Noninvasive Detection of Ischemic Vascular Damage in a Pig Model of Liver Donation After Circulatory Death.

BACKGROUND AND AIMS: Liver graft quality is evaluated by visual inspection prior to transplantation, a process highly dependent on the surgeon's experience. We present an objective, noninvasive, quantitative way of assessing liver quality in real time using Raman spectroscopy, a laser-based tool for analyzing biomolecular composition. APPROACH AND RESULTS: A porcine model of donation after circulatory death (DCD) with normothermic regional perfusion (NRP) allowed assessment of liver quality premortem, during warm ischemia (WI) and post-NRP. Ten percent of circulating blood volume was removed in half of experiments to simulate blood recovery for DCD heart removal. Left median lobe biopsies were obtained before circulatory arrest, after 45 minutes of WI, and after 2 hours of NRP and analyzed using spontaneous Raman spectroscopy, stimulated Raman spectroscopy (SRS), and staining. Measurements were also taken in situ from the porcine liver using a handheld Raman spectrometer at these time points from left median and right lateral lobes. Raman microspectroscopy detected congestion during WI by measurement of the intrinsic Raman signal of hemoglobin in red blood cells (RBCs), eliminating the need for exogenous labels. Critically, this microvascular damage was not observed during WI when 10% of circulating blood was removed before cardiac arrest. Two hours of NRP effectively cleared RBCs from congested livers. Intact RBCs were visualized rapidly at high resolution using SRS. Optical properties of ischemic livers were significantly different from preischemic and post-NRP livers as measured using a handheld Raman spectrometer. CONCLUSIONS: Raman spectroscopy is an effective tool for detecting microvascular damage which could assist the decision to use marginal livers for transplantation. Reducing the volume of circulating blood before circulatory arrest in DCD may help reduce microvascular damage.

Transcranial bioimpedance measurement in horses: a pilot study.

OBJECTIVE: This pilot study aimed to evaluate the feasibility of transcranial bioimpedance (TCBI) measurement and variability of TCBI values in healthy conscious horses and to study effects of body position and time on TCBI in anaesthetized horses. STUDY DESIGN: Prospective, observational study. ANIMALS: A total of four research horses and 16 client-owned horses presented for surgery. METHODS: After establishing optimal electrode position using computed tomography scans of cadaver heads, TCBI [described using impedance at zero frequency, R0, (Ω)] was measured in four conscious, resting horses to investigate the feasibility and changes in TCBI over time (80 minutes). Data were compared using a paired t test. TCBI was then measured throughout anaesthesia (duration 92 ± 28 minutes) in 16 horses in dorsal and lateral recumbency. Data were analysed using a general linear model; gamma regression was chosen as a model of characteristic impedance [Zc; (Ω)] against time. Data are presented as mean ± standard deviation. RESULTS: No change in R0 was seen in conscious horses (age = 15.3 ± 7.3 years, body mass = 512 ± 38 kg) over 80 minutes. The technique was well tolerated and caused no apparent adverse effects. In 16 horses (age = 7.4 ± 4.7 years; body mass = 479 ± 134 kg) anaesthetized for 92 ± 28 minutes, Zc fell during anaesthesia, decreasing more in horses in lateral recumbency than in horses in dorsal recumbency (p = 0.008). There was no relationship between Zc and body mass or age. CONCLUSIONS AND CLINICAL RELEVANCE: TCBI is readily measured in horses. TCBI did not change with time in conscious horses, but decreased with time in anaesthetized horses; this change was greater in horses in lateral recumbency, indicating that TCBI changes in anaesthetized horses may be related to the effects of recumbency, general anaesthesia, surgery or a combination of these factors.

Comparison of Brody's formula with respiratory gas analysis for estimating oxygen consumption in anaesthetized dogs.

