Sound detection by the American lobster (Homarus americanus).

Although many crustaceans produce sounds, their hearing abilities and mechanisms are poorly understood, leaving uncertainties regarding whether or how these animals use sound for acoustic communication. Marine invertebrates lack gas-filled organs required for sound pressure detection, but some of them are known to be sensitive to particle motion. Here, we examined whether the American lobster (Homarus americanus) could detect sound and subsequently sought to discern the auditory mechanisms. Acoustic stimuli responses were measured using auditory evoked potential (AEP) methods. Neurophysiological responses were obtained from the brain using tone pips between 80 and 250 Hz, with best sensitivity at 80-120 Hz. There were no significant differences between the auditory thresholds of males and females. Repeated controls (recordings from deceased lobsters, moving electrodes away from the brain and reducing seawater temperature) indicated the evoked potentials' neuronal origin. In addition, AEP responses were similar before and after antennules (including statocysts) were ablated, demonstrating that the statocysts, a long-proposed auditory structure in crustaceans, are not the sensory organs responsible for lobster sound detection. However, AEPs could be eliminated (or highly reduced) after immobilizing hairfans, which cover much of lobster bodies. These results suggest that these external cuticular hairs are likely to be responsible for sound detection, and imply that hearing is mechanistically possible in a wider array of invertebrates than previously considered. Because the lobsters' hearing range encompasses the fundamental frequency of their buzzing sounds, it is likely that they use sound for intraspecific communication, broadening our understanding of the sensory ecology of this commercially vital species. The lobsters' low-frequency acoustic sensitivity also underscores clear concerns about the potential impacts of anthropogenic noise.

Effects of Prior Experience on Shelter-Seeking Behavior of Juvenile American Lobsters.

Shelter-seeking behaviors are vital for survival for a range of juvenile benthic organisms. These behaviors may be innate or they may be affected by prior experience. After hatching, American lobsters Homarus americanus likely first come into contact with shelter during the late postlarval (decapodid) stage, known as stage IV. After the subsequent molt to the first juvenile stage (stage V), they are entirely benthic and are thought to be highly cryptic. We hypothesized that postlarval (stage IV) experience with shelter would carry over into the first juvenile stage (stage V) and reduce the time needed for juveniles to locate and enter shelters (sheltering). We found some evidence of a carryover effect, but not the one we predicted: stage V juveniles with postlarval shelter experience took significantly longer to initiate sheltering. We also hypothesized that stage V juveniles would demonstrate learning by relocating shelters more quickly with immediate prior experience. Our findings were mixed. In a maze, juveniles with immediate prior experience were faster to regain visual contact with shelter, suggesting that they had learned the location of the shelter. In contrast, there was no significant effect of immediate prior experience on time to initiate sheltering in an open arena, or in the maze after juveniles had regained visual contact. We conclude that very young (stage V) juvenile lobsters modify their shelter-seeking behavior based on prior experiences across several timescales. Ecologically relevant variation in habitat exposure among postlarval and early juvenile lobsters may influence successful recruitment in this culturally and commercially important fishery species.

Sex matters in massive parallel sequencing: Evidence for biases in genetic parameter estimation and investigation of sex determination systems.

Using massively parallel sequencing data from two species with different life history traits, American lobster (Homarus americanus) and Arctic Char (Salvelinus alpinus), we highlight how an unbalanced sex ratio in the samples and a few sex-linked markers may lead to false interpretations of population structure and thus to potentially erroneous management recommendations. Here, multivariate analyses revealed two genetic clusters separating samples by sex instead of by expected spatial variation: inshore and offshore locations in lobster, or east and west locations in Arctic Char. To further investigate this, we created several subsamples artificially varying the sex ratio in the inshore/offshore and east/west groups and then demonstrated that significant genetic differentiation could be observed despite panmixia in lobster, and that FST values were overestimated in Arctic Char. This pattern was due to 12 and 94 sex-linked markers driving differentiation for lobster and Arctic Char, respectively. Removing sex-linked markers led to nonsignificant genetic structure in lobster and a more accurate estimation of FST in Arctic Char. The locations of these markers and putative identities of genes containing or nearby the markers were determined using available transcriptomic and genomic data, and this provided new information related to sex determination in both species. Given that only 9.6% of all marine/diadromous population genomic studies to date have reported sex information, we urge researchers to collect and consider individual sex information. Sex information is therefore relevant for avoiding unexpected biases due to sex-linked markers as well as for improving our knowledge of sex determination systems in nonmodel species.

