Sachit Butail

Three-dimensional reconstruction of the fast-start swimming kinematics of densely schooling fish

Information transmission via non-verbal cues such as a fright response can be quantified in a fish school by reconstructing individual fish motion in three dimensions. In this paper, we describe an automated tracking framework to reconstruct the full-body trajectories of densely schooling fish using two-dimensional silhouettes in multiple cameras. We model the shape of each fish as a series of elliptical cross sections along a flexible midline. We estimate the size of each ellipse using an iterated extended Kalman filter. The shape model is used in a model-based tracking framework in which simulated annealing is applied at each step to estimate the midline. Results are presented for eight fish with occlusions. The tracking system is currently being used to investigate fast-start behaviour of schooling fish in response to looming stimuli.

Collective Response of Zebrafish Shoals to a Free-Swimming Robotic Fish

In this work, we explore the feasibility of regulating the collective behavior of zebrafish with a free-swimming robotic fish. The visual cues elicited by the robot are inspired by salient features of attraction in zebrafish and include enhanced coloration, aspect ratio of a fertile female, and carangiform/subcarangiform locomotion. The robot is autonomously controlled with an online multi-target tracking system and swims in circular trajectories in the presence of groups of zebrafish. We investigate the collective response of zebrafish to changes in robot speed, achieved by varying its tail-beat frequency. Our results show that the speed of the robot is a determinant of group cohesion, quantified through zebrafish nearest-neighbor distance, which increases with the speed of the robot until it reaches . We also find that the presence of the robot causes a significant decrease in the group speed, which is not accompanied by an increase in the freezing response of the subjects. Findings of this study are expected to inform the design of experimental protocols that leverage the use of robots to study the zebrafish animal model.

Reconstructing the flight kinematics of swarming and mating in wild mosquitoes

We describe a novel tracking system for reconstructing three-dimensional tracks of individual mosquitoes in wild swarms and present the results of validating the system by filming swarms and mating events of the malaria mosquito Anopheles gambiae in Mali. The tracking system is designed to address noisy, low frame-rate (25 frames per second) video streams from a stereo camera system. Because flying A. gambiae move at 1–4 m s−1, they appear as faded streaks in the images or sometimes do not appear at all. We provide an adaptive algorithm to search for missing streaks and a likelihood function that uses streak endpoints to extract velocity information. A modified multi-hypothesis tracker probabilistically addresses occlusions and a particle filter estimates the trajectories. The output of the tracking algorithm is a set of track segments with an average length of 0.6–1 s. The segments are verified and combined under human supervision to create individual tracks up to the duration of the video (90 s). We evaluate tracking performance using an established metric for multi-target tracking and validate the accuracy using independent stereo measurements of a single swarm. Three-dimensional reconstructions of A. gambiae swarming and mating events are presented.

Sociality Modulates the Effects of Ethanol in Zebra Fish

BACKGROUND The complex social behavior exhibited by zebra fish is often leveraged in preclinical studies to investigate whether and how psychoactive compounds modulate inter individual interactions. Due to theoretical and methodological constraints, previous studies on the effects of ethanol (EtOH) on social behavior focused on homogeneous groups in which all individuals were treated, thereby limiting the possibility of isolating all the intervening variables. METHODS To identify how a social group affects the individual response to EtOH, we quantified the behavior of a single treated individual (acute 0.00, 0.25, 0.50, and 1.00% concentration/volume) swimming together with a group of untreated subjects or alone. A novel in-house-developed automated tracking system was utilized to extract the trajectories of each subject and analyze individual and social behavior. Specifically, we characterized the locomotion of each individual, the cohesion and degree of alignment of the group of untreated subjects, and the interaction between treated and untreated subjects. RESULTS Individual response to high EtOH concentrations varied depending on the presence or absence of conspecifics. Specifically, EtOH-exposed subjects swam faster when group-tested than in isolation. Remarkably, the presence of the exposed individual substantially influenced the behavior of the untreated subjects. Thus, untreated subjects swam faster when the treated individual was exposed to intermediate EtOH concentrations, without varying their cohesion and degree of alignment. No change in the distance between treated and untreated subjects was found; however, the likelihood that the swimming direction of the treated individual anticipated the response of the group was influenced by EtOH concentration. CONCLUSIONS Our results demonstrate the feasibility of exposing a single individual to EtOH and test it together with untreated subjects. This approach has the potential to unravel the social determinants of individual response to alcohol, by enabling us to dissociate EtOH exposure from sociality.

