Grimace Scales Research Articles

Development of the calf grimace scale for pain and stress assessment in castrated Angus beef calves

Grimace scales have been used to assess pain in various animal species. This study aimed to develop the calf grimace scale (CGS), evaluate its responsiveness and the effect of external factors (change of environment and dam separation, and restraint) on CGS. Sixty-nine Angus calves, 6–8 weeks old, were randomly allocated into castrated (n = 34) and sham castrated (n = 35) groups. Images were extracted from videos pre- (M1-M4), during- (M5), and post-castration/sham castration (M6, M7). Six facial action units (FAUs) were identified: ear position, orbital tightening, tension above the eye, nostril dilation, straining of chewing muscle, and mouth opening. Final CGS median scores increased after castration (P < 0.001) for both non-restrained (M7 versus M2) and restrained (M6 versus M3) calves, indicating scale responsiveness. Final CGS median scores increased (P < 0.001) when calves were subjected to external factors before castration (M1 [baseline] versus M2 and M3). However, there was no difference (P > 0.05) in CGS median scores before and after sham castration, regardless of restraint (M3 versus M6, and M2 versus M7), indicating that the external factors may have reached a maximum effect. The CGS is composed of six FAUs, responsive to acute pain and can identify stress unrelated to pain.

Enhancing the Horse Grimace Scale (HGS): Proposed updates and anatomical descriptors for pain assessment

The use of grimace scales enables the clinical identification of changes in the facial expressions of animals caused by pain. The Horse Grimace Scale (HGS) is one such tool, comprising a pain coding system based on facial expressions and assessing six Facial Action Units (FAUs). Each FAU is accompanied by descriptions and anatomical details to assist the evaluator. However, the morphological descriptions for certain FAUs in the HGS are not sufficiently detailed, potentially hindering accurate interpretation. This study is an analytical investigation aimed at enhancing the morphoanatomical details in the HGS and providing raters with more comprehensive materials for pain evaluation in horses using this scale.To achieve this, detailed anatomical analyses were conducted using established references in veterinary anatomy. Initially, we propose substituting the term 'ear' with 'auricle' or 'pinna' and replacing 'area above the eye' with 'supraorbital region' for anatomical accuracy. Additionally, we introduce detailed morphoanatomical descriptions that identify specific landmarks, with the goal of ensuring more consistent application of the HGS and reducing interpretation variability. Furthermore, this study provides an explanation of the muscles involved in the investigated FAUs. These adjustments on the descriptions and evaluations remain unverified, however it is anticipated that the descriptive enhancements lead us to understand that higher interobserver reliability can be achieved for each of the FAUs.

Refining pain management in mice by comparing multimodal analgesia and NSAID monotherapy for neurosurgical procedures

While neurosurgical interventions are frequently used in laboratory mice, refinement efforts to optimize analgesic management based on multimodal approaches appear to be rather limited. Therefore, we compared the efficacy and tolerability of combinations of the non-steroidal anti-inflammatory drug carprofen, a sustained-release formulation of the opioid buprenorphine, and the local anesthetic bupivacaine with carprofen monotherapy. Female and male C57BL/6J mice were subjected to isoflurane anesthesia and an intracranial electrode implant procedure. Given the multidimensional nature of postsurgical pain and distress, various physiological, behavioral, and biochemical parameters were applied for their assessment. The analysis revealed alterations in Neuro scores, home cage locomotion, body weight, nest building, mouse grimace scales, and fecal corticosterone metabolites. A composite measure scheme allowed the allocation of individual mice to severity classes. The comparison between groups failed to indicate the superiority of multimodal regimens over high-dose NSAID monotherapy. In conclusion, our findings confirmed the informative value of various parameters for assessment of pain and distress following neurosurgical procedures in mice. While all drug regimens were well tolerated in control mice, our data suggest that the total drug load should be carefully considered for perioperative management. Future studies would be of interest to assess potential synergies of drug combinations with lower doses of carprofen.

