Routine Husbandry Procedures Research Articles

Training Laboratory Rabbits to Refine Routine Husbandry Procedures.

Non-aversive handling and training techniques for laboratory animals are required to facilitate experimental and routine husbandry procedures, improving both animal welfare and scientific quality. Clicker training was utilized to develop training protocols for rabbits to refine stressful routine husbandry procedures usually associated with lifting (i.e., being picked up from the floor)/restraining (i.e., being held in the arms of a human) them. Thirteen female New Zealand White rabbits were trained over three weeks. All rabbits learned the predefined goal behaviors: they followed the target stick, jumped onto the weighing scale, entered a transport box, and reared while placing their front paws onto the trainer's hand. In addition, ten animals jumped from the floor onto the sitting trainer's lap and allowed the trainer to lift their paws off the surface while sitting on the trainer's lap. For some individuals, the protocols had to be adapted by additional interim steps. At the end of the training, the rabbits reliably showed the expected goal behaviors, even after short and long training breaks. With few exceptions, a familiar person other than the trainer could elicit the goal behaviors from the rabbits (generalization), though further sessions were required for generalization. In the voluntary approach test, the rabbits preferred interacting with the trainer in the 1st trial but spent as much time with an unfamiliar person as with the trainer in the 2nd trial. The behavioral observations suggested that picking the rabbits up with the transport box, as described in the protocol, instead of restraining them with the scruff of their neck and lifting them on the arm, was less aversive. All in all, the training protocols were feasible and can serve as a refinement strategy in laboratory animal facilities. In the interest of animal welfare, the training protocols should be applied wherever possible.

Comparison of Tricaine Methanesulfonate (MS-222) and Alfaxalone Anesthesia in Zebrafish (Danio rerio).

The research use of zebrafish has risen exponentially over the past decade while anesthetic options have remained largely unchanged.6 ricaine methanesulfonate (MS-222) is widely accepted as an anesthetic for routine husbandry procedures, however it has limitations and safety concerns. 11 A greater variety of effective anesthetic options for surgical procedures would be advantageous for the research community. Adult zebrafish were randomly assigned to one of the following groups (n = 10, 5 males and 5 females): 200 mg/L MS-222; 6-, 10-, 13-, and 16-mg/L alfaxalone, and control. All zebrafish in the MS-222 group reached a surgical plane of anesthesia within 95 ± 32 s. By contrast, only 2 of 10, 1 of 10, 0 of 10, and 0 of 4 of the 6, 10, 13, and 16 mg/L alfaxalone groups, respectively, reached a surgical plane of anesthesia within the allotted 10-min period. Recovery time was also significantly slower in the alfaxalone groups as compared with MS-222, with some fish taking greater than 10 min to recover. In addition, 33 of 34 zebrafish (the 16 mg/L group was not completed due to safety concerns) in the alfaxalone groups lost opercular movements for greater than one minute during their anesthetic event and had to be removed to the recovery tank. The results demonstrated that alfaxalone was unable to provide a reliable and safe surgical plane of anesthesia at any of the drug doses tested. Therefore, we recommend alfaxalone not be used as an anesthetic for painful procedures on zebrafish and conclude that MS-222 remains a more viable anesthetic for immersion anesthesia in zebrafish.

Arterial oxygenation and acid-base status before and during oxygen supplementation in captive European bison (Bison bonasus) immobilized with etorphine-acepromazine-xylazine.

