Remote monitoring of electroencephalogram, electrocardiogram, and behavior during controlled atmosphere stunning in broilers: implications for welfare.

This study examined the welfare implications of euthanizing broilers with 3 gas mixtures relevant to the commercial application of controlled atmosphere stunning (CAS). Birds were implanted/equipped with electrodes to measure brain activity (electroencephalogram, EEG) and heart rate. These signals were recorded using a purpose-built telemetry-logging system, small enough to be worn by each bird in a spandex backpack. The birds were euthanized in a scaled-down CAS apparatus consisting of a conveyor belt passing through 2 compartments. Three gas environments were applied (8 birds per treatment): 1) anoxia (N(2) with <2% residual O(2), in both compartments), 2) hypercapnic anoxia (N(2) with 30% CO(2) and <2% residual O(2), in both compartments), and 3) a 2-phase approach with a hypercapnic hyperoxygenated anesthetic phase (40% CO(2), 30% O(2), and 30% N(2), in the first compartment, 80 s) followed by a second euthanasia phase (80% CO(2) in air, in the second compartment). All 3 CAS approaches effectively achieved nonrecovery states, and time to loss of consciousness for each bird was determined by visual determination of isoelectric EEG and by calculation of the correlation dimension of the EEG. Hypercapnic anoxia resulted in rapid unconsciousness and death; both anoxic treatments were associated with early onset prolonged wing flapping and sustained tonic convulsions as displayed in the electrophysiological recordings. These responses were seen in the period when consciousness remained a possibility. Hypercapnic hyperoxygenation (the 2-phase approach) was associated with respiratory disruption, but this treatment eliminated initial clonic convulsions in the stunning process, and tonic convulsions were not seen. These results suggest that the presence of O(2) in the first stage of CAS is associated with an absence of potentially distressing behavioral responses. The respiratory discomfort associated with hypercapnic hyperoxygenation is an issue. We propose that this may be compensated by a more gradual induction to unconsciousness, which eliminates the impact of other potentially negative experiences.

N150 in amygdalar ERPs in the rat: is there modulation by anticipatory fear?

The hypothesis was tested whether the amygdalar N150 of rats, a slow, negative component in the event-related potential from the lateral amygdala, is sensitive to a state of anxious anticipation. A conditioning procedure was applied in which a series of six auditory stimuli was followed by a shock when presented alone, but not when the auditory stimuli were preceded by a visual stimulus. Heart rate recordings confirmed that the auditory stimulus train induced a state of increasing anticipatory fear and that this condition was modulated by the visual stimulus. During behavioral training, a N150 appeared in the amygdalar event-related potential evoked by the auditory stimuli, replicating previous findings. However, the amplitude of the N150 was not affected by whether or not the visual stimulus had been presented before. These results failed to support the idea that the N150 is related to the expectancy of an aversive event. An alternative interpretation, emphasizing the increase in arousal and attention that is inherent to aversive learning, is discussed.

Are there different mechanisms of synchronization in the course of spike-wave discharges (SWDs) burst development in WAG/Rij rats?

In WAG/Rij rats the pair linear correlation r was calculated for bipolar recordings in fronto-temporal, fronto-occipital and occipito-temporal zones of both hemispheres as well as in paleocerebellar cortex (culmen). It was shown that development of SWD bursts resulted in interhemispheric decreases of correlation between the right occipito-temporal cortical region on one side, and left fronto-temporal on the contralateral side. Towards the end of SWD, we found an increased interhemispheric correlation between left fronto-temporal and right fronto-occipital cortical zones, as well as, between both fronto-temporal zones. Paleocerebellum correlates at a weak to moderate level during different periods of SWD burst generation.

Behavioral transitions modulate hippocampal electroencephalogram correlates of open field behavior in the rat: support for a sensorimotor function of hippocampal rhythmical synchronous activity.

