In this article, adverse events are defined as events that lead to significant injury or illness, unrelieved pain or distress, or the death of an animal, excluding those caused by IACUC-approved research procedures. The ability to identify possible adverse events is an essential step in planning for risk prevention and mitigation. Using data from news sources and the Animal and Plant Health Inspection Service (APHIS) website, this article provides guidance to research facilities regarding various types of adverse events to consider in risk management plans because these events might occur or have occurred and, in some cases, have been documented as noncompliances at APHIS-regulated research facilities. APHIS classifies noncompliances as 'direct' when they currently (at the time of the inspection) have a serious or severe adverse effect on the health and wellbeing of animals. Not all direct noncompliances are associated with adverse events, and not all adverse events are documented as direct noncompliances (for example, a past adverse event that does not currently affect the wellbeing of animals is not a direct noncompliance). However, because APHIS does not require reporting of adverse events, the information regarding direct noncompliances was the only APHIS data available to study adverse events at research facilities. Direct noncompliances documented by APHIS were 4% of all documented noncompliances from 2010 through 2014. The greatest number of direct noncompliances was in the category of veterinary care issues (44%), followed by animal husbandry issues (34%), 'other issues' (15%), and physical plant issues (7%). The category of other issues included events due to human error, equipment failures, and accidents.

Black-tailed prairie dogs (Cynomys ludovicianus) are kept in zoological collections, maintained as companion pets, and are tested in field and laboratory settings. Biochemical analysis for routine health and research purposes can be performed by using point-of-care (POC) testing; however, analyzer- and species-specific reference intervals need to be determined. In this prospective study, 50 captive-raised sexually intact prairie dogs (16 females, 34 males) underwent plasma biochemical analysis by using a veterinary POC biochemical analyzer. We used a manufacturer-predetermined profile of 14 analytes: albumin, ALP, ALT, amylase, BUN, calcium, creatinine, glucose, potassium, sodium, phosphorus, total bilirubin, total protein and globulin. A subset of 17 samples was tested concurrently for the same 14 analytes by using a reference laboratory analyzer, and we determined RI for the POC analyzer for these 14 biochemical analytes. Sex had a significant effect on albumin and creatinine values, which were higher in females than males, and on ALT, which was lower in females. In addition, age had an effect on 9 plasma analytes: juvenile animals had higher plasma concentrations of albumin, ALP, ALT, BUN, and glucose than adult animals, whereas adults had higher concentrations of creatinine, sodium, total protein, and globulins. Only calcium and BUN had acceptable analytical agreement between the POC and reference analyzers. The reference intervals determined in this study can aid clinicians and researchers performing POC plasma biochemical analysis in prairie dogs, providing that they consider potential analyzer-, sex-, and age-related effects.

Silastic capsules are frequently used to study the physiologic effects of estrogen exposure in animal models. The Office of Laboratory Animal Welfare requires the sterilization of nonpharmaceutical-grade compounds before use. We compared 2 commonly used terminal sterilization methods—ionizing radiation (IR) and ethylene oxide (EO)—for their utility in sterilizing silastic capsules containing 0.05 or 0.1 mg 17β-estradiol (E2). E2-specific ELISA demonstrated that serum estrogen levels did not differ between mice implanted with 0.05-mg E2 capsules that were sterilized with IR or EO and those implanted with nonsterilized capsules. Likewise, mammary gland morphology and progesterone receptor expression and proliferation in mammary epithelium were similar among mice treated with E2 capsules, regardless of sterilization method, and pregnant day 15 mice. In addition, IR-sterilized 0.1-mg E2 pellets provided high serum E2. We conclude that neither ionizing radiation nor ethylene oxide degraded E2 or the cellulose matrix, suggesting that these methods of sterilization are appropriate to provide effective sterile hormone capsules for animal research.

The physical form of the diet fed to laboratory animals should be evaluated to reduce experimental variations and confounding factors. This 14-d study evaluated the effects of diet form (pelleted or extruded) on intracage ammonia concentrations, feed disappearance, body weight, cage weight, and the degree of cage soilage and whether these effects were influenced by strain or stock or sex. Mice (C57BL/6, ICR, and nude; age, 4 wk) were randomly assigned to 4 treatment groups representing pelleted and extruded diets from each of 2 vendors (pelleted diet groups, P1 and P2; extruded diet groups, E1 and E2). Intracage ammonia concentrations depended on strain or stock, diet, and day and were higher in cages housing nude mice that consumed P1. Diet type did not affect the weight of mice at the end of the study. Feed disappearance was dependent on diet type and mouse strain or stock and was greatest in the cages of mice that consumed P1. In addition, the greatest feed disappearance was seen with ICR mice, whereas the least was seen with C57BL/6 mice. Cages housing male nude mice had greater cage soilage than those housing female nude mice. The degree of cage soilage was influenced by diet type and day also. These results show that diet form and mouse strain or stock significantly affect intracage ammonia concentrations, feed disappearance, cage weight, and the degree of cage soilage.

