Comparative Genome Sequence Analysis of Actinobacillus pleuropneumoniae Serovar 8 Isolates From Norway, Denmark, and the United Kingdom Indicates Distinct Phylogenetic Lineages and Differences in Distribution of Antimicrobial Resistance Genes.

Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a disease of major impact on pig health, welfare, and productivity globally. Serovar 8 (APP) is the predominant clinical serovar in Norway and the United Kingdom (UK), and has been isolated from clinical cases in Denmark. The primary objective of this study was to characterize the genetic variability of isolates of A. pleuropneumoniae APP8 in the Norwegian population. The secondary objectives were to determine the within-host variability of APP8; to compare the APP8 bacterial populations in Norway, Denmark, and the UK, including antimicrobial resistance (AMR) gene profiles and to assess the effect of national differences in antimicrobial drug use and restricted animal movement on the occurrence of resistance. Isolates of APP8 from the UK (n=67), Denmark (n=22), and Norway (n=123) collected between 1983 and 2020 were compared using whole genome sequencing. To investigate genetic variability within individual hosts, an additional 104 APP8 isolates from the lungs of six Norwegian pigs were compared. Very low within-host variation was observed (≤ 2 single nucleotide polymorphisms). The phylogeny of 123 Norwegian APP8 isolates from 76 herds revealed some within-herd genetic variation, but substantial geographical clustering. When inferring the relatedness of the three international APP8 collections, the topology highlighted the existence of two distinct monophyletic branches characterized by the Norwegian and UK isolates, respectively. Three Danish isolates were scattered across the UK branch, whereas the remaining 19 Danish isolates clustered in two monophyletic groups nested in the Norwegian branch. Coalescence analysis, performed to estimate the divergences from a common ancestor, indicated a last common ancestor several centuries ago. The phylogenetic analyses also revealed striking differences in occurrence of AMR genes, as these were 23-times more prevalent among the UK isolates than among the Norwegian isolates. An increased understanding of the effects of population strategies is helpful in surveillance and control of infectious diseases.

A descriptive study of acute outbreaks of respiratory disease in Norwegian fattening pig herds.

BACKGROUND: Respiratory diseases are major health concerns in the pig production sector worldwide, contributing adversely to morbidity and mortality. Over the past years there was a rise in reported incidents of respiratory disease in pigs in Norway, despite population wide freedom from Aujeszky´s disease, porcine reproductive and respiratory syndrome, porcine respiratory corona virus and enzootic pneumonia. The main objective of this study was to investigate acute outbreaks of respiratory disease in conventional Norwegian fattening pig herds. The study included 14 herds. In seven herds with reported outbreaks of acute respiratory disease, data on clinical signs was recorded and samples for laboratory examination were collected. Diagnostic protocols were compared by parallel analysis of clinically healthy pigs from seven non-outbreak herds. RESULTS: The most commonly reported clinical signs were sudden deaths and dyspnea. An average compartment morbidity of 60%, mortality of 4% and case fatality of 9% was recorded in the outbreak herds. Post-mortem examinations revealed acute lesions resembling porcine pleuropneumonia in all 28 pigs investigated from the outbreak herds and in 2 of the 24 (8%) pigs from the non-outbreak herds. Chronic lesions were recorded in another 2 pigs (8%) from the non-outbreak herds. Actinobacillus pleuropneumoniae serovar 8 was isolated from lungs and/or pleura from all tested pigs (n = 28) in the outbreak herds, and from 2 out of 24 pigs (8%) in the non-outbreak herds, one pig with an acute and another pig with a chronic infection. No other significant bacterial findings were made. Seroconversion to A. pleuropneumoniae antibodies was detectable in all outbreak herds analyzed and in six out of seven non-outbreak herds, but the risk ratio for seroconversion of individual pigs was higher (risk ratio 2.3 [1.50- 3.43 95% CI; P < 0.001]) in the outbreak herds. All herds tested positive for porcine circovirus type 2 and negative for influenza A viruses on oral fluid RT-qPCR. CONCLUSION: The main etiological pathogen found during acute outbreaks of respiratory disease was A. pleuropneumoniae serovar 8. All pigs from outbreak herds had typical lesions of acute porcine pleuropneumonia, and only A. pleuropneumoniae serovar 8 was identified. Co-infections were not found to impact disease development.