OBJECTIVE: To determine agreement in oxygen consumption (V˙O2) values calculated using Sykes' formula V˙O2 = (FiO2 - Fe'O2) * V˙E (where Fi and Fe are the inspired and end-tidal fractional concentrations of O2, respectively, and V˙E is minute volume) with values derived using Brody's formula (V˙O2 = 10 kg3/4). It was hypothesized that the two methods would not yield statistically significant differences in calculated values. STUDY DESIGN: Prospective, clinical, pilot study. ANIMALS: A total of 22 client-owned dogs. METHODS: Dogs undergoing surgery were anaesthetized with either isoflurane or sevoflurane. The V˙E, FiO2 and Fe'O2 were measured during mechanical ventilation of the lungs (tidal volume 10 mL kg-1; respiratory rate: 8-12 breaths minute-1). Oesophageal temperature was maintained between 37.0 °C and 38.5 °C. Values for V˙O2 derived by Sykes' and Brody's methods were compared and agreement was determined using Bland-Altman analysis. RESULTS: Mean V˙O2 values were 4.67 ± 0.51 mL kg-1 minute-1 and 5.32 ± 1.69 mL kg-1 minute-1 calculated using Brody's formula and Sykes' equation, respectively. There was greater variability in the values obtained from Sykes' equation. The Bland-Altman plot revealed a proportional error with correlation but poor agreement between values. CONCLUSIONS AND CLINICAL RELEVANCE: Both methods yielded V˙O2 values of approximately 5 mL kg-1minute-1 with no statistically significant differences between the two methods.

Modulation of spinal shape with growth following implantation of a novel surgical implant.

PURPOSE: First, to determine whether scoliosis development could be limited or reversed by growth when a novel modular hinged implant was fixed to the convexity of a scoliosis created by contralateral rib and laminar tethering and unilateral rib resection in a sheep model. Second, to assess the effect and performance of the implant in normal non-tethered sheep. METHODS: At 5 weeks, 20 Scottish Blackface lambs underwent surgery to create a right sided scoliosis by (i) tethering the left lamina of T5-L1 and the left lower six ribs and (ii) resecting a segment of their right lower six ribs [1, 2]. Twelve weeks later, through an antero-lateral thoracotomy, a mobile bi-planar hinged implant was inserted onto the right side of the spine of eight animals (group 1). For comparison, 12 sheep were tethered only but had no implant insertion (group 2). In addition, seven had no tethering but were implanted (group 3) and normal growth patterns were observed in five that had no surgery (group 4). Curve progression was assessed by plain radiography and CT over a 1-year period. RESULTS: Before implant insertion the trial animals had a scoliosis of 35º ± 16º and a lordosis of 44º ± 20º (n = 8, mean ± SD). Surgery immediately reduced these values to 25º ± 14º, p < 0.01 and 35º ± 18º, p < 0.001, with scoliosis continuing to decrease during the next three months. Spinal flexibility was retained. In the un-tethered sheep, a scoliosis of 10º ± 6º was created on the opposite side to the implant (p < 0.05) with no significant change in alignment in the sagittal plane (1º ± 6º). The implant did not cause any adverse effect on growth or affect neurological function. CONCLUSIONS: In the un-tethered animals the effect of the implant was to create a scoliotic deformity and in the tethered to improve deformity while maintaining spinal motion. We believe that the results are promising and that devices of similar construct may be of use in children with scoliosis, potentially changing current methods of clinical care.

Creation of an ovine model of progressive structural lordo-scoliosis using a unilateral laminar tether.

PURPOSE: To characterise changes in vertebral dimension in an ovine model of scoliosis and determine whether a reproducible curvature could be created that would be suitable for future testing of curve modifying surgical implants. METHODS: At a mean age of 5 weeks, 28 Scottish blackface sheep were anaesthetised. A 4 mm braided synthetic tape was laid under the left lamina of T5 and L1 and tightened to 'hand' tension. A scoliosis was then created by binding the six lowest ribs on the same side just distal to their rib angles and resecting a segment from each of the opposite ribs. Radiographs were taken at 4 weekly intervals, and CT images at 2, 5 and 7 months post tethering, to determine multi-planar curve progression. 20 animals were assessed at age 3 months, 12 at 41 weeks and 10 at 1 year with comparisons to five control animals. RESULTS: A significant bi-planar deformity was produced in all animals (scoliosis 32 ± 13º and lordosis 53 ± 21º 2 months post tethering; mean ± SD, n = 22). During the next 5 months the scoliosis decreased slightly (p = 0.08) but the sagittal deformity remained static: 21 ± 18° and 53 ± 19°, respectively (n = 12). The values at 7 months were associated with a mean 6 ± 4º rotational deformity. There was approximately twice as much growth in the right anterior aspects of the apical vertebrae as in the left posterior. CONCLUSIONS: With appropriate care it was possible to obtain a reproducible curvature in immature sheep. The methods described are suitable for use in studies of growth modulation and other spinal devices.

