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Research organizations should be proactive in regularly evaluating and refining their animal care and use programs in order to advance animal welfare and minimize distress. Pigs are often used in research, but few empirical studies have examined optimal husbandry and research use practices for pigs in a research environment. We developed the Pig Welfare Working Group (PWWG) to address the need for more formal guidelines on the management and use of pigs in research. The PWWG was a stakeholder focus group whose goal was to identify challenges and opportunities relevant to improving animal welfare through collaboration, knowledge sharing, and inclusive decision-making. Through consensus building, the PWWG developed 12 recommendations for behavioral management, housing, research procedures, transportation, and rehoming programs. The recommendations were rolled out across the contract research organization, business units, sites, and countries. Follow up will be conducted regularly to assess welfare, monitor progress toward implementing the recommendations, and recognize and reward participants making changes at their site.

Cynomolgus macaques may undergo surgical procedures for scientific and veterinary purposes. Recognition and assessment of pain using validated tools is a necessary first step for adequately managing pain in these primates. Grimace scales are one means of assessing the occurance of acute pain using action units such as facial expressions and posture. The aim of this study was to create and validate a Cynomolgus Macaque Grimace Scale (CMGS). Cynomolgus macaques (n = 43) were video recorded before and after a surgical procedure. Images were extracted from videos at timepoints at which breakthrough pain might be expected based on analgesic pharmacokinetics. Using the CMGS images were scored by 12 observers blinded to animal identification, times, and conditions. To validate the tool, detailed behavioral analyses emphasizing changes to baseline activity ethograms were compared to grimace scores. Four action units were identified related to potential pain including orbital tightening, brow lowering, cheek tightening, and hunched posture. The CMGS tool was found to have moderate inter- (ICC average action unit mean ± SD: 0.67 ± 0.28) and good intra- (ICC single mean ± SD: 0.79 ± 0.14) observer reliability. Grimace scores increased significantly (p < 0.0001) in the first four post-operative timepoints compared to baseline, correlating with behavioral findings (rho range = 0.22-0.35, p < 0.001). An analgesic intervention threshold was determined and should be considered when providing additional pain relief. The CMGS was shown to be a reliable and valid tool; however, more research is needed to confirm external validity. This tool will be highly valuable for refining analgesic protocols and acute peri-procedural care for cynomolgus macaques., (© 2023. The Author(s).)

Nonaversive or low stress handling techniques can reduce fear and stress in research rodents, ultimately improving study data quality. Uptake of low stress handling has been slow in the USA and Canada. In this study we explored the understanding, experience, and attitudes toward low stress handling of rats and mice in laboratory animal professionals from the USA (US) and Canada (CA). Participants ( n = 40) were recruited for a standardized interview and job categories were divided into veterinary/PhD level roles (doctoral level; DL) and non-veterinary/non-PhD level roles (non-doctoral level, NDL) (US: 23, DL: 9, NDL: 14; CA: 17, DL: 8, and NDL: 9). Interviews were transcribed and analyzed using NVIVO. Two research assistants independently coded themes for each question and consolidated responses based on commonality. Laboratory animal professionals understood the benefits of low stress handling and training techniques with rats and mice, stating reduced stress, better data, and improved welfare, with CA participants more likely to mention animal welfare as a benefit, and DL more likely to mention improved research data and reduced stress. Participants across demographic groups indicated improved job satisfaction and decreased stress as the positive impacts low stress handling would have on their positions. The primary perceived barriers to low stress handling implementation were researcher attitudes, the time needed to implement and use these techniques, and training personnel to use the techniques properly and consistently. To promote refinement of handling of rats and mice, more educational opportunities on the benefits and implementation of low stress handling techniques need to be provided to laboratory animal professionals, as well as to researchers., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 O'Malley, Hubley, Moody and Turner.)

Laboratory animal professionals (LAP) may experience situations that contribute to compassion fatigue (CF). The goal of this research was to better understand CF in LAP in and across employment categories. Surveys were distributed through LAP organizational listservs in the European Union (EU), China, and Japan, and results were analyzed to identify CF prevalence, personal and work-related factors, coping mechanisms, and beneficial work-support programs. Independent χ²-tests compared personal and work-related factors and feelings of CF. Feelings of CF and coping mechanisms were compared with personality scores using independent sample t tests. There were 302 respondents from the EU, 39 from China, and 77 from Japan. Over half of respondents from the EU (52%) and China (56%) reported experiencing CF (52%), with fewer (32%) reporting CF in Japan. No major differences were found based on employer type. Personality scores were significantly related to feelings of CF and preferred coping mechanisms. Work-related factors that contributed to feelings of CF in over half of respondents included staffing levels, workplace relationships, and availability of programs geared to address CF. Across regions, talking to someone, physical activity, getting away from work, and self-care were effective coping mechanisms in over 50% of respondents. Fewer than 30% of respondents indicated that their place of employment had CF support programs, and even fewer (8% to 28%) indicated that these programs were helpful. The study results suggest that to be effective, employer CF programs for LAP should consider providing quiet places at work and programs for self-care, promoting physical and mental health and social support systems, and establishing opportunities to memorialize animals.

Pigs are common research models and are strong animals that can be difficult to restrain. Improper restraint can put pigs and research personnel at risk for injury and induce stress, which can affect research outcomes. This study aimed to refine restraint techniques for research pigs using habituation and operant conditioning. Forty-four (22 males, 22 females; 4 months old, ~8.1 kg) Göttingen minipigs were randomly assigned to a control (C: no interventions) or a treatment group (T). Pigs in the T group received 3 min training sessions 3 days/week for the first 14 d after arrival. Training sessions included human socialization and habituation to a hammock sling for blood collection. Blood collection occurred on day 13 for all pigs by novel technicians. Pigs were placed in the sling, blood was collected from the radial vein, and serum cortisol levels were determined (ug/dL). Pig behavior was recorded and scored for duration of time spent struggling (s) and vocalizing (s). Novel human approach tests occurred on day 12, before blood collection, and day 14, after blood collection. Pigs were scored on latency to touch the human (s) and duration of time spent in contact with the human (s). Pig weight was taken upon arrival and on day 15. Separate linear models were fitted for response variables struggle duration in sling, serum cortisol, latency to touch human, time spent in contact with human, and body weight. Fixed effects were treatment and sex. Prior to blood collection, there was no difference in response to a novel human ( P > 0.05) but after blood collection, T pigs were quicker to approach (estimate: -5.352, SE: 1.72, P = 0.003) and spent more time in contact with the novel human (estimate: 3.091, SE: 1.448, P = 0.039). T pigs also had lower cortisol levels during blood collection (estimate: -2.36, SE: 0.657, P = 0.001). There was no difference in behavior while in the sling ( P > 0.05). The results of the study suggest that even small investments in habituation and training pigs to study procedures is beneficial in reducing stress and improving human-animal relationships, but more time would be beneficial to promote calmer behavior in the sling., Competing Interests: All authors were employed by company Charles River Laboratories. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors declare that this study received funding from Charles River. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication., (Copyright © 2022 O'Malley, Hubley, Tambadou and Turner.)