Effects of pain on the facial expressions of goat kids

ABSTRACT Pain assessment is essential in laboratory and farm animals. Grimace scales have been used for this purpose since 2010. The aim of the present study was to investigate how pain (due to disease presence) affects the main facial expressions of goat kids. For this purpose, 60 goat kids aged 1 to 20 days were included in the study. An experienced veterinarian graded the animals’ pain based on his experience (pain 0, 1, and 2), and two photographs (frontal and lateral) were taken. The following measurements were recorded: Height and width of the palpebral fissure, mouth angle, angles of the nose in profile and front. Pain level affected the height (0.83 and 1.29 cm, pain level 0 and 2 respectively, p < 0.001) and width of the palpebral fissure (1.85 and 2.35 cm, pain level 0 and 2 respectively, p < 0.001), which increased at pain level 2. Thus, narrowing of the eye was not observed when pain level increased. The angle of the mouth increased at pain level 2 (39.2 and 41.0 degrees, pain level 0 and 2 respectively, p = 0.013), and the frontal angle of the nose decreased at pain level 2 (93.5 and 85.0 degrees, pain level 0 and 2 respectively, p = 0.009).

Pain Assessment in Cattle during Castration Using Facial Expressions as a Promising Tool

Abstract Pain is a significant welfare issue in farm animals including cattle, impacting their wellbeing and normal behaviors. The frequent occurrence of pain in cattle is due to husbandry procedures, injuries, and diseases. Cattle, as a prey species, avoid expressing pain which makes pain identification a challenging issue. Rapid pain recognition is crucial to its effective treatment. Several pain assessment tools have been developed, and some of them rely on observing alterations in animal behaviors such as activity, locomotion, feeding, play, and grooming. Moreover, changes in physiological parameters including blood biomarkers and heart rate have been used as indicators of pain. Furthermore, grimace scales, which rely on evaluating facial expressions of animals, have been developed and proven to be accurate pain assessment tool in multiple species. To reliably identify pain in cattle, it is preferred to employ a combination of diagnostic methods, as no single approach can stand alone in assessing pain accurately. Mitigation strategies become imperative when pain is anticipated during husbandry procedures such as castration. Prompt application of treatment strategies is paramount to avoid transition of acute to chronic pain which is hard to be treated. Information © The Author 2024

Role of induced nitric oxide synthases in orofacial nociception/discomfort after dental tooth bleaching with hydrogen peroxide

ObjectiveTo evaluate the role of induced nitric oxide synthase (iNOS) in nociception/orofacial discomfort in rats submitted to tooth whitening with hydrogen peroxide (H₂O₂). Design:Wistar rats were divided into three groups (n = 24/group): a sham group not submitted to whitening treatment, a saline group submitted to whitening treatment, and a test group submitted to whitening treatment and blockade of iNOS with aminoguanidine 50 mg/kg/day. After 24 and 48 h, and 7 days, the animals were euthanized to collect trigeminal ganglia and maxillae to histomorphometric analysis (size of neuronal bodies and percentage of pulp area filled by vessels) and behavior/nociception (Grimace scales, scratching and biting counting, weight loss and nociception assay). ANOVA-1- or − 2-way tests were used (p < 0.05, GraphPadPrism 5.0). Results:The aminoguanidine-treated group showed a reduction in nociceptive threshold in the masseteric region (p < 0.001), Grimace scale scores (p < 0.001), number of scratching (p = 0.011) and body mass loss (p = 0.007). After 24 and 48 h of tooth bleaching, the saline group showed a significant increase in the mean area of the blood vessels (p = 0.020) and iNOS immunostaining in odontoblasts (p = 0.002) and non-odontoblasts cells (p = 0.025). Aminoguanidine reversed both increases. Tooth bleaching reduced the mean area of neuronal bodies, and aminoguanidine significantly reversed it (p = 0.019), but an increase in GFAP immunostaining in neuronal bodies did not reduce after seven-days or after aminoguanidine treatment (p = 0.003). Conclusion:iNOS blockage by aminoguanidine plays an important role in nociception and orofacial discomfort by control of inflammation in dental pulp after tooth bleaching with hydrogen peroxide (H₂O₂) 35%.