Electrocardiographic indicators of excitability in horses for predicting recovery quality after general anaesthesia.

OBJECTIVE: To determine whether preoperative electrocardiographic measurements linked to sympathetic nervous activity could be used to predict recovery quality after general anaesthesia in horses. STUDY DESIGN: Prospective, clinical study. ANIMALS: Eighteen adult client-owned horses. METHODS: The electrocardiogram (ECG) was recorded presurgery in horses under three standard conditions: stabled unattended; with a groom whilst being led along a standard course; alone in the induction box. After surgery, each animal's recovery quality was scored by eight experienced anaesthetists or technicians using Donaldson's scoring system (2000). The digitized ECG recordings were analyzed for T-wave morphology, mean heart rate (HR), HR variability (HRV) and HRV derivatives including mean R-R interval, standard deviation of normal intervals (SDNN) and the root mean square of successive intervals (RMSSD), low (LF) and high frequency (HF) activity and the LF/HF ratio in both fast Fourier transformed and autoregressive spectra. Correlations between ECG variables and recovery score were examined using Spearman's rank correlation. RESULTS: There was no significant correlation between preanaesthetic ECG measurements and recovery quality. CONCLUSIONS: Predictions of recovery quality after general anaesthesia in horses cannot be based on preanaesthetic ECG variables. CLINICAL RELEVANCE: Measures other than those based on the ECG should be investigated as predictors of recovery quality.

Tracking Ovine Pulmonary Adenocarcinoma Development Using an Experimental Jaagsiekte Sheep Retrovirus Infection Model.

Ovine pulmonary adenocarcinoma (OPA) is an infectious, neoplastic lung disease of sheep that causes significant animal welfare and economic issues throughout the world. Understanding OPA pathogenesis is key to developing tools to control its impact. Central to this need is the availability of model systems that can monitor and track events after Jaagsiekte sheep retrovirus (JSRV) infection. Here, we report the development of an experimentally induced OPA model intended for this purpose. Using three different viral dose groups (low, intermediate and high), localised OPA tumour development was induced by bronchoscopic JSRV instillation into the segmental bronchus of the right cardiac lung lobe. Pre-clinical OPA diagnosis and tumour progression were monitored by monthly computed tomography (CT) imaging and trans-thoracic ultrasound scanning. Post mortem examination and immunohistochemistry confirmed OPA development in 89% of the JSRV-instilled animals. All three viral doses produced a range of OPA lesion types, including microscopic disease and gross tumours; however, larger lesions were more frequently identified in the low and intermediate viral groups. Overall, 31% of JSRV-infected sheep developed localised advanced lesions. Of the sheep that developed localised advanced lesions, tumour volume doubling times (calculated using thoracic CT 3D reconstructions) were 14.8 ± 2.1 days. The ability of ultrasound to track tumour development was compared against CT; the results indicated a strong significant association between paired CT and ultrasound measurements at each time point (R2 = 0.799, p < 0.0001). We believe that the range of OPA lesion types induced by this model replicates aspects of naturally occurring disease and will improve OPA research by providing novel insights into JSRV infectivity and OPA disease progression.

The construction and evaluation of a device for mechanomyography in anaesthetized Göttingen minipigs.