Modulation of shark prey capture kinematics in response to sensory deprivation.

The ability of predators to modulate prey capture in response to the size, location, and behavior of prey is critical to successful feeding on a variety of prey types. Modulating in response to changes in sensory information may be critical to successful foraging in a variety of environments. Three shark species with different feeding morphologies and behaviors were filmed using high-speed videography while capturing live prey: the ram-feeding blacktip shark, the ram-biting bonnethead, and the suction-feeding nurse shark. Sharks were examined intact and after sensory information was blocked (olfaction, vision, mechanoreception, and electroreception, alone and in combination), to elucidate the contribution of the senses to the kinematics of prey capture. In response to sensory deprivation, the blacktip shark demonstrated the greatest amount of modulation, followed by the nurse shark. In the absence of olfaction, blacktip sharks open the jaws slowly, suggestive of less motivation. Without lateral line cues, blacktip sharks capture prey from greater horizontal angles using increased ram. When visual cues are absent, blacktip sharks elevate the head earlier and to a greater degree, allowing them to overcome imprecise position of the prey relative to the mouth, and capture prey using decreased ram, while suction remains unchanged. When visual cues are absent, nurse sharks open the mouth wider, extend the labial cartilages further, and increase suction while simultaneously decreasing ram. Unlike some bony fish, neither species switches feeding modalities (i.e. from ram to suction or vice versa). Bonnetheads failed to open the mouth when electrosensory cues were blocked, but otherwise little to no modulation was found in this species. These results suggest that prey capture may be less plastic in elasmobranchs than in bony fishes, possibly due to anatomical differences, and that the ability to modulate feeding kinematics in response to available sensory information varies by species, rather than by feeding modality.

Odor tracking in sharks is reduced under future ocean acidification conditions.

Recent studies show that ocean acidification impairs sensory functions and alters the behavior of teleost fishes. If sharks and other elasmobranchs are similarly affected, this could have significant consequences for marine ecosystems globally. Here, we show that projected future CO2 levels impair odor tracking behavior of the smooth dogfish (Mustelus canis). Adult M. canis were held for 5 days in a current-day control (405 ± 26 µatm) and mid (741 ± 22 µatm) or high CO2 (1064 ± 17 µatm) treatments consistent with the projections for the year 2100 on a 'business as usual' scenario. Both control and mid CO2 -treated individuals maintained normal odor tracking behavior, whereas high CO2 -treated sharks significantly avoided the odor cues indicative of food. Control sharks spent >60% of their time in the water stream containing the food stimulus, but this value fell below 15% in high CO2 -treated sharks. In addition, sharks treated under mid and high CO2 conditions reduced attack behavior compared to the control individuals. Our findings show that shark feeding could be affected by changes in seawater chemistry projected for the end of this century. Understanding the effects of ocean acidification on critical behaviors, such as prey tracking in large predators, can help determine the potential impacts of future ocean acidification on ecosystem function.

Multisensory integration and behavioral plasticity in sharks from different ecological niches.

The underwater sensory world and the sensory systems of aquatic animals have become better understood in recent decades, but typically have been studied one sense at a time. A comprehensive analysis of multisensory interactions during complex behavioral tasks has remained a subject of discussion without experimental evidence. We set out to generate a general model of multisensory information extraction by aquatic animals. For our model we chose to analyze the hierarchical, integrative, and sometimes alternate use of various sensory systems during the feeding sequence in three species of sharks that differ in sensory anatomy and behavioral ecology. By blocking senses in different combinations, we show that when some of their normal sensory cues were unavailable, sharks were often still capable of successfully detecting, tracking and capturing prey by switching to alternate sensory modalities. While there were significant species differences, odor was generally the first signal detected, leading to upstream swimming and wake tracking. Closer to the prey, as more sensory cues became available, the preferred sensory modalities varied among species, with vision, hydrodynamic imaging, electroreception, and touch being important for orienting to, striking at, and capturing the prey. Experimental deprivation of senses showed how sharks exploit the many signals that comprise their sensory world, each sense coming into play as they provide more accurate information during the behavioral sequence of hunting. The results may be applicable to aquatic hunting in general and, with appropriate modification, to other types of animal behavior.

Reef odor: a wake up call for navigation in reef fish larvae.

