Pain perception, aversion and fear in fish.

There is now compelling evidence that teleost fish possess similar nociceptive processing systems to those found in terrestrial vertebrates. Noxious stimulation of these nociceptors--specialised pain receptors -in the skin around the snout of fish generates neural activity that can be electrophysiologically recorded, and induces a number of behavioural and physiological changes. To determine whether changes in behaviour are more than simple responses to the noxious stimulation it is necessary to demonstrate that higher order cognitive processes such as mental state or 'affective state' are involved. However, quantifying the 'motivational affected state' of an animal--a concept encompassing not just pain but also fear, hunger, thirst and pleasure - is difficult owing to its subjectivity. Recent empirical work is beginning to test these concepts in fish, and we review a number of these studies and suggest how these general methodologies could be used to further our understanding of fish cognition and the capacity for fish to experience mental states such as fear or suffering.

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title: Do fishes have nociceptors? Evidence for the evolution of a vertebrate sensory system
abstract: Nociception is the detection of a noxious tissue–damaging stimulus and is sometimes accompanied by a reflex response such as withdrawal. Pain perception, as distinct from nociception, has been demonstrated in birds and mammals but has not been systematically studied in lower vertebrates. We assessed whether a fish possessed cutaneous nociceptors capable of detecting noxious stimuli and whether its behaviour was sufficiently adversely affected by the administration of a noxious stimulus. Electrophysiological recordings from trigeminal nerves identified polymodal nociceptors on the head of the trout with physiological properties similar to those described in higher vertebrates. These receptors responded to mechanical pressure, temperatures in the noxious range (more than 40°C) and 1% acetic acid, a noxious substance. In higher vertebrates nociceptive nerves are either A–delta or C fibres with C fibres being the predominating fibre type. However, in the rainbow trout A–delta fibres were most common, and this offers insights into the evolution of nociceptive systems. Administration of noxious substances to the lips of the trout affected both the physiology and the behaviour of the animal and resulted in a significant increase in opercular beat rate and the time taken to resume feeding, as well as anomalous behaviours. This study provides significant evidence of nociception in teleost fishes and furthermore demonstrates that behaviour and physiology are affected over a prolonged period of time, suggesting discomfort.

title: Fish and welfare: Do fish have the capacity for pain perception and suffering?
abstract: Humans interact with fish in a number of ways and the question of whether fish have the capacity to perceive pain and to suffer has recently attracted considerable attention in both scientific and public fora. Only very recently have neuroanatomical studies revealed that teleost fish possess similar pain-processing receptors to higher vertebrates. Research has also shown that fish neurophysiology and behaviour are altered in response to noxious stimulation. In the light of this evidence and in combination with work illustrating the cognitive capacities of fish, it seems appropriate to respond to a recently published critique (Rose, 2002) in which it is argued that it is not possible for fish to experience fear or pain and that, therefore, they cannot suffer. Whilst we agree with the author that fish are unlikely to perceive pain in the same way that humans do, we believe that currently available evidence indicates that fish have the capacity for pain perception and suffering. As such, it would seem timely to reflect on the implications of fish pain and suffering, and to consider what steps can be taken to ensure the welfare of the fish that we exploit.

title: The Neurobehavioral Nature of Fishes and the Question of Awareness and Pain
abstract: This review examines the neurobehavioral nature of fishes and addresses the question of whether fishes are capable of experiencing pain and suffering. The detrimental effects of anthropomorphic thinking and the importance of an evolutionary perspective for understanding the neurobehavioral differences between fishes and humans are discussed. The differences in central nervous system structure that underlie basic neurobehavioral differences between fishes and humans are described. The literature on the neural basis of consciousness and of pain is reviewed, showing that: (1) behavioral responses to noxious stimuli are separate from the psychological experience of pain, (2) awareness of pain in humans depends on functions of specific regions of cerebral cortex, and (3) fishes lack these essential brain regions or any functional equivalent, making it untenable that they can experience pain. Because the experience of fear, similar to pain, depends on cerebral cortical structures that are absent from fish brains, it is concluded that awareness of fear is impossible for fishes. Although it is implausible that fishes can experience pain or emotions, they display robust, nonconscious, neuroendocrine, and physiological stress responses to noxious stimuli. Thus, avoidance of potentially injurious stress responses is an important issue in considerations about the welfare of fishes.