USING SHEEP FACIAL GRIMACE SCALE, INFRARED THERMOGRAPHY AND CORTISOL HORMONE TO MEASURE PAIN IN SHEEP INFECTED WITH MASTITIS DISEASE

The purpose of this study was to apply sheep pain facial expression scales (SPFES) to sheep infected with mastitis, measuring peripheral temperatures and serum cortisol hormone. Twenty-three healthy ewes and 23 infected ewes with clinical mastitis were included in this research from January 2022 to April 2023. Images were taken from each ewe to detect facial grimace scales of ewes. Nasal, eye and ear temperatures were collected using infrared thermal imaging camera. Moreover, serum cortisol hormone was collected from the blood. It was found that there was a significant decrease (p&lt;0.01) in nasal, eye and ear temperatures of infected ewes in comparison to healthy ewes. Additionally, cortisol hormone was found significantly (p&lt;0.01) higher in infected ewes at day 1 and day 7 compared to non-infected animals. The changes in facial expressions for all the determined five regions were significantly (p&lt;0.01) different than healthy ewes. The total pain score was significantly (p&lt;0.05) higher on day 1 and day 7, compared to healthy ewes. It was concluded that sheep facial grimace scale was a useful and practical tool for measuring pain in sheep. In addition, cortisol from the serum and peripheral temperatures were found to be useful pain indicators.

Nesting behavior is associated with body weight and grip strength loss in mice suffering from experimental arthritis

Objective animal health evaluation is essential to determine welfare and discomfort in preclinical in vivo research. Body condition scores, body weight, and grimace scales are commonly used to evaluate well-being in murine rheumatoid arthritis (RA) and osteoarthritis experiments. However, nest-building, a natural behavior in mice, has not yet been evaluated in wild type (WT) or genetically modified rodents suffering from collagen antibody-induced arthritis (CAIA). To address this, we analyzed nesting behavior in WT mice, calcitonin gene-related peptide alpha-deficient (αCGRP-/-) mice, and calcitonin receptor-deficient (Calcr-/-) mice suffering from experimental RA compared to healthy control (CTRL) groups of the same genotypes. CAIA was induced in 10–12-week-old male mice, and clinical parameters (body weight, grip strength, clinical arthritis score, ankle size) as well as nesting behavior were assessed over 10 or 48 days. A slight positive association between the nest score and body weight and grip strength was found for animals suffering from CAIA. For the clinical arthritis score and ankle size, no significant associations were observed. Mixed model analyses confirmed these associations. This study demonstrates that clinical effects of RA, such as loss of body weight and grip strength, might negatively affect nesting behavior in mice. Assessing nesting behavior in mice with arthritis could be an additional, non-invasive and thus valuable health parameter in future experiments to monitor welfare and discomfort in mice. During severe disease stages, pre-formed nest-building material may be provided to animals suffering from arthritis.

An automated rat grimace scale for the assessment of pain.

Pain is a complex neuro-psychosocial experience that is internal and private, making it difficult to assess in both humans and animals. In pain research, animal models are prominently used, with rats among the most commonly studied. The rat grimace scale (RGS) measures four facial action units to quantify the pain behaviors of rats. However, manual recording of RGS scores is a time-consuming process that requires training. While computer vision models have been developed and utilized for various grimace scales, there are currently no models for RGS. To address this gap, this study worked to develop an automated RGS system which can detect facial action units in rat images and predict RGS scores. The automated system achieved an action unit detection precision and recall of 97%. Furthermore, the action unit RGS classifiers achieved a weighted accuracy of 81-93%. The system's performance was evaluated using a blast traumatic brain injury study, where it was compared to trained human graders. The results showed an intraclass correlation coefficient of 0.82 for the total RGS score, indicating that the system was comparable to human graders. The automated tool could enhance pain research by providing a standardized and efficient method for the assessment of RGS.

Facial Image-Based Automatic Assessment of Equine Pain

Recognition of pain in animals is essential for their welfare. However, since there is no verbal communication, this assessment depends solely on the ability of the observer to locate visible or audible signs of pain. The use of grimace scales is proven to be efficient in detecting the pain visually, but the assessment quality depends on the level of training of the assessor and the validity is not easily ensured. There is a clear need for automating the pain assessment process. This work provides a system for pain prediction in horses, based on grimace scales. The pipeline automatically determines the quantitative pose of the equine head and finds facial landmarks before classification, proposing a novel scale-normalisation approach for equine heads. The pain estimation is achieved for each facial region of interest separately, following the clinical pain estimation procedure. We introduce a database of horse images, annotated by professional veterinarians for training and assessment. We also propose a data augmentation method to alleviate the data scarcity issues, which relies on generating realistic 3D equine face models based on 2D annotated images. We show that the data augmentation method improves the performance of both quantitative pose estimation and landmark detection. Our results establish a strong baseline for automatic equine pain estimation.