Research primates may undergo surgical procedures making effective pain management essential to ensure good animal welfare and unbiased scientific data. Adequate pain mitigation is dependent on whether veterinarians, technicians, researchers, and caregivers can recognize and assess pain, as well as the availability of efficacious therapeutics. A survey was conducted to evaluate primate veterinary approaches to pain assessment and alleviation, as well as expressed challenges for adequately managing primate pain. The survey ( n = 93 respondents) collected information regarding institutional policies and procedures for pain recognition, methods used for pain relief, and perceived levels of confidence in primate pain assessment. Results indicated that 71% ( n = 60) of respondents worked at institutions that were without formal experimental pain assessment policies. Pain assessment methods were consistent across respondents with the majority evaluating pain based on changes in general activity levels (100%, n = 86) and food consumption (97%, n = 84). Self-reported confidence in recognizing and managing pain ranged from slightly confident to highly confident, and there was a commonly expressed concern about the lack of objective pain assessment tools and science-based evidence regarding therapeutic recommendations of analgesics for research primates. These opinions correspond with significant gaps in the primate pain management literature, including limited specific pharmacokinetic data and efficacy testing for commonly used analgesics in research primate species as well as limited research on objective and specific measures of pain in research primates. These results demonstrate that there are inconsistencies in institutional policies and procedures surrounding pain management in research primates and a lack of objective pain assessment methods. Demonstrating the gaps and challenges in primate pain management can inform guideline development and suggest areas for future research.

Mental well-being (MWB) and diversity, equity, and inclusiveness (DEI) continue to be critical within the veterinary profession but there is less information regarding how professional associations around the world tackle these issues. A mixed-method study including an international online survey in English ( n = 137 responses via snowball sampling), fourteen interviews, and two webinars was used to identify the availability and impact of MWB and DEI support programs for veterinarians. Survey results showed that more veterinary organizations designated MWB and DEI challenges (54%, n = 43/79 and 58%, n = 45/78, respectively) as a key priority area than veterinary clinics (26%, n = 15/57 and 33%, n = 19/57, respectively). Whereas, MWB support programs were available in a moderate number of mainly English-speaking countries, DEI support programs were available in only a few countries and focused primarily on specific groups, with an unknown impact due to their recent implementation. Universally, survey respondents believed activities for specific groups, such as MWB webinars, training, and awareness campaigns, as well as MWB/DEI helplines and DEI peer-to-peer support programs had a high impact (median 3.5-4/5) yet were underemployed by both veterinary organization and veterinary clinics. Further feedback from respondents during focused interviews indicated that requiring initial and continuing training as well as tailored group activities would be most beneficial to improve MWB/DEI throughout the veterinary professional career. There are many areas of the intersection between MWB and DEI that remain to be elucidated in the future studies. Having a sufficient sample size, improving accessibility, and addressing varying cultural perceptions are the main challenges, as seen in our study. To truly address MWB and DEI disparities, change is also needed in veterinary workplace culture and environment. In conclusion, raising awareness for an inclusive profession, including increasing openness and acceptance to enhance DEI and destigmatizing MWB challenges, is needed to ensure a thriving, modern veterinary profession., Competing Interests: Authors WJ and NDB were employed by Federation of Veterinarians of Europe. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Timmenga, Jansen, Turner and De Briyne.)

Industrial food animal production practices are efficient for producing large quantities of milk, meat, and eggs for a growing global population, but often result in the need to alter animals to fit a more restricted environment, as well as creating new animal welfare and health problems related to animal confinement in high densities. These practices and methods have become normalized, to the extent that veterinarians and others embedded in these industries rarely question the ethical challenges associated with raising animals in this fashion. Moral 'lock-in' is common with those working in food animal industries, as is the feeling that it is impossible to effect meaningful change. Animal welfare issues associated with the industrialization of food animal production are 'wicked problems' that require a multi- and transdisciplinary approach. We argue that veterinarians, as expert animal health and welfare advocates, should be critical stakeholders and leaders in discussions with producers and the food animal sector, to look for innovative solutions and technology that will address current and future global sustainability and food security needs. Solutions will necessarily be different in different countries and regions, but ethical issues associated with industrial food animal production practices are universal.

For many years, researchers, veterinarians, animal ethics committees, and regulators have focused on minimizing pain and distress as a primary goal of refinement when working with animals in science. More recent publications as well as a shift in animal ethics and public opinion have emphasized promotion of positive affective states, culminating in the concept of positive animal welfare. Robust measures are required to know when positive animal welfare states are occurring, and a number of measures are proposed and discussed. Regardless of whether there are newer methods available that focus exclusively on measuring positive affective states, consistent consideration of research animal behavioral programs, refinement, and adopting periodic stand-alone animal welfare assessments for all species involved will help to push the care and practices of research animals towards an increased focus on positive animal welfare., (© The Author(s) 2020. Published by Oxford University Press on behalf of the National Academies of Sciences, Engineering, and Medicine. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Research animals are important for scientific advancement, and therefore, their long-term welfare needs to be monitored to not only minimize suffering, but to provide positive affective states and experiences. Currently, there is limited guidance in countries around the world on cumulative and experimental endpoints. This paper aims to explore current opinions and institutional strategies regarding cumulative use and endpoints through a scoping survey and review of current regulations and welfare assessment tools, and ultimately to provide recommendations for assessment of cumulative and lifetime use of research animals. The survey found that only 36% of respondents indicated that their institution had cumulative use endpoint policies in place, but these policies may be informal and/or vary by species. Most respondents supported more specific guidelines but expressed concerns about formal policies that may limit their ability to make case-by-case decisions. The wide diversity in how research animals are used makes it difficult for specific policies to be implemented. Endpoint decisions should be made in an objective manner using standardized welfare assessment tools. Future research should focus on robust, efficient welfare assessment tools that can be used to support planning and recommendations for cumulative endpoints and lifetime use of research and teaching animals.

Pain causes behavioral, autonomic, and neuroendocrine changes and is a common cause of animal welfare compromise in farm animals. Current societal and ethical concerns demand better agricultural practices and improved welfare for food animals. These guidelines focus on cattle, sheep, and pigs, and present the implications of pain in terms of animal welfare and ethical perspectives, and its challenges and misconceptions. We provide an overview of pain management including assessment and treatment applied to the most common husbandry procedures, and recommendations to improve animal welfare in these species. A cost-benefit analysis of pain mitigation is discussed for food animals as well as the use of pain scoring systems for pain assessment in these species. Several recommendations are provided related to husbandry practices that could mitigate pain and improve farm animal welfare. This includes pain assessment as one of the indicators of animal welfare, the use of artificial intelligence for automated methods and research, and the need for better/appropriate legislation, regulations, and recommendations for pain relief during routine and husbandry procedures.

Various pharmaceutical products have been derived from horse blood and urine for over a century. Production of biologics and therapeutics from these samples is a niche industry and often occurs in regions with little regulation or veterinary oversight. To ensure good welfare of horses maintained for these purposes, guidance has been developed to support the industry.