A clear relationship exists between moment-to-moment behavioral elements and hippocampal rhythmical synchronous activity (RSA) (theta rhythm). However, behavioral elements are not isolated events but are part of behavioral sequences in a context of behavioral activity. By concurrently monitoring open field behavior and hippocampal EEG, EEG correlates of open field behavior in relation to preceding and following behavior were studied in Sprague Dawley rats to determine whether the behavioral context influences EEG correlates of behavior. Results show that preceding and subsequent behavioral patterns influenced the spectral power correlates of behavior. RSA power was increased when a "type 1 behavior" (voluntary movement) preceded the behavior compared with when a "type 2 behavior" (automatic movement, awake immobility) preceded it. The modulating effect of behavioral transitions was shown for several types of behaviors, and systematic modulation of hippocampal EEG correlates of behavior was demonstrated. The present report shows that the strong and systematic relationship between hippocampal RSA and behavior is modulated by the behavioral-sequential context. Thus, in addition to the well established relationship between RSA and motor activity, a second nonmotor process seems to contribute to hippocampal RSA. A likely candidate is a sensory process, which is in accordance with theories on the sensorimotor function of hippocampal RSA.

The relationship between hippocampal EEG theta activity and locomotor behaviour in freely moving rats: effects of vigabatrin.

The relationship between hippocampal electroencephalogram (EEG) theta activity and locomotor speed in both spontaneous and forced walking conditions was studied in rats after vigabatrin injection (500 mg/kg i.p.). Vigabatrin increased the percentage of time that rats spent being immobile. During spontaneous walking in the open field, the speed of locomotion was increased by vigabatrin, while theta peak frequency was decreased. Vigabatrin also reduced the theta peak frequency during forced (speed controlled) walking. There was only a weak positive correlation (r=0.22) between theta peak frequency and locomotor speed for the saline condition. Furthermore, vigabatrin abolishes the weak relationship between speed of locomotion and theta peak frequency. Vigabatrin and saline did not differ in the slope of the regression line, but showed different offset points at the theta peak frequency axis. Thus, other factors than speed of locomotion seem to be involved in determination of the theta peak frequency.

Audiogenic seizures associated with a cortical spreading depression wave suppress spike-wave discharges in rats.

To study the role of the cortex and sub-cortical structures in the generation of epileptic spike-wave discharges in more detail, cortical and striatal activity was eliminated by the induction of spreading depression in a non-invasive way. EEG and DC potentials were recorded from the cortex and striatum of WAG/Rij rats. Several of these rats show two forms of generalised epilepsy: spontaneously occurring non-convulsive absence seizures, together with convulsive audiogenic seizures. The latter can be evoked by a brief sound stimulation, provoking a fit of wild running, which is regarded as the first phase of an audiogenic seizure. In a majority of fits the cortical DC potential does not show main changes, while the spontaneously occurring spike-wave discharges are briefly suppressed for some minutes. In a minority of fits, however, audiogenic seizures are associated with a spreading depression wave, clearly expressed in the cortical DC potential. This wave is bilaterally initiated in the cortex and propagates to the caudate nucleus of the striatum. In these cases spontaneously occurring spike-wave discharges are fully suppressed for about 1 h. It is suggested that cortical spreading depression, triggered by a short audiogenic seizure, induces a long-lasting suppression of spike-wave discharges. These results are in line with the concept that spike-wave discharges are originally initiated in the cortex, as proposed by the 'cortical focus' theory. The precise role of the striatum remains less clear, although this structure seems not to play a pivotal role in spike-wave generation.

Neuronal activities underlying the electroencephalogram and evoked potentials of sleeping and waking: implications for information processing.

