Assessing Susceptibility to Carbon Dioxide Gas in Three Rat Strains Using the Loss of Righting Reflex.

Overdose of carbon dioxide gas (CO2) is a common euthanasia method for rodents; however, CO2 exposure activates nociceptors in rats at concentrations equal to or greater than 37% and is reported to be painful in humans at concentrations equal to or greater than 32.5%. Exposure of rats to CO2 could cause pain before loss of consciousness. We used 2 standardized loss of righting reflex (LORR) methods to identify CO2 concentrations associated with unconsciousness in Wistar, Long-Evans, and Sprague-Dawley rats (n = 28 animals per strain). A rotating, motorized cylinder was used to test LORR while the rat was being exposed to increasing concentrations of CO2. LORR was defined based on a 15-second observation period. The 2 methods were 1) a 1-Paw assessment (the righting reflex was considered to be present if one or more paws contacted the cylinder after the rat was positioned in dorsal recumbency), and 2) a 4-Paw assessment (the righting reflex was considered to be present if all 4 paws contacted the cylinder after the rat was positioned in dorsal recumbency). Data were analyzed with Probit regression, and dose-response curves were plotted. 1-Paw EC95 values (CO2 concentration at which LORR occurred for 95% of the population) were Wistar, 27.2%; Long-Evans, 29.2%; and Sprague-Dawley, 35.0%. 4-Paw EC95 values were Wistar, 26.2%; Long-Evans, 25.9%, and Sprague-Dawley, 31.1%. Sprague-Dawley EC95 values were significantly higher in both 1- and 4-Paw tests as compared with Wistar and Long-Evans rats. No differences were detected between sexes for any strain. The 1-Paw EC95 was significantly higher than the 4-Paw EC95 only for Sprague-Dawley rats. These results suggest that a low number of individual rats from the strains studied may experience pain during CO2 euthanasia.

Use of Loss of Righting Reflex to Assess Susceptibility to Carbon Dioxide Gas in Three Mouse Strains.

Exposure to CO2 gas is a common rodent euthanasia method. CO2 activates nociceptors in rats and is painful to humans at concentrations equal to or greater than 32.5% The concentration of CO2 at which rodents become unconsciousness is inadequately defined. We used loss of righting reflex (LORR) to identify the concentration at which CO2 caused loss of consciousness in C57Bl/6, CD1 and 129P3J mice (16 females and 16 males per strain). We used a custom built, rotating, motorized cylinder to determine LORR as CO2 concentrations were increased. Two LORR assessment methods were used: 1) a 1-Paw assessment in which the righting reflex was considered to be present if one or more paws contacted the cylinder after rotation into dorsal recumbency and 2) a 4-Paw assessment in which the righting reflex was considered to be present only if all 4 paws contacted the cylinder. LORR test data were analyzed with Probit regression and dose response curves were plotted. 1-Paw EC95 values (CO2 concentration at which LORR occurred for 95% of the population) were: C57Bl/6; 30.7%, CD1; 26.2%, 129P3J; 20.1%. The EC95 for C57Bl/6 was significantly higher than that of the 129P3J mice, with no significant differences between other strains. Four-Paw EC95 values were: C57Bl/6; 22.8%, CD1; 25.3%, 129P3J; 20.1%. Values for 129P3J mice were significantly lower than those of CD1 mice), with no significant difference between other strains. The EC95 varied significantly between 1-Paw and 4-Paw methods only for C57Bl/6 mice. These results suggest a potential for nociception and pain to occur in some individuals of some mouse strains during CO2 euthanasia.

Intrahepatic Injection of Sodium Pentobarbital as an Alternative to Intraperitoneal Injection for the Euthanasia of Rats (Rattus norvegicus).