Little research has been conducted to examine the influence of various methods of providing nest materials-such as dispersing them, providing them as single units, or clustering them-on the behavior and welfare of group-housed mice. In this study, 6 wk-old C57BL/6NCrl mice were housed 3 per cage and randomized into 1 of 3 nest-material groups: 1) one facial tissue per cage (control; female mice, 3 cages; male mice, 3 cages); 2) an 8-g 'puck' of compressed nesting material and a facial tissue (females, 3 cages; males, 3 cages); or 3) 8 g of dispersed paper strips and a facial tissue (females, 3 cages; males, 3 cages). Mouse behavior (agonistic, stereotypic, nesting), physical examination data, and nest scores were evaluated over 16 d. The results showed that mice in the puck and control groups spent more time manipulating nest materials after cage changes than did mice in the paper-strip group. Average nest scores were highest in the paper-strip group compared with controls and puck cages. Female cages with pucks showed no barbering, whereas all other female mice cages demonstrated barbering. Overall, nest pucks may provide a time-consuming activity for mice and may help protect female C57BL/6 mice from barbering. However, more research is needed to replicate and expand these study results.

Objectives: In many jurisdictions, ethical concerns require surrogate humane endpoints to replace death in small animal models of acute lung injury. Heterogenous selection and reporting of surrogate endpoints render interpretation and generalizability of findings between studies difficult. We aimed to establish expert-guided consensus among preclinical scientists and laboratory animal veterinarians on selection and reporting of surrogate endpoints, monitoring of these models, and the use of analgesia., Design: A three-round consensus process, using modified Delphi methodology, with researchers who use small animal models of acute lung injury and laboratory animal veterinarians who provide care for these animals. Statements on the selection and reporting of surrogate endpoints, monitoring, and analgesia were generated through a systematic search of MEDLINE and Embase. Participants were asked to suggest any additional potential statements for evaluation., Setting: A web-based survey of participants representing the two stakeholder groups (researchers, laboratory animal veterinarians). Statements were rated on level of evidence and strength of support by participants. A final face-to-face meeting was then held to discuss results., Subjects: None., Interventions: None., Measurements and Main Results: Forty-two statements were evaluated, and 29 were rated as important, with varying strength of evidence. The majority of evidence was based on rodent models of acute lung injury. Endpoints with strong support and evidence included temperature changes and body weight loss. Behavioral signs and respiratory distress also received support but were associated with lower levels of evidence. Participants strongly agreed that analgesia affects outcomes in these models and that none may be necessary following nonsurgical induction of acute lung injury. Finally, participants strongly supported transparent reporting of surrogate endpoints. A prototype composite score was also developed based on participant feedback., Conclusions: We provide a preliminary framework that researchers and animal welfare committees may adapt for their needs. We have identified knowledge gaps that future research should address., Competing Interests: Drs. McGinn, Fergusson, Barron, Liu, and Lalu institutions received funding from Canadian Institutes of Health Research (CIHR). Dr. Kristof received funding from Ottawa Hospital Research Institute (travel costs). Dr. Downey received support for article research from the National Institutes of Health. Dr. Brown received support for article research from CIHR. Dr. dos Santos disclosed that she is funded by CIHR. Dr. Lalu is supported by The Ottawa Hospital Anesthesia Alternate Funds Association and the University of Ottawa Junior Research Chair in Innovative Translational Research. All of work was completed at the Ottawa Hospital Research Institute in Ottawa, ON, Canada. The remaining authors have disclosed that they do not have any potential conflicts of interest., (Copyright © 2020 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.)

Compassion fatigue (CF) is a topic of increasing concern because it can affect the mental wellbeing of caregivers, including those caring for or using research animals. If unaddressed, compassion fatigue may adversely impact the quality of life for personnel working with animals in research settings and may influence their decision to remain in the field. This study used a cross-sectional anonymous online questionnaire to 1) examine compassion fatigue in individuals working with research animals in Canada and the US; 2) better understand how personal and work-related factors may influence feelings of CF; 3) assess coping mechanisms used to deal with CF; and 4) determine the beneficial components of a CF support program. A questionnaire was sent to laboratory animal professionals in Canada and the US via email listserves to survey the general population of laboratory animal workers and personnel working for a large North American contract research organization (CRO). A total of 422 responses were received and analyzed ( n = 154 from the general population, n = 268 from the CRO). Most participants were female (73%, 309/422); 66% (101/154) and 69% (184/268) of the general laboratory animal science respondents and the CRO respondents, respectively, reported experiencing feelings of CF. Survey participants indicated that the most influential work-related factors associated with feelings of CF were understaffing, close relationships with experimental animals, a lack of resources for coping with CF, poor relationships with superiors, and lack of training in managing CF. Respondents indicated that the most influential personal factors contributing to feelings of CF were poor mental and physical health. The most commonly reported beneficial coping mechanisms were talking to a trusted individual, getting away from work, practicing self-care strategies, increasing opportunities for physical activity, and owning or caring for companion animals.

: The use of carbon dioxide (CO 2 ) for stunning and killing animals is considered to compromise welfare due to air hunger, anxiety, fear, and pain. Despite decades of research, no alternatives have so far been found that provide a safe and reliable way to induce unconsciousness in groups of animals, and also cause less distress than CO 2 . Here, we revisit the current and historical literature to identify key research questions that may lead to the identification and implementation of more humane alternatives to induce unconsciousness in mice, rats, poultry, and pigs. In addition to the evaluation of novel methods and agents, we identify the need to standardise the terminology and behavioural assays within the field. We further reason that more accurate measurements of consciousness state are needed and serve as a central component in the assessment of suffering. Therefore, we propose a roadmap toward improving animal welfare during end-of-life procedures., Competing Interests: The authors declare no conflict of interest.

Farber disease (FD) is a rare lysosomal storage disorder (LSD) characterized by systemic ceramide accumulation caused by a deficiency in acid ceramidase (ACDase). In its classic form, FD manifests with painful lipogranulomatous nodules in extremities and joints, respiratory complications, and neurological involvement. Hepatosplenomegaly is commonly reported, and severe cases of FD cite liver failure as a cause of early death. Mice homozygous for an orthologous patient mutation in the ACDase gene (Asah1 P361R/P361R ) recapitulate the classical form of human FD. In this study, we demonstrate impaired liver function and elevation of various liver injury markers in Asah1 P361R/P361R mice as early as 5 weeks of age. Histopathology analyses demonstrated significant formation and recruitment of foamy macrophages, invasion of neutrophils, progressive tissue fibrosis, increased cell proliferation and death, and significant storage pathology within various liver cell types. Lipidomic analyses revealed alterations to various lipid concentrations in both serum and liver tissue. A significant accumulation of ceramide and other sphingolipids in both liver and hepatocytes was noted. Sphingolipid acyl chains were also altered, with an increase in long acyl chain sphingolipids coinciding with a decrease in ultra-long acyl chains. Hepatocyte transcriptome analyses revealed significantly altered gene transcription. Molecular pathways related to inflammation were found activated, and molecular pathways involved in lipid metabolism were found deactivated. Altered gene transcription within the sphingolipid pathway itself was also observed. The data presented herein demonstrates that deficiency in ACDase results in liver pathology as well as sphingolipid and gene transcription profile changes that lead to impaired liver function.

