Intraperitoneal injection of sodium pentobarbital has the potential to elicit pain in adult rats (Rattus norvegicus).

An effective and pain-free killing method is required to achieve the goal of euthanasia, a "good death". Overdose of sodium pentobarbital (PB) by intraperitoneal (IP) injection is a widely accepted technique in laboratory rats, but questions remain regarding pain associated with administration. As PB rapidly causes sedation and loss of consciousness, most studies have relied on indirect evidence of pain. The objective of this study was to assess pain associated with IP PB using an appropriate vehicle control. Adult male and female Sprague Dawley (SD) and female Wistar rats (N = 84) were block randomised by sex and strain to receive one of three treatments: 1) 800 mg/kg PB (pH 11), 2) saline or 3) vehicle controls (pH 11 or 12.5). Behavior (Rat Grimace Scale (RGS), writhing, back arching) was evaluated at baseline, before loss of righting reflex (LORR, PB group), and at 80s, 151s and 10 min post-injection (PI; saline and vehicle control groups). In the PB group, mean time to LORR was 78 ± 7.9 seconds. In the vehicle control groups, RGS scores were increased at 151s PI (SD: p = 0.0002, 95%CI 0.73 to 0.20) from baseline, as was relative frequency of writhing (SD: p < 0.0001; Wistar; p = 0.0004). RGS scores remained elevated 10 mins PI (SD: p = 0.0005, 95%CI 0.71 to 0.18; Wistar: p = 0.0234, 95%CI 0.91 to 0.07) but the relative frequency of writhing did not (p > 0.999). The RGS scores and the relative frequency of writhing remained low in the PB and saline groups (p > 0.05). These results show that, vehicle controls for IP PB result in signs associated with pain, pain may not be experienced following IP PB when LORR occurs quickly, and that the effects of PB limit behavioral pain assessments.

CCL22 is a biomarker of cartilage injury and plays a functional role in chondrocyte apoptosis.

BACKGROUND: Osteoarthritis (OA) is one of the leading causes of disability worldwide. Previous history of knee injury is a significant risk factor for OA. It has been established that low-level chronic inflammation plays a pivotal role in the onset and pathogenesis of OA. The primary aim of this research was to determine if a history of knee joint injury is associated with systemic inflammation. A secondary aim was to determine if systemic inflammation is related to knee pain and joint structure. METHODS: Differences in serum cytokine association networks, knee joint structural changes (MRI), and self-reported pain (i.e., Knee Injury and Osteoarthritis Outcome Score Pain subscale, KOOSPAIN and Intermittent and Constant Osteoarthritis Pain score, ICOAP) between individuals who had sustained a youth (aged 15-26 years) sport-related knee injury 3-10 years previously and age- and sex-matched controls were examined. Proteins of interest were also examined in an OA rat model. RESULTS: Cytokine association networks were found to differ significantly between study groups, yet no significant associations were found between networks and KOOSPAIN or MRI-defined OA. A group of cytokines (MCP1/CCL2, CCL22 and TNFα) were differentially associated with other cytokines between study groups. In a pre-clinical rat OA model, serum CCL22 levels were associated with pain (r = 0.255, p = 0.045) and structural changes to the cartilage. CCL22 expression was also observed in human OA cartilage and furthermore, CCL22 induced apoptosis of isolated human chondrocytes. DISCUSSION: These results suggest that CCL22 may be an early factor in the onset/pathogenic process of cartilage degeneration and/or related to pain OA.

Forced-air pre-warming prevents peri-anaesthetic hypothermia and shortens recovery in adult rats.

General anaesthesia disrupts thermoregulation in mammals, which can cause hypothermia. Decreases in core body temperature of 1℃ cause significant postoperative complications in humans, and peri-anaesthetic hypothermia in mice increases data variability, which can potentially increase animal use. In rats, the impact of different temperature management strategies on the incidence and severity of hypothermia, and the accuracy of different temperature measurement methods, is unknown. Eighteen adult male and female SD rats were block-randomized to one of three treatment groups: no-warming (NW), limited-warming (LW, heat pad during anaesthesia), and pre-warming (PW, warm air exposure before anaesthesia, followed by heat pad). Anaesthesia (isoflurane) duration was for 40 min. Core body temperature (intra-abdominal telemetric temperature capsule) was recorded during anaesthesia and recovery. During anaesthesia, rectal, skin, and tail temperatures were also recorded. In the PW group, core temperature was maintained during anaesthesia and recovery. By contrast, the NW group was hypothermic (11% temperature decrease) during anaesthesia. The LW group showed a decrease in temperature during recovery. Recovery to sternal recumbency was significantly faster in the PW (125 [70-186] s, P = 0.0003) and the LW (188 [169-420] s, P = 0.04) groups than in the NW group (525 [229-652] s). Rectal temperature underestimated core temperature (bias -0.90℃, 95% limits of agreement -0.1 to 1.9℃). Skin and tail temperatures showed wide 95% limits of agreement, spanning 6 to 15℃, respectively. The novel strategy of PW was effective at maintaining core temperature during and after anaesthesia. Rectal temperature provided an acceptable proxy for core body temperature.

