Shoals in troubled waters? The impact of rising temperatures on metabolism, foraging, and shoaling behavior in mixed-species shoals.

Rising water temperatures across aquatic habitats, in the current global climate change scenario, can directly affect metabolism and food intake in fish species. This can potentially alter their physiological, behavioral, and shoaling properties. In the current study, we examined the effects of high temperatures on metabolism, foraging, and shoaling in tropical fish. Mixed-species (comprising flying barbs, zebrafish, and gambusia) and single-species (flying barbs and zebrafish) shoals were conditioned for 45 days to three kinds of temperature regimes: the current temperature regime (CTR), in which shoals were maintained at water temperature of 24°C (i.e., the current mean temperature of their habitat), the predicted temperature regime (PTR) at 31°C (i.e., simulating conditions projected for their habitat in 2100), and the dynamic temperature regime (DTR), which experienced daily temperature fluctuations between 24 and 31°C (i.e., resembling rapid temperature changes expected in their natural environments). We found species-specific responses to these temperature regimes. Flying barbs exhibited significantly lower body weight at PTR but maintained consistent muscle glycogen content across all temperature regimes. In contrast, zebrafish and gambusia displayed significantly elevated muscle glycogen content at PTR, with similar body weights across all three temperature regimes. Cohesion within flying barb shoals and cohesion/polarization in mixed-species shoals decreased significantly at PTR. Shoals exposed to DTR exhibited intermediate characteristics between those conditioned to CTR and PTR, suggesting that shoals may be less impacted by dynamic temperatures compared to prolonged high temperatures. This study highlights species-specific metabolic responses to temperature changes and their potential implications for larger-scale shoal properties.

River zebrafish combine behavioral plasticity and generalized morphology with specialized sensory and metabolic physiology to survive in a challenging environment.

Phenotypes that allow animals to detect, weather, and predict changes efficiently are essential for survival in fluctuating environments. Some phenotypes may remain specialized to suit an environment perfectly, while others become more plastic or generalized, shifting flexibly to match current context or adopting a form that can utilize a wide range of contexts. Here, we tested the differences in behavior, morphology, sensory and metabolic physiology between wild zebrafish (Danio rerio) in highly variable fast-flowing rivers and still-water sites. We found that river zebrafish moved at higher velocities than did still-water fish, had lower oxygen demands, and responded less vigorously to small changes in flow rate, as we might expect for fish that are well-suited to high-flow environments. River zebrafish also had less streamlined bodies and were more behaviorally plastic than were still-water zebrafish, both features that may make them better-suited to a transitional lifestyle. Our results suggest that zebrafish use distinct sensory mechanisms and metabolic physiology to reduce energetic costs of living in fast-flowing water while relying on morphology and behavior to create flexible solutions to a challenging habitat. Insights on animals' reliance on traits with different outcomes provide a framework to better understand their survival in future environmental fluctuations.

Correlations begin at home: drivers of co-occurrence patterns in personality and cognitive ability in wild populations of zebrafish.

Aquatic habitats are extremely dynamic, with constantly changing ecological factors, which has now been exacerbated due to human-induced rapid environmental change. In such variable environments, it becomes essential to understand how personality and cognition in organisms affect the adaptability of individuals to different habitat conditions. To test this, we studied how personality-related traits as well as cognitive ability differ between populations of wild-caught zebrafish (Danio rerio) from habitats that differed in various environmental factors. We measured emergence into a novel environment as an indicator of boldness, and performance in a spatial task inferred from feeding latencies in a maze over repeated trials to assess learning and memory, as an indicator of cognitive ability. We found that personality affects cognition and although bolder fish are better learners, they show poorer retention of memory across populations. Although personality and cognitive ability varied between habitats, the patterns of their correlations remained similar within each population. However, the individual traits (such as sex and size) that were drivers of personality and cognition differed between the habitats, suggesting that not only do behavioral traits vary between populations, but also the factors that are important in determining them. Personality and cognitive ability and the correlations between these traits determine how well an organism performs in its habitat, as well as how likely it is to find new habitats and adapt to them. Studying these across wild zebrafish populations helps predict performance efficiencies among individuals and also explains how fish adapt to extremely dynamic environments that can lead to variation in behavioral traits and correlations between them. This study not only sheds light on the drivers of interindividual variation and co-occurrence patterns of personality and cognition, but also individual and population factors that might have an effect on them.