The use of CO 2 inhalation with different gas chamber fill rates has not been evaluated for euthanasia in commercial meat rabbits. Our objectives were to evaluate the behavioral and physiologic responses of rabbits (pre-weaned to adult) when exposed to gradual- and fast-fill rates of CO 2 , and to determine the time to onset of insensibility and death. Cull rabbits ( n = 81) were randomly assigned to either a gradual-fill chamber displacement rate of 28% volume change/min ( n = 42) or a fast-fill chamber displacement rate of 58% volume change/min ( n = 39). The fast-fill rate resulted in a more rapid loss of sensibility at a lower CO 2 chamber concentration and in a faster death than for gradual-fill. There were minimal differences in behavioral responses between fill rates with no clear signs of distress. These findings indicate that CO 2 at the studied displacement rates is suitable for commercial meat rabbit euthanasia.

Background: There is a critical need to develop appropriate on-farm euthanasia methods for poultry species. Euthanasia methods should affect the brain first causing insensibility, followed by cardiorespiratory arrest. Neck or cervical dislocation methods, either manual (CD) or mechanical (MCD), are reported to cause a prolonged time to loss of sensibility and death with inconsistent results upon application, especially MCD methods. However, there is limited information on cervical dislocation in turkeys. The overall objective of this study was to assess the welfare implications of CD and a newly developed MCD device for euthanasia of cull turkeys in comparison with intravenous (IV) pentobarbital sodium (1 mL/4.5 kg), the gold standard euthanasia method. Time to death using electroencephalographic (EEG) and behavioural responses were monitored in eight and eighteen week-old turkeys for five minutes after each euthanasia method application. Spectral analyses of EEG responses and onset of isoelectric EEGs were compared to baseline EEG recordings of birds under anesthesia and behavioural responses were studied among euthanasia treatments. A significant decrease in brain activity frequencies analysis and isoelectric EEG were recorded as time of brain death., Results: All turkeys euthanized with IV pentobarbital sodium presented a rapid and irreversible decrease in the EEG activity at approximately 30s post-injection with minimal behavioural responses. CD and MCD methods caused EEG responses consistent with brain death at approximately 120 s and 300 s, respectively. Additionally, isoelectric EEGs resulted in all pentobarbital sodium and CD groups, but only in 54 and 88% of the eight and eighteen week-old turkeys in the MCD groups, respectively. There were few clear patterns of behavioural responses after CD and MCD application. However, cessation of body movement and time to isoelectric EEG after CD application were positively correlated., Conclusions: Use of CD and MCD resulted in a prolonged time to death in both age groups of turkeys. MCD application presents a number of welfare risks based on electroencephalographic and behavioural findings. Intravenous pentobarbital sodium induced rapid brain death, but possesses several on-farm limitations. To develop improvements in cervical dislocation methods, further investigations into combined or alternative methods are required to reduce the prolonged time to insensibility and death.

Background: Surgical castration is a painful procedure, performed routinely on commercial pig farms to prevent boar taint and reduce aggression. The objectives of this study were to assess the efficacy of 0.04 mg/kg buprenorphine (BUP) in reducing pain in castrated piglets, using behavioral indicators and vocalization analysis. This study also sought to further validate the Piglet Grimace Scale (PGS) as a pain assessment tool. A pilot study first assessed the safety of BUP or 0.2 mg/kg butorphanol administration to piglets (n = 4 per treatment). When no side effects were noted with BUP, ten litters of 5-day old piglets (n = 60 total, 15 per treatment group) were used, and randomly assigned to one of four possible treatments: BUP (castrated or uncastrated), saline, or sham. Treatments were administered as an intramuscular injection 20 min prior to surgical castration. Piglets were video recorded 1 h pre-procedure, post-castration for 8 h and for another hour, 24 h post-procedure. Behaviors were scored continuously for the first 15 min of each hour and 511 still-images of piglet faces were scored using the PGS. Vocalizations were recorded from each piglet at three points in the study: at initial handling, injection, and castration., Results: Butorphanol caused some piglets to become groggy and vomit and was not further evaluated. BUP-castrated piglets demonstrated significantly fewer pain behaviors and less facial grimacing compared to saline-treated pigs (P < 0.0001 and P = 0.0073, respectively). There was no difference between the pain behaviours displayed by BUP-castrated piglets compared to BUP-uncastrated and sham piglets (P = 0.9986 and P = 0.7484). There was also no difference in PGS score between BUP-castrated and BUP-uncastrated piglets (P = 0.9376). Piglets in the BUP-castrated group produced vocalizations of similar frequency, amplitude, power, and energy to saline-treated piglets., Conclusions: Buprenorphine was highly effective in alleviating castration-associated pain behaviors and facial grimacing in piglets, without causing any obvious side effects. Its administration did not reduce piglet vocalizations at the time of castration. The PGS corresponded well to piglet pain behaviors and has utility as a pain assessment tool.

Background: Rabbits are cecotrophic, hindgut-fermenters that rely heavily on their gastrointestinal microbiota for optimal digestion of plant-based diets. Dysbiosis, caused by disruption of the gastrointestinal microbiota, is known to predispose rabbits to rabbit enteritis complex (REC), a major cause of morbidity and mortality. The objectives of this study were to describe the fecal microbiota of domestic rabbits from a variety of settings (commercial meat, companion, laboratory, and shelter) and to identify how factors such as age, season, and routine antimicrobial use affect the fecal microbiota composition., Results: A total of 86 pooled commercial meat, 54 companion, 14 pooled laboratory, and 14 shelter rabbit fecal samples were evaluated using 16S rRNA gene sequencing of the V4 region. In all sample types, the predominant bacterial phylum was Firmicutes. Other commonly identified phyla (composing ≥ 1% of the total microbiota composition) were Verrucomicrobia, Proteobacteria, and Bacteroidetes. Significant differences in composition were noted between commercial, companion, laboratory, and shelter rabbit samples for proportions of Verrucomicrobia (P < 0.01), Proteobacteria (P < 0.01), and Lentisphaerae (P = 0.01) within the total microbiota. Within the commercial meat rabbit samples, significant differences between the microbiota composition of growers (n = 42) and does (n = 44) were limited to one unclassified Firmicutes (P = 0.03) and no differences were identified at the phylum level. Significant differences were present between fecal samples taken from rabbits during the summer (n = 44) compared to the winter (n = 42), with Firmicutes (P = 0.04), Verrucomicrobia (P = 0.03), Proteobacteria (P = 0.02), Deinococcus-Thermus (P = 0.04), Armatimonadates (P = 0.003), and Actinobacteria (P = 0.03) forming significantly different proportions of the microbiota. The only significant difference in composition between those farms that routinely reported antimicrobial use and those that did not was in one unclassified Bacteroidetes (P < 0.05) and no differences were identified at the phylum level., Conclusions: Rabbit husbandry and diet, in addition to season, significantly influence the fecal microbiota composition of domestic rabbits, while age of the rabbit post-weaning has minimal impact.

