Effects of Acetic Acid and Morphine in Shore Crabs, Carcinus maenas: Implications for the Possibility of Pain in Decapods.

Noxious chemicals, coupled with morphine treatment, are often used in studies on pain in vertebrates. Here we show that injection of morphine caused several behavioural changes in the crab, Carcinus maenas, including reduced pressing against the sides of the enclosure and more rubbing and picking at the mouth parts and, at least for a short time, more defensive displays. Subsequent injection of acetic acid into one rear leg caused rubbing of the injected leg and the injected leg was held vertically off the ground. These activities directed at or involving the specific leg are consistent with previous observations of directed behaviour following noxious stimuli and are consistent with the idea that decapods experience pain. Further, acetic acid but not injection of water induced autotomy of the injected leg in these animals. Because autotomy is temporally associated with directed behaviour, it is possible that the autotomy is a pain-related response. Acetic acid is clearly a noxious substance when applied to decapods. However, morphine had no effect on the activities associated with acetic acid injection and thus there is no evidence for an analgesic effect. Further, the injection of acetic acid did not interfere with behavioural effects of morphine. The activities directed towards the site of injection are like those observed with injection, or with external application, of various noxious substances and the present study adds to a growing body of knowledge about possible pain in decapods.

Trade-Offs between Avoidance of Noxious Electric Shock and Avoidance of Bright Light in Shore Crabs Are Consistent with Predictions of Pain.

The suggestion that decapod crustaceans might experience pain has been dismissed by some authors who claim decapods only respond to noxious stimuli by nociceptive reflexes. Because reflexes do not require complex neuronal processing, but pain does, demonstrating reflex responses to noxious stimuli would not support the case for pain. Here, we report an experiment in which shore crabs are repeatedly placed in a light area (20 trials), but the animals can avoid the light by moving to a dark shelter. However, some crabs received an electric shock of 6 or 12 volts each time they entered the shelter. Those receiving either level of shock swiftly reduced their use of shelters and remained in the light. However, the magnitude of shelter avoidance was influenced by the brightness of the arena and the intensity of the shock. Shelter use was subsequently reduced to a greater extent if the shock level was high and the light intensity low. That is, crabs traded their avoidance of shock for their avoidance of bright light. Further, these animals showed avoidance learning and demonstrated activities suggesting anxiety, such as contact with the tank wall in the light area and increased latency to enter shelters when making the decision to enter the shelter if they had received shock in earlier trials. These results fulfil three key behavioural criteria for pain and, thus, are consistent with the idea that decapods can experience pain.

The Effects of Caustic Soda and Benzocaine on Directed Grooming to the Eyestalk in the Glass Prawn, Palaemon elegans, Are Consistent with the Idea of Pain in Decapods.

Acceptance of the possibility of pain in animals usually requires that various criteria are fulfilled. One such criterion is that a noxious stimulus or wound would elicit directed rubbing or grooming at the site of the stimulus. There is also an expectation that local anaesthetics would reduce these responses to damage. These expectations have been fulfilled in decapod crustaceans but there has been criticism of a lack of replication. Here, we report an experiment on the effects of a noxious chemical, sodium hydroxide, applied to one eyestalk of the glass prawn. This caused an immediate escape tail-flick response. It then caused nipping and picking with the chelipeds at the treated eyestalk but much less so at the alternative eyestalk. Prior treatment with benzocaine also caused an immediate tail-flick and directed behaviour, suggesting that this agent is aversive. Subsequently, however, it reduced the directed behaviour caused by caustic soda. We thus demonstrated responses that are consistent with the idea of pain in decapod crustaceans.

Behavioural Indicators of Pain and Suffering in Arthropods and Might Pain Bite Back?

Pain in response to tissue damage functions to change behaviour so that further damage is minimised whereas healing and survival are promoted. This paper focuses on the behavioural criteria that match the function to ask if pain is likely in the main taxa of arthropods. There is evidence consistent with the idea of pain in crustaceans, insects and, to a lesser extent, spiders. There is little evidence of pain in millipedes, centipedes, scorpions, and horseshoe crabs but there have been few investigations of these groups. Alternative approaches in the study of pain are explored and it is suggested that studies on traumatic mating, agonistic interactions, and defensive venoms might provide clues about pain. The evolution of high cognitive ability, sensory systems, and flexible decision-making is discussed as well as how these might influence the evolution of pain-like states.

