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.     

Analgesia in phasic and tonic pain tests in a pharmacological model of autotomy

Self-mutilation or self-injurious behaviour is a well known behavioural disorder in humans. The proposition that this behaviour in animals is a response to chronic pain of peripheral nerve injury has been met with controversy. In the present study a pharmacological model, which produces no sensory or motor loss was used to study how autotomy is related to pain. In a group of rats autotomy was induced by amphetamine in phenoxybenzamine and reserpine treated animals. The pain tests, both phasic and tonic were then performed. The results of this study showed that a total analgesia was produced in both phasic and tonic pain tests, in animals that exhibited autotomy. Injection of naloxone in these animals prevented autotomy. A correlation between autotomy and no pain is suggested in this pharmacological model of autotomy.     

Nociceptive reactions during stimulation of immunity in rats with various individual resistance to stress

Various behavioural nociceptive reactions and individual resistance against stress were studied under conditions of stimulation of the immune processes by various techniques. The research problems included a study of influence of the immune stimulation with preparation "Imunofan" upon pain responses depending on individual resistance of animals to a stress, and the obtained results were compared with similar data in natural model of immune activation. To reveal central immune regulation of nociceptive reactions, imunofan was injected into brain ventricles. The work was carried out in 43 "Wistar" adult male rats. Free "open field" behaviour of animals was recorded to define a stress-resistance. Following nociceptive reactions, tail-flick to thermal stimuli; start, escape, jumping and vocalization to electrical skin stimulation, were studied. It was shown that intramuscular injection of imunofan (0.01 ml, 0.005% solution) depressed an active behaviour of animals in open field and reduced pain thresholds. This hyperalgesia was much higher in non-resistant rats in comparison to the resistant ones. Similar results were obtained in natural activation of immunity caused by operative procedure necessary for injection of imunofan into ventricles. Intracerebroventricular injections were accompanied by stronger and more complex changes of pain sensitivity.     

Are We Looking in the Wrong Place? Implications for Behavioural-Based Pain Assessment in Rabbits (Oryctolagus cuniculi) and Beyond?

Background

Successful observation of behaviour depends upon knowing both which behaviours to look for and focusing on the appropriate areas of the body to observe them. Behaviour based scoring systems have become increasingly widely used to assess animal pain and distress. Although studies are available demonstrating which behaviours need to be observed, there has been little attempt to assess how effectively observers apply such information when viewing an animal''s behaviour.

Methodology/Principal Findings

This study used historical video recordings of New Zealand white rabbits (Oryctolagus cuniculi) considered to be experiencing varying degrees of post-operative pain to assess the pattern of observation and the ability to assess pain exhibited by both experienced and inexperienced human participants (n = 151). Eye tracking equipment was used to identify how quickly, how frequently, for how long different areas of the rabbit''s body were attended to by the participants. Simple visual analgoue scoring was used to assess the pain experienced in each sequence. The results demonstrate that irrespective of their experience or gender, observers focus first, more frequently and for longer on the face, compared to the abdomen, ears, back and hindquarters of the rabbit and that participants were poor at identifying rabbits in pain. Observing the back and hindquarters was correlated with ‘correct’ assessments and observing the face was correlated with ‘incorrect’ assessments.

Conclusions

In conclusion, irrespective of experience and gender, observers focused on the face when using behaviour to assess pain and were unable to effectively identify rabbits in pain. Focusing on the face is unlikely to be effective when using behavioural indicators of pain since they involve other body areas. Alternatively, if animals exhibit pain-related facial expressions, then it could improve our ability to assess pain. In addition, these results have potential implications for the use of behaviour to assess how rabbits and potentially other species feel.     

Acute pain and a motivational pathway in adult rats: influence of early life pain experience

Background

The importance of neonatal experience upon behaviour in later life is increasingly recognised. The overlap between pain and reward pathways led us to hypothesise that neonatal pain experience influences reward-related pathways and behaviours in adulthood.

