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The creation of species-specific valid tools for pain assessment is essential to recognize pain and determine the requirement and efficacy of analgesic treatments. This study aimed to assess behaviour and investigate the validity and reliability of an acute pain scale in pigs undergoing orchiectomy. Forty-five pigs aged 38±3 days were castrated under local anaesthesia. Behaviour was video-recorded 30 minutes before and intermittently up to 24 hours after castration. Edited footage (before surgery, after surgery before and after rescue analgesia, and 24 hours postoperatively) was analysed twice (one month apart) by one observer who was present during video-recording (in-person researcher) and three blinded observers. Statistical analysis was performed using R software and differences were considered significant when p<0.05. Intra and inter-observer agreement, based on intra-class correlation coefficient, was good or very good between most observers (>0.60), except between observers 1 and 3 (moderate agreement 0.57). The scale was unidimensional according to principal component analysis. The scale showed acceptable item-total Spearman correlation, excellent predictive and concurrent criterion validity (Spearman correlation ≥ 0.85 between the proposed scale versus visual analogue, numerical rating, and simple descriptive scales), internal consistency (Cronbach's α coefficient >0.80 for all items), responsiveness (the pain scores of all items of the scale increased after castration and decreased after intervention analgesia according to Friedman test), and specificity (> 95%). Sensitivity was good or excellent for most of the items. The optimal cut-off point for rescue analgesia was ≥ 6 of 18. Discriminatory ability was excellent for all observers according to the area under the curve (>0.95). The proposed scale is a reliable and valid instrument and may be used clinically and experimentally to assess postoperative acute pain in pigs. The well-defined cut-off point supports the evaluator´s decision to provide or not analgesia.

Abstract Background There is an association between overweight status and life span in kenneled dogs, but a similar association has not been reported for pet dogs. Objectives To examine the effects of being overweight in middle age on the life span of neutered client-owned dogs. Animals Fifty-thousand seven-hundred eighty seven middle-aged neutered client-owned dogs attending a network of approximately 900 veterinary hospitals across North America. Methods Retrospective case-control study. For each of 12 breeds, groups of dogs aged between 6.5 and 8.5 years were identified as being in “overweight” or “normal” body condition. Within each breed and sex, differences in life span between dogs in normal body condition and overweight body condition in the 2 groups were then analyzed by Cox proportional hazards models. Results For all breeds, instantaneous risk of death for dogs in overweight body condition was greater than those in normal body condition throughout the age range studied, with hazard ratios ranging from 1.35 (99.79% confidence interval [CI] 1.05-1.73) for German Shepherd dog to 2.86 (99.79% CI 2.14-3.83) for Yorkshire Terrier. In all breeds, median life span was shorter in overweight compared with normal weight dogs, with the difference being greatest in Yorkshire Terriers (overweight: 13.7 years, 99.79% CI 13.3-14.2; normal: 16.2 years, 99.79% CI 15.7-16.5) and least in German Shepherd dogs (overweight: 12.1 years, 99.79% CI 11.8-12.4; normal: 12.5 years, 99.79% CI 12.2-12.9). Conclusions and Clinical Importance Veterinary professionals should consider promoting healthy body condition for dogs, particularly from midlife onward.

Surgical castration of males is carried out on a large scale in the US swine industry and the pain resulting from this procedure can be assessed using the Unesp-Botucatu pig composite acute pain scale (UPAPS). We aim to propose a short version of UPAPS based on the behaviors best-ranked by a random forest algorithm. We used behavioral observations from databases of surgically castrated pre-weaned and weaned pigs. We trained a random forest algorithm using the pain-free (pre-castration) and painful (post-castration) conditions as target variable and the 17 UPAPS pain-altered behaviors as feature variables. We ranked the behaviors by their importance in diagnosing pain. The algorithm was refined using a backward step-up procedure, establishing the Short UPAPS. The predictive capacity of the original and short version of the UPAPS was estimated by the area under the curve (AUC). In refinement, the algorithm with the five best-ranked behaviors had the lowest complexity and predictive capacity equivalent to the algorithm with all behaviors. The AUC of Short UPAPS (89.62%) was statistically equivalent ( p  = 0.6828) to that of UPAPS (90.58%). In conclusion, the proposed Short UPAPS might facilitate the implementation of a standard operating procedure to monitor and diagnose acute pain post-castration in large-scale systems.