OBJECTIVE: To devise a method for assessing evoked muscle strength on nerve stimulation [mechanomyography (MMG)] in the anaesthetized minipig. STUDY DESIGN: Prospective observational. ANIMALS: Sixty male Göttingen minipigs weighing 10.5-26.0 kg. METHODS: After cadaveric studies, a limb fixation device was constructed which allowed the twitch responses of the pelvic limb digital extensor muscles to be measured by force-displacement transduction in response to supramaximal train-of-four (TOF) stimulation of the common peroneal nerve. The device was tested in 60 minipigs weighing 10.5-26.0 kg positioned in dorsal recumbency. RESULTS: The technique recorded the MMG of the common peroneal-pelvic limb digital extensor nerve-muscle unit for up to 12 hours during which twitch height remained constant in 18 animals in which single twitch duration was <300-500 ms. In 42, in which twitch duration was >300-500 ms, 2 Hz nerve stimulation caused progressive baseline elevation (reverse fade) necessitating a modified signal capture method for TOF ratio (TOFR) computation. However, T1 was unaffected. The mean (range) of the TOFR in pigs with reverse fade was 1.2 (1.1-1.3). CONCLUSIONS AND CLINICAL RELEVANCE: The technique allowed MMG recording in unparalysed pigs in response to TOF nerve stimulation and revealed a hitherto unreported complication of MMG monitoring using TOF in animals: reverse fade. This complicated TOFR calculation.

Rapid and complete bioavailability of antidotes for organophosphorus nerve agent and cyanide poisoning in minipigs after intraosseous administration.

STUDY OBJECTIVE: Management of chemical weapon casualties includes the timely administration of antidotes without contamination of rescuers. Personal protective equipment makes intravenous access difficult but does not prevent intraosseous drug administration. We therefore measured the systemic bioavailability of antidotes for organophosphorus nerve agent and cyanide poisoning when administered by the intraosseous, intravenous, and intramuscular routes in a small study of Göttingen minipigs. METHODS: Animals were randomly allocated to sequentially receive atropine (0.12 mg/kg by rapid injection), pralidoxime (25 mg/kg by injection during 2 minutes), and hydroxocobalamin (75 mg/kg during 10 minutes) by the intravenous or intraosseous route, or atropine and pralidoxime by the intramuscular route. Plasma concentrations were measured for 6 hours to characterize the antidote concentration-time profiles for each route. RESULTS: Maximum plasma concentrations of atropine and pralidoxime occurred within 2 minutes when administered by the intraosseous route compared with 8 minutes by the intramuscular route. Maximum plasma hydroxocobalamin concentration occurred at the end of the infusion when administered by the intraosseous route. The mean area under the concentration-time curve by the intraosseous route was similar to the intravenous route for all 3 drugs and similar to the intramuscular route for atropine and pralidoxime. CONCLUSION: This study showed rapid and substantial antidote bioavailability after intraosseous administration that appeared similar to that of the intravenous route. The intraosseous route of antidote administration should be considered when intravenous access is difficult.

Near-fatal misuse of medical tape around an endotracheal tube connector during inhalation anesthesia in a horse.

A 7-year-old gelded Irish sports horse weighing 650 kg was anesthetized on 2 consecutive days for lavage of a septic right radio-carpal joint. On both occasions the endotracheal tube connector, which had been bound in medical tape to produce an airtight seal, functioned as a unidirectional valve during mechanical ventilation, retarding expiration, imposing positive end expiratory pressure (PEEP), and probably continuous positive airway pressure (CPAP). The equipment dysfunction was not identified on either occasion despite close inspection prompted by progressive increases in airway pressure and thoracic distension. Whilst the PEEP and CPAP exerted unexpectedly modest cardiovascular effects and the horse recovered uneventfully on both occasions, the improvisation may have proven fatal in a higher-risk subject.A 7-year-old gelded Irish sports horse weighing 650 kg was anesthetized on 2 consecutive days for lavage of a septic right radio-carpal joint. On both occasions the endotracheal tube connector, which had been bound in medical tape to produce an airtight seal, functioned as a unidirectional valve during mechanical ventilation, retarding expiration, imposing positive end expiratory pressure (PEEP), and probably continuous positive airway pressure (CPAP). The equipment dysfunction was not identified on either occasion despite close inspection prompted by progressive increases in airway pressure and thoracic distension. Whilst the PEEP and CPAP exerted unexpectedly modest cardiovascular effects and the horse recovered uneventfully on both occasions, the improvisation may have proven fatal in a higher-risk subject.