What drives mixed-species shoaling among wild zebrafish? The roles of predators, food access, abundance of conspecifics and familiarity.

Mixed-species groups occur across a wide range of faunal communities and provide several benefits to members. While zebrafish have often been observed to form mixed-species shoals with coexisting species, the factors determining their occurrence are not yet fully understood. Shoals comprising zebrafish (Danio rerio), flying barbs (Esomus danricus), and whitespots (Aplocheilus panchax) were collected from a stagnant canal at Haringhata (West Bengal, India), and using laboratory-based experiments, we deciphered likely drivers of mixed-species shoaling among zebrafish. Experiments assessing foraging efficiency revealed that the amount of food consumed by individual zebrafish in mixed shoals was comparable to the amount consumed by these individuals in conspecific shoals. Within mixed-species shoals, zebrafish individuals, despite being smaller than the other species, consumed a comparable amount of food as the other species. Shoal choice experiments revealed that under predator risk, zebrafish associate more with mixed shoals and showed comparable associations to shoals differing in the abundance of conspecifics. Furthermore, zebrafish preferred associating with familiar conspecifics over unfamiliar mixed and unfamiliar conspecific shoals. Therefore, equitable food consumption in mixed shoals, greater association with mixed shoals in the presence of predators, and familiarity were important in driving zebrafish towards mixed-species shoaling. This article has an associated First Person interview with the first author of the paper.

Sex and Population Drive Interindividual Variations in a Cognitive Task Across Three Populations of Wild Zebrafish.

Animal personality refers to the consistency of variation in behavior among individuals which may be the driving force behind variations in complex behaviors as well. Individual personality could predict how well an organism would perform in behavior and cognition related tasks, as well as survive and thrive in its environment. Therefore, we would expect inter-individual variations in many behaviors, which would persist even if habituation to the experimental setup occurs, which generally results in convergence of behavior (i.e., the difference between individuals becomes less pronounced). Our study used wild-caught zebrafish (Danio rerio) from three natural habitats with differing ecological regimes, to understand how consistency and repeatability in specific traits such as boldness, exploration, and spatial ability varies across and within populations even when habituation causes change in behavior. We found that the extent of individual variation differs between populations, with dynamic habitats showing similar repeatability. This indicates that habitat conditions are important drivers of individual variation in addition to other factors, such as sex or size of individuals within populations. Although we found that sex and size played an important role within some populations for some behaviors, in others, the variation was likely caused by other factors (for example, ecological factors such as vegetation and/or resource availability), for which we have not accounted. This study underlines the importance of studying inter-individual differences as the phenomenon that underpins multiple behavioral traits and explains the possible role of environmental and inherent factors that drive these differences.

Structural Insights into Self-Assembled Aerosol-OT Aggregates in Aqueous Media Using Atomistic Molecular Dynamics.

In water, the surfactant dioctyl sulfosuccinate (Aerosol-OT or AOT) exhibits diverse aggregate structures, ranging from micelles to lamella. An atomic-level understanding, however, of the formation and structure of these aggregates is lacking. Herein, using atomistic molecular dynamics (MD) with microsecond-long simulations, self-assembly of AOT in water is studied for concentrations of 1, 7.2, and 20 wt % at 293 K and for 7.2 wt % at 353 K. Assembly proceeds through stepwise association and dissociation of single AOT molecules, and the fusion and fission of AOT clusters. At 293 K, AOT self-assembles into either (i) spherical micelles (1 wt %), (ii) biphasic systems consisting of rod-like and prolate spheroidal micelles (7.2 wt %), or (iii) bilayers (20 wt %). We hypothesize that the observed rod-like structure is a precursor to lamellar microdomains found experimentally in biphasic dispersions. Increasing temperature to 353 K at 7.2 wt % results in a system consisting of prolate micelles but no rod-like micelles. Simulated phase behavior agrees with previously published experimental observations. Individual aggregates formed during self-assembly are identified using graph theory. Structural metrics of these aggregates like the radius of gyration, shape anisotropy, and prolateness are presented. Trends in structural metrics quantitatively reflect how shapes and sizes of AOT aggregates vary with surfactant concentration and temperature. These simulations provide deeper insight into open questions in the scientific community and demonstrate a method to generate physics-based micelle structures that can be used to rationalize experimental observations.

