Sandie Millot

Sound production in red-bellied piranhas (Pygocentrus nattereri, Kner): an acoustical, behavioural and morphofunctional study

SUMMARY Piranhas are known to be sound-producing animals. Nevertheless, the biological significance of piranha calls remains unclear because sounds have been recorded only when specimens were held by hand or trapped in a gill net. These sounds are generated by rapid contractions of sonic muscles that insert on a broad tendon surrounding ventrally the cranial sac of the swimbladder. The piranha swimbladder is thought to play an important role in sound production as an impedance-matching device and as a resonator. However, the vibratory capacities of the cranial and caudal sacs and the exact role of both sacs in sound production remain poorly understood. In this study, three sounds were each associated to a specific behaviour. The first sound (type 1) was produced during frontal display; it had numerous pulses and lasted 140!±17 ms, with a fundamental frequency of 120±4 Hz. It corresponded to the sound made by hand-held fishes. The second sound (type 2) was produced during circling and fighting behaviour; it was a single pulse lasting 36±8 ms, with a fundamental frequency of 43±10 Hz. The third sound (type 3) corresponded to chasing behaviour and comprised three to four pulses, each lasting 3±1 ms, with a fundamental frequency of 1739±18 Hz. Using a laser vibrometer to study the swimbladder displacement when stimulated at different frequencies, it was demonstrated that the first two sounds corresponded to the swimbladder mechanism. By contrast, the third sound was associated with the jaw mechanism. The vibrometer indicated that the swimbladder is a highly damping structure, simply copying the sonic muscle contraction rate. This study provides two interesting insights. First, it shows the relationships between three kinds of piranha sound and three specific behaviours. Second, using muscle stimulation at different rates, it shows which simultaneous conditions are required for production of sound in this species. Swimbladder calls were produced by a muscle contraction rate of approximately 100 Hz because this periodicity allowed the swimbladder to vibrate. At this frequency range, the contraction–relaxation cycles of the swimbladder muscles engendered wall displacements that had short amplitudes and with only a small variability between them.

Risk-taking behaviour variation over time in sea bass Dicentrarchus labrax: effects of day-night alternation, fish phenotypic characteristics and selection for growth.

Differences in bold and shy personality on sea bass Dicentrarchus labrax were investigated between a population (wild) produced from wild-brood fish and a population (selected) produced from selected-brood fish. During the experiment (112 days), fish were reared under self-feeding condition to characterize the feeding behaviour of each individual fish. Three risk-taking tests (T1, T2 and T3 of 24 h with day-night alternation) were carried out at >1 month intervals on 180 fish of each strain in order to monitor D. labrax behaviour over time and in relation to the light:dark period. A risk-taking score was evaluated via a preference choice between a safe zone (without food) and a risky zone (potentially with food) by recording the number and the duration of individual passages through an opening in an opaque divider. Results showed that fish performed passages preferentially during the night period and that wild fish were generally bolder than selected fish during T1 and T2 but showed a decrease in risk taking during T3, contrary to selected fish which showed a constant increase in their risk-taking behaviour. The phenotypic characteristics of the bold fish were different in the two strains: wild bold fish were the smallest within the wild strain and selected bold fish presented the higher growth rate within the selected strain. For both strains, these bold fish were also generally characterized by a high feed-demand activity. Fish hunger state thus seemed to be the highest motivation for risk-taking behaviour under the present conditions. Furthermore, behavioural variations over tests such as higher risk taking (number of passages) and faster exploratory responses (higher score emergence) could be interpreted as relevant indicators of the learning process and habituation. According to the results, however, no real difference in coping strategy between strains could be observed at this first stage of domestication and selection.

Demand feeding and welfare in farmed fish

Following the development of demand-feeding systems, many experiments have been conducted to explore feeding motivation and feed intake in farmed fish. This work aims to review a selection of studies in the field, focusing on three key factors, related to demand feeding and fish welfare. Firstly, we outline how demand feeders should be considered when developing feed management strategies for improving welfare in production conditions. Secondly, via laboratory demand-feeding experiments, we show self-feeding activities depend not only on feeding motivation and social organisation, but also on individual learning capacity and risk-taking behaviour. Thirdly, we report encouraging results demonstrating that when presented with two or more self-feeders containing complementary foods, fish select a diet according to their specific nutritional requirements, suggesting that demand feeders could be used to improve welfare by allowing fish to meet their nutritional needs.