On-farm euthanasia is a critical welfare issue in the poultry industry and can be particularly difficult to perform on mature turkeys due to their size. We evaluated the efficacy of two commercially available non-penetrating captive bolt devices, the Zephyr-EXL and the Turkey Euthanasia Device (TED), on 253 turkeys at three stages of production: 4-5, 10, and 15-20 weeks of age. Effectiveness of each device was measured using both ante- and post-mortem measures. Application of the Zephyr-EXL resulted in a greater success rate (immediate abolishment of brainstem reflexes) compared to the TED (97.6% vs. 89.3%, p = 0.0145). Times to last movement ( p = 0.102) and cardiac arrest ( p = 0.164) did not differ between devices. Ante- and post-mortem measures of trauma and hemorrhage were highly correlated. Skull fractures and gross subdural hemorrhage (SDH) were present in 100% of birds euthanized with both the Zephyr-EXL and TED devices. Gross SDH scores were greater in birds killed with the Zephyr-EXL than the TED ( p < 0.001). Microscopic SDH scores indicated moderate to severe hemorrhage in 92% of turkeys for the Zephyr-EXL and 96% of turkeys for the TED, with no difference between devices ( p = 0.844). Overall, both devices were highly effective inducing immediate insensibility through traumatic brain injury and are reliable, single-step methods for on-farm euthanasia of turkeys., Competing Interests: Tina M. Widowski is the Egg Farmers of Canada Chair in Poultry Welfare Research. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Guinea pigs are a commonly used model for tuberculosis vaccine research. Loss of body weight is the most frequently described humane endpoint for animals used in these studies. During a chronic study, we noted labored breathing in some tuberculosis-infected guinea pigs. To develop consistent humane endpoints for these guinea pigs, we performed an observational study using multiple clinical signs. A combination of body weight loss, labored breathing, and activity level during handling estimated the time to euthanasia within approximately 7 d. Histologic severity scores of lesions in the cranial or caudal lung lobe (or both) supported clinical endpoints. This study presents humane endpoints for the refinement of studies using guinea pigs in tuberculosis research.

Although expectations for appropriate animal care are present in most developed countries, significant animal welfare challenges continue to be seen on a regular basis in all areas of veterinary practice. Veterinary ethics is a relatively new area of educational focus but is thought to be critically important in helping veterinarians formulate their approach to clinical case management and in determining the overall acceptability of practices towards animals. An overview is provided of how veterinary ethics are taught and how common ethical frameworks and approaches are employed-along with legislation, guidelines and codes of professional conduct-to address animal welfare issues. Insufficiently mature ethical reasoning or a lack of veterinary ethical sensitivity can lead to an inability or difficulty in speaking up about concerns with clients and ultimately, failure in their duty of care to animals, leading to poor animal welfare outcomes. A number of examples are provided to illustrate this point. Ensuring that robust ethical frameworks are employed will ultimately help veterinarians to "speak up" to address animal welfare concerns and prevent future harms., Competing Interests: The authors declare no conflict of interest.

The commercial meat rabbit industry is without validated on-farm euthanasia methods, potentially resulting in inadequate euthanasia protocols. We evaluated blunt force trauma (BFT), a mechanical cervical dislocation device (MCD), and a non-penetrating captive bolt device (NPCB) for euthanasia of pre-weaned kits, growers, and adult rabbits. Trials were conducted on three commercial meat rabbit farms using 170 cull rabbits. Insensibility was assessed by evaluating absence of brainstem and spinal reflexes, rhythmic breathing, and vocalizations. Survey radiographs on a subsample of rabbits ( n = 12) confirmed tissue damage prior to gross dissection and microscopic evaluation. All 63 rabbits euthanized by the NPCB device were rendered immediately and irreversibly insensible. The MCD device was effective in 46 of 49 (94%) rabbits. Method failure was highest for BFT with euthanasia failures in 13 of 58 (22%) rabbits. Microscopically, brain sections from rabbits killed with the NPCB device had significantly more damage than those from rabbits killed with BFT ( p = 0.001). We conclude that BFT is neither consistently humane nor effective as a euthanasia method. MCD is an accurate and reliable euthanasia method generally causing clean dislocation and immediate and irreversible insensibility, and the NPCB device was 100% effective and reliable in rabbits >150 g., Competing Interests: The authors declare no conflict of interest.

A common dilemma faced by all animal bioethics committees arises when exceptions are proposed to the use of analgesics in painful procedures. The committee and researcher must weigh the possible confounding effects of including additional drugs (analgesics) in their treatment regimen against the moral obligation to perform humane research. Often neglected in these considerations are the potential confounding effects of unrelieved pain and consistency with pain-relieving practices in human medicine. In this review, we summarize what is currently known regarding the molecular and physiologic effects of pain and analgesics in common animal models used across several therapeutic areas. This work is intended to help provide guidance and assurance that a comprehensive approach has been taken when contemplating how pain relief will be applied in animal research protocols.

Antimicrobial resistance (AMR) in zoonotic (e.g. Salmonella spp.), pathogenic, and opportunistic (e.g. E. coli) bacteria in animals represents a potential reservoir of antimicrobial resistant bacteria and resistance genes to bacteria infecting humans and other animals. This study evaluated the prevalence of E. coli and Salmonella enterica, and the presence of associated AMR in commercial meat, companion, research, and shelter rabbits in Canada. Associations between antimicrobial usage and prevalence of AMR in bacterial isolates were also examined in commercial meat rabbits. Culture and susceptibility testing was conducted on pooled fecal samples from weanling and adult commercial meat rabbits taken during both summer and winter months (n=100, 27 farms), and from pooled laboratory (n=14, 8 laboratory facilities), companion (n=53), and shelter (n=15, 4 shelters) rabbit fecal samples. At the facility level, E. coli was identified in samples from each commercial rabbit farm, laboratory facility, and 3 of 4 shelters, and in 6 of 53 companion rabbit fecal samples. Seventy-nine of 314 (25.2%; CI: 20.7-30.2%) E. coli isolates demonstrated resistance to >1 antimicrobial agent. At least one E. coli isolate resistant to at least one antimicrobial agent was present in samples from 55.6% of commercial farms, and from 25% of each laboratory and shelter facilities, with resistance to tetracycline being most common; no resistance was identified in companion animal samples. Salmonella enterica subsp. was identified exclusively in pooled fecal samples from commercial rabbit farms; Salmonella enterica serovar London from one farm and Salmonella enterica serovar Kentucky from another. The S. Kentucky isolate was resistant to amoxicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, streptomycin, and tetracycline, whereas the S. London isolate was pansusceptible. Routine use of antimicrobials on commercial meat rabbit farms was not significantly associated with the presence of antimicrobial resistant E. coli or S. enterica on farms; trends towards resistance were present when resistance to specific antimicrobial classes was examined. E. coli was widely prevalent in many Canadian domestic rabbit populations, while S. enterica was rare. The prevalence of AMR in isolated bacteria was variable and most common in isolates from commercial meat rabbits (96% of the AMR isolates were from commercial meat rabbit fecal samples). Our results highlight that domestic rabbits, and particularly meat rabbits, may be carriers of phenotypically antimicrobial-resistant bacteria and AMR genes, possibly contributing to transmission of these bacteria and their genes to bacteria in humans through food or direct contact, as well as to other co-housed animal species., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Appropriate and timely on-farm euthanasia is the responsibility of the producer, working together with their herd veterinarian. Unfortunately, validated methods for euthanasia of commercial meat rabbits are lacking and there are few educational materials available for producer training. Because euthanasia must be performed in a timely fashion to minimise suffering, it is critical to ensure that methods used are aesthetic, humane and effective. We surveyed Canadian meat rabbit producers for current on-farm euthanasia practices as well as attitudes towards the methods they employed and thoughts on novel euthanasia techniques. Surveys were distributed with a response rate of 26 per cent (n=26). Blunt force trauma was the most common euthanasia method used (54 per cent), followed by assisted manual cervical dislocation (31 per cent). Half of producers admitted to not having a euthanasia method in place for all age groups of rabbits, instead electing to let sick and injured rabbits die on their own. While some producers reported feeling highly skilled and satisfied with their current euthanasia method, 58 per cent reported concerns with their current method and 42 per cent desired alternative methods to be developed. Responses to additional questions on training and awareness of euthanasia resources indicated that veterinarians are not part of on-farm euthanasia planning for meat rabbits., Competing Interests: Competing interests: None declared., (© British Veterinary Association (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