Wild Zebrafish Sentinels: Biological Monitoring of Site Differences Using Behavior and Morphology.

Environmental change poses a devastating risk to human and environmental health. Rapid assessment of water conditions is necessary for monitoring, evaluating, and addressing this global health danger. Sentinels or biological monitors can be deployed in the field using minimal resources to detect water quality changes in real time, quickly and cheaply. Zebrafish (Danio rerio) are ideal sentinels for detecting environmental changes due to their biomedical tool kit, widespread geographic distribution, and well-characterized phenotypic responses to environmental disturbances. Here, we demonstrate the utility of zebrafish sentinels by characterizing phenotypic differences in wild zebrafish between two field sites in India. Site 1 was a rural environment with flowing water, low-hypoxic conditions, minimal human-made debris, and high iron and lead concentrations. Site 2 was an urban environment with still water, hypoxic conditions, plastic pollution, and high arsenic, iron, and chromium concentrations. We found that zebrafish from Site 2 were smaller, more cohesive, and less active than Site 1 fish. We also found sexually dimorphic body shapes within the Site 2, but not the Site 1, population. Advancing zebrafish sentinel research and development will enable rapid detection, evaluation, and response to emerging global health threats.

Temporal Patterns in Interactions Across Same- and Mixed-Sex Wild Zebrafish Dyads.

Animals communicate with each other through a variety of behavioral interactions, many of which are often complex due to the interplay of several ecological factors. Observations on dyadic interactions can help throw light on the more complex interactions observed among group living organisms and can help in understanding mechanisms of behaviors related to mating strategies, dominance hierarchies, and decision-making. This study focused on the assessment of several generally observed interactions among dyads of different sexes (female-female, male-male, and male-female) in wild zebrafish (Danio rerio). Temporal dynamics of these interactive behaviors were observed in 45 dyads across 3 time intervals of the day. We used generalized linear mixed models to investigate the effect of time, sex of dyad, and their interaction on specific behaviors. While the frequency of occurrence of some behaviors showed clear variation across time intervals of the day, these were further found to depend on the composition of the dyad. Contrary to previous reports, we found that same-sex dyads are equally aggressive and aggressive interactions did not vary temporally. Mating-associated interactions, as expected, were significantly higher in mixed-sex dyads and declined significantly from early morning to afternoon. Interestingly, we also found some mating-associated interactions in same-sex dyads. A fine line exists between social and mating-associated interactions in many organisms and so we speculate that these interactions could also be social interactions and not mating-related behavior. Our findings shed light on complex interactive behaviors among zebrafish, that are likely to be affected by time as well as sex composition of interacting individuals and thus has important implications for groups varying in sex ratios in the wild.

Group size and aquatic vegetation modulates male preferences for female shoals in wild zebrafish, Danio rerio.

Shoaling decisions in the wild are determined by a combination of innate preferences of the individual along with the interplay of multiple ecological factors. In their natural habitat as well as in the laboratory, zebrafish is a shoaling fish. Here, we investigate the role of group size and associated vegetation in shaping shoaling preferences of wild male zebrafish. We studied the association preference of males to groups of female shoals in a multi-choice test design. We found that males made greater proportion of visits to an 8-female group compared to 2 and 4-female groups. However, males spent similar proportions of time across the three female-containing groups. When artificial vegetation was incorporated along with female number as an additional factor, we found that males prefer high and moderately vegetated patches compared to low or no-vegetation groups, irrespective of the number of females in these patches. Based on experiments using a novel multi-choice design, our results show that preference for group size can change due to interaction of two separate factors. This work is a first attempt to understand the role of aquatic flora in determining shoaling preferences in zebrafish, using an experimental paradigm consisting of a gradation in female and vegetation densities.

Bolder and Brighter? Exploring Correlations Between Personality and Cognitive Abilities Among Individuals Within a Population of Wild Zebrafish, Danio rerio.