Abstract Background Androgen-dependent cystinuria has been described in intact male dogs. Castration has been recommended to reduce urinary cystine excretion. Hypothesis/Objectives Urinary cystine/creatinine concentration will significantly decrease in dogs with suspected androgen-dependent (Type III) cystinuria after castration and will be associated with a lack of sonographic evidence of urolithiasis. Animals Six intact male dogs with cystine uroliths. Methods Prospective, observational study. Urinary amino acid/creatinine concentrations and abdominal ultrasound were evaluated at baseline (T0), 30 days (T30), and 90 days (T90) after castration and urolith removal. No dietary change was recommended unless urolith recurrence was noted. DNA was evaluated for the cystinuria-associated marker for Type III cystinuria. Results The median decline in urinary cystine/creatinine (uCys/creat) concentration from T0 to T30 was 577 nmol/mg (IQR, 415–969; p = 0.03), from T0 to T90 was 798 nmol/mg (IQR, 580–989; p = 0.03) and from T30 to T90 was 34 nmol/mg (IQR, 19.5–443; p = 0.03). Two dogs had small (2.5–3 mm) uroliths at T30, which resolved in one of them at T90. Both dogs with ultrasonographic evidence of recurrence at T30 were positive for the cystinuria-associated genetic marker, although no signs of lower urinary tract disease were noted in either dog. Three of 6 enrolled dogs were homozygous positive for the genetic marker. Conclusions and Clinical Importance Castration should be considered for intact male dogs with cystine urolithiasis.

Bovine pain assessment relies on validated behavioral scales related to normal and pain-related behaviors. This study investigated the reliability and applicability of real-time and video-recorded pain assessment, and their agreement, in young, adult bulls undergoing surgical castration. Ten Nelore and nine Angus bulls underwent general anesthesia and surgical castration. Three-minute real-time observations and simultaneous videos were recorded at − 48 h (M0), before sedation, under fasting (M1), after surgery, 3 h after sternal recumbency (M2), after rescue analgesia (M3) and at 24 h (M4). Animals received morphine (after M2), dipyrone (after M3), and flunixin meglumine after surgical castration (M4). Two trained evaluators assessed real-time (n = 95) and video-recorded time-points (n = 95) using the Unesp-Botucatu Cattle Pain Scale (UCAPS). Both assessment methods inferred ‘very good’ reliability (≥ 0.81) with minimal bias, however, video-recorded assessment (4.33 ± 2.84) demonstrated slightly higher scores compared to real-time (3.08 ± 2.84). The results from this study suggest that UCAPS can be used in real-time or video-recorded to assess pain and guide analgesic therapy in cattle.

Abstract Background The majority of cystine uroliths occur in intact male dogs. Androgen-dependent (Type III) cystinuria is considered the most common cause. Objectives Identify dog breeds in which castration is likely to decrease the risk of cystine uroliths, the potential effect of delaying castration on cystine urolith formation, and urolith recurrence frequency. Animals Records of 5477 dogs with cystine uroliths and comparison groups without cystine uroliths (263 938 dogs with non-cystine uroliths and 44 491 dogs from a hospital population). Methods In this case-control study, odds ratios and 95% confidence intervals (CIs) were calculated to identify breeds where the proportion of intact males with cystine uroliths was higher than that of intact males without cystine uroliths. The proportions of intact males forming cystine uroliths before 12, 24, and 36 months of age were calculated. Cystine urolith recurrence rates were assessed by breed in male dogs. Results Dogs with cystine uroliths were 99% male. Across 60 breeds, the median proportion of male cystine urolith formers that were intact was 98% (range, 40%–100%). When compared with dogs without cystine uroliths, intact males were overrepresented in cystine urolith formers in all breeds except 8 (Akita, Belgian Malinois, Brussels Griffon, Cane Corso, Coonhound, Newfoundland, Scottish Terrier, and Silky Terrier). Diagnosis occurred before 36 months of age in 28% (n = 1328) of intact male cystine urolith formers. Cystine uroliths recurred in 5.0% (n = 255) of males; 81% (n = 207) were intact males. Conclusions and Clinical Importance Androgens likely play a role in the development of cystine uroliths across many dog breeds.