A clinical audit cycle of post-operative hypothermia in dogs.

OBJECTIVES: Use of clinical audits to assess and improve perioperative hypothermia management in client-owned dogs. METHODS: Two clinical audits were performed. In Audit 1 data were collected to determine the incidence and duration of perioperative hypothermia (defined as rectal temperatures <37·0°C). The results from Audit 1 were used to reach consensus on changes to be implemented to improve temperature management, including re-defining hypothermia as rectal temperature <37·5°C. Audit 2 was performed after 1 month with changes in place. RESULTS: Audit 1 revealed a high incidence of post-operative hypothermia (88·0%) and prolonged time periods (7·5 hours) to reach normothermia. Consensus changes were to use a forced air warmer on all dogs and measure rectal temperatures hourly post-operatively until temperature ≥37·5°C. After 1 month with the implemented changes, Audit 2 identified a significant reduction in the time to achieve a rectal temperature of ≥37·5°C, with 75% of dogs achieving this goal by 3·5 hours. The incidence of hypothermia at tracheal extubation remained high in Audit 2 (97·3% with a rectal temperature <37·5°C). CLINICAL SIGNIFICANCE: Post-operative hypothermia was improved through simple changes in practice, showing that clinical audit is a useful tool for monitoring post-operative hypothermia and improving patient care. Overall management of perioperative hypothermia could be further improved with earlier intervention.

The relationship between the Rat Grimace Scale and mechanical hypersensitivity testing in three experimental pain models.

BACKGROUND: The assessment of facial expressions associated with pain has been used to evaluate pain in humans and has recently found application in non-human mammals. These so called 'grimace scales' have the potential to be developed into a widely accepted non-invasive method of measuring pain in laboratory rodents. Currently, common methodologies to assess pain rely on nociceptive tests that assess stimulus evoked withdrawal responses. These tests, however, are limited to the assessment of a reflexive response without an affective component. This study aimed to use the recently developed Rat Grimace Scale (RGS) and assess its relationship with a conventional nociceptive test (the application of von Frey filaments). METHODS: Fifty-two adult, male Wistar rats were randomized to one of five treatment groups: intraplantar carrageenan, intraplantar complete Freund's adjuvant (CFA), plantar incision, anaesthetic control and saline injection control. The RGS and response to mechanical hypersensitivity testing was evaluated at predetermined time points before and after treatment until withdrawal responses returned to baseline levels. RESULTS: The RGS score significantly increased in all pain models. The peak RGS score also coincided with the development of paw hypersensitivity. However, mechanical hypersensitivity persisted after RGS scores returned to baseline. CONCLUSION: This study confirms that the three pain models induce pain in rodents and showed that peak pain coincided with peak mechanical hypersensitivity. However, mechanical hypersensitivity remained once pain subsided, mimicking the human experience of CFA injection. These findings further our understanding of the roles of, and relationship between, these assays in the assessment of nociception and pain.

Sound Pressure Levels in 2 Veterinary Intensive Care Units.

BACKGROUND: Intensive care units (ICUs) in human hospitals are consistently noisy environments with sound levels sufficient to substantially decrease sleep quality. Sound levels in veterinary ICUs have not been studied previously, but environmental sound has been shown to alter activity in healthy dogs. HYPOTHESIS: Veterinary ICUs, like those in human medicine, will exceed international guidelines for hospital noise. ANIMALS: NA. METHODS: Prospective, observational study performed consecutively and simultaneously over 4 weeks in 2 veterinary ICUs. Conventional A-weighted sound pressure levels (equivalent continuous level [a reflection of average sound], the sound level that is exceeded 90% of the recording period time [reflective of background noise], and maximum sound levels) were continuously recorded and the number of spikes in sound >80 dBA were manually counted. RESULTS: Noise levels were comparable to ICUs in human hospitals. The equivalent continuous sound level was higher in ICU1 than in ICU2 at every time point compared, with greatest differences observed on week day (ICU1, 60.1 ± 3.7 dBA; ICU2, 55.9 ± 2.5 dBA, P < .001) and weekend nights (ICU1, 59.9 ± 2.4 dBA; ICU2, 53.4 ± 1.7 dBA, P < .0001) reflecting a 50% difference in loudness. Similar patterns were observed for the maximum and background noise levels. The number of sound spikes was up to 4 times higher in ICU1 (162.3 ± 84.9 spikes) than in ICU2 (40.4 ± 12.2 spikes, P = .001). CONCLUSIONS AND CLINICAL IMPORTANCE: These findings show that sound in veterinary ICUs is loud enough to potentially disrupt sleep in critically ill veterinary patients.