Chemical immobilization of captive European bison (Bison bonasus) is often required for veterinary care, transportation, or husbandry practices playing an important role in conservation breeding and reintroduction of the species. We evaluated the efficiency and physiological effects of an etorphine-acepromazine-xylazine combination with supplemental oxygen in 39 captive European bison. Animals were darted with a combination of 1.4 mg of etorphine, 4.5 mg of acepromazine, and 20 mg of xylazine per 100 kg based on estimated body mass. Arterial blood was sampled on average 20 min after recumbency and again 19 min later and analyzed immediately with a portable i-STAT analyzer. Simultaneously, heart rate, respiratory rate, and rectal temperature were recorded. Intranasal oxygen was started after the first sampling at a flow rate of 10 mL.kg-1.min-1 of estimated body mass until the end of the procedure. The initial mean partial pressure of oxygen (PaO2) was 49.7 mmHg with 32 out of 35 sampled bison presenting with hypoxemia. We observed decreased respiratory rates and pH and mild hypercapnia consistent with a mild respiratory acidosis. After oxygen supplementation hypoxemia was resolved in 21 out of 32 bison, but respiratory acidosis was accentuated. Bison immobilized with a lower initial drug dose required supplementary injections during the procedure. We observed that lower mean rectal temperatures during the immobilization event were significantly associated with longer recovery times. For three bison, minor regurgitation was documented. No mortality or morbidity related to the immobilizations were reported for at least 2 months following the procedure. Based on our findings, we recommend a dose of 0.015 mg.kg-1 etorphine, 0.049 mg.kg-1 acepromazine, and 0.22 mg.kg-1 xylazine. This dose reduced the need for supplemental injections to obtain a sufficient level of immobilization for routine management and husbandry procedures in captive European bison. Nevertheless, this drug combination is associated with development of marked hypoxemia, mild respiratory acidosis, and a small risk of regurgitation. Oxygen supplementation is strongly recommended when using this protocol.

Post Mortem Study on the Effects of Routine Handling and Manipulation of Laboratory Mice.

Routine handling and manipulation of laboratory mice are integral components of most preclinical studies. Any type of handling and manipulation may cause stress and result in physical harm to mice, potentially leading to unintended consequences of experimental outcomes. Nevertheless, the pathological effects of these interventions are poorly documented and assumed to have a negligible effect on experimental variables. In that context, we provide a comprehensive post mortem overview of the main pathological changes associated with routine interventions (i.e., restraint, blood drawing, and intraperitoneal injections) of laboratory mice with an emphasis on presumed traumatic osteoarticular lesions. A total of 1000 mice from various studies were included, with 864 animals being heavily manipulated and 136 being handled for routine husbandry procedures only. The most common lesions observed were associated with blood collection or intraperitoneal injections, as well as a series of traumatic osteoarticular lesions likely resulting from restraint. Osteoarticular lesions were found in 62 animals (61 heavily manipulated; 1 unmanipulated) with rib fractures and avulsion of the dens of the axis being over-represented. Histopathology and micro-CT confirmed the traumatic nature of the rib fractures. While these lesions might be unavoidable if mice are manipulated according to the current standards, intentional training of research personnel on appropriate mouse handling and restraint techniques could help reduce their frequency and the impact on animal wellbeing as well as study reproducibility.

Moving toward Fear-Free Husbandry and Veterinary Care for Horses.

Husbandry and veterinary procedures have the potential to generate fear and stress in animals. In horses, the associated responses can pose a significant safety risk to the human personnel involved in the procedure, as well as to the animal itself. Traditionally, physical restraint, punishment, and/or threat of an aversive, have been the most common strategies used to achieve compliance from the horse. However, from a welfare perspective, this is less than ideal. This approach also has the potential for creating a more dangerous response from the horse in future similar situations. When caring for companion animals, and captive animals within zoological facilities, there has been a steady transition away from this approach, and toward strategies aimed at reducing fear and stress during veterinary visits and when undertaking routine husbandry procedures. This review discusses the current approaches to horse care and training, the strategies being used in other animal sectors, and potential strategies for improving human safety, as well as the horse's experience, during husbandry and veterinary procedures.

393 Young Scholar Award Talk: Investigation of Pain and Analgesic Strategies in Cattle Undergoing Painful Husbandry Procedures and Disease Conditions