Within populations, individual differences in behavioral and cognitive traits are dependent on the habitat and specific contexts, such as the presence of a predator or other risks. The ability to show variable responses to changing conditions can be of immense survival advantage to organisms. We studied individual differences in specific personality traits, such as boldness, exploration, and spatial ability, and the effect of these traits on learning ability and memory in the presence of a predatory threat, among wild caught zebrafish (Danio rerio). Under laboratory conditions, individuals were trained to perform a simple navigation task, and their performance, exploration, boldness traits were measured, along with learning and memory abilities under two contexts (i.e., in the presence and absence of a predator). Our results revealed that fish showed a clear decline in emergence time, exploration time, and feeding latency over trials, indicative of learning, and further tests for memory also showed memory retention. While the presence of a predator increased emergence time and latencies for navigating, indicating declines in boldness and exploration, these were found to be correlated to different personalities among the individuals and dependent on their sex. While females tended to be bolder and learned the spatial task faster, they showed lower memory retention abilities than males. Personality traits were also found to affect cognitive abilities among individuals. In general, the presence of a predator decreased performance latencies. However, bolder individuals were less affected and emerged more quickly from the refuge chamber than shy individuals. Our results point to the complex interplay of ecological context along with inherent correlations across personality traits that decide the overall personality and cognitive responses among individuals even within populations. These findings thus highlight the importance of an inclusive approach that combines personality and cognition studies for understanding variations within populations.

Collective Behavior in Wild Zebrafish.

Anthropogenic change is expected to alter environments at alarming rates. To predict the impact of modified environments on social behavior, we must study the relationship between environmental features and collective behavior in a genetically tractable model, zebrafish (Danio rerio). Here, we conducted a field study to examine the relationship between salient environmental features and collective behavior in four populations of zebrafish. We found zebrafish in flowing water formed volatile groups, whereas those in still water had more consistent membership and leadership. Groups in fast-flowing water were large (up to 2000 fish) and tightly knit with short nearest neighbor distances, whereas group sizes were smaller (11 fish/group) with more space between individual fish in still and slow-flowing water. These observations point to a possible profound role of water flow in influencing collective behavior in wild zebrafish.

Temporal and environmental drivers of fish-community structure in tropical streams from two contrasting regions in India.

Environmental and anthropogenic factors are known to drive fish community structure in aquatic systems across the world. This study investigates fish assemblages in lower order streams across contrasting landscapes in central and eastern India. We documented the species diversity of these monsoon driven lower order streams in the two regions. We also investigated the potential common environmental drivers of richness and diversity and effect of season in these tropical streams. The study was based on seasonal data on abundance of fishes and environmental parameters collected between 2015-2017 from streams in states of Madhya Pradesh and West Bengal. Species diversity were compared across regions and seasons, based on their richness (SR) as well as diversity (Shannon index H'). Drivers of overall richness and diversity were analyzed using multiple linear regression methods, based on best subset selection. Analysis of data revealed high diversity in these streams in both regions. Cyprinidae, Bagridae and Channidae were the most dominant families in both regions. Despite the geographical and local ecological differences across the regions, common environmental parameters were found to influence richness and diversity across the two regions, indicating these as being key drivers of fish community structure. Water flow was a common factor driving both richness and diversity across both regions. Our study revealed a lack of seasonal effect in structuring fish communities in tropical streams. With stream and river ecosystems facing increasing threats due to habitat alterations and water quality degradation in countries such as India, a clear understanding of regional and local drivers of community structure of aquatic fauna is crucial. These results on the role of common environmental factors across ecoregions provides baseline information for understanding their ecological roles and developing management plans for important river basins and fish conservation in future.

Color preferences affect learning in zebrafish, Danio rerio.

Animals may exhibit preference for colors that match their environment or the resources in the environment. These preferences may impact ability to learn associations with these colors and revert the associations when the reward contingency is modified. We used zebrafish Danio rerio from four populations to test if color preferences impact associative and reversal learning ability. First, we tested if preference for blue or green impact associative ability. We subjected individual fish through eight trials to associate a social stimulus with blue or green. Next, we tested if preference for red or green impact associative reversal learning ability. We trained fish in groups of three to associate a social stimulus with red or green over three trials, and reversed the reward contingency during the following session. Results showed that zebrafish preferred green over blue and domesticated fish chose green more than blue when there was a reward attached. Zebrafish also preferred red over green. Fish from one wild population learned with both colors and reversed learning only from green to red and not vice-versa. Fish from another population showed an overwhelming preference for red irrespective of what was rewarded. Domesticated fish did not show reversal learning ability.