Demyelinating polyneuropathy in goats lacking prion protein.

Studies in mice with ablation of Prnp, the gene that encodes the cellular prion protein (PrPC ), have led to the hypothesis that PrPC is important for peripheral nerve myelin maintenance. Here, we have used a nontransgenic animal model to put this idea to the test; namely, goats that, due to a naturally occurring nonsense mutation, lack PrPC . Teased nerve fiber preparation revealed a demyelinating pathology in goats without PrPC . Affected nerves were invaded by macrophages and T cells and displayed vacuolated fibers, shrunken axons, and onion bulbs. Peripheral nerve lipid composition was similar in young goats with or without PrPC , but markedly different between corresponding groups of adult goats, reflecting the progressive nature of the neuropathy. This is the first report of a subclinical demyelinating polyneuropathy caused by loss of PrPC function in a nontransgenic mammal.

The Effect of LPS and Ketoprofen on Cytokines, Brain Monoamines, and Social Behavior in Group-Housed Pigs.

Poor health is a risk factor for damaging behaviors, but the mechanisms behind this link are unknown. Injection of pigs with lipopolysaccharide (LPS) can be used to model aspects of poor health. Recent studies have shown that LPS-injected pigs perform more tail- and ear-directed behavior compared to saline-injected pigs and suggest that pro-inflammatory cytokines may play a role in these behaviors. The aims of this study were to test the effect of LPS on the social behavior of pigs and the neurotransmitters and modulators in their brains and to test the effect of a nonsteroidal anti-inflammatory drug on the effects of LPS. Fifty-two female pigs (11-12 weeks) were allocated to four treatments comprising two injections: saline-saline (SS), saline-LPS (SL), ketoprofen-saline (KS), and ketoprofen-LPS (KL). Activity was scan-sampled every 5 min for 6 h after the last injection in the pen. Social behavior was observed continuously in 10 × 15-min bouts between 8 a.m. and 5 p.m. 1 day before (baseline) and 1 and 2 days after the injection. Saliva was analyzed for cortisol and plasma for tryptophan and kynurenine. The frontal cortex, hippocampus, hypothalamus, and brain stem were sampled 72 h after the injection and analyzed for cytokines and monoamines. LPS activated the HPA axis and decreased the activity within 6 h after the injection. Ketoprofen lowered the effect of LPS on cortisol release and attenuated the behavioral signs of sickness in challenged pigs. SL pigs manipulated the ears of their pen mates significantly longer than SS pigs 2 days after the injection. LPS had no observed effect on IFN-γ, TNF-α, and IL-18. At 72 h after the injection, plasma tryptophan was depleted in SL pigs, and tryptophan and kynurenine concentrations in the frontal cortex and brain stem of SL pigs were significantly lower compared to those in SS pigs. Dopamine concentrations in the hypothalamus of SL pigs were significantly lower compared to those in SS pigs. Serotonin concentrations in the hypothalamus and noradrenaline concentrations in the hippocampus of SL pigs were significantly lower compared to those in KL pigs. In conclusion, LPS influenced the different neurotransmitters and modulators in the brain that are hypothesized to play an important role in the regulation of mood and behavior.

Disposition and effect of intra-articularly administered dexamethasone on lipopolysaccharide induced equine synovitis.

BACKGROUND: Dexamethasone is used for the intra-articular route of administration in management of aseptic arthritis in horses. Despite its widespread use there is very little quantitative data of the disposition and response to dexamethasone. The aim of this study was to investigate and describe the synovial fluid and plasma dexamethasone concentration over time and to explore the relation between synovial fluid concentration and response using clinical endpoints as response biomarkers after IA injection of dexamethasone disodium salt solution in an equine model of synovitis. RESULTS: Inflammation was induced in the radiocarpal joint of six horses by injection of 2 ng lipopolysaccharide (LPS). Two hours later either saline or dexamethasone was injected in the same joint in a two treatment cross over design. Each horse was treated once with one of the six doses dexamethasone used (0.01, 0.03, 0.1, 0.3, 1 or 3 mg) and once with saline. Dexamethasone was quantified by means of UHPLC-MS/MS. Dexamethasone disposition was characterised by means of a non-linear mixed effects model. Lameness was evaluated both objectively with an inertial sensor based system and subjectively scored using a numerical scale (0-5). Joint circumference, skin temperature over the joint and rectal temperature were also recorded. The LPS-challenge induced lameness in all horses with high inter-individual variability. Dexamethasone significantly decreased lameness compared with saline. Other variables were not statistically significant different between treatments. Objective lameness scoring was the most sensitive method used in this study to evaluate the lameness response. A pharmacokinetic/pharmacodynamic model was successfully fitted to experimental dexamethasone and lameness data. The model allowed characterization of the dexamethasone synovial fluid concentration-time course, the systemic exposure to dexamethasone after intra-articular administration and the concentration-response relation in an experimental model of synovitis. CONCLUSIONS: The quantitative data improve the understanding of the pharmacology of dexamethasone and might serve as input for future experiments and possibly contribute to maintain integrity of equine sports.