Abstract Cattle are stoic by nature and have long been subject to evolutionary pressure to mask pain to avoid becoming prey. Quantifying pain via biomarkers allows researchers to capture changes that are not easily detected by the human eye. Negative public perception of routine husbandry practices such as dehorning and castration is growing, increasing the need for the development of practices to relieve pain and suffering in cattle. The objective was to quantify pain associated with induced bacterial pneumonia, scoop and hot-iron dehorning, surgical castration, and hot-iron branding, and to investigate analgesic effectiveness. Calves were experimentally inoculated with Mannheimia haemolytica, scoop or cautery dehorned, surgically castrated, or hot-iron branded. Outcome variables were collected at baseline and following husbandry procedures and included serum cortisol; infrared thermography (IRT); mechanical nociceptive threshold (MNT); substance P; kinematic gait analysis; a visual analog scale; chute defense and behavior scoring; clinical illness score; computerized lung score; average activity and rumination; prostaglandin E2 metabolite; plasma serum amyloid A and rectal temperature. Responses were analyzed using repeated measures with calf nested in treatment designated as a random effect, and treatment, time, and their interaction designated as fixed effects. A combination of reduced activity levels, decreased force on calves’ right front limb, and increased visual analog scale pain scores all support that bacterial pneumonia in cattle is painful (P ≤ 0.01). Differences in right front force were observed in calves challenged with M. haemolytica and treated with flunixin transdermal (96.5 kg) and those given a placebo (85.5 kg), indicating that flunixin transdermal may attenuate specific pain biomarkers in cattle with respiratory disease (P < 0.01). Administration of a bupivacaine liposome suspension block at the time of dehorning and castration was as effective at controlling pain as a multi-modal approach of lidocaine and meloxicam (as evidenced by treatment differences in MNT, cortisol and gait analysis (P ≤ 0.04)). Oral meloxicam administration at branding reduced IRT differences from the branding and control site and reduced lying bouts (P < 0.01). Breed and sex effects were observed across a wide range of biomarkers and should be investigated in future pain studies. The need for long-acting analgesic options for cattle that demonstrate pain alleviation across multiple biomarkers is apparent and would be beneficial to alleviating pain from routine husbandry procedures like dehorning, castration, and branding, as well as painful disease conditions such as bovine respiratory disease.

Features of the functional state of the organism of sheep under stress

The stresses that arise during routine animal husbandry procedures, such as milking, shearing, weighing, loading and hoof care, are an actual problem in animal husbandry. The components of economic damage under stress are a decrease in overall resistance and productivity, deterioration in product quality, an increase in labor costs and funds per unit of production, an increase in animal morbidity and mortality. The studies were carried out on the basis of Belozernoe farm in the Salsky district, the Rostov region. A complex of clinical and laboratory studies was performed on 18 sheep (10 Romanov and 8 Tsigay breeds) aged 2 to 3 years, before and after a planned routine shearing. Stress in sheep caused by planned shearing was clinically manifested by general depression, in some cases, subfebrile fever, moderate tachypnea and tachycardia. It was shown that stress was accompanied by anemia, the development of erythropenia and leukopenia. Immunocompetent cells in the blood of experimental animals during stress development were analyzed. Severe lymphocytopenia was found to occur after a haircut. In addition, in the blood of experimental animals under stress, a tendency to a decrease in T-total cells was noted, which was accompanied by an increase in 0-cells, which was significant in sheep of the Tsigay breed (p0.05). When analyzing the main immunoregulatory T cells in sheep under stress, a significant increase in T-suppressors was revealed with a relatively stable level of T-helpers in the blood studied. Thus, a significant (p0.05) increase in the level of T-suppressors in sheep of Romanov and Tsigay breeds was recorded; it was by 1.15 and 1.19 times higher compared with the initial data. It should be noted that we did not find any significant differences between animals of Romanov and Tsigay breeds in stress development after shearing. The obtained data should be taken into account in the further search for methods of stress correction in sheep.