Why it hurts: with freedom comes the biological need for pain.

We argue that pain is not needed to protect the body from damage unless the organism is able to make free choices in action selection. Then pain (including its affective and evaluative aspects) provides a necessary prioritising motivation to select actions expected to avoid it, whilst leaving the possibility of alternative actions to serve potentially higher priorities. Thus, on adaptive grounds, only organisms having free choice over action selection should experience pain. Free choice implies actions must be selected following appraisal of their effects, requiring a predictive model generating estimates of action outcomes. These features give organisms anticipatory behavioural autonomy (ABA), for which we propose a plausible system using an internal predictive model, integrated into a system able to produce the qualitative and affective aspects of pain. Our hypothesis can be tested using behavioural experiments designed to elicit trade-off responses to novel experiences for which algorithmic (automaton) responses might be inappropriate. We discuss the empirical evidence for our hypothesis among taxonomic groups, showing how testing for ABA guides thinking on which groups might experience pain. It is likely that all vertebrates do and plausible that some invertebrates do (decapods, cephalopods and at least some insects).

Caution is required when considering sentience in animals: a response to the commentary by Briffa (2022) on "Hermit crabs, shells, and sentience" (Elwood 2022).

My recent review examined the complex and intimate relationship between hermit crabs and the empty gastropod shells upon which they depend for survival. Because shells come with costs as well as benefits, the crabs are highly selective about which ones they prefer to occupy. Thus, a new shell is investigated and the information that is gathered appears to be compared with their existing shell before a decision is made. This is often prolonged and complex. Crabs also fight for shells and again the information that is gathered and used to inform fight decisions is complex. In my review, I consider these and other situations with reference to the possibility of sentience (including awareness). The excellent commentary from Mark Briffa expands on aspects of sentience and invokes the use of Lloyd Morgan's Canon to avoid suggesting complex abilities when simpler explanations would suffice. I agree with this approach. However, I also suggest that when simpler explanations appear not to explain the data, then it is reasonable to consider if more complex mental abilities might be used. I also like Briffa's suggestion that the use of an apparently higher mental faculty might result in a mechanism that is simpler than a large number of apparently simple decisions used to make complex decisions.

Invader abundance and contraction of niche breadth during replacement of a native gammarid amphipod.

The introduction of non-native species to new locations is a growing global phenomenon with major negative effects on native species and biodiversity. Such introductions potentially bring competitors into contact leading to partial or total species replacements. This creates an opportunity to study novel species interactions as they occur, with the potential to address the strength of inter- and intraspecific interactions, most notably competition. Such potential has often not been realized, however, due to the difficulties inherent in detecting rapid and spatially expansive species interactions under natural field conditions. The invasive amphipod crustacean Gammarus pulex has replaced a native species, Gammarus duebeni celticus, in river and lake systems across Europe. This replacement process is at least partially driven by differential parasitism, cannibalism, and intraguild predation, but the role of interspecific competition has yet to be resolved. Here, we examine how abundance of an invasive species may affect spatial niche breadth of a native congeneric species. We base our analyses of niche breadth on ordination and factor analysis of biological community and physical parameters, respectively, constituting a summative, multidimensional approach to niche breadth along environmental gradients. Results derived from biological and environmental niche criteria were consistent, although interspecific effects were stronger using the biological niche approach. We show that the niche breadth of the native species is constrained as abundance of the invader increases, but the converse effect does not occur. We conclude that the interaction between invasive G. pulex and native G. d. celticus under natural conditions is consistent with strong interspecific competition whereby a native, weaker competitor is replaced by a superior invasive competitor. This study indicates a strong role of interspecific competition, alongside other known interactions such as differential intraguild predation, in rapid and expansive species replacements following biological invasions.

Hermit crabs, shells, and sentience.