The effect of lipopolysaccharide (LPS) on inflammatory markers in blood and brain and on behavior in individually-housed pigs.

Most of us have experienced deterioration of mood while ill. In humans, immune activation is associated with lethargy and social withdrawal, irritability and aggression; changes in social motivation could, in theory, lead to less functional interactions. This might also be the case for animals housed in close confinement. Tail biting in pigs is an example of damaging social behavior, and sickness is thought to be a risk factor for tail biting outbreaks. One possible mechanism whereby sickness may influence behavior is through cytokines. To identify possible mediators between immune activation and behavioral change, we injected 16 gilts with lipopolysaccharide (LPS; O111:B4; 1.5 µg kg-1 IV through a permanent catheter). In LPS-treated pigs, a significant increase in cortisol, TNF-α, IL-1 receptor antagonist, IL-6, and IL-8 was observed alongside decreased activity within the first 6 h after the injection. CRP was elevated at 12 and 24 h after injection, and food intake was reduced for the first 24 h after injection. Three days post-injection, LPS pigs had lower levels of noradrenaline in their hypothalamus, hippocampus and frontal cortex compared to saline-injected pigs. Pigs injected with LPS also had higher levels of the pro-inflammatory cytokine IFN-γ in their frontal cortex compared to saline-injected pigs. Thus, a low dose of LPS can induce changes in brain cytokine levels and neurotransmitter levels that persist after inflammatory and stress markers in the periphery have returned to baseline levels.

Dexmedetomidine or fentanyl? Cardiovascular stability and analgesia during propofol-ketamine total intravenous anaesthesia in experimental pigs.

OBJECTIVE: To compare cardiovascular function and response to nociception during total intravenous anaesthesia in pigs with propofol, ketamine and either dexmedetomidine or fentanyl administered as a continuous infusion. STUDY DESIGN: Blinded, randomized, balanced, crossover study ANIMALS: Eight immunocastrated male, mixed breed pigs with a mean ± standard deviation body weight of 26.4 ± 1.9 kg for dexmedetomidine and 27.5 ± 3.8 kg for fentanyl treatment. METHODS: The animals were anaesthetized twice with either propofol-ketamine-dexmedetomidine (DEX) or fentanyl (FENT). DEX was infused at 2, 4 and 8 µg kg-1 hour-1 and FENT at 25, 50 and 100 µg kg-1 hour-1. Each infusion rate was administered for 80 minutes prior to commencing the next. Heart rate (HR), 3-lead electrocardiogram, systolic, mean and diastolic arterial blood pressure (SAP, MAP, DAP) in addition to cardiac output measured by transpulmonary thermodilution was used to monitor cardiovascular function. Mechanical and electrical stimulation (nociceptive withdrawal reflex, NWR) was used to elicit nociceptive responses. Similar anaesthetic depth was determined based on the NWR response. Cardiovascular parameters were compared statistically at this time. Additionally, response to nociceptive stimulation and cardiovascular response over time were compared. RESULTS: DEX-treated pigs had significantly higher HR, SAP, DAP, MAP, systemic vascular resistance, haemoglobin concentration, content of oxygen in arterial and venous blood and oxygen delivery index than FENT-treated pigs at similar anaesthetic depth, whereas stroke volume index was significantly higher in FENT. Motoric response to mechanical nociceptive stimulation was abolished prior to any decrease in NWR response in FENT, whereas the two responses decreased more in unison in DEX. The cardiovascular response to nociception was less pronounced in DEX than in FENT. CONCLUSIONS AND CLINICAL RELEVANCE: Propofol combined with ketamine and either fentanyl or dexmedetomidine provides stable cardiovascular conditions in normovolaemic, healthy pigs. Based on cardiovascular response and depression of NWR, dexmedetomidine apparently provides superior analgesia to fentanyl.