The behavior of reef fish larvae, equipped with a complex toolbox of sensory apparatus, has become a central issue in understanding their transport in the ocean. In this study pelagic reef fish larvae were monitored using an unmanned open-ocean tracking device, the drifting in-situ chamber (DISC), deployed sequentially in oceanic waters and in reef-born odor plumes propagating offshore with the ebb flow. A total of 83 larvae of two taxonomic groups of the families Pomacentridae and Apogonidae were observed in the two water masses around One Tree Island, southern Great Barrier Reef. The study provides the first in-situ evidence that pelagic reef fish larvae discriminate reef odor and respond by changing their swimming speed and direction. It concludes that reef fish larvae smell the presence of coral reefs from several kilometers offshore and this odor is a primary component of their navigational system and activates other directional sensory cues. The two families expressed differences in their response that could be adapted to maintain a position close to the reef. In particular, damselfish larvae embedded in the odor plume detected the location of the reef crest and swam westward and parallel to shore on both sides of the island. This study underlines the critical importance of in situ Lagrangian observations to provide unique information on larval fish behavioral decisions. From an ecological perspective the central role of olfactory signals in marine population connectivity raises concerns about the effects of pollution and acidification of oceans, which can alter chemical cues and olfactory responses.

Sun Compass Orientation Helps Coral Reef Fish Larvae Return to Their Natal Reef.

Reef fish sustain populations on isolated reefs and show genetic diversity between nearby reefs even though larvae of many species are swept away from the natal site during pelagic dispersal. Retention or recruitment to natal reefs requires orientation capabilities that enable larvae to find their way. Although olfactory and acoustically based orientation has been implicated in homing when larvae are in the reef's vicinity, it is still unclear how they cope with greater distances. Here we show evidence for a sun compass mechanism that can bring the larvae to the vicinity of their natal reef. In a circular arena, pre-settlement larvae and early settlers (<24 hours) of the cardinal fish, Ostorhinchus doederleini, showed a strong SSE directional swimming response, which most likely has evolved to compensate for the locally prevailing large scale NNW current drift. When fish were clock-shifted 6 hours, they changed their orientation by ca. 180° as predicted by the tropical sun curve at One Tree Island, i.e. they used a time-compensated sun compass. Furthermore, the fish oriented most consistently at times of the day when the sun azimuth is easy to determine. Microsatellite markers showed that the larvae that had just arrived at One Tree Island genetically belonged to either the local reef population or to Fitzroy Reef located 12 kilometers to the SSE. The use of a sun compass adds a missing long-distance link to the hierarchy of other sensory abilities that can direct larvae to the region of origin, including their natal reef. Predominant local recruitment, in turn, can contribute to genetic isolation and potential speciation.

The function of bilateral odor arrival time differences in olfactory orientation of sharks.

The direction of an odor signal source can be estimated from bilateral differences in signal intensity and/or arrival time. The best-known examples of the use of arrival time differences are in acoustic orientation. For chemoreception, animals are believed to orient by comparing bilateral odor concentration differences, turning toward higher concentrations. However, time differences should not be ignored, because odor plumes show chaotic intermittency, with the concentration variance several orders of magnitude greater than the concentration mean. We presented a small shark species, Mustelus canis, with carefully timed and measured odor pulses directly into their nares. They turned toward the side stimulated first, even with delayed pulses of higher concentration. This is the first conclusive evidence that under seminatural conditions and without training, bilateral time differences trump odor concentration differences. This response would steer the shark into an odor patch each time and thereby enhance its contact with the plume, i.e., a stream of patches. Animals with more widely spaced nares would be able to resolve smaller angles of attack at higher swimming speeds, a feature that may have contributed to the evolution of hammerhead sharks. This constitutes a novel steering algorithm for tracking odor plumes.

Sharks need the lateral line to locate odor sources: rheotaxis and eddy chemotaxis.