Carbon dioxide, but not isoflurane, elicits ultrasonic vocalizations in female rats.

Gradual filling of a chamber with carbon dioxide is currently listed by the Canadian Council on Animal Care guidelines as a conditionally acceptable method of euthanasia for rats. Behavioural evidence suggests, however, that exposure to carbon dioxide gas is aversive. Isoflurane is less aversive than carbon dioxide and may be a viable alternative, though objective data are lacking for the period leading up to loss of consciousness. It has been shown that during negative states, such as pain and distress, rats produce ultrasonic vocalizations. The objective of this study was to detect ultrasonic vocalizations during exposure to carbon dioxide gas or isoflurane as an indicator of a negative state. Specialized recording equipment, with a frequency detection range of 10 to 200 kHz, was used to register these calls during administration of each agent. Nine female Sprague-Dawley rats were exposed to either carbon dioxide or isoflurane on two different occasions. All rats vocalized in the ultrasonic range (30 to 70 kHz) during exposure to carbon dioxide. When exposed to isoflurane, no calls were detected from any of the animals. The frequent occurrence of ultrasonic vocalizations during carbon dioxide exposure suggests that the common practice of carbon dioxide euthanasia is aversive to rats and that isoflurane may be a preferable alternative.

Effects of prednisone on blood lactate concentrations in healthy dogs.

BACKGROUND: Glucocorticoids affect carbohydrate and lactate metabolism. HYPOTHESIS: Administration of prednisone to healthy dogs will result in clinically relevant hyperlactatemia. ANIMALS: Twelve healthy adult Beagle dogs. METHODS: Prospective, controlled experimental study. Twelve healthy adult Beagles were divided into 2 groups (3 of each sex per group). One group served as control. The other group received 2 treatments: low, 1 mg/kg prednisone PO q24h for 2 weeks; high, 4 mg/kg prednisone PO q24h for 2 weeks. A washout period of 6 weeks separated the treatments. Blood samples were drawn for whole blood lactate measurement on day (D) 0, D4, and D14 and measured in duplicate. RESULTS: Compared with the control group, low and high groups had significantly higher blood lactate concentrations at D4 and D14. There was no difference at D0. There was no effect of time within the control group. In the low and high groups, blood lactate concentration was increased at D4 and D14 versus D0. Blood lactate concentration was greater in the high group than the low group at D14 only. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs treated with prednisone experience statistically significant increases in blood lactate concentrations, which can result in type B hyperlactatemia. In such cases, improving tissue perfusion, treatment for the commonest form of hyperlactatemia (type A) would be unnecessary.

Target-controlled infusion of propofol in dogs--evaluation of four targets for induction of anaesthesia.

Four groups of 20 dogs were anaesthetised by means of target-controlled infusions of propofol designed to achieve 2.5 microg/ml, 3.0 microg/ml, 3.5 microg/ml or 4.0 microg/ml of propofol in blood. The dogs' pulse rate and respiratory rate were recorded before premedication and induction, immediately after endotracheal intubation and three and five minutes later (times 0, 3 and 5, respectively), and their arterial blood pressure was recorded oscillometrically just before induction and at times 0, 3 and 5. The targets of 2.5, 3.0, 3.5 and 4.0 microg/ml resulted in the successful induction of anaesthesia in 13 (65 per cent), 16 (80 per cent), 20 (100 per cent) and 20 (100 per cent) of the dogs, respectively. The incidence of postinduction apnoea was 0 (0 per cent), one (5 per cent), two (10 per cent) and eight (40 per cent) at time 5 for groups 2.5, 3.0, 3.5 and 4.0 mug/ml, respectively, and its incidence at time 5 was significantly higher in the 4.0 microg/ml group (P<0.05) than in the other groups. In all the groups there was a significant (P<0.05) decrease in blood pressure between just before induction and the later measurements. Although there were no statistically significant differences between the groups in terms of inducing anaesthesia at a specific target, a target of 3.5 microg/ml appears to ensure a successful induction of anaesthesia without a significant increase in the incidence of apnoea.