RésuméMauvais usage quasi-mortel de ruban adhésif médical autour d'un raccord de tube endotrachéal durant l'anesthésie par inhalation chez un cheval. Un hongre de sport Irlandais âgé de 7 ans pesant 650 kg a été anesthésié deux journées consécutives pour le lavage d'une articulation radio-carpienne droite septique. Lors des deux instances, le raccord du tube endotrachéal, qui avait été attaché à l'aide de ruban adhésif médical pour produire une fermeture hermétique, a servi de valve unidirectionnelle durant la ventilation mécanique, retardant l'expiration, imposant une pression positive en fin d'expiration et probablement une ventilation spontanée en pression positive continue. La défaillance de l'équipement n'a pas été identifiée dans l'une ou l'autre de ces instances, malgré une inspection minutieuse suscitée par des hausses progressives de la pression des voies aériennes et un gonflement thoracique. Même si la pression positive en fin d'expiration et la ventilation spontanée en pression positive continue ont eu des effets cardiovasculaires modestes inattendus et que le cheval s'est rétabli sans incident lors des deux instances, l'improvisation aurait pu s'avérée mortelle chez un sujet présentant des risques plus élevés.(Traduit par Isabelle Vallières).

Anesthetic management of a horse with traumatic pneumothorax.

A traumatic pneumothorax and severe hemorrhage were present in a mare with a large thoracic wall defect, lung perforation, and multiple rib fractures. General anesthesia was induced to allow surgical exploration. We describe the anesthetic technique, and discuss the management of the ventilatory, hemodynamic, and metabolic disturbances encountered.

Animal welfare requirements in publishing guidelines.

Descriptions of measures taken to optimize animal welfare are often absent from scientific reports of animal experiments. One reason may be that journal guidelines inadequately compel authors to provide such information. In this study, online English language versions of the 'Guidelines to authors' (GTAs) from 54 national biomedical journals were examined for neutral (unrelated to welfare) and non-neutral keywords referring to: animal welfare; the '3Rs'; the ARRIVE (2010) guidelines, and regulations pertaining to animal experimentation. Journals were selected from nine countries (UK, US, China, Canada, India, Brazil, Germany, Japan and Australia) and seven biomedical specialties (oncology, rheumatology, surgery, pharmacology, medicine, anaesthesia and veterinary medicine). Total GTA word counts varied from 1137 to 31,609. The keyword count identified per category were expressed per myriad (10,000) of total word count. One-way analyses of variance followed by post hoc Tukey pairwise comparisons revealed greater non-neutral per myriad word counts for (a) veterinary GTAs compared with medicine, oncology, rheumatology or surgery; (b) British, compared with Australian, Canadian, German and Japanese GTAs; and (c) no differences between non-neutral categories. The English language versions of GTAs of British and veterinary medical journals contain more words associated with animal welfare, the 3Rs and the ARRIVE guidelines than those from eight other countries and six other medical specialities. The exclusion of 'national' language versions from analysis precludes attempts to identify national differences in attitudes to laboratory animal welfare.

Postoperative complications in a lamb after major surgery.