초보자 승마운동 시 완충고삐 사용을 통한 말의 스트레스 완화에 관한 연구

During equestrian exercise, communication between riders and horses is very important, and riders’ appropriate signals without any disturbance for the horses’ proper movements are required. Therefore, horses may get different stress depending on the rider’s riding balance, posture, and proficiency. This study was conducted to confirm the difference in the stress of horses between the beginner’s riding and the expert’s riding, additionally, to prove the effectiveness of buffer rein reducing stress even when beginners ride. Average heart rate (HR) was measured for five minutes in stages with a walk, and HR variability (HRV) were measured for three minutes in stages with both rising trot and sitting trot. HRV variables include root mean square of successive differences (RMSSD), standard deviation of normal to normal interval (SDNN), and ratio of low to high frequency components of a continuous series of heartbeats (low-frequency [LF]/high-frequency [HF]). The study involved 6 horses that underwent routine husbandry procedures. The average HR did not show a difference among groups in all exercise stages includes walk, rising trot, and sitting trot. RMSSDs showed lower values for the sitting trot when the beginner rides compared to the expert’s riding (p < 0.05), which means the stress was more increased when beginners ride. On the other hand, when beginners ride using buffer reins, the RMSSD was increased in all exercise stages compared to the case of using regular reins. SDNN was increased when the beginners ride using buffer reins in the rising trot (p < 0.05), which means the stress was more relieved. Ratio of low to high frequency components of a continuous series of heartbeats was not different among groups, but it showed decreased value when beginners use buffer reins rather than regular reins. In this study, it was confirmed that beginners induce more stress on horses than experts during riding exercise and that the use of the buffer reins by beginners more reduces the riding stress of horses compared to when they use regular reins.

The Neuroimmune Interface and Chronic Pain Through the Lens of Production Animals.

Communication between the central nervous system (CNS) and the immune system has gained much attention for its fundamental role in the development of chronic and pathological pain in humans and rodent models. Following peripheral nerve injury, neuroimmune signaling within the CNS plays an important role in the pathophysiological changes in pain sensitivity that lead to chronic pain. In production animals, routine husbandry procedures such as tail docking and castration, often involve some degree of inflammation and peripheral nerve injury and consequently may lead to chronic pain. Our understanding of chronic pain in animals is limited by the difficulty in measuring this pathological pain state. In light of this, we have reviewed the current understanding of chronic pain in production animals. We discuss our ability to measure pain and the implications this has on animal welfare and production outcomes. Further research into the neuroimmune interface in production animals will improve our fundamental understanding of chronic pain and better inform human clinical pain management and animal husbandry practices and interventions.

Which factors can influence the perception of pain by veterinarians and animal scientists from Brazil?

Society and animal professionals are increasingly concerned about the pain management during routine husbandry procedures in farm animals. In this paper, we aim to identify the factors that affect Brazilian veterinarians’ and animal scientists’ perceptions about the level of pain cattle experience during castration and horn removal. An online survey was conducted with these professionals, resulting in a sample size of 147 participants. Data were analyzed by descriptive statistics, Chi-squares tests, Fisher exact tests, and Mann-Whitney U tests. The results of descriptive statistics showed that the most frequently, the most and the least painful technique quoted, for castration, was scalpel, burdizzo, and immunocastration, respectively; and for horn removal, hot iron (for newborn and suckled calves) and saw/wires (for adult cattle), knife, and caustic paste, respectively. Women assessed cattle pain higher than men professionals for castration in newborn calves. Higher mean scores for the pain perception were associated to painful indicators: heart and/or respiration rate during castration in newborn and suckled calves; and defensive movements during horn removal in adult cattle. Those respondents who were more likely to perceived high level of pain also scored higher pain for (i) newborn calves when toxic agent or immunocastration were used to castrate; (ii) suckled calves dehorned/disbudded with chemical methods or dehorners; and (iii) all cattle ages when surgical methods or burdizzo were used for castration and amputation techniques or hot iron, for horn removal. Professionals that perceived higher level of pain in both painful procedures agreed more with the statements “Farm animals are not as sensitive to pain as pets” and “If there are no defensive movements, there is no need for pain mitigation use” than those that perceived lower level of pain. The pain estimation is subjective, but our results illustrate the importance of the management and individual/professional aspects on pain perception. Finally, our study highlights that improvement in pain perception by veterinary and animal scientists regarding castration and horn removal in cattle is required.

An Investigation into the Perceptions of Veterinarians towards Perioperative Pain Management in Calves.

