Tail Beat Synchronization during Schooling Requires a Functional Posterior Lateral Line System in Giant Danios, Devario aequipinnatus.

Swimming in schools has long been hypothesized to allow fish to save energy. Fish must exploit the energy from the wakes of their neighbors for maximum energy savings, a feat that requires them to both synchronize their tail movements and stay in certain positions relative to their neighbors. To maintain position in a school, we know that fish use multiple sensory systems, mainly their visual and flow sensing lateral line system. However, how fish synchronize their swimming movements in a school is still not well understood. Here, we test the hypothesis that this synchronization may depend on functional differences in the two branches of the lateral line sensory system that detects water movements close to the fish's body. The anterior branch, located on the head, encounters largely undisturbed free-stream flow, while the posterior branch, located on the trunk and tail, encounters flow that has been affected strongly by the tail movement. Thus, we hypothesize that the anterior branch may be more important for regulating position within the school, while the posterior branch may be more important for synchronizing tail movements. Our study examines functional differences in the anterior and posterior lateral line in the structure and tail synchronization of fish schools. We used a widely available aquarium fish that schools, the giant danio, Devario equipinnatus. Fish swam in a large circular tank where stereoscopic videos recordings were used to reconstruct the 3D position of each individual within the school and to track tail kinematics to quantify synchronization. For one fish in each school, we ablated using cobalt chloride either the anterior region only, the posterior region only, or the entire lateral line system. We observed that ablating any region of the lateral line system causes fish to swim in a "box" or parallel swimming formation, which was different from the diamond formation observed in normal fish. Ablating only the anterior region did not substantially reduce tail beat synchronization but ablating only the posterior region caused fish to stop synchronizing their tail beats, largely because the tail beat frequency increased dramatically. Thus, the anterior and posterior lateral line system appears to have different behavioral functions in fish. Most importantly, we showed that the posterior lateral line system played a major role in determining tail beat synchrony in schooling fish. Without synchronization, swimming efficiency decreases, which can have an impact on the fitness of the individual fish and group.

Red muscle activity in bluegill sunfish Lepomis macrochirus during forward accelerations.

Fishes generate force to swim by activating muscles on either side of their flexible bodies. To accelerate, they must produce higher muscle forces, which leads to higher reaction forces back on their bodies from the environment. If their bodies are too flexible, the forces during acceleration could not be transmitted effectively to the environment, but fish can potentially use their muscles to increase the effective stiffness of their body. Here, we quantified red muscle activity during acceleration and steady swimming, looking for patterns that would be consistent with the hypothesis of body stiffening. We used high-speed video, electromyographic recordings, and a new digital inertial measurement unit to quantify body kinematics, red muscle activity, and 3D orientation and centre of mass acceleration during forward accelerations and steady swimming over several speeds. During acceleration, fish co-activated anterior muscle on the left and right side, and activated all muscle sooner and kept it active for a larger fraction of the tail beat cycle. These activity patterns are both known to increase effective stiffness for muscle tissue in vitro, which is consistent with our hypothesis that fish use their red muscle to stiffen their bodies during acceleration. We suggest that during impulsive movements, flexible organisms like fishes can use their muscles not only to generate propulsive power but to tune the effective mechanical properties of their bodies, increasing performance during rapid movements and maintaining flexibility for slow, steady movements.

Hydrodynamics of linear acceleration in bluegill sunfish, Lepomis macrochirus.

