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Elevated ethanol concentrations in yeast affect the plasma membrane. The plasma membrane in yeast has many lipid-protein complexes, such as Pma1 (MCP), Can1 (MCC), and the eisosome complex. We investigated the response of eisosomes, MCPs, and membraneless structures to ethanol stress. We found a correlation between ethanol stress and proton flux with quick acidification of the medium. Moreover, ethanol stress influences the symporter expression in stressed cells. We also suggest that acute stress from ethanol leads to increases in eisosome size and SG number: we hypothesized that eisosomes may protect APC symporters and accumulate an mRNA decay protein in ethanol-stressed cells. Our findings suggest that the joint action of these factors may provide a protective effect on cells under ethanol stress.

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Sheep pain is an animal welfare issue monitored based on behavioral responses, including appetite. Dominant (alpha) males have priority for accessing limited feed resources, however, the effects of pain on feed interest in members of a group with defined social hierarchy are unknown. Our objective was to investigate effects of acute post-orchiectomy pain on alpha rams' interest in accessing a limited feed resource. Eighteen rams were randomly housed in pens of 3 rams. After acclimation, the first 5-d (consecutive) battery of a behavior test was performed. In this test, 180 g of the regular diet concentrate was placed in a portable trough in the center of the pen; this feed was supplemental to the diet and represented a limited, albeit strongly preferable feed resource. Rams were filmed for 5 min after the feed introduction. Hierarchical levels (alpha, beta, and gamma) were defined based on the social hierarchical index according to higher initiator and lower receptor agonistic behaviors from the social network analyses. After 15 d, a second 5-d behavioral test battery was repeated. On the following day, alpha rams were castrated. Flunixin meglumine was given immediately before surgery and a final behavioral test was performed 8 h post-orchiectomy, concurrent with an expected peak in postoperative pain. For all recordings, the latency, frequency, and duration of time that each ram had its mouth inside the feed trough were recorded, and the Unesp-Botucatu sheep acute pain scale pain scale (USAPS) was applied. The social hierarchical index was highest in alpha rams, followed by beta and gamma. The pain scores were statistically equivalent across the 11 evaluation days for beta and gamma rams, whereas there was an increase in the final evaluation for alpha. There was no difference in latency, frequency, and duration between alpha, beta, and gamma rams across evaluations. We concluded that acute post-orchiectomy pain did not decrease alpha rams' interest in accessing limited feed. Routine feeding offers a valuable chance to detect pain-related behavior using the USAPS in rams. However, dominance may confound appetite-related behaviors in assessing acute pain, as alpha rams' interest in limited feed remained unaffected by the pain.

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The aim of the present study was to compare affiliative behaviours of owned and shelter cats directed to human in a novel environment after a brief temporary absence of the person. A sample of 20 owned and 20 shelter animals were individually tested in a Secure Base Test, with three 2-min episodes: 1) cat accompanied by a person who sits on the floor inside a circle, 2) the person leaves and the animal is left alone, 3) the person returns, and sits inside the circle again. Three categories were used for coding videotapes of experimental sessions: (1) inside circle, (2) allo-rubbing and (3) tail up. Shelter animals showed more proximity maintenance and affiliative signs with a person than owned animals. Our findings suggested similarities between the secure base effect in cats and in human children: cats seek proximity and maintain contact with a person, displaying affiliative signs to the person, especially after being left alone frightened by an unfamiliar environment.

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Pigs are raised on a global scale for commercial or research purposes and often experience pain as a by product of management practices and procedures performed. Therefore, ensuring pain can be effectively identified and monitored in these settings is critical to ensure appropriate pig welfare. The Unesp-Botucatu Pig Composite Acute Pain Scale (UPAPS) was validated to diagnose pain in pre-weaned and weaned pigs using a combination of six behavioral items. To date, statistical weighting of supervised and unsupervised algorithms was not compared in ranking pain-altered behaviors in swine has not been performed. Therefore, the aim of this study was to verify if supervised and unsupervised algorithms with different levels of complexity can improve UPAPS pain diagnosis in pigs undergoing castration. The predictive capacity of the algorithms was evaluated by the area under the curve (AUC). Lower complexity algorithms containing fewer pain-altered behaviors had similar AUC (90.1-90.6) than algorithms containing five (89.18-91.24) and UPAPS (90.58). In conclusion, utilizing a short version of the UPAPS did not influence the predictive capacity of the scale, and therefore it may be easier to apply and be implemented consistently to monitor pain in commercial and experimental settings.