Simple SummaryDespite developments in animal welfare science regarding perioperative pain management in calves (Bos taurus), there are concerns that current knowledge has not been adopted in practice. Given that the perceptions of veterinarians have implications for how the welfare needs of calves are assessed and managed in practice, this study sought to quantify veterinary perceptions towards perioperative pain management in calves, including barriers to its use and whether demographic differences may influence those perceptions. A nationwide survey was electronically distributed to veterinarians registered with the Veterinary Council of New Zealand. Veterinarians largely associated multimodal pain management with the greatest reduction in perioperative pain. Most veterinarians also perceived that postprocedural pain persists beyond 24 h for disbudding and castration and did not support the use of differential treatment based on developmental age. Despite this, certain barriers were identified for their potential to inhibit the use of pain management on-farm. While demographic differences were found to influence veterinary perceptions towards perioperative pain management, the findings revealed considerable support among veterinarians for improving pain mitigation in calves. Given the opportunity, veterinarians in New Zealand would likely support strengthening the minimum provisions afforded to calves in practice and policy.While veterinarians are instrumental to the welfare of calves (Bos taurus), limited knowledge exists concerning veterinary perceptions towards perioperative pain management in calves. As a part of a larger, nationwide study investigating the perceptions of veterinarians towards calf welfare, the current work sought to quantify veterinary perceptions towards perioperative pain management, including barriers to its use, and investigate demographic influences affecting those perceptions. An electronic mixed-methods survey was completed by 104 veterinarians registered with the Veterinary Council of New Zealand. The current work revealed that most veterinarians considered a multimodal approach as the most effective method for ameliorating perioperative pain in calves, rejected the practice of differential treatment based on developmental age, and perceived that postprocedural pain persists beyond 24 h for the majority of procedures included in the survey. Despite this, veterinarians identified certain barriers that may inhibit the provision of pain mitigation on-farm, including costs, inadequate recognition of pain, and ingrained farming practices. Certain demographic effects were found to influence perceptions towards perioperative pain management, including gender, the number of years since graduation, and species emphasis. Nevertheless, the current work demonstrated considerable support among veterinarians to improve pain management protocols during routine husbandry procedures. The asymmetries that exist between the current minimum provisions of perioperative pain management and veterinary perspectives suggest that substantive improvements are necessary in order to reconcile New Zealand’s existing regulatory regime with developments in scientific knowledge.

Positive Human Contact and Housing Systems Impact the Responses of Piglets to Various Stressors.

Simple SummaryEarly life experiences such as contact with humans, maternal care and the physical environment can play a substantial role in shaping behavioural and physiological responses to stress. This experiment studied the effects of lactation housing systems and human interaction on stress in young pigs. We hypothesised that piglets handled in a positive manner and reared in loose farrowing and lactation pens with increased opportunity for interaction with their dam, greater space and more complexity in their physical environment have improved stress resilience than piglets reared in traditional farrowing crates with routine contact from stockpeople. In both housing systems, providing regular opportunities for positive human interaction reduced piglets’ fear of humans and routine husbandry procedures imposed by humans, and reduced the number of injuries obtained after weaning. However, contrary to the expected findings, piglets from loose farrowing and lactation pens were more reactive to capture by a stockperson, more fearful of novel and human stimuli, had more injuries during the lactation period and were more likely to perform behaviours which may be indicative of reduced coping after weaning. Whether these effects are specific to the loose farrowing and lactation system studied in this experiment or are reflective of other loose systems requires further research.This experiment studied the effects of lactation housing systems and human interaction on piglets’ responses to routine stressors. Forty litters of piglets were reared in either a standard farrowing crate (FC) or a loose farrowing and lactation pen (LP; PigSAFE pen) and received either routine contact with humans (C) or regular opportunities for positive human contact (+HC; 3 min of patting, stroking and scratching 5 times/week). Behavioural and physiological responses to routine husbandry procedures, weaning, novelty and humans were studied in addition to effects on piglet growth, injuries and survival. Compared to C piglets, +HC piglets vocalised for shorter durations (p = 0.018) during husbandry procedures and showed a lower intensity of escape behaviour during iron injection (p = 0.042) and oral vaccination (p = 0.026) at 3 d of age, capture at 2 wk of age (p < 0.001), and intramuscular vaccination (p = 0.005) at 3 wk of age. +HC piglets at 2 wk of age were faster than C piglets to approach (p = 0.048) and interact (p = 0.042) with a stationary unfamiliar human. Compared to LP piglets, FC piglets showed a lower intensity of escape behaviour during capture and iron administration by a stockperson at 3 d of age (p = 0.043). FC piglets at 2 wk of age were faster than LP piglets to approach (p = 0.005) and interact (p = 0.027) with a novel object and approach (p = 0.009) and interact (p = 0.008) with an unfamiliar human. FC piglets had fewer injuries than LP piglets at 2 wk of age (p = 0.004). +HC pigs had fewer injuries than C pigs after weaning (p = 0.003). After weaning there were more pigs from LP than FC observed to be upright (both stationary, p = 0.002 and walking, p = 0.024), vocalizing (p = 0.004), nosing another pig (p = 0.035) and nosing the pen floor (p = 0.038). There were no significant effects on neutrophil:lymphocyte ratios or plasma cortisol concentrations 1.5 h after weaning. However, 25 h after weaning +HC pigs had higher haptoglobin concentrations than C pigs (p = 0.002), and C/LP pigs had higher cortisol concentrations than +HC/LP and C/FC pigs (p = 0.012). There were no significant effects on piglet growth, the number of piglets born alive or the number stillborn, however there were more piglets weaned from FC than LP (p = 0.035). The results from this experiment raise questions that require further research on the ability of pigs reared in loose pens to cope with stressors such as exposure to humans, novelty, husbandry procedures and weaning. This experiment also provides evidence that regular positive human interaction reduces pigs’ fear of humans and husbandry procedures imposed by stockpeople. More research is required to determine if any of these effects are sustained long-term.