Methodology/Principal Findings

Rat pups received repeat plantar skin incisions (neonatal IN) or control procedures (neonatal anesthesia only, AN) at postnatal days (P)3, 10 and 17. When adult, rats with neonatal ‘pain history’ showed greater sensory sensitivity than control rats following acute plantar skin incision. Motivational behaviour in the two groups of rats was tested in a novelty-induced hypophagia (NIH) paradigm. The sensitivity of this paradigm to pain-induced changes in motivational behaviour was shown by significant increases in the time spent in the central zone of the arena (43.7±5.9% vs. 22.5±6.7%, p<0.05), close to centrally placed food treats, and decreased number of rears (9.5±1.4 vs. 19.2±2.3, p<0.001) in rats with acute plantar skin incision compared to naive, uninjured animals. Rats with a neonatal ‘pain history’ showed the same pain-induced behaviour in the novelty-induced hypophagia paradigm as controls. However, differences were observed in reward-related neural activity between the two groups. Two hours after behavioural testing, brains were harvested and neuronal activity mapped using c-Fos expression in lateral hypothalamic orexin neurons, part of a specific reward seeking pathway. Pain-induced activity in orexin neurons of control rats (18.4±2.8%) was the same as in uninjured naive animals (15.5±2.6%), but in those rats with a ‘pain history’, orexinergic activity was significantly increased (27.2±4.1%, p<0.01). Furthermore the extent of orexin neuron activation in individual rats with a ‘pain history’ was highly correlated with their motivational behaviour (r = −0.86, p = 0.01).

Conclusions/Significance

These results show that acute pain alters motivational behaviour and that neonatal pain experience causes long-term changes in brain motivational orexinergic pathways, known to modulate mesolimbic dopaminergic reward circuitry.     

The development of nociceptive circuits

The study of pain development has come into its own. Reaping the rewards of years of developmental and molecular biology, it has now become possible to translate fundamental knowledge of signalling pathways and synaptic physiology into a better understanding of infant pain. Research has cast new light on the physiological and pharmacological processes that shape the newborn pain response, which will help us to understand early pain behaviour and to design better treatments. Furthermore, it has shown how developing pain circuitry depends on non-noxious sensory activity in the healthy newborn, and how early injury can permanently alter pain processing.     

Effects of verapamil on behavioral and microcirculatory disorders in pain syndrome of spinal etiology

In the experiments on Wistar rats with pain syndrome of spinal origin (PSSO) caused by the generator of pathologically enhanced excitation (GPEE) in dorsal horns of the spinal cord lumbosacral segments, it was shown that the intravenous verapamil injection (1.25 mg/kg) undoubtedly decreased behaviour response and improves the state of microcirculation. The compound of 10-fold decreased dose does not affect the behaviour response and microcirculation. When PSSO exists, the intravenous injection of analgin (150 mg/kg) produced an effect on the behaviour response and does not produce any action on microcirculation. When verapamil reaches the dorsal surface of the spinal cord (GPEE area) it decreases the behaviour response and microcirculation disorders created in PSSO. The obtained data make it clear that the GPEE depression caused by the verapamil calcium channel blocker weakens PSSO and normalizes the microcirculation.     

Glial activation and pathological pain

Pain is a sensation we have all experienced. For most of us, the pain has been temporary. However, for patients with pathological pain, the pain experience is unending, with little hope for therapeutic relief. Pathological pain is characterized by an amplified response to normally innocuous stimuli, and an amplified response to acute pain. Pathological pain has long been described as the result of dysfunctional neuronal activity. While neuronal functioning is indeed altered, there is significant evidence showing that exaggerated pain is regulated by the activation of astrocytes and microglia. In exaggerated pain, astrocytes, and microglia are activated by neuronal signals including substance P, glutamate, and fractalkine. Activation of glia by these substances leads to the release of mediators that then act on other glia and neurons. These include a family of proteins called "proinflammatory cytokines" released from microglia and astrocytes. These cytokines have been shown to be critical mediators of exaggerated pain. Some patients with pathological pain also report "extra-territorial" and/or "mirror" image pain. That is, exaggerated pain is experienced not only in the area of trauma. In extra-territorial pain, pain is also perceived as arising from neighboring healthy tissues outside of the site of trauma. In the rare cases of mirror-image pain, such pain is perceived as arising from the healthy, corresponding body part on the opposite side of the body. New data suggest that activation of astrocyte communication via gap junctions may mediate such spread of pain. While traditional therapies for pathological pain have focused on neuronal targets, the following review describes glia as newly recognized mediators of exaggerated pain, and as new therapeutic targets. Moreover, the glial-neuronal interactions discussed here are likely not exclusive to pain, but rather are likely to play significant roles in other behavioral phenomena.     

