was identical in the two experiments, the change in the experimental setting led to a much poorer performance as most fish failed even the 2 versus 3 discrimination. In four additional experiments, we varied the duration of the decision time, the type of stimuli, the length of training, and whether correction was allowed in order to identify the factors responsible for the difference. None of these parameters succeeded in increasing the performance to the level of the previous study, although the group trained with three-dimensional stimuli learned the easiest numerical task. We suggest that the different results with the two experimental settings might be due to constraints on learning and that guppies might be prepared to accurately estimate patch quality during foraging but not to learn an abstract stimulus-reward association.

Global/local processing of hierarchical visual stimuli in a conflict-choice task by capuchin monkeys (Sapajus spp.).

In the last two decades, comparative research has addressed the issue of how the global and local levels of structure of visual stimuli are processed by different species, using Navon-type hierarchical figures, i.e. smaller local elements that form larger global configurations. Determining whether or not the variety of procedures adopted to test different species with hierarchical figures are equivalent is of crucial importance to ensure comparability of results. Among non-human species, global/local processing has been extensively studied in tufted capuchin monkeys using matching-to-sample tasks with hierarchical patterns. Local dominance has emerged consistently in these New World primates. In the present study, we assessed capuchins' processing of hierarchical stimuli with a method frequently adopted in studies of global/local processing in non-primate species: the conflict-choice task. Different from the matching-to-sample procedure, this task involved processing local and global information retained in long-term memory. Capuchins were trained to discriminate between consistent hierarchical stimuli (similar global and local shape) and then tested with inconsistent hierarchical stimuli (different global and local shapes). We found that capuchins preferred the hierarchical stimuli featuring the correct local elements rather than those with the correct global configuration. This finding confirms that capuchins' local dominance, typically observed using matching-to-sample procedures, is also expressed as a local preference in the conflict-choice task. Our study adds to the growing body of comparative studies on visual grouping functions by demonstrating that the methods most frequently used in the literature on global/local processing produce analogous results irrespective of extent of the involvement of memory processes.

Quantitative abilities in a reptile (Podarcis sicula).

The ability to identify the largest amount of prey available is fundamental for optimizing foraging behaviour in several species. To date, this cognitive skill has been observed in all vertebrate groups except reptiles. In this study we investigated the spontaneous ability of ruin lizards to select the larger amount of food items. In Experiment 1, lizards proved able to select the larger food item when presented with two alternatives differing in size (0.25, 0.50, 0.67 and 0.75 ratio). In Experiment 2 lizards presented with two groups of food items (1 versus 4, 2 versus 4, 2 versus 3 and 3 versus 4 items) were unable to select the larger group in any contrast. The lack of discrimination in the presence of multiple items represents an exception in numerical cognition studies, raising the question as to whether reptiles' quantitative abilities are different from those of other vertebrate groups.

The contribution of fish studies to the "number sense" debate.

Leibovich et al. propose that number sense is not innate but gradually emergent during ontogeny following experience. We argue that this hypothesis cannot be reasonably tested in humans, in which the contribution of neural maturation and experience cannot be experimentally manipulated. Studies on animals, especially fish, can more effectively provide critical insights into the innate nature of numerical abilities.

Early visual experience influences behavioral lateralization in the guppy.

Individual differences in lateralization of cognitive functions characterize both humans and non-human species. Genetic factors can account for only a fraction of the variance observed and the source of individual variation in laterality remains in large part elusive. Various environmental factors have been suggested to modulate the development of lateralization, including asymmetrical stimulation of the sensory system during ontogeny. In this study, we raised newborn guppies in an asymmetric environment to test the hypothesis that early left-right asymmetries in visual input may affect the development of cerebral asymmetries. Each fish was raised in an impoverished environment but could voluntarily observe a complex scene in a nearby compartment containing a group of conspecifics. Using asymmetric structures, we allowed some subjects to observe the complex scene with the right eye, others with the left eye, and control fish with both eyes. Among asymmetrically stimulated fish, the mirror test revealed eye dominance congruent with the direction of asymmetric stimulation, while controls showed no left-right laterality bias. Interestingly, asymmetric exposure to social stimuli also affected another aspect of visual lateralization-eye preference for scrutinizing a potential predator-but did not influence a measure of motor asymmetry. As the natural environment of guppies is fundamentally asymmetrical, we suggest that unequal left-right stimulation is a common occurrence in developing guppies and may represent a primary source of individual variation in lateralization as well as an efficient mechanism for producing laterality phenotypes that are adapted to local environmental conditions.

Male and female guppies differ in speed but not in accuracy in visual discrimination learning.

In many species, males and females have different reproductive roles and/or differ in their ecological niche. Since in these cases the two sexes often face different cognitive challenges, selection may promote some degree of cognitive differentiation, an issue that has received relatively little attention so far. We investigated the existence of sex differences in visual discrimination learning in the guppy, Poecilia reticulata, a fish species in which females show complex mate choice based on male colour pattern. We tested males and females for their ability to learn a discrimination between two different shapes (experiment 1) and between two identical figures with a different orientation (experiment 2). In experiment 3, guppies were required to select an object of the odd colour in a group of five objects. Colours changed daily, and therefore, the solution for this task was facilitated by concept learning. We found males' and females' accuracy practically overlapped in the three experiments, suggesting that the two sexes have similar discrimination learning abilities. Yet, males showed faster decision time than females without any evident speed-accuracy trade-off. This result indicates the existence of consistent between-sex differences in decision speed perhaps due to impulsivity rather than speed in information processing. Our results align with previous literature, indicating that sex differences in cognitive abilities are the exception rather than the rule, while sex differences in cognitive style, i.e. the way in which an individual faces a cognitive task, are much more common.