Methods for Pain Assessment in Calves and Their Use for the Evaluation of Pain during Different Procedures-A Review.

Simple SummaryPain recognition in calves is difficult. Pain is assessed either subjectively or objectively. Subjective pain assessment can be done using ethograms or pain scales, or by evaluation of changes of the facial expression due to pain. The problem with subjective pain assessment is that the evaluation of the amount of pain a calf is experiencing depends on the evaluation and the experience of the observer. Variables for the objective description of pain are assessment of biomarkers in the blood (e.g., cortisol or substance P), use of algometry to measure mechanical nociceptive thresholds, activity measurements by use of accelerometers and pedometers, use of infrared thermography, and the assessment of heart rate, heart rate variability, feed and water intake, or weight gain. Studies about pain recognition and pain management in calves mostly use more than one variable. Often a combination of subjective and objective measures is used to evaluate the level of pain calves are experiencing and to improve pain recognition.The evaluation and assessment of the level of pain calves are experiencing is important, as the experience of pain (e.g., due to routine husbandry procedures) severely affects the welfare of calves. Studies about the recognition of pain in calves, and especially pain management during and after common procedures, such as castration, dehorning, and disbudding, have been published. This narrative review discusses and summarizes the existing literature about methods for pain assessment in calves. First, it deals with the definition of pain and the challenges associated with the recognition of pain in calves. Then it proceeds to outline the different options and methods for subjective and objective pain assessment in calves, as described in the literature. Research data show that there are several tools suitable for the assessment of pain in calves, at least for research purposes. Finally, it concludes that for research purposes, various variables for the assessment of pain in calves are used in combination. However, there is no variable which can be used solely for the exclusive assessment of pain in calves. Also, further research is needed to describe biomarkers or variables which are easily accessible in the field practice.

Preliminary Findings on a Novel Behavioural Approach for the Assessment of Pain and Analgesia in Lambs Subject to Routine Husbandry Procedures.