Oral meloxicam is labelled for reducing pain and inflammation associated with castration in cattle in Canada, however, subcutaneous meloxicam is only labelled for pain associated with dis-budding and abdominal surgery. The aim of this project was to determine the pharmacokinetic profile of oral (PO; 1.0 mg/kg BW) and subcutaneous meloxicam (SC; 0.5 mg/kg BW), and to assess the effect of meloxicam on physiological and behavioural indicators of pain associated with knife castration in 7-8 month old calves. Twenty-three Angus crossbred beef calves (328 ± 4.4 kg BW) were randomly assigned to two treatments: PO n = 12 or SC n = 11 administration of meloxicam immediately before knife castration. Physiological parameters included salivary and hair cortisol, substance P, haptoglobin, serum amyloid-A, weight, complete blood count, scrotal and rectal temperature. Behavioural parameters included standing and lying behaviour, pen behaviour and feeding behaviour. Data were analyzed using PROC GLIMMIX (SAS), with repeated measures using mixed procedures including treatment as a fixed effect and animal and pen as a random effect. The pharmacokinetic profile of the drug including area under the curve, volume of distribution and clearance was greater (P < 0.05) in PO than SC calves. After surgery, substance P concentrations, white blood cell counts (WBC), weight and lying duration were greater (P < 0.05) in PO than SC calves, while scrotal circumference was lower (P < 0.05) in PO calves than SC calves. Although statistical differences were observed for pharmacokinetic, physiological and behavioural parameters differences were small and may lack biological relevance.

To accurately assess pain and support broadly-based analgesic protocols to mitigate swine pain, it is imperative to develop and validate a species-specific pain scale. The objective of this study was to investigate the clinical validity and reliability of an acute pain scale (UPAPS) adapted for newborn piglets undergoing castration. Thirty-nine male piglets (five days of age, 1.62 ± 0.23 kg BW) served as their own control, were enrolled in the study and underwent castration in conjunction with an injectable analgesic administered one-hour post-castration (flunixin meglumine 2.2 mg/kg IM). An additional 10, non-painful female piglets were included to account for the effect of natural behavioral variation by day on pain scale results. Behavior of each piglet was video recorded continuously at four recording periods (24 h pre-castration, 15 min post-castration, 3 and 24 h post-castration). Pre- and post-operative pain was assessed by using a 4-point scale (score 0–3) including the following six behavioral items: posture, interaction and interest in surroundings, activity, attention to the affected area, nursing, and miscellaneous behavior. Behavior was assessed by two trained blinded observers and statistical analysis was performed using R software. Inter-observer agreement was very good (ICC = 0.81). The scale was unidimensional based on the principal component analysis, all items except for nursing were representative ( r s ≥ 0.74) and had excellent internal consistency (Cronbach’s alpha ≥ 0.85). The sum of scores were higher in castrated piglets post-procedure compared to pre-procedure, and higher than in non-painful female piglets confirming responsiveness and construct validity, respectively. Scale sensitivity was good when piglets were awake (92.9%) and specificity was moderate (78.6%). The scale had excellent discriminatory ability (area under the curve > 0.92) and the optimal cut-off sum for analgesia was 4 out of 15. The UPAPS scale is a valid and reliable clinical tool to assess acute pain in castrated pre-weaned piglets.