Quantity discrimination in parental fish: female convict cichlid discriminate fry shoals of different sizes.

Numerical abilities have been found to be adaptive in different contexts, including mating, foraging, fighting assessment and antipredator strategies. In species with parental care, another potential advantage is the possibility to adjust parental behavior in relation to the numerosity of the progeny. The finding that many fish vary their parental investment in relation to brood size advocates the existence of a mechanism for appraising offspring number, an aspect that has never been directly investigated. Here we tested the ability of parental female convict cichlid (Amatitlania nigrofasciata) to discriminate between two fry groups differing in number by measuring time spent attempting to recover groups of fry experimentally displaced from the next. Females spent more time trying to recover the fry from larger groups when tested with contrasts 6 versus 12 (1:2) and 6 versus 9 fry (2:3); however, they showed no preference in the 6 versus 8 (3:4) contrast, suggesting that this task exceeds their discrimination capacity.

Relative versus absolute numerical representation in fish: Can guppies represent "fourness"?

In recent years, the use of operant conditioning procedures has shown that species as diverse as chimpanzees, honeybees, and mosquitofish can be trained to discriminate between sets containing different numbers of objects. However, to succeed in this task, subjects can use two different strategies: either select the array containing a specific number of items (an absolute numerosity rule), or select the set containing the larger (or smaller) quantity of items (a relative numerosity rule). In the latter case, subjects need not only be able to judge whether two numerosities are equal or different but also be able to order numerosities. Here, in two experiments, we address whether fish can perform both kinds of judgment by training them with specific numerosities and testing their generalization to new numerosity contrasts. In Experiment 1, subjects were initially trained to select between visual arrays of 6 and 12 shapes, and were then tested with a contrast pairing the previously trained numerosity (either 6 or 12) with a novel numerosity (respectively, 3 or 24). Spontaneously, subjects selected the novel numerosity, in accordance with a relative numerosity rule. The second experiment tested whether guppies can also learn to select one specific number against all others, if appropriately trained. Fish trained to select an array of 4 shapes against several alternatives (4 vs. 1, 4 vs. 2, 4 vs. 8, 4 vs. 10) learned to recognize the number 4 against all alternatives and proved able to generalize their discrimination to novel, more difficult contrasts (4 vs. 3 and 4 vs. 6 items). In summary, although guppies preferentially opt for relative comparisons, they can flexibly learn either relative or absolute decision criteria on numerosity stimuli, depending on the context.

Non-visual numerical discrimination in a blind cavefish (Phreatichthys andruzzii).

Over a decade of comparative studies, researchers have found that rudimentary numerical abilities are widespread among vertebrates. While experiments in mammals and birds have employed a variety of stimuli (visual, auditory and tactile), all fish studies involved visual stimuli and it is unknown whether fish can process numbers in other sensory modalities. To fill this gap, we studied numerical abilities in Phreatichthys andruzzii, a blind cave-dwelling species that evolved in the phreatic layer of the Somalia desert. Fish were trained to receive a food reward to discriminate between two groups of objects placed in opposite positions of their home tank. In Experiment 1, subjects learned to discriminate between two and six objects, with stimuli not controlled for non-numerical continuous variables that co-vary with numbers, such as total area occupied by stimuli or density. In Experiment 2, the discrimination was two versus four, with half of the stimuli controlled for continuous quantities and half not controlled for continuous quantities. The subjects discriminated only the latter condition, indicating that they spontaneously used non-numerical information, as other vertebrates tested in similar experiments. In Experiments 3 and 4, cavefish trained from the beginning only with stimuli controlled for continuous quantities proved able to learn the discrimination of quantities based on the sole numerical information. However, their numerical acuity was lower than that reported in other teleost fish tested with visual stimuli.

Numerical acuity of fish is improved in the presence of moving targets, but only in the subitizing range.

There is controversy in comparative psychology about whether on the one hand non-symbolic number estimation of small (≤4) and large numbers involves a single mechanism (an approximate number system), or whether on the other hand enumeration of the numbers 1-4 is accomplished by a separate mechanism, an object tracking system. To date, support for the latter hypothesis has come only from the different ratio-dependency of performance seen in the two numerical ranges, a reading that has been criticized on several grounds. In humans, the two-system hypothesis is supported by evidence showing that manipulation of the physical properties of the stimuli (e.g., the motion of the items) has dissimilar effects on small- and large-number discrimination. In this research, we studied this effect on guppies. Initially, fish were trained to simultaneously discriminate two numerical contrasts having the same easy ratio (0.50): one in the small-number (2 vs. 4) range and one in the large-number (6 vs. 12) range. Half of the fish were presented with moving items; the other half were shown the same stimuli without motion. Fish were then subjected to non-reinforced probe trials in the presence of a more difficult ratio (0.75: 3 vs. 4 and 9 vs. 12). Under both static and moving conditions, the fish significantly discriminated 6 versus 12, but not 9 versus 12 items. As regards small numbers, both groups learned to discriminate a 0.50 ratio, but only fish tested with moving stimuli also discriminated 3 and 4 items. This differential effect suggests that fish may possess two separate systems for small- and large-number discrimination.