Review of Rat (Rattus norvegicus), Mouse (Mus musculus), Guinea pig (Cavia porcellus), and Rabbit (Oryctolagus cuniculus) Indicators for Welfare Assessment.

The monitoring and assessment of animals is important for their health and welfare. The appropriate selection of multiple, validated, and feasible welfare assessment indicators is required to effectively identify compromises or improvements to animal welfare. Animal welfare indicators can be animal or resource based. Indicators can be collated to form assessment tools (e.g., grimace scales) or animal welfare assessment models (e.g., 5 Domains) and frameworks (e.g., 5 Freedoms). The literature contains a wide variety of indicators, with both types needed for effective animal welfare assessment; however, there is yet to be an ideal constellation of indicators for animal-based welfare assessment in small mammals such as guinea pigs (Cavia Porcellus), mice (Mus musculus), rabbits (Oryctolagus cuniculus), and rats (Rattus norvegicus). A systematic review of grey and peer-reviewed literature was performed to determine the types of animal-based welfare indicators available to identify and assess animal health and welfare in these small mammals maintained across a wide variety of conditions. The available indicators were categorised and scored against a selection of criteria, including potential ease of use and costs. This review and analysis aim to provide the basis for further research into animal welfare indicators for these species. Future applications of this work may include improvements to animal welfare assessments or schemes, guiding better management, and implementing future strategies to enable better animal welfare.

Development and validation of a cynomolgus macaque grimace scale for acute pain assessment

Cynomolgus macaques may undergo surgical procedures for scientific and veterinary purposes. Recognition and assessment of pain using validated tools is a necessary first step for adequately managing pain in these primates. Grimace scales are one means of assessing the occurance of acute pain using action units such as facial expressions and posture. The aim of this study was to create and validate a Cynomolgus Macaque Grimace Scale (CMGS). Cynomolgus macaques (n = 43) were video recorded before and after a surgical procedure. Images were extracted from videos at timepoints at which breakthrough pain might be expected based on analgesic pharmacokinetics. Using the CMGS images were scored by 12 observers blinded to animal identification, times, and conditions. To validate the tool, detailed behavioral analyses emphasizing changes to baseline activity ethograms were compared to grimace scores. Four action units were identified related to potential pain including orbital tightening, brow lowering, cheek tightening, and hunched posture. The CMGS tool was found to have moderate inter- (ICCaverage action unit mean ± SD: 0.67 ± 0.28) and good intra- (ICCsingle mean ± SD: 0.79 ± 0.14) observer reliability. Grimace scores increased significantly (p < 0.0001) in the first four post-operative timepoints compared to baseline, correlating with behavioral findings (rho range = 0.22–0.35, p < 0.001). An analgesic intervention threshold was determined and should be considered when providing additional pain relief. The CMGS was shown to be a reliable and valid tool; however, more research is needed to confirm external validity. This tool will be highly valuable for refining analgesic protocols and acute peri-procedural care for cynomolgus macaques.

Measurement properties of grimace scales for pain assessment in nonhuman mammals: a systematic review.

Facial expressions of pain have been identified in several animal species. The aim of this systematic review was to provide evidence on the measurement properties of grimace scales for pain assessment. The protocol was registered (SyRF#21-November-2019), and the study is reported according to the PRISMA guidelines. Studies reporting the development, validation, and the assessment of measurement properties of grimace scales were included. Data extraction and assessment were performed by 2 investigators, following the COnsensus-based Standards for the Selection of Health Measurement INstruments guidelines. Six categories of measurement properties were assessed: internal consistency, reliability, measurement error, criterion and construct validity, and responsiveness. Overall strength of evidence (high, moderate, and low) of each instrument was based on methodological quality, number of studies, and studies' findings. Twelve scales for 9 species were included (mice, rats, rabbits, horses, piglets, sheep or lamb, ferrets, cats, and donkeys). Considerable variability regarding their development and measurement properties was observed. The Mouse, Rat, Horse and Feline Grimace Scales exhibited high level of evidence. The Rabbit, Lamb, Piglet and Ferret Grimace Scales and Sheep Pain Facial Expression Scale exhibited moderate level of evidence. The Sheep Grimace Scale, EQUUS-FAP, and EQUUS-Donkey-FAP exhibited low level of evidence for measurement properties. Construct validity was the most reported measurement property. Reliability and other forms of validity have been understudied. This systematic review identified gaps in knowledge on the measurement properties of grimace scales. Further studies should focus on improving psychometric testing, instrument refinement, and the use of grimace scales for pain assessment in nonhuman mammals.