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Introduction: Horse racing is a major sport practiced worldwide. The environment to which horses are exposed during race meetings can influence their behavior. However, to the best of our knowledge, a method for assessing a horse's response to its surroundings during the pre- and post-race periods has not yet been reported. This study aimed to create a standard list of descriptors for use in a qualitative behavioral assessment (QBA) focused on assessing the emotional expressivity of horses before and after racing events. Materials and methods: Seventy pre- or post-race 30-second videos of horses were randomly selected from our database of 700 videos. A panel of 8 experienced equine sports medicine specialist veterinarians watched a 60 min presentation on QBA. The panel then watched all videos randomly, simultaneously, individually, continuously, and without any verbal interaction, describing the descriptors related to the emotional expressivity of the horse after each video using a method known as free-choice profiling (FCP). Results: The initial selection of descriptors was based on those indicated by more than one evaluator in the same video, or descriptors with more than 20 occurrences. The second selection was performed based on the content validity index (CVR) to select the descriptors retained in the previous step. Another panel of six veterinarians scored each of the descriptors retained for content validity on a visual scale. Interobserver reliability was estimated using the intraclass correlation coefficient (ICC) and its respective 95% confidence intervals (CI). A natural language processing (NLP) algorithm was used to analyze the behavior (positive or negative polarity) of the descriptors based on the lexicoPT package of R software. Discussion/Conclusion: NLP analysis considered the descriptors "agitated," "troubled," "restless" and "irritated" to have a negative polarity, while "focused," "relaxed" and "peaceful" had a positive polarity. In the principal component analysis (PCA), descriptors in a negative state were associated with each other and inversely associated with descriptors in a positive state. We conclude with a fixed list of descriptors to be used in a QBA to assess emotional and welfare expressivity in racehorses' pre- and post-race environments.

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BACKGROUND: Confinement of cattle imposes spatial restrictions and predisposes to aversive social encounters that can lead to contusions, wounds, pain, stress, fright, and reduced productivity. Although endogenous testosterone concentrations are linked to agonistic dominance behaviors in males, it is unknown whether decreased blood testosterone concentrations after castration alter social hierarchy rank in Nelore bulls. Therefore, in this study, we investigated the impact of the surgical would inflammation post-orchiectomy on social dynamics in a group of Nelore bulls (Bos indicus). Fourteen Nelore (Bos indicus) bulls were castrated and assessed pre- and post-surgically. Parameters evaluated were agonistic (mounting, headbutting, and fighting) and affiliative (head-play) behavior, plasma testosterone concentrations, average daily weight gain (ADG), and a score for severity of post-surgical infection. Exploratory statistics included social network analysis (SNA), hierarchy rank delta (Δ), and principal component analysis (PCA). Furthermore, statistical inferences included the Wilcoxon test, multiple logistic regression models, and Spearman's correlation. RESULTS: The social dynamic of Nelore bulls was modified after castration based on the findings of the SNA and the PCA. The moderate correlation between the postoperative inflammation level with the Δ, and the significant effect of this level in the logistic model post-castration were partially attributed to effects of pain on social relations. CONCLUSIONS: Our findings suggest the severity of post-surgical inflammation, which has an association with pain intensity, was closely associated with changes in the social hierarchy.

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We aimed to develop and validate the Unesp-Botucatu goat acute pain scale (UGAPS). Thirty goats (5 negative controls and 25 submitted to orchiectomy) were filmed for 7 min at the time points 24 h before and 2 h, 3 h (1 h after analgesia), and 24 h after orchiectomy. After content validation, according to an ethogram and literature, four blind observers analyzed the videos randomly to score the UGAPS, repeating the same assessment in 30 days. According to the confirmatory factor analysis, the UGAPS is unidimensional. Intra- and interobserver reliability was very good for all raters (Intraclass correlation coefficient ≥85%). Spearman's correlation between UGAPS versus VAS was 0.85 confirming the criterion validity. Internal consistency was 0.60 for Cronbach's α Cronbach and 0.67 for McDonald's ω. The item-total correlation was acceptable for 80% of the items (0.3-0.7). Specificity and sensitivity based on the cut-off point were 99% and 90%, respectively. The scale was responsive and demonstrated construct validity shown by the increase and decrease of scores after surgery pain and analgesia, respectively. The cut-off point for rescue analgesia is ≥3 of 10, with an area under the curve of 95.27%. The UGAPS presents content, criterion, and construct validities, responsiveness, and reliability to assess postoperative pain in castrated goats.