title: Balancing animal research with animal well-being: establishment of goals and harmonization of approaches.
abstract: A resource is provided for the creation of an institutional program that balances the scientific mission of an institution with the well-being of the animals used in support of the research. The concept of harmonizing scientific goals with animal well-being was first suggested in the early part of the twentieth century and later revitalized in the literature of the 1950s. Harmonization can best be achieved through the promotion of a team initiative. The team should include, at a minimum, the scientist, veterinarian, institutional animal care and use committee, and animal care staff. It is the responsibility of this animal research team to promote and balance the generation of scientifically valid data with animal well-being. The team must strive to minimize or eliminate non-protocol variables that could adversely affect the validity and repeatability of the experimental data. Good experimental design coupled with excellent communication between team members can often minimize or eliminate many variables and result in both better science and animal well-being. To ensure the scientific validity of experimental data, scientists must be aware of the complex nature of the environment in which their animals are maintained. To ensure repeatablity of an experiment, scientists must document and publish both the inanimate and social environments in which their animals are housed. Better documentation of environmental variables and their correlation with experimental results will promote critical knowledge about the relationships between an animal's environment, its well-being, and science.

title: Nociception in vertebrates: key receptors participating in spinal mechanisms of chronic pain in animals
abstract: Our view of vertebrate nociceptive processing is ever changing with the discovery of novel molecules that differentially affect sensory responses to noxious and innocuous stimulation and might be involved specifically in chronic pain states. In order to understand the physiology of nociception and design novel analgesics for intractable chronic pain, it is essential to uncover precisely what changes occur between a normal nociceptive processing state and hypersensitive chronic pain states in the spinal cord following different types of injury. An important area of focus for future work in this area will be the cellular and molecular mechanisms of neuronal plasticity that occur.

title: Organization of peripheral nerves and spinal roots of the Atlantic stingray, Dasyatis sabina.
abstract: 1. The sizes and numbers of axons in peripheral nerves and spinal roots were investigated in the stingray, Dasyatis sabina. 2. The axons of the dorsal and ventral roots do not mingle in peripheral nerves of this animal as they do in higher vertebrates. Thus, it was usually possible to split the peripheral nerve into two portions, one containing only dorsal root axons, the other containing only ventral root axons. This feature was useful for the analysis of certain aspects of spinal cord organization. 3. The fact that dorsal and ventral root axons were segregated in peripheral nerves enabled us to demonstrate, without experimental surgery, that the central processes of the dorsal root ganglion cells and the proximal ventral root axons were 10-20% narrower, on the average, than the distal processes of the same dorsal root ganglion cells or the distal parts of the same ventral root axons. 4. The stingray is remarkable in having very few unmyelinated axons in the dorsal roots, ventral roots, or peripheral nerves. This paucity of unmyelinated axons distinguishes the Atlantic stingrays from all other vertebrates whose roots and nerves have been examined for unmyelinated fibers. 5. Similar findings were obtained for one spotted eagle ray (Aetobatus narinari) and two cow-nose rays (Rhinoptera bonasus).

title: Anatomical and electrophysiological analysis of the trigeminal nerve in a teleost fish, Oncorhynchus 
 mykiss

abstract: The trigeminal nerve in the rainbow trout, Oncorhynchus mykiss, was examined for the presence of A-delta and C fibres. Sections of the three branches of the trigeminal nerve were found to comprise a range of fibre types including A-delta and C fibres. The size range of the cell bodies of the trigeminal ganglion reflected the fibre range since they correlated with the size range of axons in the nerve branches. Electrophysiological recordings of evoked activity from the ganglion confirmed the presence of these fibre types and the proportion of these mirrored the proportion of fibre types in the anatomical analyses. A-beta fibres were most common followed by A-delta fibres, then A-alpha fibres with C fibres being the fewest fibre type found. In higher vertebrates, A-delta and C fibres in the trigeminal nerve convey both somatosensory and nociceptive information to the brain. The evolutionary significance of these results is discussed as well as the potential for nociceptive capability in a lower vertebrate.

