Jerald Silverman

Ammonia and carbon dioxide concentrations in disposable and reusable static mouse cages

The value of a static mouse cage is partially determined by the cages ability to dissipate carbon dioxide and ammonia. The authors compared the concentrations of ammonia and carbon dioxide produced by mice housed in two types of static isolator cages: a newly introduced disposable cage and a conventional reusable cage. Female mice were housed in three disposable and three reusable cages (n = 5 per cage). After 7 d, groups that were housed in disposable cages were rehoused in fresh reusable cages and vice versa. Intracage carbon dioxide, ammonia, temperature and relative humidity were measured daily. Overall, there were no significant differences in carbon dioxide or ammonia concentrations between the cage types. Within 30 min of placing mice in cages, carbon dioxide concentrations rose to more than 10,000 ppm in both cage types and rarely dropped below 8,000 ppm during both phases of the study. Ammonia concentrations rose slowly until day 4 and then began to rise rapidly. The maximum average ammonia concentration was 710 ppm. There was a highly significant relationship between increasing levels of humidity and ammonia production in the disposable cages (r = 0.88). For the reusable cages, the correlation was not as strong (r = 0.68). Both cage types were similar in their ability to dissipate carbon dioxide and ammonia. The suggested frequency of cage changing can only be estimated; on the basis of existing literature, it seems prudent to change cages when the ammonia concentration reaches 50 ppm.

Ensuring due process in the IACUC and animal welfare setting: Considerations in developing noncompliance policies and procedures for institutional animal care and use committees and institutional officials

Every institution that is involved in research with animals is expected to have in place policies and procedures for the management of allegations of noncompliance with the Animal Welfare Act and the U.S. Public Health Service Policy on the Humane Care and Use of Laboratory Animals. We present here a model set of recommendations for institutional animal care and use committees and institutional officials to ensure appropriate consideration of allegations of noncompliance with federal Animal Welfare Act regulations that carry a significant risk or specific threat to animal welfare. This guidance has 3 overarching aims: 1) protecting the welfare of research animals; 2) according fair treatment and due process to an individual accused of noncompliance; and 3) ensuring compliance with federal regulations. Through this guidance, the present work seeks to advance the cause of scientific integrity, animal welfare, and the public trust while recognizing and supporting the critical importance of animal research for the betterment of the health of both humans and animals.—Hansen, B. C., Gografe, S., Pritt, S., Jen, K.‐L. C., McWhirter, C. A., Barman, S. M., Comuzzie, A., Greene, M., McNulty, J. A., Michele, D. E., Moaddab, N., Nelson, R. J., Norris, K., Uray, K. D., Banks, R., Westlund, K. N., Yates, B. J., Silverman, J., Hansen, K. D., Redman, B. Ensuring due process in the IACUC and animal welfare setting: considerations in developing noncompliance policies and procedures for institutional animal care and use committees and institutional officials. FASEB J. 31, 4216–4225 (2017). www.fasebj.org

The Institutional Animal Care and Use Committee

The institutional animal care and use committee (IACUC) is a federally mandated committee established in institutions that receive federal funding for research, testing, or teaching using live animals. The IACUC represents its institutions animal care and use program to the federal government, assures compliance with federal regulations governing animal use, and helps to assure the welfare of research animals. The regulations followed by the IACUC emanate from the Animal Welfare Act and the Health Research Extension Act of 1985. The IACUC must approve animal use procedures before a study begins or before a change is made to an already approved study. Noncompliance with regulatory requirements can occur from a lack of adequate training, unrealistic expectations placed on investigators or educators, cultural misunderstandings, inadequate study monitoring, or from an institutional culture that does not support compliance. Compliance can be strengthened by adequate training of all animal users, adequately monitoring ongoing studies, establishing clear and simple guidelines for compliance (such as no research or teaching that uses live animals can be performed without prior IACUC approval), and promoting an organizational culture supporting compliance. The IACUC has the authority to investigate allegations of noncompliance and, if necessary, the committee can suspend an ongoing animal activity.

Factors Influencing IACUC Decision Making: Who Leads the Discussions?

Decisions about the appropriate use of animals in research are largely made by Institutional Animal Care and Use Committees (IACUCs). Several commentators claim that scientists exert excessive influence on IACUC decisions. We studied 87 protocol reviews from 10 IACUCs to assess whether any group of participants appeared to dominate the protocol discussions. Audio recordings of the meetings were coded to capture the topics of the discussions. We found that animal research scientists made the largest total number of topic mentions while community members made the fewest. On a per person basis, chairpersons and attending veterinarians made the most mentions. Scientists presented the largest number of protocols, and the subsequent discussions tended to contain the same topics mentioned in the presentations. The large number of protocols presented by scientists and their total number of comments made during protocol discussions suggest that scientists may significantly influence IACUC decision making.

Sensory neuron development in mouse coccygeal vertebrae and its relationship to tail biopsies for genotyping

A common method of genotyping mice is via tissue obtained from tail biopsies. However, there is no available information on the temporal development of sensory neurons in the tail and how their presence or absence might affect the age for performing tail biopsies. The goals of this study were to determine if afferent sensory neurons, and in particular nociceptive neurons, are present in the coccygeal vertebrae at or near the time of birth and if not, when they first can be visualized on or in those vertebrae. Using toluidine blue neuronal staining, transmission electron microscopy, and calcitonin-related gene peptide immunostaining, we found proximal to distal maturation of coccygeal nerve growth in the C57BL/6J mouse. Single nerve bundles were first seen on postpartum day (PPD) 0. On PPD 3 presumptive nociceptive sensory nerve fibers were seen entering the vertebral perichondrium. Neural development continued through the last time point (PPD 7) but at no time were neural fibers seen entering the body of the vertebrae. The effect of age on the development of pain perception in the neonatal mouse is discussed.