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Recently, the Unesp-Botucatu sheep acute pain scale (USAPS) was created, refined, and psychometrically validated as a tool that offers fast, robust, and simple application. Evidence points to an improvement in pain diagnosis when the importance of the behavioral items of an instrument is statistically weighted; however, this has not yet been investigated in animals. The objective was to investigate whether the implementation of statistical weightings using machine learning algorithms improves the USAPS discriminatory capacity. A behavioral database, previously collected for USAPS validation, of 48 sheep in the perioperative period of laparoscopy was used. A multilevel binomial logistic regression algorithm and a random forest algorithm were used to determine the statistical weights and classify the sheep as to whether they needed analgesia or not. The quality of the classification, estimated by the area under the curve (AUC) and its 95% confidence interval (CI), was compared between the USAPS versions. The USAPS AUCs weighted by multilevel binomial logistic regression (96.59 CI: [95.02-98.15]; p = 0.0004) and random forest algorithms (96.28 CI: [94.17-97.85]; p = 0.0067) were higher than the original USAPS AUC (94.87 CI: [92.94-96.80]). We conclude that the implementation of statistical weights by the two machine learning algorithms improved the USAPS discriminatory ability.

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Our primary goal was to investigate human ability to recognize basic emotions from only the eyes of dogs in comparison to the whole face. Simultaneously, we replicated and extended previous research (Bloom et al., 2021), while validating American canine emotional facial expression photographs cross-culturally to Brazil. Participants (N = 120) viewed behaviorally-anchored photographs of three breeds. Half the participants in each condition (faces or eyes-only) viewed two-word forced choice items while the other half viewed four-word forced choice items. Participants identified target emotions from images of both dogs' faces and eyes-only at a higher rate than chance. Fear was accurately recognized more than the other emotions. When dogs are afraid, they open their eyes and expose the sclera, a conspicuous signal. Emotion identification accuracy was highest for the Rhodesian Ridgeback, who is similar in morphology to common Brazilian stray dogs (Vira-Latas Carmelo). We conjectured that Brazilians were more accustomed to seeing dogs with the Rhodesian Ridgeback morphology than the erect-eared breeds, thus increasing accuracy for this breed. Further studies with additional dog morphologies are desirable. In addition to research interest, our Canine Eyes task has the potential to become a test of individual differences in Theory of Mind with clinical applications.

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This study aimed to validate a scale for assessing acute pain in donkeys. Forty-four adult donkeys underwent castration after sedation with intravenous (IV) xylazine, induction with guaifenesin and thiopental IV, local anesthetic block, and maintenance with isoflurane. The scale was constructed from a pilot study with four animals combined with algetic behaviors described for equines. After content validation, the scale was evaluated in 40 other donkeys by three blinded and one reference evaluator, by means of edited videos referring to the preoperative and postoperative periods: before anesthesia, 3-4 h after recovery from anesthesia, 5-6 h after recovery from anesthesia (2 h after analgesia with flunixin-1.1 mg/kg, dipyrone-10 mg/kg, and morphine-0.2 mg/kg) IV, and 24 h after recovery. Content validity, sensitivity, specificity, and responsiveness of behaviors were investigated to refine the scale. Intra- and inter-evaluator reliabilities were investigated by the weighted kappa coefficient, criterion validity by comparing the scale with the visual analog scale (VAS), internal consistency by Cronbach's α coefficient, item-total correlation by the Spearman coefficient, and intervention point for rescue analgesic by the receiver operating characteristics curve and Youden index. The scale showed very good intra-evaluator reliability (0.88-0.96), good to moderate (0.56-0.66) inter-evaluator reliability, responsiveness for all items, good criterion validity vs. VAS (0.75), acceptable internal consistency (0.64), adequate item-total correlation, except for head position and direction, and according to the principal component analysis, good association among items. The accuracy of the point for rescue analgesic was excellent (area under the curve = 0.91). The rescue analgesic score was ≥ 4 of 11 points. The scale can diagnose and quantify acute pain in donkeys submitted to castration, as the instrument is reliable and valid, with a defined intervention analgesic score.