Assessment of Genetic Variability of Fish Personality Traits using Rainbow Trout Isogenic Lines

The study of inter-individual variability of personality in fish is a growing field of interest but the genetic basis of this complex trait is still poorly investigated due to the difficulty in controlling fish genetic origin and life history. When available, isogenic lines that allow performing independent tests on different individuals having identical genotype constitute a very relevant experimental material to disentangle the genetic and environmental components of behavioural individuality. We took advantage of heterozygous isogenic lines to investigate the personality in rainbow trout through the analysis of their reactions to different experimental situations. To this end, seven to ten rainbow trout isogenic lines were screened for their spatial exploratory behaviour, their flight response toward a stressor and their risk taking behaviour. Results showed that some lines seemed less sensitive to new events or environmental changes and could be defined as low responsive, while others were very sensitive and defined as high responsive. The use of isogenic lines highlighted the importance of genetic factors, in combination with life history, in the expression of personality in domesticated fish.

Behavioural Stress Responses Predict Environmental Perception in European Sea Bass (Dicentrarchus labrax)

Individual variation in the response to environmental challenges depends partly on innate reaction norms, partly on experience-based cognitive/emotional evaluations that individuals make of the situation. The goal of this study was to investigate whether pre-existing differences in behaviour predict the outcome of such assessment of environmental cues, using a conditioned place preference/avoidance (CPP/CPA) paradigm. A comparative vertebrate model (European sea bass, Dicentrarchus labrax) was used, and ninety juvenile individuals were initially screened for behavioural reactivity using a net restraining test. Thereafter each individual was tested in a choice tank using net chasing as aversive stimulus or exposure to familiar conspecifics as appetitive stimulus in the preferred or non preferred side respectively (called hereafter stimulation side). Locomotor behaviour (i.e. time spent, distance travelled and swimming speed in each tank side) of each individual was recorded and analysed with video software. The results showed that fish which were previously exposed to appetitive stimulus increased significantly the time spent on the stimulation side, while aversive stimulus led to a strong decrease in time spent on the stimulation side. Moreover, this study showed clearly that proactive fish were characterised by a stronger preference for the social stimulus and when placed in a putative aversive environment showed a lower physiological stress responses than reactive fish. In conclusion, this study showed for the first time in sea bass, that the CPP/CPA paradigm can be used to assess the valence (positive vs. negative) that fish attribute to different stimuli and that individual behavioural traits is predictive of how stimuli are perceived and thus of the magnitude of preference or avoidance behaviour.

Innovative behaviour in fish: Atlantic cod can learn to use an external tag to manipulate a self-feeder

Abstract This study describes how three individual fish, Atlantic cod (Gadus morhua L.), developed a novel behaviour and learnt to use a dorsally attached external tag to activate a self-feeder. This behaviour was repeated up to several hundred times, and over time these fish fine-tuned the behaviour and made a series of goal-directed coordinated movements needed to attach the feeder’s pull string to the tag and stretch the string until the feeder was activated. These observations demonstrate a capacity in cod to develop a novel behaviour utilizing an attached tag as a tool to achieve a goal. This may be seen as one of the very few observed examples of innovation and tool use in fish.

Self-feeding behavior changes induced by a first and a second generation of domestication or selection for growth in the European sea bass, Dicentrarchus labrax

Among the strategies that can be used to improve fish welfare in a rearing environment, domestication and/or selective breeding was proposed to minimize fish responsiveness to husbandry practices. To verify this hypothesis on a recently domesticated species, the sea bass Dicentrarchus labrax, two experiments were realized, each using two populations differing according to their level of domestication or selection. For the first experiment, we used one population produced from wild parents (Wild; initial body mass: 106 ± 3 g), and one population from parents selected for growth for one generation (Selected 1; initial body mass: 129 ± 4 g). For the second experiment, we used one population produced from parents domesticated for two generations (Domesticated; initial body mass: 72 ± 3g ), and one produced from parents selected for growth for two generations (Selected 2; initial body mass: 89 ± 4 g). The first experiment was carried out over 112 days with 240 fish (60 fish per tank, 120 fish per population), and the second one over 84 days with 200 fish (50 fish per tank, 100 fish per population). Two variables, self-feeding behavior and growth performance, were measured over the time of the experiments. After a control period, the fish were submitted twice, at three-week intervals, to an acute stress treatment consisting of draining the tank and leaving the fish out of water for one minute. Both self-feeding behavior and growth performance were altered by the acute stress treatment. During the first post-stress period, the Domesticated and Selected (1 and 2) groups showed more pronounced post-stress exposure responses than the Wild fish: they modified their feeding rhythm, their feed intake, and their growth rate. During the second post-stress period, feeding rhythm was still affected (being more diurnal with a well defined peak), but the feed intake and growth rate results showed that the Domesticated and Wild groups seemed less affected than the Selected (1 and 2) populations, which continued to express a high post-stress response. According to these results, it can be concluded that: (1) an application of two acute stress treatments, at three-week intervals, modified fish feeding behavior and growth performance; (2) the domestication process seemed to improve fish adaptation abilities to this kind of stress; and (3) the process of selection for growth led to a final, better growth, but did not seem to improve fish acute stress tolerance.