The most commonly accepted method of rat euthanasia in North America is intraperitoneal injection of sodium pentobarbital (PB). However, misinjection can occur, and intraperitoneal PB may cause pain and distress. The objective of this study was to test an alternative method of euthanasia: intrahepatic injection of PB. A pilot study was conducted to develop a method of intrahepatic injections (evaluated using CT scans and test injections), followed by a full study comparing intraperitoneal (n = 14) and intrahepatic PB injections (n = 66) in adult rats. Full study outcomes were: 1) time from injection to loss of right- ing reflex (LORR), 2) time from injection to cessation of heartbeat (CHB), 3) number of failed euthanasia attempts, and 4) confirmation of successful intrahepatic injection or misinjection via necropsy. All injections were performed by a veterinary student. CT revealed that intrahepatic injections were feasible. Times (median [range]) to LORR and CHB were faster after successful intrahepatic injections (LORR, 3 s [1 to 5 s]; CHB, 8 s [2 to 242 s]) than after intraperitoneal injections (LORR, 89.5 s [73 to 110 s], CHB: 284.5 s [237 to 423 s]). The misinjection rate was higher with intrahepatic injections (59%) than with intraperitoneal injections (29%), but intrahepatic misinjection still resulted in fast and successful euthanasia (LORR, 29 s [1 to 96 s]; CHB, 216 s [12 to 330 s]), with the injectate distributed between the intraperitoneal and intrahepatic locations. The number of failed euthanasia attempts with intrahepatic injections was low (n = 2). Intrahepatic injections show potential as an alternative to intraperitoneal injections for rat euthanasia.

Randomization, blinding, data handling and sample size estimation in papers published in Veterinary Anaesthesia and Analgesia in 2009 and 2019.

OBJECTIVE: To evaluate reporting of items indicative of bias and weak study design. STUDY DESIGN: Literature survey. POPULATION: Papers published in Veterinary Anaesthesia and Analgesia. METHODS: Reporting of randomization, blinding, sample size estimation and data exclusion were compared for papers published separated by a 10 year interval. A reporting rate of more than 95% was considered ideal. The availability of data supporting results in a publicly accessible repository was also assessed. Selected papers were randomized and identifiers removed for review, with data from 59 (57 in 2009, two in 2008) and 56 (52 in 2019, four in 2018) papers analyzed. Items were categorized for completeness of reporting using a previously published operationalized checklist. Two reviewers reviewed all papers independently. RESULTS: Full reporting of randomization increased over time from 13.6% to 85.7% [95% confidence interval (CI), 57.8-86.6%; p < 0.0001], as did sample size estimation (from 0% to 20%; 95% CI, 7.6-32.4%; p = 0.002). Reporting of blinding (49.2% and 50.0%; 95% CI, -18.3% to 20.0%; p = 1.0) and exclusions of samples/animals (39.0% and 50.0%; 95% CI, -8.8% to 30.8%; p = 0.3) did not change significantly. Data availability was low (2008/2009, zero papers; 2018/2019, two papers). None of the items studied exceeded the predetermined ideal reporting rate. CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that reporting quality remains low, with a risk of bias.

Development and Testing of a Sedation Scale for Use in Rabbits (Oryctolagus cuniculus).

In biomedical research, rabbits are commonly sedated to facilitate a variety of procedures. Developing a sedation assessment scale enables standardization of levels of sedation and comparisons of sedation protocols, and may help in predicting sedation level requirements for different procedures. The goal of this study was to develop a rabbit sedation assessment scale using a psychometric approach. We hypothesized that the sedation scale would have construct validity, good internal consistency, and reliability. In a prospective, randomized, blinded study design, 15 (8 females, 7 males) healthy 1-y-old New Zealand white rabbits received 3 intramuscular treatments: midazolam (0.5 mg/kg; n = 6); midazolam (1.5 mg/kg)-ketamine (5 mg/kg; n = 7); and alfaxalone (4 mg/kg)-dexmedetomidine (0.1 mg/kg)-midazolam (0.2 mg/kg; n = 3). One rabbit received 2 treatments. A sedation scale was developed by using psychometric methods, with assessment performed by 6 independent raters who were blind to treatment. Final sedation scale items included posture, palpebral reflex, orbital tightening, lateral recumbency, loss of righting reflex, supraglottic airway device placement, toe pinch, and general appearance. The scale showed construct validity, good to very good interrater reliability for individual items (6 raters; intraclass correlation coefficient, 0.671 to 0.940), very good intrarater reliability (5 raters; intraclass correlation coefficient, 0.951 to 0.987), and excellent internal consistency (Cronbach α, 0.947). The sedation scale performed well under the conditions tested, suggesting that it can be applied in a wider range of settings (different populations, raters, sedation protocols).

A Pharmacokinetic-Pharmacodynamic Study of Intravenous Midazolam and Flumazenil in Adult New Zealand White-Californian Rabbits (Oryctolagus cuniculus).