The low amplitude, high frequency waves of the electroencephalogram (EEG) indicative of wakefulness, are produced by a summation of potentials of thalamocortical neurons, which fire in a "tonic mode" of depolarization. In this mode, the transfer of information from the peripheral sense organs to the sensory cortex is facilitated, due to a tonic lowering of the discharge threshold of thalamocortical neurons. The transfer decreases during drowsiness when thalamocortical units are more hyperpolarized and have higher thresholds. In this state, neurons fire synchronously in a "burst mode," which is expressed in EEG spindling. During slow wave sleep sensory blocking reaches a maximum, when thalamocortical cells are yet more deeply hyperpolarized, although that what still passes to the cortex allows a shallow, subconscious, evaluation. The collective burst firing is more irregular, which results in large and slow EEG waves. In contrast, during rapid eye movement (REM) sleep the depolarized tonic mode of firing commonly associated with waking, is again reached. Similar to EEG-patterns, the architecture of evoked potentials is dependent on the state of alertness. During waking, components in event related potentials (ERP) are moderate in amplitude, while during slow wave sleep larger waves are visible. This is caused by more synchronized unit responses with sharper phases of excitations and inhibitions, which results from increased hyperpolarizations. In contrast, visual ERPs belonging to REM sleep closely resemble those of wakefulness. In analyzing unit responses of thalamocortical neurons, it appeared that neuronal excitations are expressed in negative components of the ERP, while inhibitory neuronal activities are associated with positivity. Transient phenomena in the EEG, such as ERP waves, spindles and spike-wave discharges, are the expression of synaptic potentials in superficial cortical layers, where numerous synapses of afferent thalamocortical fibers are localized on the apical dendrites of deeper lying pyramidal neurons. It is suggested that the morphology of these EEG components is primarily due to the discharge characteristics of thalamocortical relay cells, whereby excitations underly negative waves and inhibitions positive waves. The notion of a general correspondence between thalamocortical neuronal activities and the polarity of transients in the cortical surface EEG, allows prudent speculations regarding components of ERPs. Two examples are given: the contingent negative variation (CNV) and the P300 of an ERP which can be elicited by an infrequent stimulus.(ABSTRACT TRUNCATED AT 400 WORDS)

Absence epilepsy and the level of vigilance in rats of the WAG/Rij strain.

In man, a relationship exists between sleep-wake states and absence epilepsy. During wakefulness, spike-wave discharges predominantly occur when the level of vigilance is not high, while during sleep they have a preference to occur during slow-wave sleep. During this latter type of sleep, spike-wave discharges prevail in periods where slow-wave sleep is light. In a series of experiments, the WAG/Rij rat model for absence epilepsy was characterized with respect to the relationships between the level of vigilance, sleep-wake states and the occurrence of spike-wave discharges. In the first experiment, continuous recordings were made for a period of 48 h and a clear circadian rhythm was established for the number of spike-wave discharges. A maximum appeared during the middle of the dark period of the rat, whereas a minimum was detected directly after the onset of the light period, the time period during which deep slow-wave sleep predominates. The relationship of spike-wave discharges with states of vigilance was elaborated in a second study. Spike-wave discharges were mainly found during light slow-wave sleep, during passive wakefulness and in transition phases from sleep to wakefulness. During REM sleep no spike-wave discharges were found. In the last three experiments, the level of alertness was enhanced by various procedures as photostimulation, a learning task and deprivation of REM sleep. In all cases, an increase of alertness decreased the amount of epilepsy.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of remacemide and its metabolite FPL 12495 on spike-wave discharges, electroencephalogram and behaviour in rats with absence epilepsy.

The effects of the anti-convulsant drug remacemide and one of its active metabolites FPL 12495 were examined in a genetic model for generalized absence epilepsy, the WAG/Rij strain of rats. Number, mean and total duration of spike-wave discharges were measured following oral administration of remacemide and FPL 12495, together with parameters of background electroencephalographic activity (EEG) and spontaneous behaviour in the recording cage. A decrease in the number of the spike-wave discharges was found after remacemide administration. At the highest dose there was near total suppression of the spike-wave discharges. There were no important effects on behaviour and on spectral content of the background EEG, suggesting that remacemide has little side effects. A decrease in the number of spike-wave discharges was also found after FPL 12495 gavage and there was a prolongation of the mean duration. Behavioural changes were only noticed after the highest dose. These were accompanied by changes in the spectral content and particularly by an increase in the amplitude of the delta and the high beta frequencies, together with a decrease in the spindle frequency range. FPL 12495 appeared to be more potent that remacemide in all its effects. The effects of mainly FPL 12495 are uncommon in the sense that so far no other investigated drug shows a decrease in the number together with an increase in the mean duration of the discharges. It seems that in contrast to other anti-epileptic drugs, FPL 12495 exerts a differential action on the two commonly distinguished mechanisms controlling number and duration.