The effect of acute alcohol administration on auditory evoked potentials in altricial nestlings during the development of natural behavior]

Acute ethanol influence on field L auditory evoked potentials (AEP) was studied in 4-8-days-old altricial nestlings of pied flycatcher. Nestlings were presented with tone pips related with the realization of natural behaviour (2.0 and 5.0 kHz) and bearing no meaning for the behaviour of the young of the age under study (3.0 kHz). Ethanol ingestion was found to reduce the maturity index (MI) of AEP in response to "behavioural" but not to control frequencies; this effect was first observed at day 5, when nestlings eyes opened and defence behaviour appeared, while previously formed feeding behaviour was significantly modified. During the next 2 days alcohol had a greater effect upon the AEP in response to 2.0 kHz tone pips, related with feeding behaviour of increasing complexity than upon the AEP in response to 5.0 kHz, related with the defence behaviour that remained relatively constant. The previous data concerning the effect of alcohol on unit activity are used to support the view that MI increase during the early postembryonic ontogeny is due to the involvement of neurons with newly formed behavioural specializations into the subserving of new behavioural patterns while the decrease of MI under alcohol is due to the depression of activity in these neurons.     

The pain-adaptation model: a discussion of the relationship between chronic musculoskeletal pain and motor activity

Articles describing motor function in five chronic musculoskeletal pain conditions (temporomandibular disorders, muscle tension headache, fibromyalgia, chronic lower back pain, and postexercise muscle soreness) were reviewed. It was concluded that the data do not support the commonly held view that the pain of these conditions is maintained by some form of tonic muscular hyperactivity. Instead, it seems clear that in these conditions the activity of agonist muscles is often reduced by pain, even when this does not arise from the muscle itself. On the other hand, pain causes small increases in the level of activity of the antagonist. As a consequence of these changes, force production and the range and velocity of movement of the affected body part are often reduced. To explain how such changes in the behaviour come about, we propose a neurophysiological model based on the phasic modulation of excitatory and inhibitory interneurons supplied by high-threshold sensory afferents. We suggest that the "dysfunction" that is characteristic of several types of chronic musculoskeletal pain is a normal protective adaptation and is not a cause of pain.     

Evaluation of EMLA Cream for Preventing Pain during Tattooing of Rabbits: Changes in Physiological, Behavioural and Facial Expression Responses

Background

Ear tattooing is a routine procedure performed on laboratory, commercial and companion rabbits for the purpose of identification. Although this procedure is potentially painful, it is usually performed without the provision of analgesia, so compromising animal welfare. Furthermore, current means to assess pain in rabbits are poor and more reliable methods are required. The objectives of this study were to assess the physiological and behavioural effects of ear tattooing on rabbits, evaluate the analgesic efficacy of topical local anaesthetic cream application prior to this procedure, and to develop a scale to assess pain in rabbits based on changes in facial expression.

Methodology/Principal Findings

In a crossover study, eight New Zealand White rabbits each underwent four different treatments of actual or sham ear tattooing, with and without prior application of a topical local anaesthetic (lidocaine/prilocaine). Changes in immediate behaviour, heart rate, arterial blood pressure, serum corticosterone concentrations, facial expression and home pen behaviours were assessed. Changes in facial expression were examined to develop the Rabbit Grimace Scale in order to assess acute pain. Tattooing without EMLA cream resulted in significantly greater struggling behaviour and vocalisation, greater facial expression scores of pain, higher peak heart rate, as well as higher systolic and mean arterial blood pressure compared to all other treatments. Physiological and behavioural changes following tattooing with EMLA cream were similar to those in animals receiving sham tattoos with or without EMLA cream. Behavioural changes 1 hour post-treatment were minimal with no pain behaviours identifiable in any group. Serum corticosterone responses did not differ between sham and tattoo treatments.

Conclusions

Ear tattooing causes transient and potentially severe pain in rabbits, which is almost completely prevented by prior application of local anaesthetic cream. The Rabbit Grimace Scale developed appears to be a reliable and accurate way to assess acute pain in rabbits.     