The identification and assessment of pain in sheep under field conditions are important, but, due to their stoic nature, are fraught with many challenges. In Australia, various husbandry procedures that are documented to cause pain are routinely performed at lamb marking, including ear tagging, castration, mulesing, and tail docking. This study evaluated the validity of a novel methodology to assess pain in lambs: qualitative behavioural assessment (QBA) was used to compare the behavioural expression of control lambs (CONTROL) with that of lambs subject to these procedures that received either a saline placebo 15 min before procedures (PLACEBO), or were administered meloxicam 15 min before procedures in addition to the standard analgesic Tri-Solfen at the time of procedures, as per the manufacturer's recommendations (ANALGESIC TREATMENT; AT). In terms of behavioural expression, it was expected that: CONTROL ≠ PLACEBO, AT = CONTROL, and PLACEBO ≠ AT. Video footage of the 6-8-week-old lambs (n = 10 for each treatment) was captured approximately 1.5 h postprocedure and was presented, in a random order, to 19 observers for assessment using the Free-Choice Profiling (FCP) approach to QBA. There was significant consensus (p < 0.001) among the observers in their assessment of the lambs, with two main dimensions of behavioural expression explaining 69.2% of the variation. As expected, observers perceived differences in the demeanour of lambs in the first dimension, scoring all lambs subject to the routine husbandry procedures as significantly more 'dull' and 'uneasy' compared to the control lambs (p < 0.05). Contrary to expectations, the results also suggested that analgesic treatment did not provide relief at the time of observation. Further investigations to validate the relationship between behavioural expression scores and pain are necessary, but these results suggest that painful husbandry procedures alter the behavioural expression of lambs and these differences can be captured using QBA methodology.

Automated and Continuous Monitoring of Animal Welfare through Digital Alerting.

A primary goal in preclinical animal research is respectful and responsible care aimed toward minimizing stress and discomfort while enhancing collection of accurate and reproducible scientific data. Researchers use hands-on clinical observations and measurements as part of routine husbandry procedures or study protocols to monitor animal welfare. Although frequent assessments ensure the timely identification of animals with declining health, increased handling can result in additional stress on the animal and increased study variability. We investigated whether automated alerting regarding changes in behavior and physiology can complement existing welfare assessments to improve the identification of animals in pain or distress. Using historical data collected from a diverse range of therapeutic models, we developed algorithms that detect changes in motion and breathing rate frequently associated with sick animals but rare in healthy controls. To avoid introducing selec- tion bias, we evaluated the performance of these algorithms by using retrospective analysis of all studies occurring over a 31-d period in our vivarium. Analyses revealed that the majority of the automated alerts occurred prior to or simultaneously with technicians' observations of declining health in animals. Additional analyses performed across the entire duration of 2 studies (animal models of rapid aging and lung metastasis) demonstrated the sensitivity, accuracy, and utility of automated alerting for detecting unhealthy subjects and those eligible for humane endpoints. The percentage of alerts per total subject days ranged between 0% and 24%, depending on the animal model. Automated alerting effectively complements standard clinical observations to enhance animal welfare and promote responsible scientific advancement.

Salivary microRNAs are potential biomarkers for the accurate and precise identification of inflammatory response after tail docking and castration in piglets.

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.

Dim Light at Night and Constant Darkness: Two Frequently Used Lighting Conditions That Jeopardize the Health and Well-being of Laboratory Rodents.

The influence of light on mammalian physiology and behavior is due to the entrainment of circadian rhythms complemented with a direct modulation of light that would be unlikely an outcome of circadian system. In mammals, physiological and behavioral circadian rhythms are regulated by the suprachiasmatic nucleus (SCN) of the hypothalamus. This central control allows organisms to predict and anticipate environmental change, as well as to coordinate different rhythmic modalities within an individual. In adult mammals, direct retinal projections to the SCN are responsible for resetting and synchronizing physiological and behavioral rhythms to the light-dark (LD) cycle. Apart from its circadian effects, light also has direct effects on certain biological functions in such a way that the participation of the SCN would not be fundamental for this network. The objective of this review is to increase awareness, within the scientific community and commercial providers, of the fact that laboratory rodents can experience a number of adverse health and welfare outcomes attributed to commonly-used lighting conditions in animal facilities during routine husbandry and scientific procedures, widely considered as “environmentally friendly.” There is increasing evidence that exposure to dim light at night, as well as chronic constant darkness, challenges mammalian physiology and behavior resulting in disrupted circadian rhythms, neural death, a depressive-behavioral phenotype, cognitive impairment, and the deregulation of metabolic, physiological, and synaptic plasticity in both the short and long terms. The normal development and good health of laboratory rodents requires cyclical light entrainment, adapted to the solar cycle of day and night, with null light at night and safe illuminating qualities during the day. We therefore recommend increased awareness of the limited information available with regards to lighting conditions, and therefore that lighting protocols must be taken into consideration when designing experiments and duly highlighted in scientific papers. This practice will help to ensure the welfare of laboratory animals and increase the likelihood of producing reliable and reproducible results.