title: Measuring emotional processes in animals: the utility of a cognitive approach
abstract: Contemporary researchers regard emotional states as multifaceted, comprising physiological, behavioural, cognitive and subjective components. Subjective, conscious experience of emotion can be inferred from linguistic report in humans, but is inaccessible to direct measurement in non-human animals. However, measurement of other components of emotion is possible, and a variety of methods exist for monitoring emotional processes in animals by measuring behavioural and physiological changes. These are important tools, but they have limitations including difficulties of interpretation and the likelihood that many may be sensitive indicators of emotional arousal but not valence-pleasantness/unpleasantness. Cognitive components of emotion are a largely unexplored source of information about animal emotions, despite the fact that cognition-emotion links have been extensively researched in human cognitive science indicating that cognitive processes-appraisals of stimuli, events and situations-play an important role in the generation of emotional states, and that emotional states influence cognitive functioning by inducing attentional, memory and judgement biases. Building on this research, it is possible to design non-linguistic cognitive measures of animal emotion that may be especially informative in offering new methods for assessing emotional valence (positive as well as negative), discriminating same-valenced emotion of different types, identifying phenotypes with a cognitive predisposition to develop affective disorders, and perhaps shedding light on the issue of conscious emotional experiences in animals.

title: Deficits in hippocampus-mediated pavlovian conditioning in endogenous hypercortisolism
abstract: Elevated endogenous levels of corticosteroids cause neural dysfunction and loss, especially within the hippocampus, as well as cognitive impairment in hippocampus-mediated tasks. Because Cushing's syndrome patients suffer from hypercortisolism, they represent a unique opportunity to study the impact of elevated glucocorticoids on cognitive functions. The aim of this study was to examine the performance of Cushing's syndrome patients on trace eyeblink conditioning, a cross-species, hippocampal-mediated test of learning and memory.



Eleven Cushing's syndrome patients and 11 healthy control subjects participated in an eyeblink trace conditioning test (1000-msec trace) and a task of declarative memory for words. Salivary cortisol was collected in both the patients and the control subjects, and urinary free cortisol was collected in the patients only.



The patients exhibited fewer conditional responses and remembered fewer words, compared with the control subjects. Cortisol levels correlated with immediate and delayed declarative memory only. Conditional response correlated with delayed recall after controlling for the magnitude of unconditional response.



The integrity of the hippocampus seems to be compromised in Cushing's syndrome patients. Trace eyeblink conditioning might be useful both as a clinical tool to examine changes in hippocampus function in Cushing's disease patients and as a translational tool of research on the impact of chronic exposure of glucocorticoids.

title: Use of Physiological Telemetry as a Method of Estimating Metabolism of Fish in the Natural Environment
abstract: One of the most difficult components to measure in the energy budgets of natural fish populations is the energy released in metabolism. Telemetry of physiological correlates of metabolism, such as heart rate, can (with calibration) enable direct measurements of metabolism and simultaneous observations of behavior in the natural environment. Heart rate is a good indicator of metabolic rate for northern pike Esox lucius and was used to obtain information on metabolism, activity, and food intake in the natural environment. Mean field metabolic rates were 1.5 times standard metabolic rate. Activity metabolism accounted for 5–10% of total metabolism and was approximately an order of magnitude greater than estimates based on mean swimming speed derived from location tracking. Feeding metabolism was 15–25% of total metabolism. Even when the full metabolic rate calibration is not considered. physiological telemetry can provide useful information on patterns of energy expenditure ofdirect relevance to bio...