The beneficial effect of music has been demonstrated in many species. Although commercially available music CDs intended for use with rabbits are available, these enrichments have not been critically evaluated to determine whether they reduce distress. In this study, we used an aging colony of male rabbits to evaluate how the provision of music enrichment affected the wellbeing of the colony. After 6 mo of music enrichment, the rabbits in the colony demonstrated a significant decrease in fecal cortisol, suggesting that their stress was reduced. Six months after removal of the music enrichment, the rabbits demonstrated a significant increase in fecal cortisol and the heterophil:lymphocyte ratio, suggesting that they were stressed. These findings suggest that music enrichment with a commercially available music discs for rabbits can be used to improve the wellbeing of animals used in biomedical research.

Body condition scoring (BCS) is a simple, rapid, noninvasive tool used to assess body condition in animals. In this study, we developed and validated a diagram-based BCS for adult zebrafish (Danio rerio), a popular research model. After receiving 20 min of hands-on training regarding the scoring system, 5 people each rated 95 adult zebrafish. The fish then were euthanized and measured to establish body condition indices (BMI and the Fulton K factor). Both condition indices were highly correlated with fish width. Using correlation data and observed trends in fish width, we established expected BCS definitions. We validated the BCS definitions in 2 ways. First, we calculated the Pearson correlation coefficient between the average observed BCS and expected BCS; this statistic revealed very strong correlation between observed and expected BCS. In addition, we assessed the predictive power of BCS by using multinomial logistic regression and then applied the fitted model to evaluate the accuracy of the predictions (BCS compared with BMI, 85%; BCS compared with K factor, 61%). Finally, to determine the robustness of BCS to variation among raters, we calculated the intraclass correlation coefficient and demonstrated high interrater reliability. In conclusion, adult zebrafish BCS can be used to quickly identify animals with different body condition indices (thin to obese). In addition, the diagram-based chart is easy to use and implement accurately, with minimal training.

Sterilization of rodent feed by steam autoclaving is a common practice in many research institutions. Often we only consider the beneficial effects of this process—the reduction of microbial contamination—and forget that the high temperatures and pressures can have negative effects on diet quality. The purpose of our study was to assess both the physical and chemical changes to a standard rodent feed autoclaved at multiple sterilization temperatures and the effects of the treated diets on mice. Pelleted NIH31 rodent feed was autoclaved at 4 sterilization temperatures (230, 250, 260, and 270 °F). Feed pellet hardness and the acrylamide concentrations of the diets were tested and compared with irradiated NIH31 feed. Study diets were fed to mice for 28 d, after which tissue samples were collected for analysis of acrylamide, glycidamide (the active metabolite of acrylamide), and genotoxicity. Both feed pellet hardness and acrylamide concentration increased with increasing sterilization temperatures; however, neither affected feed intake or body weight gain. Plasma acrylamide and glycidamide were significantly elevated only in mice fed NIH31 diet autoclaved at 270 °F compared with the irradiated feed, whereas urine acrylamide and glycidamide metabolites were significantly elevated in most autoclaved diets. Liver DNA adducts, which correlate with genotoxicity, were significantly elevated in all autoclaved diets compared with the irradiated diet. Institutions that autoclave their animal diets should carefully consider the temperatures necessary to achieve feed sterilization and the type of studies in which these autoclaved diets are used.

The implementation of principles and guidelines that govern the various areas of research in an educational institution is one of the key factors in international recognition of its research integrity and value. The privilege of conducting research using animal subjects requires adherence to international regulations and standards governing the humane care and use of laboratory animals. The IACUC at our university deemed it critical to have an animal care and use training program to raise researchers' understanding and knowledge. Our IACUC recently designed a training program in the principles of laboratory animal science and the ethical issues involved in animal use. The present study aimed to measure the effect of such training on scientists' attitudes and practice. During 4 successive training courses, the participants (n = 100; 72% women and 28% men) were surveyed twice through self-administered questionnaire—before starting and after completing the course. Questions focused on ethical consideration for care and use of animals in research, ethical committees, international guidelines for humane care of animals, and 3Rs concepts and their interpretation. The results revealed that the scientists' knowledge and awareness increased effectively after the completion of the training courses. They understood the 3Rs concepts of replacement, reduction, and refinement; recognized the importance of standardization of animal handling on scientific results; and were able to distinguish between different ethical committees and their roles. Overall, training leads to standardization of animal care and use practices that are vital for the reproducibility of results fundamental to quality scientific research.