Hermit crabs have an intimate relationship with gastropod shells and show numerous activities by which they locate, select, and change shells in different contexts. They gather information about new shells and update information about their existing shells. This involves integration of different sensory modalities, memory-formation, and comparison of the overall value of each shell. Crabs also fight to get shells from other crabs, and again they gather information about the shell qualities and the opponent. Attacking crabs monitor their fight performance, and defenders are influenced by attacker activities, and both crabs are influenced by the gain or loss that might be made by swapping shells. Swapping shells involves the defender being naked for a short period. Leaving a shell also occurs if the shell is experimentally fixed in place or buried in sand or if small electric shocks are applied to the abdomen, and the quality of the current shell is traded-off against escaping possible asphyxiation or the aversive shocks. Hermit crabs show remarkable abilities, involving future planning, with respect to recognizing the shape and size of shells, and how they limit their passage through environmental obstructions. They also assess if shells might become available and wait for that to happen. Groups of crabs arrange themselves in size order so that orderly transfer of shells might occur down a line of crabs. These observations are discussed in the light of complex perceptual and cognitive abilities, and the possibility of sentience and awareness is discussed.

Potential Pain in Fish and Decapods: Similar Experimental Approaches and Similar Results.

I review studies that examined the possibility of pain experience in fish and note how they provided guidance on general methods that could be applied to other animals such as decapod crustaceans. The fish studies initially reported the occurrence of prolonged rocking movements in trout and rubbing of their lips if they were injected with acetic acid. Subsequent studies examined the role of morphine in reducing these activities and examined shifts in attention when responding to noxious stimuli. Various studies take up these themes in decapods. The results reported for the two taxonomic groups are remarkably similar and indicate that responses of both go beyond those expected of mere nociceptive reflex. Thus, the idea of pain cannot be dismissed by the argument that fish and decapods respond only by reflex. The responses of both clearly involve central processing, and pain experience, although not proven for either, is a distinct possibility. These studies have been the subjects of highly critical opinion pieces and these are examined and rebutted. The conclusion is that both fish and decapods should be awarded consideration for their welfare.

Flipping the parental switch: from killing to caring in male mammals.

Killing of unrelated young by sexually naïve male mammals is taxonomically widespread, but in many species, males subsequently show paternal care or at least do not harm their own young. This dramatic and important change is due to a shift in paternal state rather than to recognition of young, the mother or the location in which mating occurred. This transition from infanticidal to paternal behaviour is timed so that the inhibition of infanticide is synchronized with the birth of their own young. Ejaculation followed by cohabitation with the pregnant female causes this transition, but the precise stimuli from the female remain elusive. However, changes in social status also cause changes in infanticide. The switch from infanticide is accompanied by physiological change in the male that can be detected by both females and pups. Hormonal changes have been implicated in the switch but establishing causal links has been difficult. Recent neuroanatomical studies show that pup odours activate the vomeronasal organ and its efferent projections to induce infanticide. The emergence of paternal care depends on the inability of the vomeronasal organ to detect pup odours. In the absence of vomeronasal input, pup odours activate a conserved parental circuit and induce caregiving behaviour. An emerging picture is of complex, antagonistic circuits competing for behavioural expression, which allow for males to commit infanticide when they may benefit from such activity but ensure that they do not damage their fitness by killing their own young. However, we stress the need for more work on the neural mechanisms that mediate this process.

Discrimination between nociceptive reflexes and more complex responses consistent with pain in crustaceans.

Animals have quick-acting nociceptive reflexes that protect them from tissue damage. Some taxa have also evolved the capacity for pain. Pain appears to be linked to long-term changes in motivation brought about by the aversive nature of the experience. Pain presumably enhances long-term protection through behaviour modification based, in part, on memory. However, crustaceans have long been viewed as responding purely by reflex and thus not experiencing pain. This paper considers behavioural and physiological criteria that distinguish nociception from potential pain in this taxon. These include trade-offs with other motivational systems and prolonged motivational change. Complex, prolonged grooming or rubbing demonstrate the perception of the specific site of stimulus application. Recent evidence of fitness-enhancing, anxiety-like states is also consistent with the idea of pain. Physiological changes in response to noxious stimuli mediate some of the behavioural change. Rapid avoidance learning and prolonged memory indicate central processing rather than mere reflexes. Thus, available data go beyond the idea of just nociception. However, the impossibility of total proof of pain described in ways appropriate for our own species means that pain in crustaceans is still disputed. Pain in animals should be defined in ways that do not depend on human pain experience. This article is part of the Theo Murphy meeting issue 'Evolution of mechanisms and behaviour important for pain'.

Fish learn collectively, but groups with differing personalities are slower to decide and more likely to split.