Addressing the pain associated with disbudding and dehorning in cattle

The pain caused by disbudding or dehorning of cattle and its alleviation may be assessed by behavioural, physiological and production responses. Disbudding can be carried out by cautery or the application of a chemical paste. Cautery disbudding and amputation dehorning stimulate definite pain related behaviours during and after the procedure but caustic disbudding causes little response during the procedure though it is followed by behaviours indicative of pain. All three procedures cause definite plasma cortisol responses but the response to amputation dehorning is significantly greater than the response to cautery or chemical disbudding. It has a characteristic format; a rapid increase following dehorning to a maximum concentration within 30 min, then a decline to plateau levels, followed by a return to pretreatment values at 7-8 h. Local anaesthesia administered before disbudding or dehorning eliminates pain-related behaviour and reduces the plasma cortisol response for about 1.5 h. Following amputation dehorning the plasma cortisol concentration then increases for about 6 h before returning to pretreatment levels. When local anaesthesia and a non-steroidal anti-inflammatory drug (NSAID) are given before disbudding or dehorning the cortisol responses to these procedures are virtually eliminated. Chronic pain in the days following disbudding and dehorning is poorly understood. As a general rule, if pain relief is not available, cautery disbudding is preferable to chemical disbudding or amputation dehorning. If possible, local anaesthesia and better still local anaesthesia plus a NSAID should be used to minimise the pain caused by all three procedures.     

A positive feedback model for switching between two activities

We describe a model for the alternation between two activities. Each activity has an associated tendency, and the model performs the activity with the higher tendency. The tendencies depend on both negative and positive feedback from the performance of the activities. In general the behaviour of the model is periodic, with bout, durations that depend on the ratio of the maximum response rates. Increasing the maximum rate of a behaviour increases the magnitude of its bouts. The behaviour of the model is similar in several respects to the model developed by Ludlow (1980), which suggests that it may not be easy to distinguish between various models of feeding and drinking behaviour.     

Avoidance reactions to painful stimulation of another individual following destruction of the rat amygdala]

Experiments in male albino rats have shown that bilateral lesion of the amygdala does not prevent avoidance conditioning to pain stimulation of another rat, improvement of the reaction after a painful action of the experimental animal and its extinction after the discontinuation of pain stimulation of the partner. Amygdalectomy performed after elaboration of a conditioned avoidance reaction, improves or worsens it, depending on which of the two rivaling motivations (sensitivity to signals of the other animal or fear of an open field) relatively predominanted in the given animal before the operation. The facts obtained suggest that the amygdala may be classified as belonging to the system of brain structures, where the "interrupting function" of emotions is achieved directing the behaviour to meet the predominant need.     

Effect of previous experience in reacting to a danger signal on "open field" behavior in the rat

Modification of rats behaviour in an "hopen field" test was investigated, induced by an acoustic stimulus, previously subjected to conditioning in a shuttle chamber in experiments with possibility and impossibility of avoidance from electrical shock. It has been established that presentation of a stimulus having the meaning of a danger signal, in a new situation, significantly suppresses investigating behaviour of rats, whereas the stimulus which had not been subjected to conditioning exerts no marked effect on behaviour. The greatest suppression was observed in rats with "learned helplessness". This fact suggests that the degree of suppression of the behaviour in an open field in response to a danger signal, depends on the animal's previous experience in reacting to this signal.     

Aphids Playing Possum – Defensive or Mutualistic Response?

Paper presents the phenomenon of thanatosis or death-feigning in selected aphids species. This specific reaction was predominantly analysed on the example of aphid subfamily Lachninae. Individuals of this group were used in experiments, during which a thanatotic response was induced with various results. The response differed from prolonged thanatosis, lasting for several minutes (Eulachnus rileyi), through shrinking behaviour (e. g. in Stomaphis graffii) to non-responsive species such as Cinara (Schizolachnus) pineti. The large interspecies variation of observed responses can be linked to other defensive mechanisms existing in the studied species, as well as to their mutualistic relationship with ants. The behaviour of shrinking is hypothesized to be the mutualistic response, developed from thanatosis, and being adapted to transportation by ant workers.     

Indirect Acquisition of Pain-Related Fear: An Experimental Study of Observational Learning Using Coloured Cold Metal Bars

Background

Previous research has demonstrated that pain-related fear can be acquired through observation of another’s pain behaviour during an encounter with a painful stimulus. The results of two experimental studies were presented, each with a different pain stimulus, of which the aim was to investigate the effect of observational learning on pain expectancies, avoidance behaviour, and physiological responding. Additionally, the study investigated whether certain individuals are at heightened risk to develop pain-related fear through observation. Finally, changes in pain-related fear and pain intensity after exposure to the feared stimulus were examined.

