Deep Learning for Face Detection and Pain Assessment in Japanese macaques (Macaca fuscata).

Facial expressions have increasingly been used to assess emotional states in mammals. The recognition of pain in research animals is essential for their well-being and leads to more reliable research outcomes. Automating this process could contribute to early pain diagnosis and treatment. Artificial neural networks have become a popular option for image classification tasks in recent years due to the development of deep learning. In this study, we investigated the ability of a deep learning model to detect pain in Japanese macaques based on their facial expression. Thirty to 60 min of video footage from Japanese macaques undergoing laparotomy was used in the study. Macaques were recorded undisturbed in their cages before surgery (No Pain) and one day after the surgery before scheduled analgesia (Pain). Videos were processed for facial detection and image extraction with the algorithms RetinaFace (adding a bounding box around the face for image extraction) or Mask R-CNN (contouring the face for extraction). ResNet50 used 75% of the images to train systems; the other 25% were used for testing. Test accuracy varied from 48 to 54% after box extraction. The low accuracy of classification after box extraction was likely due to the incorporation of features that were not relevant for pain (for example, background, illumination, skin color, or objects in the enclosure). However, using contour extraction, preprocessing the images, and fine-tuning, the network resulted in 64% appropriate generalization. These results suggest that Mask R-CNN can be used for facial feature extractions and that the performance of the classifying model is relatively accurate for nonannotated single-frame images.

Housing relocation does not have to induce a significant stress response in captive Japanese macaques (Macaca fuscata).

BACKGROUND: Previous studies suggest that housing relocation may be stressful for captive non-human primates. Our study investigated the stress levels of Japanese macaques during a housing relocation by measuring their daily fecal cortisol metabolites, which are often used as an indicator of stress. METHODS: Ten adult Japanese macaques, single-housed for research purposes, were relocated to a new facility. Fecal samples were collected daily for 7 days. Cortisol metabolite concentrations were determined via enzyme immunoassay. RESULTS: No significant differences in cortisol metabolite levels were found in 7 days, but concentration levels showed that the highest median was associated to the relocation day. CONCLUSIONS: The minimal cortisol metabolite increase suggests that there was a slight activity increase in the hypothalamic-pituitary-adrenal axis. Techniques encouraging cooperation of the monkeys, the short time duration of the relocation, and consistency in the environment may have contributed to the minimal stress levels observed.

Investigating subtle changes in facial expression to assess acute pain in Japanese macaques.

Changes in facial expression provide cues for assessing emotional states in mammals and may provide non-verbal signals of pain. This study uses geometric morphometrics (GMM) to explore the facial shape variation in female Japanese macaques who underwent experimental laparotomy. Face image samples were collected from video footage of fourteen macaques before surgery and 1, 3, and 7 days after the procedure. Image samples in the pre-surgical condition were considered pain-free, and facial expressions emerging after surgery were investigated as potential indicators of pain. Landmarks for shape analysis were selected based on the underlying facial musculature and their corresponding facial action units and then annotated in 324 pre-surgical and 750 post-surgical images. The expression of pain is likely to vary between individuals. Tightly closed eyelids or squeezed eyes and lip tension were the most commonly observed facial changes on day 1 after surgery (p < 0.01974). A good overall inter-rater reliability [ICC = 0.99 (95% CI 0.75-1.0)] was observed with the method. The study emphasizes the importance of individualized assessment and provides a better understanding of facial cues to pain for captive macaque care.

Comparative study between S-(+)-ketamine­midazolam and fentanyl­droperidol in black-tufted marmosets (Callithrix penicillata) / Estudo comparativo entre S-(+)cetamina-midazolam e fentanil-droperidol em saguis-de-tufo-preto (Callithrix penicillata)

Sedative and antinociceptive effects of two anesthetic protocols in black-tufted marmosets were compared in this study. Twenty-six marmosets underwent chemical immobilization for physical examination, blood sampling, tattooing, and microchipping. Animals were randomly treated with S-(+)-ketamine (10 mg/kg) and midazolam (1 mg/kg) (KM) or fentanyl (12.5 µg/kg) and droperidol (625 µg/kg) (FD) given by intramuscular injection. Heart and respiratory rates were recorded. Sedation, antinociception, muscle relaxation, posture, auditory, and visual responses were evaluated using a scoring system. Sedation in KM was achieved faster (p < 0.001) and lasted for a shorter period of time (p = 0.0009). KM was similar to FD in its cardiorespiratory effects, auditory and visual responses. Both protocols promoted adequate sedation to allow manipulation. Animals in KM assumed lateral recumbency while animals in FD maintained a quadrupedal posture during evaluation. FD produced less intense sedation and muscle relaxation but a higher degree of antinociception compared to KM and is suitable for procedures that require analgesia in black-tufted marmosets.(AU)