The cardiovascular status of isoflurane-anaesthetized horses with and without dexmedetomidine constant rate infusion evaluated at equivalent depths of anaesthesia.

OBJECTIVE: To compare cardiac index and oxygen extraction at equivalent depths of anaesthesia between isoflurane-anaesthetized horses and horses anesthetized with isoflurane and dexmedetomidine CRI. STUDY DESIGN: Sequential, blinded, randomized, balanced, crossover study. ANIMALS: Eight horses weighing a mean ± standard deviation of 478 ± 58 kg. METHODS: Horses were premedicated with 0.03 mg kg(-1) acepromazine intramuscularly (IM) and 8 µg kg(-1) dexmedetomidine intravenously (IV). Anaesthesia was induced with 2.5 mg kg(-1) ketamine and 0.1 mg kg(-1) midazolam IV and maintained with isoflurane in oxygen and air. Horses were mechanically ventilated. Fractional concentration of end-tidal isoflurane (Fe'Iso) was stabilized at 1.7% with a CRI of 0.9% NaCl (IsoNaCl), or at 1.1% with 1.75 µg kg(-1)  hour(-1) dexmedetomidine (IsoDex). Mean arterial blood pressure was maintained above 60 mmHg by dobutamine infusion. Following nociceptive electrical stimulation, Fe'Iso was stabilized at a 0.1% lower concentration and nociceptive stimulation was repeated. This procedure was continued until the horse moved. Fe'Iso values prior to the concentration at which movement occurred were considered to indicate equivalent depths of anaesthesia between treatments. Cardiac index and oxygen extraction were compared at equivalent depths of anaesthesia using a paired Student's t-test. RESULTS: Cardiac index differed between IsoNaCl at 61 ± 12 mL kg(-1)  minute(-1) and IsoDex at 48 ± 10 mL kg(-1)  minute(-1) (p = 0.047). In addition, oxygen extraction differed between IsoNaCl at 3.4 ± 0.8 mL kg(-1)  minute(-1) and IsoDex at 4.5 ± 0.5 mL kg(-1)  minute(-1) (p = 0.0042). Two horses receiving IsoNaCl were administered dobutamine at equivalent depths of anaesthesia (7.0 and 28.8 µg kg(-1)  hour(-1) , respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Cardiovascular function in horses receiving isoflurane and 1.75 µg kg(-1)  minute(-1) dexmedetomidine is more compromised than in horses receiving a higher concentration of isoflurane and 0.9% NaCl CRI.

Antinociceptive effect of buprenorphine and evaluation of the nociceptive withdrawal reflex in foals.

OBJECTIVE: To elicit and evaluate the NWR (nociceptive withdrawal reflex) in 2 and 11 day old foals, to investigate if buprenorphine causes antinociception and determine if the NWR response changes with increasing age. The effect of buprenorphine on behaviour was also evaluated. STUDY DESIGN: Prospective, experimental cross-over trial. ANIMALS: Nine Norwegian Fjord research foals. METHODS: Buprenorphine, 10 µg kg(-1) was administered intramuscularly (IM) to the same foal at 2 days and at 11 days of age. The NWR and the effect of buprenorphine were evaluated by electromyograms recorded from the left deltoid muscle following electrical stimulation of the left lateral palmar nerve at the level of the pastern. Mentation, locomotor activity and respiratory rate were recorded before and after buprenorphine administration. RESULTS: We were able to evoke the NWR and temporal summation in foals using this model. Buprenorphine decreased the root mean square amplitude following single electrical stimulation (p < 0.001) in both age groups, and increased the NWR threshold following single electrical stimulation in 2 day old foals (p = 0.0012). Repeated electrical stimulation at 2 Hz was more effective to elicit temporal summation compared to 5 Hz (p < 0.001). No effect of age upon the NWR threshold was found (p = 0.34). Sedation when left undisturbed (11 occasions), increased locomotor activity when handled (9 occasions) and tachypnea (13 occasions) were common side-effects of buprenorphine. CONCLUSION AND CLINICAL RELEVANCE: These findings indicate that buprenorphine has antinociceptive effect in foals. Opioid side effects often recognized in adult horses also occur in foals.