Methods

During observational acquisition, healthy female participants watched a video showing coloured cold metal bars being placed against the neck of several models. In a differential fear conditioning paradigm, one colour was paired with painful facial expressions, and another colour was paired with neutral facial expressions of the video models. During exposure, both metal bars with equal temperatures (-25° or +8° Celsius) were placed repeatedly against participants’ own neck.

Results

Results showed that pain-related beliefs can be acquired by observing pain in others, but do not necessarily cause behavioural changes. Additionally, dispositional empathy might play a role in the acquisition of these beliefs. Furthermore, skin conductance responses were higher when exposed to the pain-associated bar, but only in one of two experiments. Differential pain-related beliefs rapidly disappeared after first-hand exposure to the stimuli.

Conclusions

This study enhances our understanding of pain-related fear acquisition and subsequent exposure to the feared stimulus, providing leads for pain prevention and management strategies.     

An experimental study examining the anti-predator behaviour of Sabine’s gulls (Xema sabini) during breeding

Anti-predatory behaviour is widespread among a broad range of animal taxa, including birds. Nest defence is not without risk, however, and parent birds face a trade-off between the survival of their offspring and the risk of injury or mortality to themselves. This study focused on the anti-predator behaviour of the Sabines gull (Xema sabini), a ground-nesting, Arctic breeder. Specifically, we quantified the gulls behavioural response towards natural predators, a human intruder, experimental predator decoys, and a non-predatory decoy. Neither the distance at which nesting Sabines gulls first reacted to natural predators, nor the relative intensity of their response, differed with incubation stage or predator type. However, response behaviour was highly variable among pairs. In response to decoys, Sabines gulls responded strongly towards predatory species, compared with a non-predatory species. The distance at which they first swooped at a human intruder was also variable, and there was no seasonal trend. Sabines gulls were often joined in nest defence by conspecifics, Arctic terns, and shorebirds nesting nearby, although the number of conspecifics involved in attacks was not related to the proximity of neighbouring nests.     

The interrelation between individual behavioral characteristics and the indices of brain energy metabolism in rats]

In rats with active type of behaviour in "open field" and "forced swimming" tests in response to weak stress (handling) both the rate of local blood flow (RLBF) and free oxygen tension level (pO2) in the brain are increased, and in rats with passive type of behaviour RLBF is increased, but the pO2 level is decreased. The character of pO2 level changes in the brain under stress is significantly (r = -0.74, p less than 0.001) connected with the level of depressiveness (time of passive swimming) and is nonsignificantly connected with the level of the motor activity. Indices of the active type of behaviour (the number of crossed squares, rearings, comings out to the center of the field and the time of extinction of the motor activity) positively correlate with succinate dehydrogenase (SDG) activity and negatively with NADH-dehydrogenase (NADH-DG) activity and the index of the passive type of behaviour (time of passive swimming) positively correlates with NADH-DG activity and negatively--with SDG activity.     

Is Altered Central Pain Processing Related to Disease Stage in Chronic Pancreatitis Patients with Pain? An Exploratory Study

Background

The most dominant feature in chronic pancreatitis is intense abdominal pain. Changes in spinal and/or supraspinal central nervous system pain processing due to visceral nociceptive input play an important role in this pain. How altered pain processing is related to disease stage still needs study.

Methodology/Principal Findings

Sixty chronic pancreatitis patients were compared to 15 healthy controls. Two subgroups of pancreatitis patients were defined based on the M-ANNHEIM severity index of chronic pancreatitis; i.e. moderate and severe. Pain detection and tolerance thresholds for pressure and electric stimuli were measured in six selected dermatomes (C5, T4, T10, L1, L4 and T10BACK). In addition, the conditioned pain modulation response to cold pressor task was determined. These measures were compared between the healthy controls and chronic pancreatitis patients. Severe pancreatitis patients showed lower pain thresholds than moderate pancreatitis patients or healthy volunteers. Healthy controls showed a significantly larger conditioned pain modulation response compared to all chronic pancreatitis patients taken together.

Conclusions/Significance

The present study confirms that chronic pancreatitis patients show signs of altered central processing of nociception compared to healthy controls. The study further suggests that these changes, i.e. central sensitization, may be influenced by disease stage. These findings underline the need to take altered central pain processing into account when managing the pain of chronic pancreatitis.