Flumazenil, a competitive GABAA receptor antagonist, is commonly used in rabbits to shorten sedation or postanesthetic recovery after benzodiazepine administration. However, no combined pharmacokinetic (PK) and pharmacodynamic (PD) data are available to guide its administration in this species. In a prospective, randomized, blinded, crossover study design, the efficacy of IV flumazenil (FLU; 0.05 mg/kg) or saline control (SAL; equal volume) to reverse the loss of righting reflex (LORR) induced by IV midazolam (1.2 mg/kg) was investigated in 15 New Zealand white rabbits (2.73 to 4.65 kg, 1 y old). Rabbits were instrumented with arterial (central auricular artery) and venous (marginal auricular vein) catheters. After baseline blood sampling, IV midazolam was injected (T0). Flumazenil or saline (FLU/SAL) was injected 30 s after LORR. Arterial blood samples were collected at 1 and 3 min after midazolam injection, and at 1, 3, 6, 10, 15, 21, 28, 36, 45 and 60 min after injection with flumazenil. Plasma samples for midazolam, 1-OH-midazolam and flumazenil were analyzed using high performance liquid chromatography-high-resolution mass spectrometry and the time to return of righting reflex (ReRR) was compared between groups (Wilcoxon test). FLU terminal half-life, plasma clearance and volume of distribution were 26.3 min [95%CI: 23.3 to 29.3], 18.74 mL/min/kg [16.47 to 21.00] and 0.63 L/kg [0.55 to 0.71], respectively. ReRR was 25 times faster in rabbits treated with FLU (23 [8 to 44] s) compared with SAL (576 [130 to 1141] s; 95%CI [425 to 914 s]). Return of sedation (lateral recumbency) occurred in both groups (7/13 in FLU; 12/13 in SAL) with return of LORR in a few animals (4/13 in FLU; 7/13 in SAL) at 1540 [858 to 2328] s. In the population and anesthesia protocol studied, flumazenil quickly and reliably reversed sedation induced by midazolam injection. However, the potential return of sedation after flumazenil administration warrants careful monitoring in the recovery period.

Assessment of an Electronic Mechanical Sensory Threshold Testing Device (RatMet) in Wistar rats (Rattus norvegicus).

von Frey (vF) monofilaments are used to quantify mechanical hypersensitivity and nociception in rodents; however, this method of testing has been criticized due to inconsistencies in testing methods, filament properties, and nonlinearity. This study compared withdrawal thresholds measured by using vF monofilaments with those of a novel mechanical threshold testing device currently in development (RatMet) in a carrageenan inflammatory model in 9- to 11-wk-old male Wistar rats. Rats were randomly assigned to assessment of mechanical hypersensitivity after intraplantar carrageenan injection by using either vF monofilaments (n = 10) or the RatMet device equipped with 3 sizes of probe tips (0.9 mm [RM0.9], n = 15; 0.5 mm [RM0.5], n = 11; and 0.09 mm [RM0.09], n = 11). All RatMet probe sizes and vF monofilaments identified a reduction in withdrawal threshold after treatment. Systematic differences in threshold were identified between vF and both RM0.9 and RM0.5 groups; RM0.09 did not differ from vF. Withdrawal thresholds showed linear relationships with probe diameter, square root of probe diameter, and area of the RatMet probes. In contrast, exponential relationships were observed with the vF monofilaments. Furthermore, none of the RatMet probe results differed in accuracy when comparing a single test with the averages of 3 or 5 tests per time point. Overall, the RatMet device measurements have construct validity even when the number of testing replicates is low. These data indicate that the RatMet device produces data comparable with those from vF monofilaments, with the potential for a shortened testing period without a decrease in accuracy.

Hypothermia During General Anesthesia Interferes with Pain Assessment in Laboratory Rats (Rattus norvegicus).