The Utility of Grimace Scales for Practical Pain Assessment in Laboratory Animals.

Simple SummaryGrimace scales for laboratory animals were first reported ten years ago. Yet, despite their promise as pain assessment tools it appears that they have not been implemented widely in animal research establishments for clinical pain assessment. We discuss potential reasons for this based on the knowledge gained to date on their use and suggest avenues for further research, which might improve uptake of their use in laboratory animal medicine.Animals’ facial expressions are widely used as a readout for emotion. Scientific interest in the facial expressions of laboratory animals has centered primarily on negative experiences, such as pain, experienced as a result of scientific research procedures. Recent attempts to standardize evaluation of facial expressions associated with pain in laboratory animals has culminated in the development of “grimace scales”. The prevention or relief of pain in laboratory animals is a fundamental requirement for in vivo research to satisfy community expectations. However, to date it appears that the grimace scales have not seen widespread implementation as clinical pain assessment techniques in biomedical research. In this review, we discuss some of the barriers to implementation of the scales in clinical laboratory animal medicine, progress made in automation of collection, and suggest avenues for future research.

Grimace Scores: Tools to Support the Identification of Pain in Mammals Used in Research.

Simple SummaryThe ability to identify and assess pain is paramount in animal research to address the ‘refinement’ principle of the 3Rs (Reduction, Refinement, Replacement), satisfy public acceptability of animal use in research and address ethical and legal obligations. Many physiological, behavioural and physical pain assessments are commonly used, but all have their limitations. Grimace scales are a promising adjunctive behavioural pain assessment technique in some mammalian species used in research. This paper reviews the extant literature studying pain assessment techniques in general, and grimace scales specifically, in animal research. The results indicate that the grimace scale technique is simple and able to be used spontaneously at the ‘cage side’, is non-invasive in its application, highly repeatable, reliable between interobserver and intraobserver applications and easy to train and use. The use of grimace scales should be more frequently considered as an important parameter of interest in research and animal wellbeing. Further research into the use of grimace scales is required to develop scales for a wider range of animal species, increase applicability in studies specifically related to pain assessment and for further validation of the technique.The 3Rs, Replacement, Reduction and Refinement, is a framework to ensure the ethical and justified use of animals in research. The implementation of refinements is required to alleviate and minimise the pain and suffering of animals in research. Public acceptability of animal use in research is contingent on satisfying ethical and legal obligations to provide pain relief along with humane endpoints. To fulfil this obligation, staff, researchers, veterinarians, and technicians must rapidly, accurately, efficiently and consistently identify, assess and act on signs of pain. This ability is paramount to uphold animal welfare, prevent undue suffering and mitigate possible negative impacts on research. Identification of pain may be based on indicators such as physiological, behavioural, or physical ones. Each has been used to develop different pain scoring systems with potential benefits and limitations in identifying and assessing pain. Grimace scores are a promising adjunctive behavioural technique in some mammalian species to identify and assess pain in research animals. The use of this method can be beneficial to animal welfare and research outcomes by identifying animals that may require alleviation of pain or humane intervention. This paper highlights the benefits, caveats, and potential applications of grimace scales.

Facial expressions of pain in cats: the development and validation of a Feline Grimace Scale

Grimace scales have been used for pain assessment in different species. This study aimed to develop and validate the Feline Grimace Scale (FGS) to detect naturally-occurring acute pain. Thirty-five client-owned and twenty control cats were video-recorded undisturbed in their cages in a prospective, case-control study. Painful cats received analgesic treatment and videos were repeated one hour later. Five action units (AU) were identified: ear position, orbital tightening, muzzle tension, whiskers change and head position. Four observers independently scored (0–2 for each AU) 110 images of control and painful cats. The FGS scores were higher in painful than in control cats; a very strong correlation with another validated instrument for pain assessment in cats was observed (rho = 0.86, p < 0.001) as well as good overall inter-rater reliability [ICC = 0.89 (95% CI: 0.85–0.92)], excellent intra-rater reliability (ICC > 0.91), and excellent internal consistency (Cronbach’s alpha = 0.89). The FGS detected response to analgesic treatment (scores after analgesia were lower than before) and a cut-off score was determined (total pain score > 0.39 out of 1.0). The FGS is a valid and reliable tool for acute pain assessment in cats.