INTRODUCTION: Anaesthesia in lambs undergoing experimental surgery may develop problems associated with age-related immune incompetency: a postoperative complication in a 3 week old Scottish blackface lamb after spinal surgery is presented. CASE HISTORY AND MANAGEMENT: Both lamb and ewe were in good condition. The ewe was vaccinated against Clostridium perfringens and Clostridium tetani 5 weeks pre-partum. There were no apparent problems with the lamb's intake of colostrum. Pre-anaesthetic medication was intramuscular medetomidine (10 µg kg(-1)). Anaesthesia was induced and maintained with sevoflurane in oxygen. Morphine (0.5 mg kg(-1)), meloxicam (0.6 mg kg(-1)) and ketamine (1 mg kg(-1) followed by 10 µg kg(-1) minute(-1)) were administered intravenously (IV) for perioperative analgesia. Atracurium (0.5 mg kg(-1) IV, followed by 0.17 mg kg(-1) injected when the first twitch of the four, train-of four count was palpated) was used to improve muscle relaxation. The lamb's trachea was intubated and the lungs mechanically ventilated to maintain normocapnia. Intrathecal morphine (0.2 mg kg(-1)), IV meloxicam (0.3 mg kg(-1)) and edrophonium (0.5 mg kg(-1)) were administered before recovery. Operative and initial recovery periods were unremarkable. Three hours after surgery the lamb became depressed. Tachycardia (180-250 beats minute(-1)), tachypnoea (30 breaths minute(-1)), poor peripheral perfusion and cold pelvic limb extremities were present mimicking severe pain, and/or hypovolaemic shock. Analgesics - morphine (total dose 1.3 mg kg(-1)) - and IV fluid therapy boluses - crystalloids (300 mL), colloids (120 mL) and fresh whole blood (60 mL) - failed to ameliorate clinical signs and so the lamb was euthanized 10 hours after surgery. Post-mortem findings supported a possible diagnosis of peracute Clostridium perfringens enterotoxaemia. CONCLUSION: Clostridium perfringens enterotoxaemia should be considered when clinical signs of severe pain and/or hypovolaemic shock fail to respond to analgesics and fluid resuscitation in lambs after major surgery.

Quantitative and qualitative comparison of three scoring systems for assessing recovery quality after general anaesthesia in horses.

OBJECTIVE: To assess the reproducibility and repeatability of two commonly used recovery quality scoring systems and compare them with those of a novel system based on a greater number of objective criteria. ANIMALS: The video-recorded recoveries of ten client-owned horses selected from all recovery recordings taken between September 2005 and March 2006 at the Royal (Dick) School of Veterinary Studies. MATERIALS AND METHODS: A digital versatile disc (DVD) was produced using edited video recordings of ten horses recovering from general anaesthesia. Twelve experienced equine anaesthetists (raters) studied the DVD on three occasions, and scored the recovery quality of each horse using one of three scoring systems (P, D or E) on each occasion. The process was repeated 6 months later (t = 6) to measure intra-observer reliability (repeatability). At first use (t = 0) raters were asked to comment on the advantages and disadvantages of each system. RESULTS: Inter-rater variability was limited for each system: at each observation period raters accounted for 0.3-4.4% variation. System P was insensitive to differences between recoveries. In system D, score variability increased as recovery quality deteriorated. Intra-rater variability varied with system: using system P, raters provided consistent scores between the observation periods for some, but not all horses ('horse' and 'rater' accounted for 9.7% and 1.9% of variation respectively). Raters were less consistent between t = 0 and t = 6 using system D, but each horse was scored with similar consistency. System E produced little variation at the level of horse (1.0%) and rater (1.9%). Raters broadly agreed on the principle advantages and disadvantages of the three systems. CONCLUSIONS AND CLINICAL RELEVANCE: The systems examined showed reliability and reproducibility but practicality and simplicity of use appeared to be inextricably linked with imprecision. Further work is required to produce a suitable recovery quality scoring system.

Feeding management before gastrointestinal studies in pigs.

Pigs are used to model humans in gastrointestinal (GI) studies because of their comparable size, physiology and behaviour: both are monogastric omnivores. A porcine surgical model for testing novel, tethered ultrasound capsule endoscopes (USCE) requires a clean, motile small intestine. Recommendations for human GI tract preparation before the mechanically similar process of video capsule endoscopy describe using oral purgatives, while high-carbohydrate drinks are recommended before colorectal surgery. Reports of the GI preparation of pigs exist but lack technical details, that is, administration, efficacy and side effects. This report details feeding a high-energy liquid diet to 11 female pigs undergoing surgery and USCE which was readily accepted and easily administered, and which produced a clean, motile small intestine and caused no detectable physiological/behavioural abnormalities.

A study of the correlation between objective and subjective indices of recovery quality after inhalation anaesthesia in equids.