Paradoxical sleep deprivation and the immobility response in the rat: effects of desipramine and phentolamine.

A series of experiments was conducted on the presumption that instrumental deprivation of paradoxical sleep (PS) has an effect comparable with that of antidepressant drug treatment in a behavioral paradigm, Porsolt's forced swim test. After long-term PS deprivation, we studied the duration of immobility, which can be markedly reduced by antidepressant drugs, using both the platform and the pendulum technique. In addition to a small common effect for PS deprivation, differences confirming the platform-pendulum controversy were also detected. Nonspecific platform effects are considered to explain these differences. In a second experiment it was shown that the small PS deprivation effect can be enlarged by desipramine treatment, suggesting similarities in the underlying mechanisms. In the third experiment, just before the end of the deprivation, phentolamine, a drug that blocks the rebound of PS, was administered. It reduced the effect of pendulum PS deprivation, suggesting that PS propensity is an important factor in the reduction of duration of immobility.

Deprivation of paradoxical sleep and intracranial self-stimulation.

Effects of paradoxical sleep deprivation upon intracranial self-stimulation behavior were studied. After stable response rates to electrical brain reward were obtained, rats were assigned to an experimental group in which they were deprived of paradoxical sleep with the pendulum technique for 72 h, to a pendulum control group and to a home-cage control group. In baseline, postdeprivation, and postrecovery sessions, rate-intensity functions for intracranial self-stimulation were determined. Partially in contrast to the literature, no change in the response rate or threshold for brain stimulation was found. The question was raised whether factors accompanying the different paradoxical sleep deprivation techniques rather than paradoxical sleep deprivation itself were responsible for these behavioral differences.

Sleep deprivation and spike-wave discharges in epileptic rats.

The effects of sleep deprivation were studied on the occurrence of spike-wave discharges in the electroencephalogram of rats of the epileptic WAG/Rij strain, a model for absence epilepsy. This was done before, during and after a period of 12 hours of near total sleep deprivation. A substantial increase in the number of spike-wave discharges was found during the first 4 hours of the deprivation period, whereas in the following deprivation hours epileptic activity returned to baseline values. Immediately after termination of deprivation, a decrease in the number of spike-wave discharges parallelled a rebound of rapid eye movement (REM) sleep and deep non-REM sleep. An initial increase in epileptic activity has also been reported during sleep deprivation of humans. This initial increase as well as the epileptogenic effects during the course of the sleep deprivation and during the recovery period after sleep deprivation can be interpreted in terms of changes in sleep-wake states. Although the epilepsy-provoking mechanisms are not yet understood, an explanation is suggested based on changes of transitions between sleep-wake states and shifts in level of synchronization.

Differential effects of ketamine on gating of auditory evoked potentials and prepulse inhibition in rats.

Schizophrenic patients suffer from deficits in information processing. Patients show both a decrease in P50 gating [assessed in the conditioning-testing (C-T) paradigm] and prepulse inhibition (PPI), two paradigms that assess gating. These two paradigms might have a related underlying neural substrate. Gating, as measured in both the C-T paradigm (the gating of a component of the auditory evoked potential (AEP)], and PPI can easily be measured in animals as well as in humans. This offers the opportunity to model these information processing paradigms in animals in order to investigate the effects of neurotransmitter manipulations in the brain. In order to validate the animal model for disturbances in AEP gating, d-amphetamine (0.5 and 1 mg/kg, i.p.) was administered. Gating of an AEP component was changed due to injection of d-amphetamine (1 mg/kg) in the same way as seen in schizophrenic patients: both the amplitude to the conditioning click and the gating were significantly reduced. Next, the effect of the N-methyl-D-aspartate (NMDA) antagonist ketamine (2.5 and 10 mg/kg, i.p.) was investigated to assess its effects in the two gating paradigms. It was found that ketamine (10 mg/kg) did not affect gating as measured with components of the AEP. However, ketamine (10 mg/kg) disrupted PPI of the startle response to the extent that prepulse facilitation occurred. Firstly, it is concluded that AEP gating was disrupted by d-amphetamine and not by ketamine. Secondly, PPI and the C-T paradigm reflect distinct inhibitory sensory processes, since both paradigms are differentially influenced by ketamine.