Accurate pain assessment methods are necessary to ensure animal welfare and reliable data collection in animal research. The Rat Grimace Scale (RGS), a facial expression pain scale, allows effective identification of pain. However, the potential confounds of this method remain mostly unexplored. General anesthesia, which is used in many laboratory procedures, suppresses thermoregulation and results in hypothermia. We investigated the effects of isoflurane-induced hypothermia on RGS scores. Twenty (10 male and 10 female) Sprague-Dawley rats each received 30 min of anesthesia, followed by 30 min of observation after the return of sternal recumbency. Rats were randomized to receive warming with an electric heating pad or no warming during both periods. Unwarmed rats became hypothermic within 15 min after isoflurane exposure began and returned to normothermia within 15 min after returning to sternal recumbency. Warmed rats did not deviate from the normothermic range. The RGS scores of unwarmed rats were significantly higher than baseline levels for 3 h after anesthesia and were higher than those of warmed rats at 5 and 180 min after anesthesia. Hypothermia resulted in a larger proportion of rats crossing a predetermined analgesic intervention threshold. Our findings show that hypothermia induced by isoflurane anesthesia presents a confound to accurate RGS scoring. These results emphasize the importance of maintaining normothermia to avoid inflated pain scores and to obtain accurate pain assessment.

Hypoventilation following oxygen administration associated with alfaxalone-dexmedetomidine-midazolam anesthesia in New Zealand White rabbits.

OBJECTIVE: To investigate the relationship between oxygen administration and ventilation in rabbits administered intramuscular alfaxalone-dexmedetomidine-midazolam. STUDY DESIGN: Prospective, randomized, blinded study. ANIMALS: A total of 25 New Zealand White rabbits, weighing 3.1-5.9 kg and aged 1 year. METHODS: Rabbits were anesthetized with intramuscular alfaxalone (4 mg kg-1), dexmedetomidine (0.1 mg kg-1) and midazolam (0.2 mg kg-1) and randomized to wait 5 (n = 8) or 10 (n = 8) minutes between drug injection and oxygen (100%) administration (facemask, 1 L minute-1). A control group (n = 9) was administered medical air 10 minutes after drug injection. Immediately before (PREoxy/air5/10) and 2 minutes after oxygen or medical air (POSToxy/air5/10), respiratory rate (fR), pH, PaCO2, PaO2, bicarbonate and base excess were recorded by an investigator blinded to treatment allocation. Data [median (range)] were analyzed with Wilcoxon, Mann-Whitney U and Kruskal-Wallis tests and p < 0.05 considered significant. RESULTS: Hypoxemia (PaO2 < 88 mmHg, 11.7 kPa) was observed at all PRE times: PREoxy5 [71 (61-81) mmHg, 9.5 (8.1-10.8) kPa], PREoxy10 [58 (36-80) mmHg, 7.7 (4.8-10.7) kPa] and PREair10 [48 (32-64) mmHg, 6.4 (4.3-8.5) kPa]. Hypoxemia persisted when breathing air: POSTair10 [49 (33-66) mmHg, 6.5 (4.4-8.8) kPa]. Oxygen administration corrected hypoxemia but was associated with decreased fR (>70%; p = 0.016, both groups) and hypercapnia (p = 0.016, both groups). Two rabbits (one per oxygen treatment group) were apneic (no thoracic movements for 2.0-2.5 minutes) following oxygen administration. fR was unchanged when breathing air (p = 0.5). PaCO2 was higher when breathing oxygen than air (p < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Early oxygen administration resolved anesthesia-induced hypoxemia; however, fR decreased and PaCO2 increased indicating that hypoxemic respiratory drive is an important contributor to ventilation using the studied drug combination.

Prewarming Followed by Active Warming is Superior to Passive Warming in Preventing Hypothermia for Short Procedures in Adult Rats (Rattus norvegicus) Under Isoflurane Anesthesia.

General anesthesia is a common procedure in laboratory rats; however, it impairs thermoregulation, rapidly leading to hypothermia as warm core blood is distributed to the cooler periphery. The protective strategy of prewarming before the onset of anesthesia delays hypothermia, but only for a short period. This prospective, randomized, cross-over, experimental study in adult male and female SD rats (n = 8) was designed to compare passive (fleece blanket) and active (temperature controlled heating pad) warming. Initial treatment order was randomized, with a cross-over after a minimum 5 d washout period. Both groups underwent a period of prewarming in a warming box to increase core temperature by 1% (median 0.4 °C). At completion of prewarming, general anesthesia was induced and maintained for 30 min with isoflurane carried in oxygen. Core temperature was monitored for a further 30 min after anesthesia. Active warming resulted in higher core temperatures during anesthesia. During passive warming, hypothermia occurred after approximately 30 min of anesthesia and continued into recovery. In contrast, active warming prevented hypothermia. Prewarming followed by passive warming delayed hypothermia for approximately 30min, but active warming was more effective at maintaining normothermia both during and after general anesthesia.