Feeding in murky waters: acclimatization and landmarks improve foraging efficiency of zebrafish (Danio rerio) in turbid waters.

Fish inhabiting human-dominated ecosystems are prone to altered sensory environments in which they must live and function. Increased turbidity is one such change that they must deal with. We tested whether an increase in water turbidity and the presence of visual landmarks (coloured stones) affect the foraging efficiency of wild zebrafish. We also tested the influence of extended exposure to differing turbidity levels on the subsequent foraging efficiency of acclimatized individuals. Feeding latency (time taken to find food) increased significantly with increase in turbidity levels from a minimum of 4 s to ca 300 s. However, extended exposure of fish to varying levels of turbidity decreased feeding latencies in acclimatized conditions, indicating that acclimatization to the immediate visual environment plays an important role in determining foraging success. Most significantly, we found that feeding latencies in turbid conditions decreased significantly if visual landmarks were present. This demonstrates that zebrafish use visual landmark cues to navigate to foraging sites when visibility is impaired. This study has important implications on the role of behavioural plasticity and spatial learning in animals that allow them to cope with altered sensory environments such as episodes of enhanced turbidity that could be natural or anthropogenic.

Temporal patterns and sex differences in dyadic interactions in a wild zebrafish population.

Male-female interactions in several group living organisms, including some fish species tend to be dynamic and play a key role in determining their mating and courtship behavior. Laboratory-bred zebrafish (Danio rerio) strains are one of the most widely used model systems in various fields of biology. While research on wild zebrafish behavior is gaining ground, our knowledge about their mating ecology and mating strategies is still limited. We investigated diel temporal patterns in inter-sex dyadic interactions among wild zebrafish and the occurrence of behavioral dimorphism in their interactive behaviors. We observed randomly paired male and female individuals at three distinct time intervals (early morning, morning and afternoon sessions) in the day and collected occurrence data for six discrete inter-individual interactive behaviors that were associated with mating, aggression, and display. We used generalized linear mixed models to examine the effect of time, sex of the individual and presence of oviposition substrate on these behavioral traits. We found a higher incidence of mating-associated behavior during the early parts of the day which declined by the afternoon. These mating-associated behaviors were also dependent on the presence of gravel substrate for egg-laying compared to other behaviors. This work is the first of its kind that details patterns in behavioral dimorphism between sexes in zebrafish. Our results throw light on the complex dynamics of male-female interactions in a group living externally fertilizing species and can have implications in designing experiments involving behavioral testing of zebrafish which is increasingly being done in toxicological studies and laboratory breeding.

Repeatability in boldness and aggression among wild zebrafish (Danio rerio) from two differing predation and flow regimes.

Although flexibility in behavior is adaptive, this flexibility is limited, and the extent of variation and consistency of a trait could depend on the environment. In this study, we investigated repeatability in risk-taking during feeding among individuals and agonistic interactions among dyads of wild zebrafish, Danio rerio, collected from two habitats that differed in predation and flow regimes. We measured boldness as the latency to emerge from a shelter and feed in the presence of predators. We tested this for each individual from the two populations repeatedly across seven trials. We assessed aggression by subjecting size- and sex-matched pairs of fish to dyadic contests repeatedly across seven trials. Individuals from the high-predation stream population were bolder than individuals from the low-predation stagnant water population. Males were bolder than females, and in the low-predation population, larger individuals took greater risks to feed than smaller individuals. The high-predation stream population showed lower inter- and intraindividual variation in boldness than the low-predation stagnant water population. Further, both populations showed significant repeatability in risk-taking during feeding. The high-predation stream habitat fish were more aggressive than low-predation stagnant-water fish. Male dyads from the low-predation stagnant-water population were significantly more aggressive than female dyads. Most fish from the low-predation stagnant water population did not show aggressive behavior, resulting in low between-dyad but high within-dyad variation in aggression. The difference in behavioral responses between the populations and consistency in these traits within individuals is discussed in the light of confounding role of ecological and state-dependent factors. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Population, sex and body size: determinants of behavioural variations and behavioural correlations among wild zebrafish Danio rerio.