In their natural habitat, fish rarely swim steadily. Instead they frequently accelerate and decelerate. Relatively little is known about how fish produce extra force for acceleration in routine swimming behavior. In this study, we examined the flow around bluegill sunfish Lepomis macrochirus during steady swimming and during forward acceleration, starting at a range of initial swimming speeds. We found that bluegill produce vortices with higher circulation during acceleration, indicating a higher force per tail beat, but they do not substantially redirect the force. We quantified the flow patterns using high speed video and particle image velocimetry and measured acceleration with small inertial measurement units attached to each fish. Even in steady tail beats, the fish accelerates slightly during each tail beat, and the magnitude of the acceleration varies. In steady tail beats, however, a high acceleration is followed by a lower acceleration or a deceleration, so that the swimming speed is maintained; in unsteady tail beats, the fish maintains the acceleration over several tail beats, so that the swimming speed increases. We can thus compare the wake and kinematics during single steady and unsteady tail beats that have the same peak acceleration. During unsteady tail beats when the fish accelerates forward for several tail beats, the wake vortex forces are much higher than those at the same acceleration during single tail beats in steady swimming. The fish also undulates its body at higher amplitude and frequency during unsteady tail beats. These kinematic changes likely increase the fluid dynamic added mass of the body, increasing the forces required to sustain acceleration over several tail beats. The high amplitude and high frequency movements are also likely required to generate the higher forces needed for acceleration. Thus, it appears that bluegill sunfish face a trade-off during acceleration: the body movements required for acceleration also make it harder to accelerate.

The effects of lateral line ablation and regeneration in schooling giant danios.

Fish use multiple sensory systems, including vision and their lateral line system, to maintain position and speed within a school. Although previous studies have shown that ablating the lateral line alters schooling behavior, no one has examined how the behavior recovers as the sensory system regenerates. We studied how schooling behavior changes in giant danios, Devario aequipinnatus, when their lateral line system is chemically ablated and after the sensory hair cells regenerate. We found that fish could school normally immediately after chemical ablation, but that they had trouble schooling 1-2 weeks after the chemical treatment, when the hair cells had fully regenerated. We filmed groups of giant danios with two high-speed cameras and reconstructed the three-dimensional positions of each fish within a group. One fish in the school was treated with gentamycin to ablate all hair cells. Both types of neuromasts (canal and superficial) were completely ablated after treatment, but fully regenerated after 1 week. We quantified the structure of the school using nearest neighbor distance, bearing, elevation, and the cross-correlation of velocity between each pair of fish. Treated fish maintained a normal position within the school immediately after the lateral line ablation, but could not school normally 1 or 2 weeks after treatment, even though the neuromasts had fully regenerated. By 4-8 weeks post-treatment, the treated fish could again school normally. These results demonstrate that the behavioral recovery after lateral line ablation is a longer process than the regeneration of the hair cells themselves.

Detection of artificial water flows by the lateral line system of a benthic feeding cichlid fish.

The mechanosensory lateral line system of fishes detects water motions within a few body lengths of the source. Several types of artificial stimuli have been used to probe lateral line function in the laboratory, but few studies have investigated the role of flow sensing in benthic feeding teleosts. In this study, we used artificial flows emerging from a sandy substrate to assess the contribution of flow sensing to prey detection in the peacock cichlid, Aulonocara stuartgranti, which feeds on benthic invertebrates in Lake Malawi. Using a positive reinforcement protocol, we trained fish to respond to flows lacking the visual and chemical cues generated by tethered prey in prior studies with A. stuartgranti Fish successfully responded to artificial flows at all five rates presented (characterized using digital particle image velocimetry), and showed a range of flow-sensing behaviors, including an unconditioned bite response. Immediately after lateral line inactivation, fish rarely responded to flows and the loss of vital fluorescent staining of hair cells (with 4-di-2-ASP) verified lateral line inactivation. Within 2 days post-treatment, some aspects of flow-sensing behavior returned and after 7 days, flow-sensing behavior and hair cell fluorescence both returned to pre-treatment levels, which is consistent with the reported timing of hair cell regeneration in other vertebrates. The presentation of ecologically relevant water flows to assess flow-sensing behaviors and the use of a positive reinforcement protocol are methods that present new opportunities to study the role of flow sensing in the feeding ecology of benthic feeding fishes.

The effect of light intensity on prey detection behavior in two Lake Malawi cichlids, Aulonocara stuartgranti and Tramitichromis sp.