Agreement between invasive and oscillometric arterial blood pressure measurements using the LifeWindow multiparameter monitor and two cuff sizes in anesthetized adult horses.

OBJECTIVE: To assess agreement between oscillometric noninvasive blood pressure (NIBP) measurements using LifeWindow monitors (LW9xVet and LW6000V) and invasive blood pressure (IBP). To assess the agreement of NIBP readings using a ratio of cuff width to mid-cannon circumference of 25% and 40%. STUDY DESIGN: Prospective, randomized clinical study. ANIMALS: A total of 43 adult horses undergoing general anesthesia in dorsal recumbency for different procedures. METHODS: Anesthetic protocols varied according to clinician preference. IBP measurement was achieved after cannulation of the facial artery and connection to an appropriately positioned transducer connected to one of two LifeWindow multiparameter monitors (models: LW6000V and LW9xVet). Accuracy of monitors was checked daily using a mercury manometer. For each horse, NIBP was measured with two cuff widths (corresponding to 25% or 40% of mid-cannon bone circumference), both connected to the same monitor, and six paired IBP/NIBP readings were recorded (at least 3 minutes between readings). NIBP values were corrected to the relative level of the xiphoid process. A Bland-Altman analysis for repeated measures was used to assess bias (NIBP-IBP) and limits of agreement (LOAs). RESULTS: The 40% cuff width systolic arterial pressure [SAP; bias 7.9 mmHg, LOA -26.6 to 42.3; mean arterial pressure (MAP): bias 4.9 mmHg, LOA -28.2 to 38.0; diastolic arterial pressure (DAP): bias 4.2 mmHg, LOA -31.4 to 39.7)] performed better than the 25% cuff width (SAP: bias 26.4 mmHg, LOA -21.0 to 73.9; MAP: bias 15.7 mmHg, LOA -23.8 to 55.2; DAP: bias 10.9 mmHg, LOA -33.2 to 54.9). CONCLUSIONS AND CLINICAL RELEVANCE: Using the LifeWindow multiparameter monitor in anesthetized horses, the 40% cuff width provided better agreement with IBP; however, both cuff sizes and both monitor models failed to meet American College of Veterinary Internal Medicine Consensus Statement Guidelines.

Review of Intraperitoneal Injection of Sodium Pentobarbital as a Method of Euthanasia in Laboratory Rodents.

Euthanasia is one of the most commonly performed procedures in biomedical research, involving tens of millions of animals in North America and Europe every year. The use of sodium pentobarbital, injected intraperitoneally, for killing rodents is described as an acceptable technique by the AVMA and CCAC euthanasia guidelines. This drug and route are recommended over inhalant anesthetics, carbon dioxide, and physical methods for ethical and aesthetic reasons as well as efficiency. However, a growing body of evidence challenges the efficacy and utility of intraperitoneal pentobarbital. This methodology has been described as inconsistent and may induce pain and stress. With these considerations in mind, a review of the literature is needed to assess the evidence surrounding this killing method, the associated welfare implications, and potential for refinement.

Evaluating Intrahepatic and Intraperitoneal Sodium Pentobarbital or Ethanol for Mouse Euthanasia.

Intraperitoneal (IP) injection of sodium pentobarbital (PB) is an accepted method of euthanasia for mice. However, this method has important drawbacks, including the potential for pain or misinjection. The objective of this prospective, randomized, blinded study was to determine whether intrahepatic (IH) injection of PB is more effective than IP delivery for mouse euthanasia. Secondary objectives were to: 1) determine whether IP ethanol (ET) is a suitable alternative to PB and 2) study the effect of isoflurane anesthesia on euthanasia with either PB or ET. Eighty adult CD1 mice were randomly assigned to 6 different treatment groups, were euthanized by using IP or IH injections of either PB or ET, and were either anesthetized or conscious before injection. Variables of interest were: 1) misinjection rates (based on necropsy evaluation), 2) time from injection to apnea and 3) time to cessation of heartbeat (CHB). The misinjection rate for IH injections was 93% (28/30). Two successful IH injections resulted in death within 4 s, but this method cannot be recommended due to the possibility for intrathoracic injection ( n = 4). In nonanesthetized mice, time to apnea and CHB was significantly shorter with IP ET (apnea: 72.5 s [median], CHB: 115 s) than with IP PB (apnea: 136 s, CHB: 176 s). Anesthesia at time of injection was associated with a shorter CHB time for IP PB. These data show the difficulty in achieving successful IH injections in mice, but confirm that IP ET is a viable and potentially superior alternative to IP PB. Lastly, anesthesia can shorten time to death after IP injection of PB.