This study (1) investigated variation among populations and the effects of sex and body size on boldness, activity and shoal-association tendency among wild zebrafish, and (2) tested for existence of correlations between behaviours, controlling for sex and body size. Individuals across four natural populations were tested for general activity in a novel situation, number of predator inspections undertaken and tendency to associate with a conspecific shoal in the presence of predators. Results showed a significant effect of population on boldness with a population from high-predation habitat being bolder than populations from low-predation habitats. Males showed significantly higher tendencies than females to associate with a conspecific shoal in the presence of predators. Further, a negative relationship was found between activity and boldness only within two low-predation populations. Individual body size had a strong effect on the activity-boldness relationship within the low-predation population from flowing water habitat. Smaller fish were bolder and less active while larger fish were more cautious and active. Overall, the results indicated that while population-level behavioural responses might be shaped by predation pressure, state-dependent factors could determine behavioural correlations among individuals within populations.

Divergences in learning and memory among wild zebrafish: Do sex and body size play a role?

Given its diverse ecological distribution, zebrafish has great potential for investigations on the effect of habitat characteristics on cognition. Studies were conducted on four wild-caught zebrafish populations to understand the role of native habitat, sex, and body size in determining learning through a novel task associated with a food reward. The habitat variables, namely, the relative abundances of zebrafish and predatory fish and the substrate and vegetation diversity, were quantified during fish sampling. Fish were subjected to a novel task to find a food reward in a maze over successive training trials followed by a test for memory. Performances of subjects were based on time taken to find the food reward and number of mistakes made during trials, and tests for memory. The experiments revealed significant differences in learning rates and memory across populations. Males made significantly fewer mistakes than females only within two populations. No relationship between performance and body size was observed. The differences in learning and memory among wild zebrafish could be due to differences in predation, complexity, and stability of the native habitats. These findings suggest the possible role of multiple interacting factors in determining learning and memory among populations and point to a need for incorporating effects of several factors in future studies.

Social learning in a maze? Contrasting individual performance among wild zebrafish when associated with trained and naïve conspecifics.

Social learning facilitates informed decision making about foraging, mating and anti-predatory tactics among animals. We investigated the occurrence of social learning through performance in a spatial task among wild-caught zebrafish. Individual fish (demonstrators) were trained through a novel food finding task in a maze for 8days. Demonstrators were paired with naïve individuals (observers) and subjected to trials through maze for 4days followed by removal of the demonstrators and further training of observers for 4 more days. Paired naïve individuals were subjected to trials through the maze in similar fashion separately and the performance of observers were compared with theirs. Our results showed that observers associated with knowledgeable conspecifics did not perform the task better than naïve-paired individuals. Performances across trials improved for both sets while number of mistakes committed increased indicating no learning. The presence of a demonstrator could have increased the observer's activity, increasing the chances for the observer to come in contact with the stimulus. Performance of observers and naïve-paired fish were probably affected by social distraction. Sex and body-size of the dyads (demonstrator-observer pairs and naïve pairs) could also have interfered with information transfer among individuals.

Risk-Taking During Feeding: Between- and Within-Population Variation and Repeatability Across Contexts Among Wild Zebrafish.

Behavioral responses are shaped by a complex interplay of extrinsic factors, such as predation and flow conditions, and intrinsic properties such as metabolic state. We investigated variations in boldness among four populations of zebrafish collected from widely varying habitat conditions across India. We measured feeding latencies of fish in four contexts: in a novel environment; in an environment with refuge and predators; in the presence of a shoal; and in the presence of a shoal and predators. The population from a high-flow and high-predation habitat was significantly bolder than populations from low-predation and low-flow/stagnant habitats. Individuals were significantly bolder in the presence of shoal and predators than in the other contexts. In general, males were significantly bolder than females. Smaller individuals took greater risks to feed than larger individuals only within two populations. The high-predation, high-flow habitat fish also showed significantly lower between- and within-individual variation in boldness than all the low-predation and low-flow populations. Furthermore, we found significantly repeatable responses across contexts within all populations, indicating behavioral consistency. Differences in response to variations in experimental contexts and stress and individual differences in metabolic rates could possibly explain the observed between- and within-individual variation among populations.