title: The past explains the present: Emotional adaptations and the structure of ancestral environments
abstract: Present conditions and selection pressures are irrelevant to the present design of organisms and do not explain how or why organisms behave adaptively, when they do. To whatever non-chance extent organisms are behaving adaptively, it is 1) because of the operation of underlying adaptations whose present design is the product of selection in the past, and 2) because present conditions resemble past conditions in those specific ways made developmentally and functionally important by the design of those adaptations. All adaptations evolved in response to the repeating elements of past environments, and their structure reflects in detail the recurrent structure of ancestral environments. Even planning mechanisms (such as “consciousness”), which supposedly deal with novel situations, depend on ancestrally shaped categorization processes and are therefore not free of the fast. In fact, the categorization of each new situation into evolutionarily repeating classes involves another kind of adaptation, the emotions, which match specialized modes of organismic operation to evolutionarily recurrent situations. The detailed statistical structure of these iterated systems of events is reflected in the detailed structure of the algorithms that govern emotional state. For this reason, the system of psychological adaptations that comprises each individual meets the present only as a version of the past.

title: Cognitive ornithology: the evolution of avian intelligence
abstract: Comparative psychologists interested in the evolution of intelligence have focused their attention on social primates, whereas birds tend to be used as models of associative learning. However, corvids and parrots, which have forebrains relatively the same size as apes, live in complex social groups and have a long developmental period before becoming independent, have demonstrated ape-like intelligence. Although, ornithologists have documented thousands of hours observing birds in their natural habitat, they have focused their attention on avian behaviour and ecology, rather than intelligence. This review discusses recent studies of avian cognition contrasting two different approaches; the anthropocentric approach and the adaptive specialization approach. It is argued that the most productive method is to combine the two approaches. This is discussed with respects to recent investigations of two supposedly unique aspects of human cognition; episodic memory and theory of mind. In reviewing the evidence for avian intelligence, corvids and parrots appear to be cognitively superior to other birds and in many cases even apes. This suggests that complex cognition has evolved in species with very different brains through a process of convergent evolution rather than shared ancestry, although the notion that birds and mammals may share common neural connectivity patterns is discussed.

title: Similar antagonism of morphine analgesia by MIF-1 and naloxone in Carassius auratus

abstract: Prolyl-leucyl-glycinamide (MIF-1), the C-terminal tripeptide of oxytocin, and naloxone were administered intracranially (IC) to goldfish (Carassius auratus) in doses of 0.001, 0.01, 0.1, 1.0 and 10.0 mg/kg and compared to a diluent control group for their ability to reduce the effects of morphine (30 mg/kg IC) in an assay measuring analgesia to electric shock. Threshold levels of pain were determined by the voltage necessary to produce an agitated swimming response (ASR). Both MIF-1 and naloxone were found to significantly reduce the analgesic effects of morphine when compared to the diluent control group. Similar dose-response curves in an apparent sine-wave pattern were noted with both MIF-1 and naloxone when comparisons were made both at 20 minutes after administration of morphine and over the entire 150 minutes of the experiment. The results support the evidence that MIF-1 can act as an opiate antagonist.

title: Pharmacokinetics of morphine in fish: winter flounder (Pseudopleuronectes americanus) and seawater-acclimated rainbow trout (Oncorhynchus mykiss).
abstract: We made a single intraperitoneal (IP) injection of morphine sulfate (40 mg/kg) into winter flounder and seawater acclimated rainbow trout at 10 degrees C and then followed its disposition by measuring the change in plasma morphine concentration for 100 h using a morphine specific ELISA. Disposition also was followed for 6h after a single IV injection of 7.5mg morphine sulfate in winter flounder. Plasma morphine reached a maximum within an hour post-injection IP and then decreased in a bi-exponential fashion with a rapid distribution phase followed by a slower elimination phase. The disposition was slower in flounder than in trout even though the fish were held at the same temperature. For example, plasma clearance was 76 mL h(-)(1) kg(-)(1) in the flounder but was almost twice as much in the trout (153 mL h(-)(1) kg(-)(1)) and mean residence time was 27.9h in the flounder but was 7.0 h in the trout. The present study is the first comprehensive pharmacokinetic analysis for any analgesic in an ectotherm, and our results show that: 1) significant intra-specific variation exists between fishes: and 2) the disposition of morphine in fish is approximately one order of magnitude slower than it is in mammals. These differences may be due in part to mass specific differences in cardiac output.