O presente estudo comparou os efeitos cardiorrespiratórios, sedativos e antinociceptivos de dois protocolos anestésicos em saguis-de-tufo-preto (Callithrix penicillata). Vinte e seis saguis foram submetidos à contenção química para exame físico, coleta de sangue, tatuagem de identificação e microchip. Os animais foram tratados aleatoriamente com a associação de S-(+)-cetamina (10 mg/kg) e midazolam (1 mg/kg) (KM) ou fentanil (12,5 µg/kg) e droperidol (625 µg/kg) (FD), administrados por injeção intramuscular. Foram avaliadas frequência cardíaca, frequência respiratória, sedação, antinocicepção, relaxamento muscular, postura e resposta ao estímulo auditivo e visual. A sedação em KM foi alcançada mais rapidamente (p <0,001) e teve um tempo hábil mais curto (p = 0,0009). KM foi semelhante a FD nos efeitos cardiorrespiratórios, respostas auditivas e visuais. Os dois protocolos promoveram sedação adequada para manipulação. Os animais do grupo KM permaneceram em decúbito lateral durante a avaliação, enquanto os animais em FD mantiveram postura quadrupedal. FD resultou em sedação e relaxamento muscular de menor intensidade, porém com maior escore de antinocicepção em comparação com KM, sendo adequada para procedimentos que requerem analgesia em saguis-de-tufo-preto.(AU)

Vasectomy of golden-headed lion tamarins (Leontopithecus chrysomelas) using local anesthesia after sedation with dexmedetomidine-ketamine-meperidine.

The study evaluated the combination of ketamine, dexmedetomidine, and meperidine for vasectomy in golden-headed lion tamarins. Lidocaine infiltration was required for intraoperative analgesia and atipamezole was used at the end of the procedure. The protocol promoted satisfactory sedation and analgesia with a short recovery time in tamarins.

Primate veterinarians' knowledge and attitudes regarding pain in macaques.

BACKGROUND: Assessment of pain in macaques is challenging. The aims of this study were (1) to investigate current knowledge and attitudes of primate veterinarians concerning acute pain in macaques; (2) to synthesise current knowledge and opinion to facilitate pain assessment. A primary question of interest was whether more confident individuals differ in their knowledge and attitudes from less-confident individuals. METHODS: An online survey was conducted amongst primate veterinarians serving both laboratories and zoos/sanctuaries. The questionnaire consisted of demographic information, attitudes towards pain, pain rating and analgesics, pain recognition and confidence in recognising pain and sources of information used. RESULTS AND CONCLUSIONS: There was generally good use of analgesia by respondents. More confident individuals reported that they recognise pain both behaviourally and in facial expressions, rated all pain signs more highly and used more analgesics. Specialist support networks aimed at increasing veterinarian confidence in macaque pain assessment could be beneficial.

Early reversal of ketamine/dexmedetomidine chemical immobilization by atipamezole in golden-headed lion tamarins (Leontopithecus chrysomelas).

BACKGROUND: A smooth and rapid recovery from anesthesia allowing safe release is desirable, especially for wild species. This study describes the clinical effects of the combination of dexmedetomidine and ketamine and the partial reversal with atipamezole in golden-headed lion tamarins. METHODS: Dexmedetomidine 10 µg kg-1 and ketamine 15 mg kg-1 were administered to 45 golden-headed lion tamarins undergoing vasectomy. Following surgery, animals were assigned to three groups: control (SAL; 0.9% NaCl), atipamezole 20 µg kg-1 (ATI20), and atipamezole 40 µg kg-1 (ATI40). RESULTS AND CONCLUSIONS: All animals presented great scores of sedation and muscle relaxation during the procedure. Recovery in the control group was smooth and uneventful. Salivation, muscle tremors, and head movements were observed in ATI 20 and ATI40. The administration of atipamezole did not change total recovery times (ATI20 69 ± 23 minutes; ATI40 72 ± 45 minutes; SAL 57 ± 23 minutes).