Adverse reactions to equine-derived F(ab')2 -antivenin in 54 dogs envenomated by Vipera berus berus.

OBJECTIVE: To describe the occurrence of acute and delayed adverse reactions following the administration of an equine-derived F(ab')2 -antivenin in dogs with Vipera berus berus envenomation. DESIGN: Prospective study (2006-2009). SETTING: Private small animal clinics and university small animal teaching hospital in Norway. ANIMALS: Fifty-four dogs with Vipera berus berus envenomation treated with F(ab')2 -antivenin. INTERVENTIONS: Administration of equine-derived antivenin for treatment of Vipera berus berus envenomation in dogs. MEASUREMENTS AND MAIN RESULTS: Acute adverse reactions to antivenin administration occurred in 4/54 dogs (7%), and facial swelling occurred in 2/54 dogs (4%), sudden respiratory distress in 1 dog, and vomiting occurred in 1 dog. The adverse reactions were self-limiting in 3 of the dogs, while 1 dog was treated with corticosteroids to reduce the facial swelling that occurred during administration of antivenin. No delayed adverse reactions believed to be associated with the antivenin administration were noted. CONCLUSIONS: An occurrence of adverse effects comparable to the numbers reported in human medicine (10%) should be expected when administering F(ab')2 -antivenin to dogs with Vipera berus berus envenomation. The results are also comparable with rates of adverse effects reported with antivenin administration for crotalid-envenomated dogs. Further research is required to determine whether there is a relationship between adverse and the number of vials administered or repeated use in the same patient.

Factors affecting mechanical (nociceptive) thresholds in piglets.

OBJECTIVE: To evaluate the stability and repeatability of measures of mechanical (nociceptive) thresholds in piglets and to examine potentially confounding factors when using a hand held algometer. STUDY DESIGN: Descriptive, prospective cohort. ANIMALS: Forty-four piglets from four litters, weighing 4.6 ± 1.0 kg (mean ± SD) at 2 weeks of age. METHODS: Mechanical thresholds were measured twice on each of 2 days during the first and second week of life. Data were analyzed using a repeated measures design to test the effects of behavior prior to testing, sex, week, day within week, and repetition within day. The effect of body weight and the interaction between piglet weight and behaviour were also tested. Piglet was entered into the model as a random effect as an additional test of repeatability. The effect of repeated testing was used to test the stability of measures. Pearson correlations between repeated measures were used to test the repeatability of measures. Variance component analysis was used to describe the variability in the data. RESULTS: Variance component analysis indicated that piglet explained only 17% of the variance in the data. All variables in the model (behaviour prior to testing, sex, week, day within week, repetition within day, body weight, the interaction between body weight and behaviour, piglet identity) except sex had a significant effect (p < 0.04 for all). Correlations between repeated measures increased from the first to the second week. CONCLUSIONS AND CLINICAL RELEVANCE: Repeatability was acceptable only during the second week of testing and measures changed with repeated testing and increased with increasing piglet weight, indicating that time (age) and animal body weight should be taken into account when measuring mechanical (nociceptive) thresholds in piglets. Mechanical (nociceptive) thresholds can be used both for testing the efficacy of anaesthetics and analgesics, and for assessing hyperalgesia in chronic pain states in research and clinical settings.

The influence of a continuous rate infusion of dexmedetomidine on the nociceptive withdrawal reflex and temporal summation during isoflurane anaesthesia in dogs.