A comparison of the effect of short-acting and long-acting cloxacillin-based dry-cow therapy on somatic cell counts after calving in cows also given internal teat sealants

AIM: To compare, in cows treated with an internal teat sealant, the effect of short-acting and long-acting cloxacillin-based dry-cow therapy on somatic cell counts (SCC) after calving.METHODS: Cows from a spring-calving, pasture-based dairy farm in the Manawatu-Whanganui region of New Zealand were randomly allocated to receive either a short-acting cloxacillin and ampicillin dry-cow therapy and internal teat sealant (n=291) or a long-acting cloxacillin and ampicillin dry-cow therapy and internal teat sealant (n=288) at the end of lactation. Cows were managed on-farm with routine husbandry procedures through the dry period and following calving. A multivariable logistic regression model was used to determine the association between length of action of dry-cow therapy and the proportion of cows with a SCC >150,000 cells/mL at the first herd test after calving.RESULTS: Age of cow, mean SCC for the preceding season and interval from calving to the first post-calving herd test were all associated with the proportion of cows with an individual SCC >150,000 cells/mL at the first herd test (p<0.001) Treatment with the short-acting dry-cow therapy was not associated with decreased odds of cows having a SCC >150,000 cells/mL at the first herd test compared with treatment with long-acting dry-cow therapy (OR=0.724; 95% CI=0.40–1.30).CONCLUSIONS AND CLINICAL RELEVANCE: In this herd, which routinely used internal teat sealants, the use of short-acting cloxacillin-based dry-cow therapy did not result in an increased proportion of cows with elevated SSC post-calving. This was a single farm, single year study but indicates that in this herd, changing from a long-acting to a short-acting antimicrobial may have no impact on the prevalence of subclinical mastitis.

Trauma as a Cause for Hepatopathy in Newborn Göttingen Minipigs.

Routine husbandry procedures during animal toxicity studies can result in incidental pathological changes. We report on trauma-induced hepatopathy in newborn Göttingen minipigs. Sixty-four neonatal minipigs were allocated to 13- and 26-week treatment arms. There was a 4-week recovery period for both arms. The animals were divided into 2 treatment groups and a vehicle group and were dosed 3 times daily by direct oral administration using a syringe. During the feeding procedure in the first weeks, the animals had to be handled firmly. After 13 weeks, randomly distributed foci of degeneration/necrosis and focal congestion and/or hemorrhage were found in the livers of several animals from all groups. Reduced incidence and severity were evident in the recovery phase, and the lesions were absent after 26 weeks. These changes were considered as related to the manual handling of the animals. Knowledge of these findings is crucial for interpretation of studies utilizing newborn minipigs.

Olfaction variation in mouse husbandry and its implications for refinement and standardization: UK survey of animal scents.

Olfaction plays a crucial role in mouse communication, providing information about genetic identity, physiological status of conspecifics and alerting mice to potential predators. Scents of animal origin can trigger physiological and behavioural responses that could affect experimental responses and impact positively or negatively on mouse welfare. Additionally, differing olfactory profiles could help explain variation in results between laboratories. A survey was sent to animal research units in the UK to investigate potential transfer of scents of animal origin during routine husbandry procedures, and responses were obtained from animal care workers and researchers using mice in 51 institutions. The results reveal great diversity between animal units regarding the relevant husbandry routines covered. Most [71%] reported housing non-breeding male and female mice in the same room, with 76% reporting that hands were not washed and gloves not changed between handling male and female mice. The most commonly reported species housed in the same facility as mice was the rat (91%), and 41% of respondents were aware that scents from rats could affect mice. Changing of gloves between handling mice and other species was reported by 79% of respondents. Depending on the aspect considered, between 18 and 33% of respondents believed human and non-human animal odours would strongly affect mouse physiology, behaviour or standardization, while approximately 32-54% believed these effects would be weak. This indicates uncertainty regarding the significance of these factors. Understanding and controlling these practices could reduce unwanted variability in experimental results and maximize welfare.