Comparison between total intravenous anesthesia with propofol and intermittent bolus of tiletamine-zolazepam in capuchin monkey (Sapajus apella) / Comparação entre anestesia total intravenosa com propofol e bolus intermitente de tiletamina-zolazepam em macaco-prego (Sapajus apella)

Dissociative anesthesia results in stressful and long recovery periods in monkeys and use of injectable anesthetics in medical research has to be refined. Propofol has promoted more pleasure wake up from anesthesia. The objectives of this study were to investigate the use of intravenous anesthetic propofol, establishing the required infusion rate to maintain surgical anesthetic level and comparing it to tiletamine-zolazepam anesthesia in Sapajus apella. Eight healthy capuchin monkeys, premedicated with midazolam and meperidine, were anesthetized with propofol (PRO) or tiletamine-zolazepam (TZ) during 60 minutes. Propofol was infused continually and rate was titrated to effect and tiletamine-zolazepam was given at 5mg/kg IV bolus initially and repeated at 2.5mg/kg IV bolus as required. Cardiopulmonary parameters, arterial blood gases, cortisol, lactate and quality and times to recovery were determined. Recovery quality was superior in PRO. Ventral recumbency (PRO = 43.0±21.4 vs TZ = 219.3±139.7 min) and normal ambulation (PRO = 93±27.1 vs TZ = 493.7±47.8 min) were faster in PRO (p<0.05). Cardiopulmonary effects did not have marked differences between groups. Median for induction doses of propofol was 5.9mg/kg, varying from 4.7 to 6.7mg/kg, Mean infusion rate was 0.37±0.11mg/kg/min, varying during the one-hour period. In TZ, two animals required three and five extra doses. Compared to tiletamine-zolazepam, minor post-anesthetic adverse events should be expected with propofol anesthesia due to the faster and superior anesthetic recovery. (AU)

A anestesia dissociativa em primatas resulta em recuperação anestésica lenta e estressante, e, portanto, o uso de anestesia injetável em pesquisas médicas precisa ser refinado. Por outro lado, o propofol promove recuperação mais suave. Os objetivos desse estudo foram investigar o uso do anestésico intravenoso propofol, estabelecer a taxa de infusão contínua necessária para manter anestesia cirúrgica, e comparar tal técnica com a dissociativa tiletamina-zolazepam em Sapajus apella. Oito macacos-prego saudáveis foram pré-medicados com midazolam e meperidina, e posteriormente anestesiados com propofol (PRO) ou tiletamina-zolazepam (TZ) durante 60 minutos. O propofol foi administrado em infusão contínua, e a taxa foi titulada ao efeito, já a tiletamina-zolazepam foi administrada em 5mg/kg IV como bolus inicial, e repiques de 2,5mg/kg IV conforme necessário. Os parâmetros cardiopulmonares, hemogasometria arterial, cortisol, e lactato, além da qualidade e duração da recuperação anestésica foram determinados. A qualidade da recuperação anestésica foi superior em PRO. O tempo para atingir decúbito ventral (PRO = 43,0±21,4 vs TZ = 219,3±139,7 min) e ambulação normal (PRO = 93±27,1 vs TZ = 493,7±47,8 min) foram mais rápidos em PRO (p<0,05). As variáveis cardiopulmonares não diferiram entre os grupos. A mediana para dose de indução com propofol foi de 5,9mg/kg, variando de 4,7 a 6,7mg/kg. A taxa de infusão contínua média de propofol foi de 0,37±0,11mg/kg/min, variando ao longo dos 60 minutos. Em TZ, dois animais necessitaram de três e cinco repiques. Comparado à tiletamina-zolazepam, menos efeitos adversos pós-anestésicos devem ser esperados com o propofol, devido à recuperação mais suave e rápida.(AU)

Continuous infusion of propofol or intermittent bolus of tiletamine-zolazepam in squirrel monkeys (Saimiri sciureus).