Two sand-dwelling cichlids from Lake Malawi (Aulonocara stuartgranti, Tramitichromis sp.) that feed on benthic invertebrates, but have different lateral line phenotypes, use lateral line and/or visual cues to detect prey under light versus dark conditions. The current study examined how ecologically relevant variation in light intensity [0-800 lux (lx)] influences detection of prey (mobile, immobile) in each species by analyzing six behavioral parameters. Both species fed at light intensities ≥1 lx and trends in behavior among light intensities were informative. However, prey type and/or time of day (but not light intensity) predicted all four parameters analyzed with generalized linear mixed models in A. stuartgranti, whereas the interaction of light intensity and time of day predicted three of these parameters in Tramitichromis sp. Data suggest that the critical light intensity is 1-12 lx for both species, that the integration of visual and lateral line input explains differences in detection of mobile and immobile prey and behavioral changes at the transition from 1 to 0 lx in A. stuartgranti, and that Tramitichromis sp. likely uses binocular vision to locate prey. Differences in the sensory biology of species that exploit similar prey will have important implications for the trophic ecology of African cichlid fishes.

Sensory basis for detection of benthic prey in two Lake Malawi cichlids.

The adaptive radiations of African cichlids resulted in a diversity of feeding morphologies and strategies, but the role of sensory biology in prey detection and feeding ecology remains largely unexplored. Two endemic Lake Malawi cichlid genera, Tramitichromis and Aulonocara, feed on benthic invertebrates, but differ in lateral line morphology (narrow and widened lateral line canals, respectively) and foraging strategy. The hypothesis that they use their lateral line systems differently was tested by looking at the relative contribution of the lateral line system and vision in prey detection by Tramitichromis sp. and comparing results to those from a complementary study using Aulonocara stuartgranti (Schwalbe et al., 2012). First, behavioral trials were used to assess the ability of Tramitichromis sp. to detect live (mobile) and dead (immobile) benthic prey under light and dark conditions. Second, trials were run before, immediately after, and several weeks after chemical ablation of the lateral line system to determine its role in feeding behavior. Results show that Tramitichromis sp. is a visual predator that neither locates prey in the dark nor depends on lateral line input for prey detection and is thus distinct from A. stuartgranti, which uses its lateral line or a combination of vision and lateral line to detect prey depending on light condition. Investigating how functionally distinctive differences in sensory morphology are correlated with feeding behavior in the laboratory and determining the role of sensory systems in feeding ecology will provide insights into how sensory capabilities may contribute to trophic niche segregation.

Feeding in the dark: lateral-line-mediated prey detection in the peacock cichlid Aulonocara stuartgranti.

The cranial lateral line canal system of teleost fishes is morphologically diverse and is characterized by four patterns. One of these, widened lateral line canals, has evolved convergently in a wide range of teleosts, including the Lake Malawi peacock cichlids (Aulonocara), and has been attributed to its role in prey detection. The ability to study Aulonocara in the laboratory provides an opportunity to test the hypothesis that their reported ability to feed on invertebrate prey living in sandy substrates in their natural habitat is the result of lateral-line-mediated prey detection. The goal of this study was to determine whether Aulonocara stuartgranti could detect hydrodynamic stimuli generated by tethered brine shrimp (visualized using digital particle image velocimetry) under light and dark conditions, with and without treatment with cobalt chloride, which is known to temporarily inactivate the lateral line system. Fish were presented with six pairs of tethered live and dead adult brine shrimp and feeding behavior was recorded with HD digital video. Results demonstrate that A. stuartgranti: (1) uses the same swimming/feeding strategy as they do in the field; (2) detects and consumes invertebrate prey in the dark using its lateral line system; (3) alters prey detection behavior when feeding on the same prey under light and dark conditions, suggesting the involvement of multiple sensory modalities; and (4) after treatment with cobalt chloride, exhibits a reduction in their ability to detect hydrodynamic stimuli produced by prey, especially in the dark, thus demonstrating the role of the lateral line system in prey detection.