There is considerable interest in refining laboratory rodent environments to promote animal well-being, as well as research reproducibility. Few studies have evaluated the long term impact of enhancing rodent environments with resources and additional cagemates. To that end, male and female Sprague Dawley (SD) rats were housed singly ( n = 8/sex), in pairs ( n = 16/sex), or in groups of four ( n = 16/sex) for five months. Single and paired rats were housed in standard cages with a nylon chew toy, while group-housed rats were kept in double-wide cages with two PVC shelters and a nylon chew toy and were provided with food enrichment three times weekly. Animal behaviour, tests of anxiety (open field, elevated plus maze, and thermal nociception), and aspects of animal physiology (fecal corticoid levels, body weight, weekly food consumption, organ weights, and cerebral stress signaling peptide and receptor mRNA levels) were measured. Significant differences were noted, primarily in behavioural data, with sustained positive social interactions and engagement with environmental resources noted throughout the study. These results suggest that modest enhancements in the environment of both male and female SD rats may be beneficial to their well-being, while introducing minimal variation in other aspects of behavioural or physiologic responses., Competing Interests: The authors declare no conflict of interest.

Infectious enterocolitis is a significant cause of mortality in meat rabbits. Disease risk is enhanced by intensive rearing practices and poor on-farm biosecurity. This investigation was undertaken in farmed meat rabbits during an Ontario-wide outbreak of enteritis with high mortality to determine the prevalence of causative agents. A survey evaluating on-farm biosecurity practices was also conducted to identify potential means of pathogen contamination and zoonotic risks. Gross and microscopic pathology evaluations combined with microbiologic testing were conducted on 95 rabbits over spring and winter months. Escherichia coli and Clostridium spiroforme were most commonly associated with enteritis in rabbits regardless of age or season and lesions were significantly more severe in mature does ( P < 0.0001). The survey results demonstrated a lack of consistent on-farm biosecurity practices. The infectious nature of enteric disease of rabbits combined with poor biosecurity practices may contribute to disease transmission within and between farms.

Despite several shortcomings, MS222 is the most commonly used chemical agent for euthanasia of zebrafish. Although lidocaine hydrochloride has some advantages over MS222, its effectiveness as a euthanasia agent for zebrafish is unknown. Larvae at 9 to 16 d postfertilization were exposed to 250 mg/L MS222 or 400, 500, 600, 700, 800, 900, or 1000 mg/L lidocaine and observed for cessation of heartbeat. Adult zebrafish were exposed to 250 mg/L MS222 or 400, 500, or 600 mg/L lidocaine; times to loss of righting reflex, cessation of opercular movement, and complete recovery; body length; aversive behavior; and gross and microscopic evidence of acute toxicity were evaluated. The heartbeat was not lost from any larvae in any group, regardless of drug or dosage. For adults, time to loss of righting reflex was greatest in the 500-mg/L lidocaine group. Opercular movement ceased earlier in all lidocaine groups compared with the MS222 group. Fish in the 500-mg/L lidocaine group were smaller than those in other groups. Fewer fish in the lidocaine groups displayed aversive behavior (erratic swimming and piping) compared with the MS222 group. No fish in the lidocaine hydrochloride groups (n = 30) recovered from euthanasia, whereas one fish in the MS222 group did (n = 10). Neither the MS222 nor lidocaine groups showed any gross or histologic changes suggestive of acute toxicity. Our results suggest that lidocaine hydrochloride may be an effective alternative chemical euthanasia agent for adult zebrafish but should not be used in larval fish.

The increasing occurrence of antimicrobial resistance among bacteria is a global problem that requires the development of alternative techniques to eradicate these superbugs. Herein, we used a combination of thermosensitive biocompatible polymer and gold nanorods to specifically deliver, preserve and confine heat to the area of interest. Our data demonstrates that this technique can be used to kill both Gram positive and Gram negative antimicrobial resistant bacteria in vitro. Our approach significantly reduces the antimicrobial resistant bacteria load in experimentally infected wounds by 98% without harming the surrounding tissues. More importantly, this polymer-nanocomposite can be prepared easily and applied to the wounds, can generate heat using a hand-held laser device, is safe for the operator, and does not have any adverse effects on the wound tissue and healing process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Few studies have evaluated the long-term effects of providing environmental resources to mice. This consideration is important given that mice are often maintained in vivaria for months. We evaluated the effects of providing simple cage resources (wood wool, cotton nesting material, a plastic tunnel, and oat cereal) compared with standard housing (solid-bottom cage with hardwood chips) to group-housed adult male and female C57BL/6 and BALB/c mice (n = 20/sex/strain/group) over 6 mo to determine whether these resources had a lasting effect on animal physiology, anatomy, and behavior. Body weights increased in all groups over time but were proportionately higher in male and female BALB/c mice housed in resource-supplemented environments. Throughout the study, adding environmental resources had no effect on hematology and lymphocyte subsets, fecal corticoid metabolite levels, response to LPS injection, or dendritic spine length or density. Strain- or sex×environmentspecific changes occurred in dark-light activity and thermal nociceptive responses. Dominant agonistic behaviors, abnormal conspecific sexual behaviors, and social nonagonistic behaviors demonstrated sex and strain×environment interactions such that fewer maladaptive social behaviors were noted in mice that were provided with environmental resources. This association was particularly evident in male mice of both strains in resource-supplemented environments. A small but significant increase in brain weight:body weight ratios occurred in mice in resource-supplemented environments. Under the conditions evaluated here, consistent use of simple environmental resources had a positive long-term effect on the behavioral wellbeing of male and female BALB/c and C57BL/6 mice yet minimally affected other aspects of murine physiology and neuroanatomy.