Odor plumes are complex, dynamic, three-dimensional structures used by many animals to locate food, mates, home sites, etc. Yet odor itself has no directional properties. Animals use a variety of different senses to obtain directional information. Since most odor plumes are composed of dispersing odor patches and dissipating vorticity eddies, aquatic animals may localize odor sources by simultaneous analysis of chemical and hydrodynamic dispersal fields, a process referred to as eddy chemotaxis. This study examines the contributions of olfaction, mechanoreception and vision to odor source localization in a shark, the smooth dogfish Mustelus canis. Two parallel, turbulent plumes were created in an 8 m flume: squid rinse odor and seawater control. Minimally turbulent ;oozing' sources of odor and seawater control were physically separated from sources of major turbulence by placing a brick downstream from each oozing source, creating two turbulent wakes, one or the other flavored with food odor. This created four separate targets for the sharks to locate. Animals were tested under two light conditions (fluorescent and infrared) and in two sensory conditions (lateral line intact and lateral line lesioned by streptomycin). Intact animals demonstrated a preference for the odor plume over the seawater plume and for the source of odor/turbulence (the brick on the odor side) over the source of the odor alone (the odor-oozing nozzle). Plume and target preference and search time were not significantly affected by light condition. In the light, lesioning the lateral line increased search time but did not affect success rate or plume preference. However, lesioned animals no longer discriminated between sources of turbulent and oozing odor. In the dark, search time of lesioned animals further increased, and the few animals that located any of the targets did not discriminate between odor and seawater plumes, let alone targets. These results demonstrate for the first time that sharks require both olfactory and lateral line input for efficient and precise tracking of odor-flavored wakes and that visual input can improve food-finding performance when lateral line information is not available. We distinguish between rheotaxis: orientation to the large-scale flow field (olfaction, vision and superficial lateral line), eddy chemotaxis: tracking the trail of small-scale, odor-flavored turbulence (olfaction and lateral line canals), and pinpointing the source of the plume (lateral line canals and olfaction).

Smelling home can prevent dispersal of reef fish larvae.

Many marine fish and invertebrates show a dual life history where settled adults produce dispersing larvae. The planktonic nature of the early larval stages suggests a passive dispersal model where ocean currents would quickly cause panmixis over large spatial scales and prevent isolation of populations, a prerequisite for speciation. However, high biodiversity and species abundance in coral reefs contradict this panmixis hypothesis. Although ocean currents are a major force in larval dispersal, recent studies show far greater retention than predicted by advection models. We investigated the role of animal behavior in retention and homing of coral reef fish larvae resulting in two important discoveries: (i) Settling larvae are capable of olfactory discrimination and prefer the odor of their home reef, thereby demonstrating to us that nearby reefs smell different. (ii) Whereas one species showed panmixis as predicted from our advection model, another species showed significant genetic population substructure suggestive of strong homing. Thus, the smell of reefs could allow larvae to choose currents that return them to reefs in general and natal reefs in particular. As a consequence, reef populations can develop genetic differences that might lead to reproductive isolation.

Unraveling the nature of individual recognition by odor in hermit crabs.

Individual recognition is a key element in the social life of many invertebrates. However, most studies conducted so far document that several species are capable of a "binary" discrimination among conspecifics, but not of a "true individual recognition." Our objective was to learn more about the mechanisms that underlie individual recognition by odor in hermit crabs by individuating some of its properties. Using Pagurus longicarpus Say 1817 as a model species, we conducted four series of experiments in which the response of every test crab (the "receiver") to the different odor treatments (emitted by a "sender") was evaluated from its investigative behavior toward an empty, high-quality shell. After having excluded the possibility that crabs chemically recognize familiar/unfamiliar shells and/or shells of high/low quality, we explored whether the receivers discriminate odors from two familiar senders and whether this discrimination also occurs with unfamiliar crabs. We also asked whether crabs form an association between the odor of a familiar sender and some of its relevant attributes, i.e., rank, size, and shell quality. Finally, the shells inhabited by familiar individuals were manipulated to modify the association between odor and shell quality. Results showed that: (1) there is no odor specific of a rank; (2) individual crabs discriminate their own odor from the odor of other individuals; (3) they can chemically discriminate between larger crabs inhabiting higher-quality shells and smaller crabs inhabiting lower-quality shells, provided that these crabs are familiar to them; (4) they associate the odor of an individual crab with the quality of the shell it inhabits; and (5) this association quickly changes when social partners switch to shells of different quality. These results indicate that the nature of chemical recognition in P. longicarpus is more refined than a simple binary system. The receiver appears able to associate a type of information from the sender with memories of past experiences, therefore suggesting the hermit crab's potential for relatively high-order knowledge about conspecifics.

The olfactory pathway for individual recognition in the American lobster Homarus americanus.

Individual recognition in the lobster Homarus americanus (Milne-Edwards), is based on detection of urine pheromones via chemoreceptors of the lateral antennular flagellum. The specific sensory pathway mediating this recognition is not known. Most of the chemoreceptor cells of this flagellum are found in the unimodal aesthetasc sensilla and project specifically to the glomeruli of the olfactory lobe in the brain. Additional chemoreceptor cells are located among mechanoreceptor cells in bimodal sensilla, including the guard hairs; they do not project to the olfactory lobe. This neuroanatomy suggested that aesthetascs were essential to all complex chemosensory tasks until it was shown that spiny lobsters Panulirus argus can still perform complex food odor discrimination and localization tasks without aesthetascs. Here, we demonstrate that the aesthetascs of H. americanus contain the chemoreceptors necessary for individual recognition of familiar opponents. In contrast to intact and guard hair-shaved animals, lobsters with aesthetascs removed did not recognize previous opponents as shown by second encounters statistically similar in length and aggression to first-encounter fights. Non-aesthetasc chemosensory pathways were incapable of rescuing opponent recognition. Subsequent lesion of all remaining chemoreceptor cells (by immersion in distilled water) abolished recognition and renewed fighting.