Three experiments were conducted to evaluate the effects of band and knife castration on behavioral and physiological indicators of chronic pain in beef calves at 3 different ages (36 calves/age group): 1 wk of age (Exp. 1, 4 ± 1.1 d of age, 43 ± 1.1 kg BW), 2 mo of age (Exp. 2, 63 ± 2.3 d of age, 92 ± 1.7 kg BW), and 4 mo of age (Exp. 3, 125 ± 4.6 d of age, 160 ± 3.4 kg BW). In each experiment calves were randomly assigned to either sham (CT), band (BA) or knife (KN) castration. Experiments 1, 2, and 3 ended when the testicles of banded calves had sloughed off (68, 49, and 42 d, respectively). Animal BW and rectal temperature were recorded weekly over the experimental period. Salivary cortisol, substance P, haptoglobin, scrotal area temperature using infrared thermography, visual evaluation of swelling (5-point scale), and gait stride length were collected on d -1 and immediately before castration and weekly thereafter until the end of the study. Hair samples were collected 1 d prior to and 28 d after castration and at the end of the study for cortisol concentration. Standing and lying behaviors were recorded over a 28-d period immediately after castration. No differences ( > 0.10) were observed in salivary cortisol, substance P, haptoglobin, or hair cortisol among castration methods for any of the 3 ages. No changes in behavior were observed in calves castrated at 1 wk or 2 mo of age. In 4 mo-old-calves, BA spent less time lying ( < 0.01) than CT and KN calves. Also, the average duration of lying time for BA calves was greater ( < 0.05) than for CT calves. Both, 1-wk- and 2-mo-old calves had inflammation in the scrotal area lasting 7 d after KN castration, whereas inflammation was observed for up to 14 d in 4-mo-old calves. Swelling in BA calves lasted for 21 to 28 d in the 2 younger groups of calves, whereas in 4-mo-old calves swelling was observed until d 35 postcastration. Knife- and band-castrated calves did not exhibit indicators of chronic pain or distress when the procedures were performed in calves younger than 2 mo of age. Therefore, pain mitigation should be used when castrating to improve animal welfare, especially when castrations are performed in calves older than 2 mo of age independent of the method of castration.

Abstract The present study aimed to investigate whether acute pain associated with castration and tail docking of male piglets may modulate the expression of salivary microRNAs (miRNAs) and to explore their potential use as biomarkers. Thirty-six healthy 4-d-old piglets (Hermitage × Duroc) were randomly assigned to three groups: the first group (12 piglets) has been pretreated with anesthetic and anti-inflammatory drugs (ANA) and then castrated and tail docked; the second one (12 piglets) has been castrated and tail docked without any drugs (CONV); the third one (12 piglets) has been only handled (SHAM). Saliva was collected 10 min before (control group) and 30 to 45 min after the procedures. Salivary cortisol has been quantified. The expression concentrations of seven miRNAs, namely miR-19b, miR-27b-3p, miR-215, miR-22-3p, miR-155-5p, hsa-miR-365-5p, and hsa-miR-204, were measured and assessed as potential biomarkers of pain by quantitative Polimerase Chain Reaction using TaqMan probes. The area under the receiver operating curve (AUC) was used to evaluate the diagnostic performance of miRNAs. The concentration of salivary cortisol increased after treatment in CONV and ANA, while no significant variation was observed in the SHAM group. The comparative analysis demonstrated that the concentrations of salivary miR-19b (P = 0.001), miR-27b (P = 0.042), and miR-365 (P < 0.0001) were significantly greater in CONV as compared with pretreatment. The AUC of pretreatment vs. CONV and CONV vs. ANA were excellent for miR-19b and miR-365 and fair for miR-27b. Combining two miRNAs, namely miR-19b and miR-365, in a panel increased the efficiency of distinguishing between pre- and post-treatment groups. No differences have been identified between SHAM and ANA groups. mRNA potential targets of differentially expressed-miRNA were investigated, and genes related to pain and inflammation were identified: miR-19b potentially modulates TGF-beta and focal adhesion pathways, miR-365 regulates cytokines expression (i.e., IL-1, Tumor Necross Factor-alpha, and IL-8 cytokine), and miR-27b regulates macrophage inflammatory protein pathways (i.e., MIP1-beta). In conclusion, we demonstrated that the abundance of miR-19b, miR-27b, and miR-365 increases in the saliva of piglets castrated and tail docked without the administration of pain-relieving drugs. Further studies are needed to assess their potential during routine husbandry procedures and to extend their assessment in other stressful events, such as weaning or chronic pain.