title: Investigating fear in domestic rainbow trout, Oncorhynchus mykiss, using an avoidance learning task
abstract: The capacity of rainbow trout to experience fear was assessed using an avoidance learning task. Each of 13 fish was placed individually into a two-chambered shuttle tank where it could be subjected to the putative frightening stimulus of a plunging dip net in either chamber. The fish could escape from the stimulus by swimming through a doorway to the other chamber. The fish escaped from the plunging net by swimming through the doorway, some on the first occasion and all after a few exposures. Each fish was then presented with a neutral stimulus of a light that went on 10 s before the net plunged into the water. Over a 5-day period, all fish learned to avoid the plunging net by swimming through the doorway when the light was illuminated. All fish showed evidence of longer-term memory by showing the learned avoidance response on the first occasion they were tested after 7 days of no testing. Whereas the escape responses to the plunging net were immediate and reflexive-like, the avoidance responses to the light going on were delayed a few seconds and more deliberate in nature. This evidence suggests that trout can experience fear and that they can learn to avoid frightening stimuli. It implies that they are sentient animals, more complex than previously thought.

title: Avoidance learning in goldfish (Carassius auratus) and trout (Oncorhynchus mykiss) and implications for Pain perception
abstract: This paper investigates spatially cued behavioural responses of two species of fish to an acute noxious stimulus and demonstrates response elasticity. Typical avoidance responses to a nociceptive stimulus were used to test (1) if fish learn to avoid spatial areas associated with a potentially noxious stimulus, (2) learning and memory improves with increased stimulus intensity and (3) a supposedly innate reflex behavioural responses change depending on the circumstances. Electric shocks with two different intensities, low (2.5 V and 3 V for trout and goldfish, respectively) and high (25 V and 30 V for trout and goldfish, respectively), were administered directly to the skin to stimulate cutaneous nociceptors. Goldfish (n = 8) demonstrated spatially cued shock avoidance and an increase in stimulus voltage, significantly improved shock-avoidance learning and memory. However, trout (n = 8) demonstrated shock-avoidance learning but no significant stimulus discrimination and little information retention. The presence of a conspecific significantly changed this behavioural response to a noxious stimulus. Trout were willing to remain in the vicinity of the conspecific while being subjected to low intensity shock stimuli previously shown to elicit avoidance. Goldfish tended to leave this area yet remain in the mid-tank area, adjacent to the stimulating zone, rather than at the end of the tank. These results suggest that shock avoidance in fish is not purely a reflex action. Fish were prepared to change the supposedly innate avoidance reaction according to a change in circumstances, an important concept in the ongoing debate on pain perception in fish.

title: Through Our Eyes Only?: The Search for Animal Consciousness
abstract: Through your eyes only miss Halsey moves her foot bees do it thinking ahead feeling our way balance of evidence.

title: Poultry welfare: Science or subjectivity?
abstract: 1. Most people in the developed world agree on what 'animal welfare' is, although it is impossible to give it a precise scientific definition. 2. The argument is made that animal welfare is all to do with the feelings of animals and not the primary needs that these feeling have evolved to protect. 3. Acceptance of subjective feelings as a legitimate subject for scientific investigation has a long and well-established history in science. This acceptance was interrupted by the rise of Behaviorism in the 20th century, but now seems to be re-established. 4. Subjective feelings cannot be studied directly. However, in the animal welfare debate, indirect evidence on feelings is extremely useful, and methods for obtaining this indirect evidence are described. 5. The poultry species are capable of feeling several states of suffering including fear, frustration and pain. A start has been made to elucidate these states and the conditions that cause them, but much remains to be done. Recent evidence suggests that the poultry species may also be capable of experiencing pleasure. 6. It is concluded that, although poultry welfare is all to do with the subjective feelings of the birds, it is possible to be objective and scientific about these feelings. Investigation into poultry welfare, therefore, really is science rather than subjectivity.

title: Reactions of Juvenile Pacific Salmon to Light
abstract: When given a choice between light and dark areas, schools of chum or pink salmon fry remain in the light, sockeye fry prefer the dark and coho fry show no marked preference for either. Newly emerge...