OBJECTIVE: To investigate an infusion of propofol for anesthesia in comparison to tiletamine-zolazepam anesthesia, evaluating physiological variables and recovery in squirrel monkeys. STUDY DESIGN: Prospective non-blinded randomized study. ANIMALS: Eight healthy squirrel monkeys (Saimiri sciureus), aged 3 years and weighing 0.340-0.695 kg. METHODS: Premedication was intramuscular midazolam (0.5 mg) and meperidine (4 mg). Anesthesia was induced with intravenous (IV) propofol (4 mg kg(-1)  minute(-1) ) and maintained with propofol starting at 0.4 mg kg(-1)  minute(-1) (PRO, n = 4) or IV tiletamine-zolazepam (5 mg kg(-1) ) and maintained with supplementary doses of TZ (TZ, n = 4). Cardiopulmonary variables were measured continuously. Arterial blood gases and lactate concentration were measured at the end of anesthesia. Quality and times of recovery were determined. Repeatedly measured data for significant differences were tested between groups with t-test and within groups by anova. RESULTS: Median time for induction of anesthesia in PRO was 180 seconds. Mean maintenance infusion rate of propofol was 0.43 ± 0.05 mg kg(-1)  minute(-1) , varying during the 1 hour period. One monkey died after administration of TZ; others required 1, 4, or 8 supplemental doses. Cardiopulmonary variables were similar between groups, but hypotension was recorded. Recovery times to ventral recumbency in PRO (32 ± 17 minutes) and TZ (84 ± 11 minutes) and normal ambulation in PRO (58 ± 22 minutes) and TZ (358 ± 109minutes) were significantly different (p < 0.05). Recovery quality was superior in PRO, with less ataxia and fewer unsuccessful attempts to stand. Lactate concentration was not different between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Cardiopulmonary variables were similar between protocols, aside from the higher incidence of hypotension in PRO, indicating that further studies with a larger number of animals are required. Compared to tiletamine-zolazepam, propofol anesthesia provided faster and superior anesthetic recovery in these animals.

Continuous infusion of propofol or intermittent bolus of tiletamine-zolazepam in feline night monkeys (Aotus infulatus).

BACKGROUND: Although commonly used, dissociative anesthesia results in unsatisfactory and long recovery periods. The intravenous anesthetic propofol seems to be the most advantageous solution. The objectives of this study were to investigate the required infusion rate of propofol to maintain surgical anesthesia and to compare it to tiletamine-zolazepam in Aotus infulatus. METHODS: Eight healthy feline night monkeys were anesthetized with propofol (PRO) or tiletamine-zolazepam (TZ) during 60 minutes. Cardiopulmonary parameters, arterial blood gases and lactate and quality and times to recovery were determined. RESULTS: Mean infusion rate of propofol was 0.53 ± 0.10 mg/kg/minute. Cardiopulmonary effects did not show marked differences between groups. Times for hanging, ventral recumbency, and normal ambulation were lower in PRO. Establishment of desirable anesthetic depth was easier, recovery quality was superior, and lactate levels were lower in PRO. CONCLUSIONS: Compared with tiletamine-zolazepam, minor post-anesthetic adverse events should be expected with propofol anesthesia, as well as faster and better anesthetic recovery.

Effects of the Crotalus durissus terrificus snake venom on hepatic metabolism and oxidative stress.

Snake venoms present different action mechanisms because of their complex composition, represented mainly by toxins and enzymes. This work aimed to investigate the effects of the Crotalus durissus terrificus(Cdt) venom in the liver. Wistar rats were inoculated intraperitoneally with saline (control) or Cdt venom. After 3, 4, or 6 h, the following parameters were analyzed: (a) hepatic function, (b) oxidative stress parameters, and (c) the metabolism of alanine in the isolated perfused liver. Plasma activities of alanine aminotransferase and aspartate aminotransferase and hepatic glutathione S-transferase and catalase presented significant elevation in rats inoculated with 300 µg ⋅ kg(-1) Cdt venom. Liver lipoperoxidation was enormously increased by venom doses of 100, 200, and 300 µg ⋅kg(-1) , whereas glutathione S-transferase was not changed. Perfused livers from rats inoculated with 1500 µg ⋅kg(-1) venom showed increased production of lactate, pyruvate, and ammonia when alanine was the metabolic substrate. These results demonstrate that the Cdt venom can produce several changes in hepatocytes. The causes of the changes are possibly related to the disequilibrium in the redox homeostasis but also to specific needs of the poisoned organism, for example, an increased supply of lactate and pyruvate in response to an increased activity of the Cori cycle.