OBJECTIVE: To examine the influence of a low dose dexmedetomidine infusion on the nociceptive withdrawal reflex and temporal summation in dogs during isoflurane anaesthesia. STUDY DESIGN: Prospective experimental blinded cross-over study. ANIMALS: Eight healthy mixed breed dogs, body weight Mean ± SD 26.5 ± 8.4 kg and age 25 ± 16 months. METHODS: Anaesthesia was induced with propofol and maintained with isoflurane (Fe'ISO 1.3%) delivered in oxygen and air. After stabilization, baseline recordings (time 0) were obtained, then a dexmedetomidine bolus (1 µg kg(-1) IV) followed by a continuous rate infusion (1 µg kg(-1) hour(-1)) or saline placebo were administered. At times 10, 30 and 60 minutes after the initial bolus, electrical stimulations of increasing intensity were applied over the lateral plantar digital nerve, and administered both as single and as repeated stimuli. The resulting reflex responses were recorded using electromyography. Data were analysed using a multivariable linear regression model and a Kruskal Wallis test for single stimulation data, and repeated measures anova and paired t-test for repeated stimulation data. RESULTS: The AUC for the stimulus-response curves after single stimulation were similar for both treatments at time 0. At times 10, 30 and 60 the AUCs for the stimulus-response curves were significantly lower with dexmedetomidine treatment than with placebo. Temporal summation was evident in both treatments at times 0, 10, 30 and 60 starting from a stimulation intensity of 10 mA. The magnitude of temporal summation was smaller in dexmedetomidine than in placebo treated dogs at time 10, 30 and 60, but not at time 0. CONCLUSIONS: During isoflurane anaesthesia, low dose dexmedetomidine suppresses the nociceptive reflex responses after single and repeated electrical stimulation. CLINICAL RELEVANCE: This experimental study confirms previous reports on its peri-operative efficacy under clinical conditions, and further indicates that dexmedetomidine might reduce the risk of post-operative chronic pain development.

Effects of isoflurane upon minimum alveolar concentration and cerebral cortex depression in pigs and goats: an interspecies comparison.

This study compared the effects of isoflurane in pigs (n=10 Yorkshire-Landrace cross) and dairy goats (n=10) by evaluation of electroencephalographic (EEG) burst suppression thresholds (BST) in the cerebral cortex and minimum alveolar concentration (MAC) values in the spinal cord. The study also investigated whether individual MAC values can predict the effects of isoflurane on the cerebral cortex. MAC values and BST/MAC ratios were significantly different between species. Inhibition of movement by isoflurane may be less effective in pigs than in goats. No significant correlation was found between individual MAC and BST values, indicating that in single animals the individual MAC poorly reflects the cerebrocortical depressant effect of isoflurane in pigs and goats.

Trace classical conditioning in rainbow trout (Oncorhynchus mykiss): what do they learn?

There are two main memory systems: declarative and procedural memory. Knowledge of these two systems in fish is scarce, and controlled laboratory studies are needed. Trace classical conditioning is an experimentally tractable model of declarative memory. We tested whether rainbow trout (Oncorhynchus mykiss) can learn by trace conditioning and form stimulus-stimulus, as opposed to stimulus-response, associations. We predicted that rainbow trout trained by trace conditioning would show appetitive behaviour (conditioned response; CR) towards the conditioned stimulus (CS; light), and that the CR would be sensitive to devaluation of the unconditioned stimulus (US; food). The learning group (L, N = 14) was trained on a CS + US contingency schedule with a trace interval of 3.4 s. The control group (CtrL, N = 4) was kept on a completely random schedule. The fish that learnt were further trained as either an experimental (L, N = 6) or a memory control (CtrM, N = 3) group. The L group had the US devalued. The CtrM group received only food. No fish in the CtrL group, but nine fish from the L group conditioned to the light. When tested, five L fish changed their CRs after US devaluation, indicating learning by stimulus-stimulus association of the light with the food. CtrM fish retained their original CRs. To the best of our knowledge, this experiment is the first to show that rainbow trout can learn by trace classical conditioning. The results indicate that the fish learnt by 'facts-learning' rather than by reflex acquisition in this study.

Pharmacokinetics of morphine after intramuscular injection in common goldfish Carassius auratus and Atlantic salmon Salmo salar.