Endogenous opioid peptides in brain and pituitary of rats with absence epilepsy.

The level of opioid peptides in several brain areas and in the pituitary was estimated in WAG/Rij rats, which are considered to be a genetic animal model for human absence epilepsy. In comparison with three groups of non-epileptic controls, these epileptic rats had an elevated level of the proenkephalin-derived peptide Met-enkephalin-Arg6-Gly7-Leu8 in the mesencephalon and striatum, while the level of the prodynorphin-derived peptide alpha-neoendorphin was increased in the striatum and hippocampus. In addition various age- and/or strain-related changes in these peptide levels were found in the hippocampus, thalamus, striatum, frontal cortex and neurointermediate lobe of the pituitary. No difference in the hypothalamic beta-endorphin level were found between epileptic and non-epileptic rats, though strain- and/or age-related changes in the peptide content were detected in both lobes of the pituitary. The increased level of proenkephalin and prodynorphin opioid peptides in brain structures, essential for the appearance of spike-wave discharges, suggests that these opioid systems, but not proopiomelanocortin one, may play a role in absence epilepsy.

Ictal stimulus processing during spike-wave discharges in genetic epileptic rats.

In the present experiment it was investigated whether and to what extent auditory information processing is possible during the presence of spike-wave discharges in rats. To that end, WAG/Rij rats which are an animal model for absence epilepsy, were provided with cortical electrodes for the registration of the electroencephalogram (EEG). The animals were first trained in an appetitively motivated conditioning paradigm to learn to discriminate between two auditory stimuli with equal duration and frequency but with different intensities. Next, the stimuli were presented in the test phase in pseudorandom order during spike-wave discharges. The reactivity of the ongoing EEG was analysed. It was found that the presentation of the reinforced stimulus induced a larger number of aborted spike-wave discharges than the non-reinforced stimulus, regardless of the intensity of the stimuli. This implies that during generalised spike-wave discharges the brain is still capable of evaluating the meaning of an ictally presented stimulus. It also shows that sensory, attentional and mnemonic processes are at least partially intact during the occurrence of a spike-wave discharge. The results of the present study are largely in agreement with results on human spike-wave activity-related cognitive disturbances. Moreover, they may lead to a refinement of the concept of epileptic consciousness and may emphasise the heuristic value of rodent models for studying both ictal and interictal information processing.

Paradoxical sleep deprivation does not affect neuronal excitability in the rat hippocampus.

Before, during and after paradoxical sleep deprivation, field potentials, evoked in the CA1 region and in the fascia dentata of the hippocampus by means of paired pulse stimulation, were measured. Paradoxical sleep deprivation was applied for 3 days, using the platform and the pendulum technique. No changes were observed on evoked field potential amplitude, population spike or paired pulse depression during and after the deprivation period. These results suggest that the neuronal excitability in the rat hippocampus, measured with the evoked potential technique, does not change as a result of paradoxical sleep deprivation.

Thalamic multiple-unit activity underlying spike-wave discharges in anesthetized rats.