Development of the ultrastructure of sonic muscles: a kind of neoteny?

BackgroundDrumming muscles of some sound-producing fish are ‘champions’ of contraction speed, their rate setting the fundamental frequency. In the piranha, contraction of these muscles at 150 Hz drives a sound at the same frequency. Drumming muscles of different not closely related species show evolutionary convergences. Interestingly, some characters of sonic muscles can also be found in the trunk muscles of newly hatched larvae that are able to maintain tail beat frequencies up to 100 Hz. The aim of this work was to study the development of sound production and sonic and epaxial muscles simultaneously in the red bellied piranhas (Pygocentrus nattereri) to seek for possible common characteristics.ResultsCall, pulse and period durations increased significantly with the fish size, but the call dominant frequencies decreased, and the number of pulses and the call amplitude formed a bell curve. In epaxial muscles, the fibre diameters of younger fish are first positioned in the graphical slope corresponding to sonic muscles, before diverging. The fibre diameter of older fish trunk muscles was bigger, and the area of the myofibrils was larger than in sonic muscles. Moreover, in two of the biggest fish, the sonic muscles were invaded by fat cells and the sonic muscle ultrastructure was similar to the epaxial one. These two fish were also unable to produce any sound, meaning they lost their ability to contract quickly.ConclusionsThe volume occupied by myofibrils determines the force of contraction, the volume of sarcoplasmic reticulum sets the contraction frequency, and the volume of mitochondria sets the level of sustained performance. The functional outcomes in muscles are all attributable to shifts in the proportions of those structures. A single delay in the development restricts the quantity of myofibrils, maintains a high proportion of space in the sarcoplasm and develops sarcoplasmic reticulum. High-speed sonic muscles could thus be skeletal muscles with delayed development. This hypothesis has the advantage that it could easily explain why high-speed sonic muscles have evolved so many times in different lineages.

Sedentary behaviour establishment in O-group common sole Solea solea: a laboratory video-tracking study

Spontaneous swimming activity of 0-group common sole (Solea solea) was evaluated using a video-tracking system under laboratory conditions. An experiment was conducted during two consecutive days on individuals sampled in June, July, September and November (2004) in a coastal nursery ground (Pertuis Charentais, Bay of Biscay, France). The measured behavioural variables were: distance travelled and frequency of occurrence of burying, swimming and immobility. 0-group sole showed a relatively clear circadian activity in line with the artificial light conditions (day, twilight and night). Swimming activity of 0-group sole decreased drastically from June to July, thereafter remaining at a very low level in September and November. Such important changes reflect the transition between the exploratory behaviour of the post-colonization period and a well established sedentary behaviour remaining until the onset of winter. These results highlight the potential limitation in habitat use capacities of 0-group sole once settled in coastal nursery grounds.

Genetic variability of environmental sensitivity revealed by phenotypic variation in body weight and (its) correlations to physiological and behavioral traits

Adaptive phenotypic plasticity is a key component of the ability of organisms to cope with changing environmental conditions. Fish have been shown to exhibit a substantial level of phenotypic plasticity in response to abiotic and biotic factors. In the present study, we investigate the link between environmental sensitivity assessed globally (revealed by phenotypic variation in body weight) and more targeted physiological and behavioral indicators that are generally used to assess the sensitivity of a fish to environmental stressors. We took advantage of original biological material, the rainbow trout isogenic lines, which allowed the disentangling of the genetic and environmental parts of the phenotypic variance. Ten lines were characterized for the changes of body weight variability (weight measurements taken every month during 18 months), the plasma cortisol response to confinement stress (3 challenges) and a set of selected behavioral indicators. This study unambiguously demonstrated the existence of genetic determinism of environmental sensitivity, with some lines being particularly sensitive to environmental fluctuations and others rather insensitive. Correlations between coefficient of variation (CV) for body weight and behavioral and physiological traits were observed. This confirmed that CV for body weight could be used as an indicator of environmental sensitivity. As the relationship between indicators (CV weight, risk-taking, exploration and cortisol) was shown to be likely depending on the nature and intensity of the stressor, the joint use of several indicators should help to investigate the biological complexity of environmental sensitivity.