Teleost fish have a nociceptive system and likely perceive pain. This warrants the development of analgesic protocols both for experimental surgery and for various husbandry procedures. Morphine is the standard analgesic against which the efficacy of other analgesics is assessed, and is the analgesic that has been most used in fish. The aims of this study were to describe the pharmacokinetics of morphine after an intramuscular (i.m.) injection in common goldfish and Atlantic salmon, and to illustrate the whole-body distribution of morphine in salmon following i.m. injection of tritiated morphine. In the kinetic experiment, goldfish and salmon were respectively i.m. injected with 40 and 100 mg morphine kg(-1) in the right dorsal epaxial musculature. Blood was drawn at predetermined time points. Plasma was analysed for morphine and metabolites using liquid chromatography-mass spectrometry (LC-MS/MS). Morphine had a Tmax (time at which the maximum plasma concentration was measured) of 0.5 h in both species. The Cmax (maximum plasma concentration) showed substantial inter-individual variation, with a mean (90% CI) of 187 (167 to 199) mg l(-1) in salmon and 37 (29 to 43) mg l(-1) in goldfish, as determined by bootstrap analysis. The mean elimination half-lives were 12.5 and 13.5 h in goldfish and in salmon, respectively. The degree of metabolism to morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) was low, with levels of M3G exceeding those of M6G. The distribution study demonstrated that the levels of tritiated morphine in the anterior kidney surpassed those in the other organs. A substantial amount seemed to be excreted through the gastrointestinal tract, while little tritium activity could be detected in the central nervous system.

Somatosensory evoked potentials in the telencephalon of Atlantic salmon (Salmo salar) following galvanic stimulation of the tail.

Electric activity in the brain which is time-locked to a given stimulation of the somatosensory system can be recorded as a somatosensory evoked potential (SEP). We investigated whether a galvanic stimulation of the tail base in Atlantic salmon (Salmo salar) would elicit a SEP in the telencephalon. The telencephalon is central in learning and memory, and activity here may be a prerequisite for processing of external stimuli on a cognitive or emotional level. Anaesthetized salmon (n = 11) were subjected to craniotomy and a recording electrode was inserted into the telencephalon. The fish were given stimulations of four intensities, i.e., 2, 5, 10 and 20 mA. A SEP was elicited in the contralateral dorsal telencephalon for all intensities. This result agrees with findings in other fish species. Furthermore, there was a significant difference between the maximum peak amplitude and mean amplitude of the SEP elicited by putative non-noxious (2 mA) and putative noxious (20 mA) stimulation intensities (P < 0.01). The stronger stimulation intensities also tend to introduce longer-latencies components in the SEP. The results added to the body of literature indicates that the exteroceptive senses are represented by processing within the telencephalon of the fish.

Effect of intratesticular injection of lidocaine on cardiovascular responses to castration in isoflurane-anesthetized stallions.

OBJECTIVE: To evaluate the effect of intratesticular administration of lidocaine on cardiovascular responses and cremaster muscle tension during castration of isoflurane-anesthetized stallions. ANIMALS: 28 healthy stallions (mean +/- SD age, 4.2 +/- 2.8 years) with no testicular abnormalities that were scheduled for castration. PROCEDURE: Each horse was given acepromazine (20 microg/kg, IM), romifidine (50 microg/kg, IV), and butorphanol (20 microg/kg, IV). Anesthesia was induced with ketamine (2.5 mg/kg, IV) and midazolam (50 microg/kg, IV) and maintained with isoflurane (1.7% end-tidal concentration). After 10 minutes at a stable anesthetic plane, a needle was placed in each testicle and either no fluid or 15 mL of 2% lidocaine was injected; 10 minutes after needle placement, surgery was commenced. Pulse rate and arterial blood pressures were measured invasively at intervals from 5 minutes prior to castration (baseline) until 5 minutes after the left spermatic cord was clamped. The surgeon subjectively scored the degree of cremaster muscle tension. In 2 horses, lidocaine labeled with radioactive carbon (C(14)) was used and testicular autoradiograms were obtained. RESULTS: Compared with baseline values, castration significantly increased blood pressure measurements; intratesticular injection of lidocaine decreased this blood pressure response and cremaster muscle tension. In 2 horses, autoradiography revealed diffuse distribution of lidocaine into the spermatic cord but poor distribution into the cremaster muscle. CONCLUSIONS AND CLINICAL RELEVANCE: In isoflurane-anesthetized stallions, intratesticular injection of lidocaine prior to castration appeared to decrease intraoperative blood pressure responses and cremaster muscle tension and may be a beneficial supplement to isoflurane anesthesia.