The mycotoxin aflatoxin B1 (AFB1) may initiate cancer by causing oxidatively damaged DNA, specifically by causing 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) lesions. Base excision repair removes these lesions, with 8-oxoguanine glycosylase (OGG1) being the rate-limiting enzyme. The aim of this study was to determine the effect of ogg1 deficiency on AFB1-induced oxidatively damaged DNA and tumourigenesis. Female wild-type, heterozygous and homozygous ogg1 null mice were given a single dose of 50mg/kg AFB1 or 40 µl dimethyl sulfoxide (DMSO) ip. Neither ogg1 genotype nor AFB1 treatment affected levels of oxidised guanine in lung or liver 2h post-treatment. AFB1-treated ogg1 null mice showed exacerbated weight loss and mortality relative to DMSO-treated ogg1 null mice, but AFB1 treatment did not significantly increase lung or liver tumour incidence compared with controls, regardless of ogg1 genotype. Suspect lung masses from three of the AFB1-treated mice were adenomas, and masses from two of the mice were osteosarcomas. No osteosarcomas were observed in DMSO-treated mice. All liver masses from AFB1-treated mice were adenomas, and one also contained a hepatocellular carcinoma. In DNA from the lung tumours, the K-ras mutation pattern was inconsistent with initiation by AFB1. In conclusion, ogg1 status did not have a significant effect on AFB1-induced oxidatively damaged DNA or tumourigenesis, but deletion of one or both alleles of ogg1 did increase susceptibility to other aspects of AFB1 toxicity., (© The Author 2015. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Practical implementation of the 3Rs at national and regional levels around the world requires long-term commitment, backing, and coordinated efforts by international associations for laboratory animal medicine and science, including the International Association of Colleges of Laboratory Animal Medicine (IACLAM) and the International Council for Laboratory Animal Science (ICLAS). Together these organizations support the efforts of regional organization and communities of laboratory animal science professionals as well as the development of local associations and professional colleges that promote the training and continuing education of research facility personnel and veterinary specialists. The recent formation of a World Organization for Animal Health (OIE) Collaborating Center for Laboratory Animal Science and Welfare emphasizes the need for research into initiatives promoting laboratory animal welfare, particularly in emerging economies and regions with nascent associations of laboratory animal science.

In North America, the biomedical research community faces social and economic challenges to nonhuman primate (NHP) importation that could reduce the number of NHP available for research needs. The effect of such limitations on specific biomedical research topics is unknown. The Association of Primate Veterinarians (APV), with assistance from the Centers for Disease Control and Prevention, developed a survey regarding biomedical research involving NHP in the United States and Canada. The survey sought to determine the number and species of NHP maintained at APV members' facilities, current uses of NHP to identify the types of biomedical research that rely on imported animals, and members' perceived trends in NHP research. Of the 149 members contacted, 33 (22%) replied, representing diverse facility sizes and types. Cynomolgus and rhesus macaques were the most common species housed at responding institutions and comprised the majority of newly acquired and imported NHP. The most common uses for NHP included pharmaceutical research and development and neuroscience, neurology, or neuromuscular disease research. Preclinical safety testing and cancer research projects usually involved imported NHP, whereas research on aging or degenerative disease, reproduction or reproductive disease, and organ or tissue transplantation typically used domestic-bred NHP. The current results improve our understanding of the research uses for imported NHP in North America and may facilitate estimating the potential effect of any future changes in NHP accessibility for research purposes. Ensuring that sufficient NHP are available for critical biomedical research remains a pressing concern for the biomedical research community in North America.

Whether social isolation of adult rats under standard laboratory conditions produces significant long-term alterations in behavior and physiology is unclear. In the present study, male Sprague-Dawley rats were singly or paired-housed for 10 wk. During this period, they were tested for acquisition, extinction, and reacquisition of heroin (0.3 mg/kg)-conditioned place preference. Fecal corticoid metabolite levels were analyzed several times throughout the period of housing, and food consumption and body weight were monitored. During place conditioning, heroin induced a significant increase in locomotor activity in both singly and pair housed rats, and the resulting place preference was similar in both groups. However, singly housed rats showed increased motor reactivity to heroin on reconditioning after extinction and displayed significant reacquisition of conditioned place preference, compared with pair-housed animals. Over the 10-wk period of the study, there were no differences in body weight or food consumption between groups. Mild significant increases in relative adrenal gland weight and decreases in relative brain weight were noted in singly housed animals compared with those paired. Significant decreases in nocturnal fecal corticoid metabolite output were noted in both groups, with loss of circadian variation in fecal corticoid levels over the course of the study. These data suggest that male Sprague-Dawley rats, irrespective of single or pair housing, develop reduced hypothalamic-pituitary-adrenal axis activity over time under standard laboratory housing conditions. Single housing can enhance both this effect and sensitivity to the stimulatory and rewarding actions of heroin after withdrawal.

Leporid herpesvirus 4 (LHV4) is a novel alphaherpesvirus recently identified in domestic rabbits (Oryctolagus cuniculi). Little is known about the pathogenesis or time course of disease induced by this virus. We therefore intranasally inoculated 22 female New Zealand white rabbits with 8.4 × 10(4) CCID50 of a clinical viral isolate. Rabbits were monitored for clinical signs, viral shedding in oculonasal secretions, and development and persistence of serum antibodies. Rabbits were euthanized at 3, 5, 7, 14, and 22 d postinfection (dpi) to evaluate gross and microscopic changes. Clinical signs were apparent between 3 to 8 dpi, and included oculonasal discharge, respiratory distress, and reduced appetite, and viral shedding occurred between 2 and 8 dpi. Seroconversion was seen at 11 dpi and persisted to the end of the study (day 22). Severe necrohemorrhagic bronchopneumonia and marked pulmonary edema were noted by 5 dpi and were most severe at 7 dpi. Pulmonary changes largely resolved by 22 dpi. In addition, multifocal splenic necrosis was present at 5 dpi and progressed to submassive necrosis by 7 dpi. Eosinophilic herpesviral intranuclear inclusion bodies were detected in the nasal mucosa, skin, spleen, and lung between 3 to 14 dpi. LHV4 is a pathogen that should be considered for rabbits that present with acute respiratory disease. LHV4 infection can be diagnosed based on characteristic microscopic changes in the lungs and spleen and by virus isolation. Serum antibody levels may be used to monitor viral prevalence in colonies.

We found that carprofen and meloxicam under 3 environmental conditions (ambient dark, ambient light, and 4 °C) remained stable for at least 7 d. We then evaluated the oral pharmacokinetics of meloxicam (20 mg/kg) and carprofen (10 mg/kg) in male C57BL/6 mice after oral gavage or administration in the drinking water. Mice did not drink meloxicam-medicated water but readily consumed carprofen-medicated water, consuming an average of 14.19 mL carprofen-medicated water per 100 g body weight daily; mice drank more during the dark phase than during the light phase. Plasma analyzed by HPLC (meloxicam) and tandem mass spectrometry (carprofen) revealed that the peak meloxicam and carprofen concentrations were 16.7 and 20.3 μg/mL and occurred at 4 and 2 h after oral gavage, respectively. Similar blood levels were achieved after 12 h access to the carprofen-medicated water bottle. At 24 h after oral gavage, the drugs were not detectable in plasma. Meloxicam plasma AUC, elimination half-life, apparent volume of distribution, and apparent oral clearance were 160.4 mg/L × h, 7.4 h, 0.36 L/kg, and 0.125 mL/h × kg, respectively. Carprofen plasma AUC, elimination half-life, apparent volume of distribution, and apparent oral clearance were 160.8 mg/L × h, 7.4 h, 0.42 L/kg, and 0.062 mL/h × kg, respectively. No gross or microscopic evidence of toxicity was seen in any mouse. Our findings indicate that carprofen can be administered in drinking water to mice and that medicated water bottles should be placed 12 to 24 h prior to painful procedures.