OBJECTIVE: To examine the association between objective and subjective descriptors used for assessing recovery quality in horses after anaesthesia. STUDY DESIGN: Prospective clinical study. ANIMALS: Two hundred and seventy-six equids (110 mares, 85 entire males and 81 geldings), ASA 1-5, weighing 50-850 kg and aged 1 month - 25 years. METHODS: Recoveries after general anaesthesia were assisted with head and tail ropes by two anaesthetists. One scored dichotomous objective descriptors (DOD) of recovery. Two dichotomous objective scales (DOS) were then generated from those descriptors. The same individual also scored overall recovery quality using a visual analogue scale (VAS). The second anaesthetist scored recovery (good or bad) using a dichotomous subjective scale (DSS). Each DOD, the DSS and VAS were compared with each other using Pearson's chi-square test. DOSs were compared to the DSS using Wilcoxon's test and to the VAS using a Spearman's correlation test. RESULTS: Most DODs were associated (p < 0.05) with DSS and VAS. The DSS was not associated with resting/not resting in sternal recumbency (p = 0.535) nor with the time spent in sternal recumbency (p = 0.09). VAS and DSS scores were strongly associated (p < 0.0001). The two DOSs were in agreement with DSS (p < 0.0001) and negatively correlated to VAS (r(1)(2) = 0.38, r(2)(2) = 0.34, respectively, p-value <0.0001). CONCLUSION: Objective descriptors were linked closely with the subjective evaluations of recovery quality except for the presence or absence of a sternal recumbency phase and its duration. CLINICAL RELEVANCE: These components may not be essential in recovery scoring systems. The DOS were in agreement with DSS and VAS and could be a useful tool for further studies on recoveries.

The bispectral index during recovery from halothane and sevoflurane anaesthesia in horses.

OBJECTIVE: To record the bispectral index (BIS) when horses moved during either halothane or sevoflurane anaesthesia and when they made volitional movements during recovery from these anaesthetics. STUDY DESIGN: Randomized prospective clinical study. ANIMALS: Twenty-five client-owned horses undergoing surgery aged 8.8 (+/- 5.3; 1-19) years (mean +/- SD; range). METHODS: Baseline BIS values were recorded before pre-anaesthetic medication (BIS(B)) and during anaesthesia (BIS(A)) maintained with halothane (group H; n = 12) or sevoflurane (group S; n =13) at approximately 0.8-0.9 x minimum alveolar concentrations (MAC). Bispectral indices were recorded during the surgery when unexpected movement occurred (BIS(MA)), during recovery when the first movement convincingly associated with consciousness was observed (BIS(M1)) and once sternal recumbency was achieved (BIS(ST)). RESULTS: No significant difference in BIS(M1) was found between halothane- (85 +/- 7; 75-93) and sevoflurane- (87 +/- 10; 70-98) anaesthetized horses although BIS(A) was significantly (p = 0.0002) lower in group S (62 +/- 7; 53-72) than group H (74 +/- 7; 60-84). Differences between BIS(M1) and BIS(A) were significant in sevoflurane (p = 0.00001) and halothane recipients (p = 0.002) but were greater in group S (25 +/- 9; 4-38) compared with group H (12 +/- 10; -9-25). In six of eight horses, BIS(MA) values ranged between those recorded during anaesthesia and at first movement. CONCLUSIONS AND CLINICAL RELEVANCE: Bispectral indices appear to approximate levels of unconsciousness, suggesting that monitoring the BIS may assist equine anaesthesia. However, it does not predict intra-operative movement.

Opioid analgesia in horses.

Opioid analgesics have been the foundation of human pain management for centuries, and their value in animals has increased since it was proposed that it is the veterinarian's duty to alleviate pain whenever it may occur. Compared with other domesticated species, the horse has benefitted less from the increased understanding of opioid pharmacology in animals, because early literature was overlooked and later work, which examined adverse side effects rather than analgesia, concluded that analgesic and excitatory doses were irreconcilably close. More recent studies have indicated a widening role for opioid analgesics in equine pain management, and radioligand studies have revealed a basis for the equine response pattern to opioid analgesics.

An Anglocentric History of Anaesthetics and Analgesics in the Refinement of Animal Experiments.