Temperature monitoring during critical care provides important data required to guide treatment delivery. Body temperature is an easily quantified clinical parameter that can yield much information concerning the health of an animal. In research settings, temperature has been adopted as a means to judge humane endpoints. Therefore, reliable, noninvasive, and inexpensive methods for temperature monitoring are becoming a necessity in research laboratories. This study aimed to determine the accuracy and agreement of using an infrared camera as an alternative method of temperature measurement in mice and to compare the accuracy of this noninvasive method with established subcutaneous, intraperitoneal, and rectal techniques. Measurement of body-surface temperature by using an infrared camera was compared with these 3 established methods in male NMRI nude mice (n = 10; age, 10 mo); data were obtained 3 times daily over 14 d. Subcutaneous temperatures were measured remotely by using a previously implanted subcutaneous temperature transponder, after which temperature was measured by using noncontact infrared thermometry and a rectal probe. Measurements from intraperitoneal data loggers were obtained retrospectively. The data show that using an infrared camera provides a simple, reliable method for measuring body temperature in male NMRI nu/nu mice that minimizes handling and is minimally invasive. Whether infrared thermometry is a useful method for measuring body temperature in furred mice warrants further investigation.

Current recommendations for assessing animal wellbeing during euthanasia suggest that measuring neuroendocrine hormones—such as ACTH, noradrenaline, and adrenaline—is preferable to measuring corticosterone and blood glucose because of the sensitivity of neuroendocrine hormones to the acute stress associated with rapid methods of euthanasia. However, these neuroendocrine hormones can be stimulated in ways that confound interpretation of welfare assessment in euthanasia studies. Although this property does not negate the usefulness of neuroendocrine hormones as tools of assessment, it is important to differentiate the stress associated with the induction of anesthesia before the loss of consciousness (an animal wellbeing concern) with the physiologic responses that occur after the loss of consciousness (not an animal wellbeing concern). In this study, rats were anesthetized by using a ketamine–xylazine combination. Once the rats achieved a surgical plane of anesthesia, they were exposed to O₂, CO₂, or isoflurane, followed by terminal blood collection to assess concentrations of ACTH, noradrenaline, corticosterone, and blood glucose. Compared with animals exposed to O₂ or isoflurane, rats exposed to CO₂ had significant increases in their serum concentrations of ACTH and noradrenaline, but blood glucose and corticosterone did not differ between groups. These findings indicate that noradrenaline and ACTH should be used with caution to assess animal wellbeing when the method of euthanasia might confound that assessment.

This study compared behavioral and physiologic changes in Sprague–Dawley and Brown Norway rats that were euthanized by using a 30% volume displacement rate of CO₂ in either their home cage or an induction chamber; rats euthanized in the home cage were hypothesized to demonstrate a higher level of animal wellbeing. No significant differences were detected in the physiologic responses to home cage versus induction chamber euthanasia groups. A few strain-related behavioral differences occurred. The number of digs per second was higher in Brown Norway compared with Sprague–Dawley rats when in the home cage, where a digging substrate was present. Rearing frequency was higher in both Brown Norway and Sprague–Dawley rats in the induction chamber compared with the home cage. This study demonstrated that although strainspecific differences were associated with the process of euthanasia, there were no significant differences between the treatment groups of home cage compared with induction chamber. This finding suggests that—from the perspective of a rat—either the home cage or an induction chamber can be used for euthanasia, with likely extension of this conclusion to use of either method to the induction of anesthesia.

Due to similarities in placentation, guinea pigs can be used as models of human cytomegalovirus infection, but they must be free of guinea pig cytomegalovirus. Many commercial guinea pig colonies are enzootically infected with guinea pig cytomegalovirus, which can be transmitted vertically as well as horizontally through saliva, vaginal secretions, and milk. These characteristics make its eradication in a commercial setting challenging. Because embryo transfer technology in guinea pigs is in its infancy, it is not generally a viable option for obtaining animals free of guinea pig cytomegalovirus. However, a combination of hysterectomy rederivation and testing by PCR assay and serology of both dams and offspring from an enzootically infected colony produced offspring free of guinea pig cytomegalovirus.