Farber disease (FD) is a severe inherited disorder of lipid metabolism characterized by deficient lysosomal acid ceramidase (ACDase) activity, resulting in ceramide accumulation. Ceramide and metabolites have roles in cell apoptosis and proliferation. We introduced a single-nucleotide mutation identified in human FD patients into the murine Asah1 gene to generate the first model of systemic ACDase deficiency. Homozygous Asah1(P361R/P361R) animals showed ACDase defects, accumulated ceramide, demonstrated FD manifestations and died within 7-13 weeks. Mechanistically, MCP-1 levels were increased and tissues were replete with lipid-laden macrophages. Treatment of neonates with a single injection of human ACDase-encoding lentivector diminished the severity of the disease as highlighted by enhanced growth, decreased ceramide, lessened cellular infiltrations and increased lifespans. This model of ACDase deficiency offers insights into the pathophysiology of FD and the roles of ACDase, ceramide and related sphingolipids in cell signaling and growth, as well as facilitates the development of therapy., (Copyright © 2013 The Authors. Published by John Wiley and Sons, Ltd on behalf of EMBO.)

The effect of chronic daily orogastric gavage with water (5 mL/kg) on behavior and physiology was evaluated in male Sprague-Dawley rats. Treatment groups included: unmanipulated control, restraint control, dry gavage, and gavage, with all rats singly housed (n = 9 or 10 per group). In addition, a group of pair-housed rats (n = 18) was included to determine whether social housing affected response to gavage. Weekly body weights and food consumption were recorded as well as use of a nylon chew toy for enrichment. Feces were collected biweekly at the end of the light and dark phases for fecal corticoid metabolite determinations. After 28 d of treatment, animals underwent conditioned place preference testing to evaluate sensitivity to motivational properties of the anxiolytic drug chlordiazepoxide (5.6 mg/kg SC). Brain and paired adrenal gland weights were collected at necropsy. Week 2 total fecal corticosterone levels were elevated in all groups and attributed to a fire alarm accidentally tripped during building renovations. No differences occurred in body weight or food consumption between any groups. All groups used a nylon chew toy given for enrichment and demonstrated mild preference for the drug-associated chamber. Fecal weights and corticoid metabolite levels were similar between all groups at week 4 and showed normal diurnal variation. No biologically significant variations were noted in brain or paired adrenal gland to body weight ratios. We conclude that orogastric gavage of aqueous solutions at 5 mL/kg does not negatively affect the welfare of laboratory rats acclimated to handling.

Obesity is an increasingly important health issue in both humans and animals and has been highly correlated as a risk factor for hyperglycemic conditions in humans. Naturally occurring obesity has been extensively studied in nonhuman primates with a focus on the development of biomarkers for characterizing overweight individuals and tracking the progression of obesity to conditions such as type 2 diabetes mellitus. Animal models have provided a basic understanding of metabolism and carbohydrate physiology, and continue to contribute to ongoing research of obesity and its adverse health effects. This review focuses on spontaneous obesity in rhesus and cynomolgus macaques as a model for human obesity and type 2 diabetes mellitus, including associated risk factors for the development of obesity and obesity-related health conditions. Little is known about preventive measures to minimize obesity while maintaining a healthy colony of macaques, and numerous complexities such as social status, feeding behaviors, timing of feeding, food distribution, and stress have been identified as contributing factors to overweight body condition in both single and group housed nonhuman primates. As in humans, increased body weight and obesity in macaques affect their overall health status. These conditions may interfere with the suitability of some animals in various studies unrelated to obesity.

Administration of substances to laboratory animals requires careful consideration and planning to optimize delivery of the agent to the animal while minimizing potential adverse experiences from the procedure. The equipment selected to deliver substances to animals depends on the length of the study and the nature of the material being administered. This selection provides a significant opportunity for refining animal treatment. Similarly, when substances are administered as solutions or suspensions, attention should be given to selection of vehicles and methods used for preparing the solutions and suspensions. The research team, veterinarian, technical personnel, and IACUC members should be aware of reasons underlying selection of equipment for substance delivery and should consider carefully how substances will be prepared and stored prior to administration to animals. Failure to consider these factors during experimental planning may result in unintentional adverse effects on experimental animals and confounded results.

Administration of substances to laboratory animals requires careful consideration and planning to optimize delivery of the agent to the animal while minimizing potential adverse experiences from the procedure. For all species, many different routes are available for administration of substances. The research team and IACUC members should be aware of reasons for selecting specific routes and of training and competency necessary for personnel to use these routes effectively. Once a route is selected, issues such as volume of administration, site of delivery, pH of the substance, and other factors must be considered to refine the technique. Inadequate training or inattention to detail during this aspect of a study may result in unintentional adverse effects on experimental animals and confounded results.

The highly conserved ANP32 proteins are proposed to function in a broad array of physiological activities through molecular mechanisms as diverse as phosphatase inhibition, chromatin regulation, caspase activation, and intracellular transport. On the basis of previous analyses of mice bearing targeted mutations of Anp32a or Anp32e, there has been speculation that all ANP32 proteins play redundant roles and are dispensable for normal development. However, more recent work has suggested that ANP32B may in fact have functions that are not shared by other ANP32 family members. Here we report that ANP32B expression is associated with a poor prognosis in human breast cancer, consistent with the increased levels of Anp32b mRNA present in proliferating wild-type (WT) murine embryonic fibroblasts and stimulated WT B and T lymphocytes. Moreover, we show that, contrary to previous assumptions, Anp32b is very important for murine embryogenesis. In a mixed genetic background, ANP32B-deficient mice displayed a partially penetrant perinatal lethality that became fully penetrant in a pure C57BL/6 background. Surviving ANP32B-deficient mice showed reduced viability due to variable defects in various organ systems. Study of compound mutants lacking ANP32A, ANP32B, and/or ANP32E revealed previously hidden roles for ANP32A in mouse development that became apparent only in the complete absence of ANP32B. Our data demonstrate a hierarchy of importance for the mammalian Anp32 genes, with Anp32b being the most critical for normal development.

A 1.5-year-old female rabbit (doe) was presented with a 3-day history of lethargy, anorexia, and mild facial swelling. The animal died shortly after examination and severe, acute hemorrhagic pneumonia was noted grossly. An alphaherpesvirus consistent with leporid herpesvirus-4 was isolated and characterized from this animal. This is the first confirmed report of the disease in Canada.

Pancreatic islet cell adenoma, oral squamous cell carcinoma, peripheral polyneuropathy, and multiple age-associated degenerative lesions were diagnosed in an aged Sprague-Dawley rat presenting with polyuria, polydypsia, dehydration, anorexia, weight loss, and posterior weakness. Microscopically, the islet cell adenoma was encapsulated by fibrous tissue and composed of packets of oval-to-polygonal monomorphic cells in a fibrovascular stroma. Immunohistochemically, the majority of cells within the mass expressed insulin. In light of the histologic and immunohistochemical findings, a diagnosis of insulinoma was made. The oral squamous cell carcinoma, grossly presenting as gingival ulceration, was composed of nests and cords of squamous epithelial cells that focally eroded and infiltrated the hard palate and resulted in degeneration of the maxillary nerve. The peripheral polyneuropathic lesions were characterized by extensive axonal degeneration and microangiopathic changes that were highly suggestive of a hypoglycemic etiopathogenesis secondary to insulinoma.