Information Flow in Animal-Robot Interactions

The nonverbal transmission of information between social animals is a primary driving force behind their actions and, therefore, an important quantity to measure in animal behavior studies. Despite its key role in social behavior, the flow of information has only been inferred by correlating the actions of individuals with a simplifying assumption of linearity. In this paper, we leverage information-theoretic tools to relax this assumption. To demonstrate the feasibility of our approach, we focus on a robotics-based experimental paradigm, which affords consistent and controllable delivery of visual stimuli to zebrafish. Specifically, we use a robotic arm to maneuver a life-sized replica of a zebrafish in a predetermined trajectory as it interacts with a focal subject in a test tank. We track the fish and the replica through time and use the resulting trajectory data to measure the transfer entropy between the replica and the focal subject, which, in turn, is used to quantify one-directional information flow from the robot to the fish. In agreement with our expectations, we find that the information flow from the replica to the zebrafish is significantly more than the other way around. Notably, such information is specifically related to the response of the fish to the replica, whereby we observe that the information flow is reduced significantly if the motion of the replica is randomly delayed in a surrogate dataset. In addition, comparison with a control experiment, where the replica is replaced by a conspecific, shows that the information flow toward the focal fish is significantly more for a robotic than a live stimulus. These findings support the reliability of using transfer entropy as a measure of information flow, while providing indirect evidence for the efficacy of a robotics-based platform in animal behavioral studies.

Live Predators, Robots, and Computer-Animated Images Elicit Differential Avoidance Responses in Zebrafish

Emotional disturbances constitute a major health issue affecting a considerable portion of the population in western countries. In this context, animal models offer a relevant tool to address the underlying biological determinants and to screen novel therapeutic strategies. While rodents have traditionally constituted the species of choice, zebrafish are now becoming a viable alternative. As zebrafish gain momentum in biomedical sciences, considerable efforts are being devoted to developing high-throughput behavioral tests. Here, we present a comparative study of zebrafish behavioral response to fear-evoking stimuli offered via three alternative methodologies. Specifically, in a binary-choice test, we exposed zebrafish to an allopatric predator Astronotus ocellatus, presented in the form of a live subject, a robotic replica, and a computer-animated image. The robots design and operation were inspired by the morphology and tail-beat motion of its live counterpart, thereby offering a consistent three-dimensional stimulus to focal fish. The computer-animated image was also designed after the live subject to replicate its appearance. We observed that differently from computer-animated images, both the live predator and its robotic replica elicited robust avoidance response in zebrafish. In addition, in response to the robot, zebrafish exhibited increased thrashing behavior, which is considered a valid indicator of fear. Finally, inter-individual response to a robotic stimulus is more consistent than that shown in response to live stimuli and animated images, thereby increasing experimental statistical power. Our study supports the view that robotic stimuli can constitute a promising experimental tool to elicit targeted behavioral responses in zebrafish.

Model-free information-theoretic approach to infer leadership in pairs of zebrafish.

Collective behavior affords several advantages to fish in avoiding predators, foraging, mating, and swimming. Although fish schools have been traditionally considered egalitarian superorganisms, a number of empirical observations suggest the emergence of leadership in gregarious groups. Detecting and classifying leader-follower relationships is central to elucidate the behavioral and physiological causes of leadership and understand its consequences. Here, we demonstrate an information-theoretic approach to infer leadership from positional data of fish swimming. In this framework, we measure social interactions between fish pairs through the mathematical construct of transfer entropy, which quantifies the predictive power of a time series to anticipate another, possibly coupled, time series. We focus on the zebrafish model organism, which is rapidly emerging as a species of choice in preclinical research for its genetic similarity to humans and reduced neurobiological complexity with respect to mammals. To overcome experimental confounds and generate test data sets on which we can thoroughly assess our approach, we adapt and calibrate a data-driven stochastic model of zebrafish motion for the simulation of a coupled dynamical system of zebrafish pairs. In this synthetic data set, the extent and direction of the coupling between the fish are systematically varied across a wide parameter range to demonstrate the accuracy and reliability of transfer entropy in inferring leadership. Our approach is expected to aid in the analysis of collective behavior, providing a data-driven perspective to understand social interactions.