Dynamic prediction of fluid responsiveness during positive pressure ventilation: a review of the physiology underlying heart-lung interactions and a critical interpretation.

OBJECTIVE: Cardiovascular responses to hypovolemia and hypotension are depressed during general anesthesia. A considerable number of anesthetized and critically ill animals may not benefit hemodynamically from a fluid bolus; therefore, it is important to have measures for accurate prediction of fluid responsiveness. Static measures of preload, such as central venous pressure, do not provide accurate prediction of fluid responsiveness, whereas dynamic measures of cardiovascular function, obtained during positive pressure ventilation, are highly predictive. This review describes key physiological concepts behind heart-lung interactions during positive pressure ventilation, factors that can modify this relationship and provides the basis for a rational interpretation of the information obtained from dynamic measurements, with a focus on pulse pressure variation (PPV). DATABASE USED: PubMed. Search items used were: heart-lung interaction, positive pressure ventilation, pulse pressure variation, dynamic index of fluid therapy, goal-directed hemodynamic therapy, dogs, cats, pigs, horses and rabbits. CONCLUSIONS: The veterinary literature suggests that targeting specific PPV thresholds should guide fluid therapy in lieu of conventional assessments. Understanding the physiology of heart-lung interactions during intermittent positive pressure ventilation provides a rational basis for interpreting the literature on dynamic indices of fluid responsiveness, including PPV. Clinical trials are needed to evaluate whether goal-directed fluid therapy based on PPV results in improved outcomes in veterinary patient populations.

A randomized clinical trial comparing butorphanol and buprenorphine within a multimodal analgesic protocol in cats undergoing orchiectomy.

OBJECTIVES: The aim of this study was to compare the effects of butorphanol and buprenorphine, as part of a multimodal analgesic protocol, on recovery and analgesia in cats undergoing orchiectomy. METHODS: In a prospective, randomized, blind clinical trial, 47 adult male cats were randomly assigned to receive either butorphanol (0.3 mg/kg, n = 24) or buprenorphine (0.02 mg/kg, n = 23) in combination with dexmedetomidine (25 µg/kg) and alfaxalone (2 mg/kg) as a single intramuscular injection for the induction of general anesthesia. Isoflurane carried in oxygen was supplemented as needed during orchiectomy. All cats received lidocaine (2 mg/kg intratesticular), meloxicam (0.3 mg/kg SC) and atipamezole (125 µg/kg IM) postoperatively. Pain and sedation scales were applied at baseline, and 2, 4 and 6 h postoperatively. Time to achieve sternal recumbency and to begin eating were also recorded. RESULTS: Pain scale scores were low and showed no difference between the treatment groups at all time points (P ⩾0.99, all time points). Four cats exceeded the analgesia intervention threshold for rescue analgesia (butorphanol, n = 3; buprenorphine, n = 1). There was no difference in sedation scores between groups at any time point (P >0.99, all time points). Significantly more cats in the buprenorphine group (n = 12) required isoflurane than in the butorphanol group (n = 2) (P = 0.0013; relative risk 6.3, 95% confidence interval [CI] 1.8-23.5). There was no significant difference in time to achieve sternal recumbency (P = 0.96, 95% CI -20 to 20) between groups or in return to eating (P = 0.48, 95% CI -92.0 to 113.5), with most cats eating within 1 h of surgery (butorphanol, 79%; buprenorphine, 83%). CONCLUSIONS AND RELEVANCE: There were no significant differences in analgesia or recovery between butorphanol and buprenorphine treatment groups as part of a multimodal injectable anesthetic protocol. Butorphanol is associated with superior depth of anesthesia, facilitating injectable anesthesia.

Influence of Rater Training on Inter- and Intrarater Reliability When Using the Rat Grimace Scale.