In epilept WAG/Rij rats, multiple unit activity coinciding with the occurrence of spike-wave discharges was recorded under neurolept anesthesia. Recordings were made in the frontal cortex and in various nuclei of the thalamus, in specific nuclei such as the ventroposterolateral, the ventroposteromedial and the ventrolateral nuclei, as well as in non-specific nuclei such as the mediodorsal nucleus, the reticular thalamic nucleus, the interanteromedial nucleus and the intralaminar nuclei (the central medial nucleus, the centrolateral nucleus and the paracentral nucleus). Rhythmic unit firing concurrent with the spike component of the cortical spike-wave discharge was observed in deep layers of the cortex and in the following thalamic nuclei: in specific nuclei, the mediodorsal and the reticular thalamic nucleus. The activity in the specific nuclei and the mediodorsal nucleus shortly preceded the peak of the spike component. The burst in the reticular thalamic nucleus occurred later than in the specific nuclei. A wave-concurrent firing pattern was observed in the centrolateral nucleus and the paracentral nucleus. Cells in the central medial nucleus and interanteromedial nucleus did not fire in a phase-locked manner. Neurons in the latter nucleus, however, were generally tonically activated during the occurrence of spike-wave discharges. It is suggested that those thalamic nucleic thought to be involved in the production of cortical spindles, and that also fire concurrently with the spike component of the spike-wave discharges, are mediated in the genesis of the latter activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Electrophysiological and pharmacological characteristics of two types of spike-wave discharges in WAG/Rij rats.

Rats of the WAG/Rij strain are commonly seen as a genetic model for generalised absence epilepsy in man. Interestingly, generalised absence epilepsy shows, in addition to the fully generalised spike-wave discharges, a second type of spike-wave discharge, which lasts for a shorter time, has a lower frequency, and a lower incidence. The originally described distinction between the two types of spike-wave discharges was mainly based on the shape, polarity and duration of the discharges. In the present study other characteristics such as the spatial and temporal distribution of the spike and wave components of the two discharges and frequency spectra were found to differ between the two types. In addition, a reciprocal regulation of the two types of spike-wave discharges by drugs affecting the dopaminergic system (haloperidol and apomorphine) was observed. The results convincingly demonstrate the difference between the two phenomena and warrant the search for neurobiological mechanisms underlying both types of spike-wave discharges.

Auditory evoked potentials from auditory cortex, medial geniculate nucleus, and inferior colliculus during sleep-wake states and spike-wave discharges in the WAG/Rij rat.

OBJECTIVE: Click auditory evoked potentials (AEP) were simultaneously recorded from the auditory cortex (ACx), the medial geniculate nucleus (MGN), and the inferior colliculus (IC) in the freely moving WAG/Rij rat, to investigate state-dependent changes of the AEP in different anatomical locations along the auditory pathway. METHODS: AEPs obtained during active (AW) and passive wakefulness (PW), slow wave sleep (SWS), rapid-eye-movement sleep (REM) and generalized spike-wave discharges (SWD; a specific trait of the WAG/Rij rat, a genetic model for absence epilepsy), were compared. RESULTS: The early components in ACx, MGN and IC were stable throughout the sleep-wake cycle and SWD, apart from a slight increase in the IC during SWD. At all three locations a prominent enlargement of a later component (i.e., N32 in IC, N33 in MGN, and N44 in ACx) was found during SWS and SWD. CONCLUSIONS: The early AEP components are not modulated by the normal sleep-wake states, and are not impaired during SWD. A strong state-dependent modulation of a later AEP component occurs at all three anatomical locations investigated. This suggests that apart from the thalamic burst firing mode, additional mechanisms must exist for the enlargement of the AEP during EEG-synchronized states at the prethalamic and cortical level.

Time course of chronic diazepam effects on the auditory evoked potential of the rat.

The time course of chronic diazepam effects on auditory evoked potentials was studied in rats. Auditory evoked potentials were elicited by background and target tones in a passive oddball paradigm. Diazepam was administered by slow release implants to establish constant blood concentrations. Recordings were made during 21 days of treatment and 9 days after treatment ceased. Diazepam increased the amplitude of the P40 component and decreased the amplitude of the P72-P102 components elicited by background tones. Diazepam increased the amplitude of the P40-P48 component and decreased that of the N58 component elicited by target tones. These effects remained constant during treatment. Diazepam further decreased the amplitude of the P102 component elicited by target tones. This effect became more distinct over time. No group differences were found 9 days after treatment. The constant drug effects on middle-latency components (P40-P48) might reflect diazepam-induced changes in sensory information processing. The decreased long-latency component (P102) might reflect a diminished attention to, or discrimination of, target tones. The time course of this effect might reflect diazepam-enhanced habituation.