How should the IACUC balance an efficient approval process with minimizing risk

the Program has unique insights about the presence of biological, chemical, or radiation hazards in feed, animal secretions, and animal waste and about the extent of potential human exposure during animal experimentation and husbandry. Although the institution carries the ultimate responsibilities for establishing and administering a functional OHSP, the IACUC is responsible for day to day oversight for all parts of the Program, including the OHSP7. Therefore, the IACUC is the best position for approval of animal use activities involving hazards6. Indeed, the IACUC Handbook7 states that the IACUC must have members with sufficient technical expertise to evaluate health risks associated with Animal Use Protocols, so the implication is that safety committees inform the IACUC review process, rather than review in parallel with the IACUC, although two-way communication is critical to ensure personnel safety. In fact, one The Guide requires that an Occupational Health and Safety Program (OHSP) be part of the animal care and use program3 and it references the Occupational Health and Safety in the Care and Use of Research Animals4 as guidance on establishment and performance of an OHSP. Examples of oversight of OHSP Program include, but are not limited to, verification of enrollment, training of individuals on Animal Use Protocols5; compliance with ancillary institutional committees such as the Institutional Biosafety, Radiation Safety, Institutional Review Board and Chemical Safety6. The IACUC is also required to review the OHSP during its semi-annual program evaluation6, which considers “some of the most important personnel issues, [..]the occupational health and safety of animal care, use, and support personnel”7 and including the “use of hazardous materials and provision of a safe working environment”2. The IACUC must report deficiencies in the OHSP to OLAW/NIH7. RESPONSE

The role of the IACUC in ensuring research reproducibility

There is a “village” of people impacting research reproducibility, such as funding panels, the IACUC and its support staff, institutional leaders, investigators, veterinarians, animal facilities, and professional journals. IACUCs can contribute to research reproducibility by ensuring that reviews of animal use requests, program self-assessments and post-approval monitoring programs are sufficiently thorough, the animal model is appropriate for testing the hypothesis, animal care and use is conducted in a manner that is compliant with external and institutional requirements, and extraneous variables are minimized. The persons comprising the village also must have a shared vision that guards against reproducibility problems while simultaneously avoids being viewed as a burden to research. This review analyzes and discusses aspects of the IACUCs “must do” and “can do” activities that impact the ability of a study to be reproduced. We believe that the IACUC, with support from and when working synergistically with other entities in the village, can contribute to minimizing unintended research variables and strengthen research reproducibility.

Should refinement 'mess with success'?

consult with the institution’s veterinarian to weigh the pros and cons of the suggested laparoscopic cholecystectomy approach. Based on available literature, the veterinarian should provide to Clark objective evidence of the advantages of the laparoscopic approach, such as less postoperative pain, faster recovery and less experimental variability. The veterinarian should also provide a scientific explanation as to why the newer approach would not affect the scientific goal of Clark’s study. Similarly, the veterinarian and IACUC should provide an opportunity for Clark to express his concerns, and Clark should give a scientific rationale to support his views. Further, we don’t think that Clark’s explanation regarding the success of his past surgical approach provides sufficient justification to disregard the veterinarian’s suggestion to use a new, less invasive approach that will minimize animal discomfort and pain. Also, the monetary cost does not constitute a scientific justification for not adapting a suggested refinement technique. If Clark is still concerned about his scientific outcome, the IACUC should effect of minimizing pain and distress in laboratory animals used for studies. According to the Animal Welfare Act, the IACUC should determine that the principal investigator has considered alternatives to procedures that may cause more than momentary or slight pain or distress to the animal (§2.31(d); ref. 5). Therefore, in our opinion, the new veterinarian at Great Eastern University was justified both in questioning Clark’s invasive approach for cholecystectomy and in suggesting the novel, alternative laparoscopy approach, which would be less invasive. The concept of refinement is a dynamic process that constantly evolves as new technologies and inventions become available. On the other hand, it is reasonable to expect resistance from an investigator when suggesting refined methods if his methods are already working well. The burden often lies with the IACUC and its institution to convince Principal Investigators and implement refinement approaches to minimize distress and discomfort in laboratory animal procedures. In this particular scenario, the IACUC committee should recommend that Clark RESPONSE

Use of the home cage as an anesthesia induction chamber

and enrichment devices, can inhibit visualization of the mouse, hindering observers’ ability to evaluate the anesthetic plane, and can also obstruct the nares, cause corneal injury or become lodged in the animal’s mouth if the transition to unconsciousness is not quick and smooth. In contrast, an induction chamber permits clear visualization and continuous monitoring of the animal. We feel that removing an animal from its home cage and inducing anesthesia within a specialized chamber is preferred. This practice minimizes stress to the animal and its cage-mates, is a safe way to anesthetize individual animals and allows animals to recover in a familiar home cage. The IACUC must weigh protocols carefully and consider the potential pain and stress experienced both by an individual animal and by other animals in the same cage, with the goal of minimizing pain and stress to all animals.

Biomedical Research Techniques

The chapter on biomedical research techniques encompasses a large number of procedures – some common – some restricted to hamsters, which are used in research with the Syrian golden hamster. The chapter is written primarily for the needs of the laboratory animal veterinarian or biomedical researcher. Each technique is either described in detail or directs the reader to an appropriate reference. Included are discussions on general handling and restraint, tooth trimming, radiology, electrocardiography, collecting biological specimens (such as blood, urine, milk, cerebrospinal fluid, etc.), injections and intubations, catheterizations, many surgical procedures (such as ovariectomy, adrenalectomy, and olfactory bulbectomy), and more.