title: Different types of fear-conditioned behaviour mediated by separate nuclei within amygdala
abstract: The amygdala has long been thought to be involved in emotional behaviour, and its role in anxiety and conditioned fear has been highlighted. Individual amygdaloid nuclei have been shown to project to various cortical and subcortical regions implicated in affective processing. Here we show that some of these nuclei have separate roles in distinct mechanisms underlying conditioned fear responses. Rats with lesions of the central nucleus exhibited reduction in the suppression of behaviour elicited by a conditioned fear stimulus, but were simultaneously able to direct their actions to avoid further presentations of this aversive stimulus. In contrast, animals with lesions of the basolateral amygdala were unable to avoid the conditioned aversive stimulus by their choice behaviour, but exhibited normal conditioned suppression to this stimulus. This double dissociation demonstrates that distinct neural systems involving separate amygdaloid nuclei mediate different types of conditioned fear behaviour. We suggest that theories of amygdala function should take into account the roles of discrete amygdala subsystems in controlling different components of integrated emotional responses.

title: Spatial cognition and its neural basis in teleost fishes
abstract: The present review is focused on recent laboratory studies revealing that the spatial behaviour of fishes is as complex and elaborate as described in land vertebrates. In addition, the data presented here indicate that the remarkable richness and plasticity of spatial behaviour in fishes are based on learning and memory mechanisms and cognitive processes that depend on particular brain circuits, possibly homologous to those identified in mammals and birds. For example, there is evidence that the fish hippocampal pallium is essential for processing and encoding complex spatial information to form map-like representations of the environment. In contrast, body-centred orientation strategies or emotional learning are subserved by different cerebral structures, such as the optic tectum, the cerebellum or the amygdalar pallium. These results that suggest a striking similarity in some cognitive processes and their neural basis between fish and land vertebrates are consistent with the possibility that these vertebrate groups share a common basic pattern of brain and behaviour organisation inherited from a common ancestor and conserved through a long history of separate evolution.

title: The effects of telencephalic pallial lesions on spatial, temporal, and emotional learning in goldfish
abstract: In mammals, the pallial amygdala is implicated in emotional learning and memory, whereas the hippocampus is involved in spatial, contextual, or relational memory. This review presents a set of experiments aimed to study the involvement of the dorsomedial and dorsolateral telencephalon of goldfish in spatial and active avoidance learning. Results showed that (1) medial lesions impaired both acquisition and retention of conditioned avoidance response in two-way active avoidance learning experiments with stimuli overlapping (emotional factor) and with an interstimuli gap (temporal and emotional factors), and (2) the medial lesion did not affect spatial learning (spatial, contextual, or relational factors). In contrast, lateral lesions did not impair conditioned avoidance response with stimuli overlapping, but affected conditioned avoidance response with an interstimuli gap and spatial learning. These results support the presence of two differentiated memory systems in teleost fish based on discrete pallial regions: emotional (dorsomedial telencephalon) and spatial/temporal or relational (dorsolateral telencephalon). Furthermore, these functional data support the homology between the medial pallium of the teleost and the pallial amygdala of land vertebrates, and between the teleost lateral pallium and the mammalian hippocampus.

title: Welfare, Stress, and the Evolution of Feelings
abstract: Publisher Summary This chapter discusses the different kinds of feelings and considers the origin and possible function of each feeling. Each of these feelings has a biological role that complements various other anatomical, physiological, and behavioral mechanisms. All have some potential for improving fitness and most are likely to have been the subject of considerable selection pressure, but some aspects of feelings are likely to be just epiphenomena of neural mechanisms. With this view that most aspects of feelings have evolved like other biological mechanisms and that they help significantly in coping and responding, a single view of welfare as the state of an individual as regards its attempts to cope with its environment becomes clearer. Feelings are an important part of the welfare of an individual and should be assessed as well as possible. Other coping procedures and effects of the environment on the individual should also be assessed. An effect on an individual that is adverse in the long term is categorized as stress. Programs for trying to evaluate and improve welfare should combine the use of experiments to assess what is important to the individual by measuring the strengths of preferences, with monitoring studies in which feelings and other aspects of welfare are assessed more directly.