Rodent grimace scales facilitate assessment of ongoing pain. Reported rater training using these scales varies considerably and may contribute to the observed variability in interrater reliability. This study evaluated the effect of training on interrater reliability with the Rat Grimace Scale (RGS). Two training sets (42 and 150 images) were prepared from acute pain models. Four trainee raters progressed through 2 rounds of training, scoring 42 images (set 1) followed by 150 images (set 2a). After each round, trainees reviewed the RGS and any problematic images with an experienced rater. The 150 images were then rescored (set 2b). Four years later, trainees rescored the 150 images (set 2c). A second group of raters (no-training group) scored the same image sets without review with the experienced rater. Inter- and intrarater reliability were evaluated by using the intraclass correlation coefficient (ICC), and ICC values were compared by using the Feldt test. In the trainee group, interrater reliability increased from moderate to very good between sets 1 and 2b and increased between sets 2a and 2b. Action units with the highest and lowest ICC at set 2b were orbital tightening and whiskers, respectively. In comparison to an experienced rater, the ICC for all trainees improved, ranging from 0.88 to 0.91 at set 2b. Four years later, very good interrater reliability was retained, and intrarater reliability was good or very good). The interrater reliability of the no-training group was moderate and did not improve from set 1 to set 2b. Training improved interrater reliability, with an associated reduction in 95%CI. In addition, training improved interrater reliability with an experienced rater, and performance was retained.

A Review of Pain Assessment Methods in Laboratory Rodents.

Ensuring that laboratory rodent pain is well managed underpins the ethical acceptability of working with these animals in research. Appropriate treatment of pain in laboratory rodents requires accurate assessments of the presence or absence of pain to the extent possible. This can be challenging some situations because laboratory rodents are prey species that may show subtle signs of pain. Although a number of standard algesiometry assays have been used to assess evoked pain responses in rodents for many decades, these methods likely represent an oversimplification of pain assessment and many require animal handling during testing, which can result in stress-induced analgesia. More recent pain assessment methods, such as the use of ethograms, facial grimace scoring, burrowing, and nest-building, focus on evaluating changes in spontaneous behaviors or activities of rodents in their home environments. Many of these assessment methods are time-consuming to conduct. While many of these newer tests show promise for providing a more accurate assessment of pain, most require more study to determine their reliability and sensitivity across a broad range of experimental conditions, as well as between species and strains of animals. Regular observation of laboratory rodents before and after painful procedures with consistent use of 2 or more assessment methods is likely to improve pain detection and lead to improved treatment and care-a primary goal for improving overall animal welfare.

Assessing analgesia equivalence and appetite following alfaxalone- or ketamine-based injectable anesthesia for feline castration as an example of enhanced recovery after surgery.

Objectives The primary study objective was to assess two injectable anesthetic protocols, given to facilitate castration surgery in cats, for equivalence in terms of postoperative analgesia. A secondary objective was to evaluate postoperative eating behavior. Methods Male cats presented to a local clinic were randomly assigned to receive either intramuscular ketamine (5 mg/kg, n = 26; KetHD) or alfaxalone (2 mg/kg, n = 24; AlfHD) in combination with dexmedetomidine (25 µg/kg) and hydromorphone (0.05 mg/kg). All cats received meloxicam (0.3 mg/kg SC) and intratesticular lidocaine (2 mg/kg). Species-specific pain and sedation scales were applied at baseline, 1, 2 and 4 h postoperatively. Time taken to achieve sternal recumbency and begin eating were also recorded postoperatively. Results Pain scale scores were low and showed equivalence between the treatment groups at all time points (1 h, P = 0.38, 95% confidence interval [CI] of the difference between group scores 0-0; 2 h, P = 0.71, 95% CI 0-0; 4 h, P = 0.97, 95% CI 0-0). Four cats crossed the threshold for rescue analgesia (KetHD, n = 1; AlfHD, n = 3). At 1 h, more cats in the KetHD (65%) group than in the AlfHD (42%) group were sedated, but statistical significance was not detected ( P = 0.15, 95% CI -1 to 0). Most AlfHD cats (88%) began eating by 1 h vs 65% of KetHD cats ( P = 0.039). Time to recover sternal recumbency did not differ between groups ( P = 0.86, 95% CI -14.1 to 11.8). Conclusions and relevance These results show that AlfHD and KetHD provide equivalent analgesia as part of a multimodal injectable anesthetic protocol. Alfaxalone is associated with an earlier return to eating.