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.

Grading facial expression is a sensitive means to detect grimace differences in orofacial pain in a rat model

Although pre-clinical models of pain are useful for defining relationships between biological mechanisms and pain, common methods testing peripheral sensitivity do not translate to the human pain experience. Facial grimace scales evaluate affective pain levels in rodent models by capturing and scoring spontaneous facial expression. But, the Rat Grimace Scale (RGS) has not assessed the common disorder of temporomandibular joint (TMJ) pain. A rat model of TMJ pain induced by jaw loading (1 hr/day for 7 days) was used to investigate the time course of RGS scores and compare them between different loading magnitudes with distinct peripheral sensitivity profiles (0N–no sensitivity, 2N–acute sensitivity, 3.5N–persistent sensitivity). In the 3.5N group, RGS is elevated over baseline during the loading period and one day after loading and is correlated with peripheral sensitivity (ρ = −0.48, p = 0.002). However, RGS is not elevated later when that group exhibits peripheral sensitivity and moderate TMJ condylar cartilage degeneration. Acutely, RGS is elevated in the 3.5N loading group over the other loading groups (p < 0.001). These findings suggest that RGS is an effective tool for detecting spontaneous TMJ pain and that spontaneous pain is detectable in rats that develop persistent TMJ sensitivity, but not in rats with acute resolving sensitivity.

Can grimace scales estimate the pain status in horses and mice? A statistical approach to identify a classifier.

Pain recognition is fundamental for safeguarding animal welfare. Facial expressions have been investigated in several species and grimace scales have been developed as pain assessment tool in many species including horses (HGS) and mice (MGS). This study is intended to progress the validation of grimace scales, by proposing a statistical approach to identify a classifier that can estimate the pain status of the animal based on Facial Action Units (FAUs) included in HGS and MGS. To achieve this aim, through a validity study, the relation between FAUs included in HGS and MGS and the real pain condition was investigated. A specific statistical approach (Cumulative Link Mixed Model, Inter-rater reliability, Multiple Correspondence Analysis, Linear Discriminant Analysis and Support Vector Machines) was applied to two datasets. Our results confirm the reliability of both scales and show that individual FAU scores of HGS and MGS are related to the pain state of the animal. Finally, we identified the optimal weights of the FAU scores that can be used to best classify animals in pain with an accuracy greater than 70%. For the first time, this study describes a statistical approach to develop a classifier, based on HGS and MGS, for estimating the pain status of animals. The classifier proposed is the starting point to develop a computer-based image analysis for the automatic recognition of pain in horses and mice.

Analgesic Effect of Local Infiltration of the Surgical Wound Containing Levobupivacaine, Ibuprofen, and Epinephrine in Rats Undergoing Laparotomy.

Inappropriate use of drugs and their combinations for analgesics has made it difficult to determine the optimal drug combinations for pain mangement. To reduce postoperative pain effectively and safely. Laparotomy was performed in an adult rat under isoflurane anesthesia. During surgery, the surgical wounds were infiltrated with 50 μL solution containing 0.3% w/v levobupivacaine, 2 mg/mL ibuprofen, and 8 mg/mL epinephrine (treatment group) over the sutured muscle wound before skin closing, and compared to infiltration of that of the normal saline (vehicle group). The 10-fold higher dose of the same combination of medications was injected systemically as a control. Postoperative pain assessed by rodent grimace scales scoring. One-way ANOVA following Dunnett multiple comparisons test was used at 95% of confidence level. There was decreased pain for the treatment group (p = 0.025, q = 4.527) and the control group (p = 0.031, q = 4.178) only 24 h after the end of the successful infiltration. The rodent GS scale scoring showed the fall in pain was started within three hours post-surgery in the treatment group. There was decreased pain in the treatment group (p = 0.048, q = 3.527) and the control group (p = 0.043, q = 3.891) only as compared to vehicle group 24 h after the end of the successful infiltration. The infiltration of the surgical wound with levobupivacaine, ibuprofen, and epinephrine combination was effective in the healing of wounds after laparotomy.