Influence of robotic shoal size, configuration, and activity on zebrafish behavior in a free-swimming environment.

In animal studies, robots have been recently used as a valid tool for testing a wide spectrum of hypotheses. These robots often exploit visual or auditory cues to modulate animal behavior. The propensity of zebrafish, a model organism in biological studies, toward fish with similar color patterns and shape has been leveraged to design biologically inspired robots that successfully attract zebrafish in preference tests. With an aim of extending the application of such robots to field studies, here, we investigate the response of zebrafish to multiple robotic fish swimming at different speeds and in varying arrangements. A soft real-time multi-target tracking and control system remotely steers the robots in circular trajectories during the experimental trials. Our findings indicate a complex behavioral response of zebrafish to biologically inspired robots. More robots produce a significant change in salient measures of stress, with a fast robot swimming alone causing more freezing and erratic activity than two robots swimming slowly together. In addition, fish spend more time in the proximity of a robot when they swim far apart than when the robots swim close to each other. Increase in the number of robots also significantly alters the degree of alignment of fish motion with a robot. Results from this study are expected to advance our understanding of robot perception by live animals and aid in hypothesis-driven studies in unconstrained free-swimming environments.

The dance of male Anopheles gambiae in wild mating swarms.

ABSTRACT An important element of mating in the malaria vector Anopheles gambiae Giles in nature is the crepuscular mating aggregation (swarm) composed almost entirely of males, where most coupling and insemination is generally believed to occur. In this study, we mathematically characterize the oscillatory movement of male An. gambiae in terms of an established individual-based mechanistic model that parameterizes the attraction of a mosquito toward the center of the swarm using the natural frequency of oscillation and the resistance to its motion, characterized by the damping ratio. Using three-dimensional trajectory data of ten wild mosquito swarms filmed in Mali, Africa, we show two new results for low and moderate wind conditions, and indicate how these results may vary in high wind. First, we show that in low and moderate wind the vertical component of the mosquito motion has a lower frequency of oscillation and higher damping ratio than horizontal motion. In high wind, the vertical and horizontal motions are similar to one another and the natural frequencies are higher than in low and moderate wind. Second, we show that the predicted average disagreement in the direction of motion of swarming mosquitoes moving randomly is greater than the average disagreement we observed between each mosquito and its three closest neighbors, with the smallest level of disagreement occurring for the nearest neighbor in seven out of 10 swarms. The alignment of the direction of motion between nearest neighbors is the highest in high wind. This result provides evidence for flight-path coordination between swarming male mosquitoes.

A jump persistent turning walker to model zebrafish locomotion

Zebrafish are gaining momentum as a laboratory animal species for the investigation of several functional and dysfunctional biological processes. Mathematical models of zebrafish behaviour are expected to considerably aid in the design of hypothesis-driven studies by enabling preliminary in silico tests that can be used to infer possible experimental outcomes without the use of zebrafish. This study is motivated by observations of sudden, drastic changes in zebrafish locomotion in the form of large deviations in turn rate. We demonstrate that such deviations can be captured through a stochastic mean reverting jump diffusion model, a process that is commonly used in financial engineering to describe large changes in the price of an asset. The jump process-based model is validated on trajectory data of adult subjects swimming in a shallow circular tank obtained from an overhead camera. Through statistical comparison of the empirical distribution of the turn rate against theoretical predictions, we demonstrate the feasibility of describing zebrafish as a jump persistent turning walker. The critical role of the jump term is assessed through comparison with a simplified mean reversion diffusion model, which does not allow for describing the heavy-tailed distributions observed in the fish turn rate.