Report on the international workshop on alternative methods for Leptospira vaccine potency testing: state of the science and the way forward.

Routine potency testing of Leptospira vaccines is mostly conducted using a vaccination-challenge test that involves large numbers of hamsters and unrelieved pain and distress. NICEATM, ICCVAM, and their international partners organized a workshop to review the state of the science of alternative methods that might replace, reduce, and refine the use of animals for veterinary Leptospira vaccine potency testing and to identify ways to advance improved alternative methods. Vaccine manufacturers were encouraged to initiate or continue product-specific validation using in vitro enzyme-linked immunosorbent assays as replacements for potency testing of four common Leptospira serogroups. Participants discussed the potential for eliminating the back-titration procedure in the hamster challenge assay, which could reduce animal use by 50% for each individual potency test. Further animal reduction may also be possible by using cryopreserved Leptospira stock to replace continual passaging through hamsters. Serology assays were identified as a way to further reduce and refine animal use but should be considered only after attempting in vitro assays. Workshop participants encouraged consideration of analgesics and use of earlier humane endpoints when the hamster vaccination-challenge potency assay is used. International harmonization of alternative potency methods was recommended to avoid duplicative potency testing to meet regionally different requirements.

Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward.

Potency testing of most human and veterinary rabies vaccines requires vaccination of mice followed by a challenge test using an intracerebral injection of live rabies virus. NICEATM, ICCVAM, and their international partners organized a workshop to review the availability and validation status of alternative methods that might reduce, refine, or replace the use of animals for rabies vaccine potency testing, and to identify research and development efforts to further advance alternative methods. Workshop participants agreed that general anesthesia should be used for intracerebral virus injections and that humane endpoints should be used routinely as the basis for euthanizing animals when conducting the mouse rabies challenge test. Workshop participants recommended as a near-term priority replacement of the mouse challenge with a test validated to ensure potency, such as the mouse antibody serum neutralization test for adjuvanted veterinary rabies vaccines for which an international collaborative study was recently completed. The workshop recommended that an in vitro antigen quantification test should be a high priority for product-specific validation of human and non-adjuvanted veterinary rabies vaccines. Finally, workshop participants recommended greater international cooperation to expedite development, validation, regulatory acceptance, and implementation of alternative test methods for rabies vaccine potency testing.

Non-animal replacement methods for veterinary vaccine potency testing: state of the science and future directions.

NICEATM and ICCVAM convened an international workshop to review the state of the science of human and veterinary vaccine potency and safety testing methods and to identify opportunities to advance new and improved methods that can further reduce, refine, and replace animal use. Six topics were addressed in detail by speakers and workshop participants and are reported in a series of six reports. This workshop report, the second in the series, provides recommendations for current and future use of non-animal methods and strategies for veterinary vaccine potency testing. Workshop participants recommended that future efforts to replace animal use give priority to vaccines (1) that use large numbers of animals per test and for which many serials are produced annually, (2) that involve significant animal pain and distress during procedures, (3) for which the functional protective antigen has been identified, (4) that involve foreign animal/zoonotic organisms that are dangerous to humans, and (5) that involve pathogens that can be easily spread to wildlife populations. Vaccines identified as the highest priorities were those for rabies, Leptospira spp., Clostridium spp., Erysipelas, foreign animal diseases (FAD), poultry diseases, and fish diseases. Further research on the identification, purification, and characterization of vaccine protective antigens in veterinary vaccines was also identified as a priority. Workshop participants recommended priority research, development, and validation activities to address critical knowledge and data gaps, including opportunities to apply new science and technology. Recommendations included (1) investigations into the relative impact of various adjuvants on antigen quantification assays, (2) investigations into extraction methods that could be used for vaccines containing adjuvants that can interfere with antigen assays, and (3) review of the current status of rabies and tetanus human vaccine in vitro potency methods for their potential application to the corresponding veterinary vaccines. Workshop participants recommended enhanced international harmonization and cooperation and closer collaborations between human and veterinary researchers to expedite progress. Implementation of the workshop recommendations is expected to advance alternative in vitro methods for veterinary vaccine potency testing that will benefit animal welfare and replace animal use while ensuring continued protection of human and animal health.

Decerebrate mammalian preparations: unalleviated or fully alleviated pain? A review and opinion.

In experimental decerebration of mammals, the cerebral cortex and thalamus are surgically or otherwise inactivated under traditional (pharmacologic) general anesthesia. Once the effects of the pharmacologic anesthesia have dissipated, the animal remains alive, but there is neither pain sensation nor consciousness. Because the Animal Welfare Act and its regulations recognize drugs as the only means to alleviate pain, it is unclear whether a decerebrate animal should be placed in U.S. Department of Agriculture (USDA) pain and distress category D (pain or distress alleviated by drugs) or E (unalleviated pain or distress). We present a rationale for including decerebrate animals in USDA category D. We also provide a general review of decerebration and suggestions for institutional animal care and use committees having to evaluate decerebration protocols.

USDA perspective on environmental enrichment for animals.

This article provides a brief historical background of the events and circumstances that led to the 1985 Animal Welfare Act (AWA) amendments. It describes the development of the regulations promulgated by the US Department of Agriculture (USDA) in 1991 as a result of these amendments, the reasoning given for the proposals, and the revisions that were made during the process. Information is included on USDA implementation of the regulations regarding exercise for dogs and environmental enhancement for nonhuman primates. Also mentioned briefly are the requirements for socialization of marine mammals and space requirements for certain other regulated warm-blooded species. These requirements apply to animal dealers (breeders and brokers), exhibitors, commercial transporters, and research facilities. The standards for exercise and environmental enhancement were different from any others previously contained in the AWA regulations, and required more research and understanding of species-specific needs by the regulated community. Finally, this article describes some of the initiatives being undertaken by the research community and USDA-Animal and Plant Health Inspection Services (APHIS)-Animal Care to provide the necessary education and guidance indicated by the violation history data.

Report and recommendations of the international workshop "Retrieval approaches for information on alternative methods to animal experiments".

In the member states of the EU and in the USA, scientists are obliged by animal welfare legislation not to conduct an animal experiment if another scientifically satisfactory method is reasonably and practicably available. To meet the regulatory obligation to use alternatives to animal experiments, scientists should consult literature and other relevant sources on alternatives prior to any experimental study on laboratory animals. It is the responsibility of the individual scientist to select the most appropriate database to obtain information on alternatives, which have been defined as methods that refine, reduce or replace animal experiments (the 3 Rs concept of Russell and Burch (1959)). Specialised information services provide support to scientists searching for publications on alternative methods. On occasion of a workshop in Berlin in November 2003, representatives of animal welfare information centres discussed currently available information sources on alternative methods, index terms for alternative methods, and search strategies based on index terms for alternative methods. ZEBET presented an investigation on the current status of indexing systems on alternative methods in established literature databases. The project analysed how the results of a search for publications on alternatives was influenced by the indexing procedure. The results of the study were exemplified by a typical search result. The results of the study indicated that the current indexing systems do not provide the required information, since not all of the relevant information is indexed under "alternative methods". The workshop participants developed recommendations for ad hoc working groups and research projects, e.g. development of suitable search strategies on alternative methods for scientists.