The importance of the lateral line in nocturnal predation of piscivorous catfish.

In a previous study we showed that nocturnal piscivorous catfish track the wake left by a swimming prey fish to locate it, following past locations to detect the present location of the prey. In a wake there are hydrodynamic as well as chemical signatures that both contain information on location and suitability of the prey. In order to determine how these two wake stimuli are utilised in prey tracking, we conducted experiments in catfish in which either the lateral line or the external gustation was ablated. We found that a functional lateral line is indispensable for following the wake of swimming prey. The frequency of attack and capture was greatly diminished and the attacks that did occur were considerably delayed when the lateral line was ablated. In contrast, catfish with ablated external taste still followed the wakes of their prey prior to attacking, albeit their attacks were delayed. The external taste sense, which was reported earlier to be necessary for finding stationary (dead) food, seems to play a minor role in the localisation of moving prey. Our finding suggests that an important function of the lateral line is to mediate wake-tracking in predatory fish.

Field Observations of Social Behavior, Shelter Use, and Foraging in the Lobster, Homarus americanus.

Over a three-year period (1978-1981) behavioral observations of the lobster, Homarus americanus, were made by snorkeling in a shallow cove. Three hundred and thirty-four (334) animals were individually marked and this was the only time they were disturbed. In summer, the resident population numbered about 30 animals. The size composition, activity patterns, and habitat use of this population are described in a companion paper (Karnofsky et al., 1989). Shelters are of prime importance in the life of the lobster. Lobsters spent most of their time in shelters, leaving only at night. They dug shelters under eelgrass, rocks, and boulders; shelter locations appeared clustered. Some animals changed shelters frequently whereas others occupied the same shelters for up to 10 weeks. Premolt behavior was characterized by multiple shelter use. Cohabitation in the same shelter occurred only during periods of pair formation: when a mature female shared a male's shelter prior to and following her molt. We report the only field evidence for such courtship cohabitation. Food foraging behavior was rare (0.35 instances/observation hour); most foraging involved live prey. Similarly, intraspecific interactions were surprisingly infrequent (0.2 instances/observation hour) and most, by far, did not involve physical contact. Although puncture wounds suggested intraspecific aggression, actual observations of escalating fights were rare. Premolt residents were involved in 65% of the interactions observed. In 70% of the interactions the larger animal won. However, smaller males and females could successfully defend their shelters against larger females. We report results from three homing experiments. The results suggest that much of the time that resident lobsters spend outside shelters is used to remain familiar with their constantly changing physical and social environment.

Natural Dynamics of Population Structure and Habitat Use of the Lobster, Homarus americanus, in a Shallow Cove.

We report the results of a nonmanipulative field study of the lobster, Homarus americanus, using long-term behavioral observations of marked individuals. We observed a freely mobile population in an open shallow cove habitat (50 m x 150 m) in Buzzards Bay, Massachusetts. Lobsters larger than 50 mm carapace length (CL) living in or entering the study site were marked individually (334 during the 19-month study). Without further manipulation, the animals were observed as long as they remained in the study site. Of the marked animals, 48% were transient, i.e., seen only once. The population was made up largely of subadults with a sex ratio of M:F = 1.8. The summer and fall resident population consisted of about 30 animals. Maximum residency was over 13 months. Half of the resident population, mostly small animals (50-59 mm CL), apparently overwintered in the site. A distinct peak in molting occurred both years in the spring at a water temperature of about 15°C. Injured animals were seen frequently (26% of the population) including a high proportion of mature resident males missing claws. Most other injured animals were transient (60%). These results suggest that the shallow cove is used as a refuge for injured mature males. Activity was strictly nocturnal with a peak 1-3 h after sunset and declining through the night. Activity levels were equal for both sexes. Overall activity was correlated with seasonal variations in water temperature (0-24°C). At times, activity was correlated more with molting (premolt activity peak) than with temperature. Behavioral interactions in this population are described in a companion paper (Karnofsky et al., 1989).