Previous histories of animal experimentation, e.g., Franco (2013) have focused on ethics, the law and the personalities involved, but not on the involvement of anaesthetics or analgesics. Given that these were major subjects of (UK) Parliamentary debates on vivisection in the mid-19th century and viewed as "indisputable refinements in animal experimentation" (Russell and Burch 1959), it seemed that an analysis of their role was overdue. This commentary has, in interweaving the history of animal experimentation in the UK with the evolution of anaesthesia, attempted to: (1) clarify the evidence for Russell and Burch's view; and (2) evaluate anaesthesia's ongoing contribution to experimental refinement. The history that emerges reveals that the withholding or misuse of anaesthetics and, or analgesics from laboratory animals in the UK has had a profound effect on scientists and indirectly on the attitudes of the British public in general, becoming a major driver for the establishment of the anti-vivisection movement and subsequently, the Cruelty to Animals Act (1876)-the world's first legislation for the regulation of animal experimentation. In 1902, the mismanaged anaesthetic of a dog in the Department of Physiology, University College London resulted in numerous events of public disorder initiated by medical students against the police and a political coalition of anti-vivisectionists, trade unionists, socialists, Marxists, liberals and suffragettes. The importance of anaesthesia in animal experiments was sustained over the following 150 years as small mammalian species gradually replaced dogs and cats as the principle subjects for vivisection. In discussing experimental refinement in their 1959 report, "The Principles of Humane Experimental Technique" Russell and Burch described anaesthetics as "… the greatest single advance in humane technique, (which) has at the same time been virtually indispensable for the advance of experimental biology". Since then, the role of anaesthetics and in particular analgesics has become an unavoidable consideration whenever animal experiments are planned and conducted. This has been accompanied by a proliferation of training and educational programmes in laboratory animal anaesthesia.

Efficacy of an organophosphorus hydrolase enzyme (OpdA) in human serum and minipig models of organophosphorus insecticide poisoning.

Objectives: Current therapeutic options for organophosphorus (OP) insecticide self-poisoning including atropine and oximes are inadequate and case fatality may exceed 20%. An OP hydrolase enzyme, OpdA, has been used for environmental cleansing of OP insecticides and prevented death in rat and non-human primate models of OP insecticide poisoning if given very quickly after exposure. We here tested OpdA's ability to break down OP insecticides in human serum and in clinically relevant minipig models of OP insecticide poisoning.Methods: Human serum was spiked with seven diverse WHO Class II OP insecticides (chlorpyrifos, quinalphos, diazinon, dimethoate, fenthion, phenthoate, and profenofos) and the effect of OpdA on degradation measured. The pharmacodynamic and clinical effects of OpdA treatment were studied in Gottingen minipigs orally poisoned with agricultural formulations of dimethoate EC40 or methyl parathion EC60; pharmacodynamic effects were also assessed in profenofos EC50-poisoned pigs.Results: OpdA effectively hydrolysed OP insecticides in human serum, with rates varying from 856 (SD 44) down to 0.107 (SD 0.01) moles of substrate hydrolysed/mole of enzyme/sec (kcat) for quinalphos and phenthoate, respectively, although at rates 2-3 log orders less than found in vitro in buffered solution. It showed clinical benefit in minipig models, reducing the dose of noradrenaline required to sustain an adequate mean arterial pressure after dimethoate (mean 0.149 [SD 0.10] µg/kg/h vs. 1.07 [SD 0.77] µg/kg/h, p < .0001) and methyl parathion (mean 0.077 [SD 0.08] µg/kg/h vs. 0.707 [SD 0.49] µg/kg/h, p < .0001) poisoning. OpdA reduced blood OP insecticide concentration and acetylcholinesterase inhibition after poisoning by dimethoate, methyl parathion, and profenofos insecticides.Conclusions:In vitro incubation of OpdA in human serum showed hydrolysis of diverse OP insecticides, although at lower rates than found in buffer solutions. This activity results in clinical and pharmacodynamic efficacy in vivo against several OP insecticides. These results support the testing of OpdA in further animal models before considering human trials to determine whether it may become an urgently required novel therapeutic agent for OP insecticide self-poisoning.