We tested zebrafish shoals to examine whether groups exhibit collective spatial learning and whether this relates to the personality of group members. To do this we trained shoals to associate a collective spatial decision with a reward and tested whether shoals could reorient to the learned location from a new starting point. There were strong indications of collective learning and collective reorienting, most likely by memorising distal cues, but these processes were unrelated to personality differences within shoals. However, there was evidence that group decisions require agreement between differing personalities. Notably, shoals with more boldness variation were more likely to split during training trials and took longer to reach a collective decision. Thus cognitive tasks, such as learning and cue memorisation, may be exhibited collectively, but the ability to reach collective decisions is affected by the personality composition of the group. A likely outcome of the splitting of groups with very disparate personalities is the formation of groups with members more similar in their personality.

Grouping promotes risk-taking in unfamiliar settings.

Acting collectively in a group provides risk-reducing benefits. Yet individuals differ in how they take risks, with some being more willing than others to approach dangerous or unfamiliar settings. Therefore, individuals may need to adjust their behaviour when in groups, either as a result of perceiving greater safety or to coordinate collective responses, the latter of which may rely on within-group dynamics biased by group composition. In zebrafish we explored how these aspects of grouping affect risk-taking behaviour by comparing solitary to group conditions and testing the ability of group-member solitary responses to predict collective responses. We focused on approach-latency towards a novel object and an unusual food to test this, for shoals of five fish. There was no indication that collective latencies are predicted by how each fish responded when alone in terms of the extremes, the variance or the mean of group-member latency towards the unusual food and the novel-object. However, fish were overall faster and less variable in their approach when shoaling. This indicates lower risk aversion by individuals in groups, presumably as a result of group safety. An interesting consequence of the overall low risk-aversion in shoals is that more risk-aversive fish adjust their behaviour more than less risk averse fish.

Aversive responses by shore crabs to acetic acid but not to capsaicin.

Nociception is the ability to encode and perceive harmful stimuli and allows for a rapid reflexive withdrawal. In some species, nociception might be accompanied by a pain experience, which is a negative feeling that allows for longer-term changes in behaviour. Different types of stimuli may affect nociceptors, but in crustaceans there is conflicting evidence about the ability to respond to chemical stimuli. This study attempts to resolve this situation by testing behavioural responses of the common shore crab, Carcinus maenas, to two chemical irritants frequently used in vertebrate pain studies (acetic acid and capsaicin). In our first experiment acetic acid, water, capsaicin or mineral oil were applied by brush to the mouth, and in a second experiment treatments were applied to the eyes. Application of acetic acid had a marked effect on behaviour that included vigorous movement of mouth parts, scratching at the mouth with the claws and attempts to escape from the enclosure. Acetic acid also caused holding down of the acid-treated eye in the socket. By contrast, capsaicin had no effect and was no different to the control treatment of mineral oil and water. These results demonstrate responsiveness to acetic acid and thus nociceptive capacity for at least some chemicals. Further, the responses that persist after application were consistent with the idea of pain, however, proof of pain is not possible in any animal.

Trade-offs between predator avoidance and electric shock avoidance in hermit crabs demonstrate a non-reflexive response to noxious stimuli consistent with prediction of pain.

Arthropods have long been thought to respond to noxious stimuli by reflex reaction. One way of testing if this is true is to provide the animal with a way to avoid the stimulus but to vary the potential cost of avoidance. If avoidance varies with potential cost then a decision making process is evident and the behaviour is not a mere reflex. Here we examine the responses of hermit crabs to electric shock within their shell when also exposed to predator or non-predator odours or to no odour. The electric shocks start with low voltage but increase in voltage with each repetition to determine how odour affects the voltage at which the shell is abandoned. There was no treatment effect on the voltage at which hermit crabs left their shells, however, those exposed to predator odours were less likely to evacuate their shells compared with no odour or low concentrations of non-predator odour. However, highly concentrated non-predator also inhibited evacuation. The data show that these crabs trade-off avoidance of electric shock with predator avoidance. They are thus not responding purely by reflex and the data are thus consistent with predictions of pain but do not prove pain.

No discrimination shock avoidance with sequential presentation of stimuli but shore crabs still reduce shock exposure.

Insights into the potential for pain may be obtained from examination of behavioural responses to noxious stimuli. In particular, prolonged responses coupled with long-term motivational change and avoidance learning cannot be explained by nociceptive reflex but are consistent with the idea of pain. Here, we placed shore crabs alternately in two halves of a test area divided by an opaque partition. Each area had a dark shelter and in one repeated small electric shocks were delivered in an experimental but not in a control group. Crabs showed no specific avoidance of the shock shelter either during these trials or in a subsequent test in which both were offered simultaneously; however they often emerged from the shock shelter during a trial and thus avoided further shock. More crabs emerged in later trials and took less time to emerge than in early trials. Thus, despite the lack of discrimination learning between the two shelters they used other tactics to markedly reduce the amount of shock received. We note that a previous experiment using simultaneous presentation of two shelters demonstrated rapid discrimination and avoidance learning but the paradigm of sequential presentation appears to prevent this. Nevertheless, the data show clearly that the shock is aversive and tactics, other than discrimination learning, are used to avoid it. Thus, the behaviour is only partially consistent with the idea of pain.

Plasticity varies with boldness in a weakly-electric fish.

BACKGROUND: The expression of animal personality is indicated by patterns of consistency in individual behaviour. Often, the differences exhibited between individuals are consistent across situations. However, between some situations, this can be biased by variable levels of individual plasticity. The interaction between individual plasticity and animal personality can be illustrated by examining situation-sensitive personality traits such as boldness (i.e. risk-taking and exploration tendency). For the weakly electric fish Gnathonemus petersii, light condition is a major factor influencing behaviour. Adapted to navigate in low-light conditions, this species chooses to be more active in dark environments where risk from visual predators is lower. However, G. petersii also exhibit individual differences in their degree of behavioural change from light to dark. The present study, therefore, aims to examine if an increase of motivation to explore in the safety of the dark, not only affects mean levels of boldness, but also the variation between individuals, as a result of differences in individual plasticity. RESULTS: Boldness was consistent between a novel-object and a novel-environment situation in bright light. However, no consistency in boldness was noted between a bright (risky) and a dark (safe) novel environment. Furthermore, there was a negative association between boldness and the degree of change across novel environments, with shier individuals exhibiting greater behavioural plasticity. CONCLUSIONS: This study highlights that individual plasticity can vary with personality. In addition, the effect of light suggests that variation in boldness is situation specific. Finally, there appears to be a trade-off between personality and individual plasticity with shy but plastic individuals minimizing costs when perceiving risk and bold but stable individuals consistently maximizing rewards, which can be maladaptive.

To breathe or fight? Siamese fighting fish differ when facing a real opponent or mirror image.

Displays are a feature of animal contest behaviour and have been interpreted as a means of gathering information on opponent fighting ability, as well as signalling aggressive motivation. In fish, contest displays often include frontal and lateral elements, which in the latter involves contestants showing their flanks to an opponent. Previous work in a range of fish species has demonstrated population-level lateralization of these displays, preferentially showing one side to their opponent. Mirrors are commonly used in place of a real opponent to study aggression in fish, yet they may disrupt the normal pattern of display behaviour. Here, using Siamese fighting fish, Betta splendens, we compare the aggressive behaviour of males to a mirror image and real opponent behind a transparent barrier. As this species is a facultative air-breather, we also quantify surface breathing, providing insights into underlying fight motivation. Consistent with previous work, we found evidence of population-level lateralization, with a bias to present the left side and use the left eye when facing a real opponent. Contrary to expectations, there were no differences in the aggressive displays to a mirror and real opponent, with positive correlations between the behaviour in the two scenarios. However, there were important differences in surface breathing, which was more frequent and of longer duration in the mirror treatment. The reasons for these differences are discussed in relation to the repertoire of contest behaviour and motivation when facing a real opponent.

Electric shock causes physiological stress responses in shore crabs, consistent with prediction of pain.

Animal pain is defined by a series of expectations or criteria, one of which is that there should be a physiological stress response associated with noxious stimuli. While crustacean stress responses have been demonstrated they are typically preceded by escape behaviour and thus the physiological change might be attributed to the behaviour rather than a pain experience. We found higher levels of stress as measured by lactate in shore crabs exposed to brief electric shock than non-shocked controls. However, shocked crabs showed more vigorous behaviour than controls. We then matched crabs with the same level of behaviour and still found that shocked crabs had stronger stress response compared with controls. The finding of the stress response, coupled with previous findings of long-term motivational change and avoidance learning, fulfils the criteria expected of a pain experience.