Three Rs Approaches in the Production and Quality Control of Fish Vaccines

The workshop on Three Rs Approaches in the Production and Quality Control of Fish Vaccines aimed a) to identify animal tests currently stipulated for the production and quality control of fish vaccines and to highlight animal welfare concerns associated with these tests; b) to identify viable options to replace, reduce, and refine animal use for fish vaccine testing; and c) to discuss the way forward and set out how the Three Rs may be implemented without jeopardizing the quality of the vaccines. The workshop participants -- experts from academia, regulatory authorities, a scientific animal welfare organization, and the fish vaccine industry -- agreed that efforts should be undertaken to replace the vaccination challenge batch potency testing with tests based on antigen quantification or antibody response tests. Regulatory requirements of questionable scientific value and relevance for the quality of fish vaccines, such as the re-testing of batches produced outside Europe, or the double-dose batch safety test, should be re-considered. As an immediate measure the design of the current animal tests should be evaluated and modified in the light of refinement and reduction, for example, the number of unprotected control fish in vaccination-challenge tests should be reduced to the minimum

Overcoming scientific barriers in the transition from in vivo to non-animal batch testing of human and veterinary vaccines

INTRODUCTION : Before release, vaccine batches are assessed for quality to evaluate whether they meet the product specifications. Vaccine batch tests, in particular of inactivated and toxoid vaccines, still largely rely on in vivo methods. Improved vaccine production processes, ethical concerns, and suboptimal performance of some in vivo tests have led to the development of in vitro alternatives. AREAS COVERED : This review describes the scientific constraints that need to be overcome for replacement of in vivo batch tests, as well as potential solutions. Topics include the critical quality attributes of vaccines that require testing, the use of cell-based assays to mimic aspects of in vivo vaccine-induced immune responses, how difficulties with testing adjuvanted vaccines in vitro can be overcome, the use of altered batches to validate new in vitro test methods, and how cooperation between different stakeholders is key to moving the transition forward. EXPERT OPINION : For safety testing, many in vitro alternatives are already available or at an advanced level of development. For potency testing, in vitro alternatives largely comprise immunochemical methods that assess several, but not all critical vaccine properties. One-to-one replacement by in vitro alternatives is not always possible and a combination of methods may be required.The Dutch Ministry of Agriculture, Nature and Food Quality, the National Institute of Public Health & the Environment and Intravacc.https://www.tandfonline.com/loi/ierv20am2022Veterinary Tropical Disease

The Three Rs: The Way Forward

This is the report of the eleventh of a series of workshops organised by the European Centre for the Validation of Alternative Methods (ECVAM), which was established in 1991 by the European Commission. ECVAM\u27s main goal, as defined in 1993 by its Scientific Advisory Committee, is to promote the scientific and regulatory acceptance of alternative methods which are of importance to the biosciences and which reduce, refine or replace the use of laboratory animals. One of the first priorities set by ECVAM was the implementation of procedures which would enable it to become well-informed about the state-of-the-art of non-animal test development and validation. and the potential for the possible incorporation of replacement alternative tests into regulatory procedures. It was decided that this would be best achieved by the organisation of ECVAM workshops on specific topics, at which small groups of invited experts would review the current status of various types of in vitro tests and their potential uses, and make recommendations about the best ways forward

The Isolated Chicken Eye test to replace the Draize test in rabbits

In 1944, Draize et al., published a paper entitled “Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes”. The Organization for Economic Co-operation and Development published their first guideline on eye irritation in 1981, using rabbits. In the early eighties the development of alternative non-animal tests to replace the Draize eye test started. The first attempts to validate alternative tests for eye irritation were considered to be relatively simple by comparing in vitro and in vivo irritation index scores. In the early nineteen-eighties, we introduced the use of isolated eyes as an alternative test for the Draize eye irritation test. What was expected to be a process of several years, however, turned out to be a decades spanning process still not fully completed. For a large part, this can be attributed to the nature of the in vivo test in rabbits, which is more complicated and compromised than originally believed. This paper describes, most chronologically, the development, performance, validation and application of the Isolated Eye Test and, in broader perspective, the international validation and acceptance of this alternative test by regulatory authorities and agencies.</p

Workaholic ferrets: Does a two-chamber consumer demand study give insight in the preferences of laboratory ferrets (Mustela putorius furo)?

Although provision of environmental enrichment is an effective tool to refine laboratory animal experiments, it is currently unknown which enrichments ferrets prefer. This study aimed to assess the suitability of a closed economy, two-chamber consumer demand set-up to determine ferrets’ preferences for selected enrichments. Twelve female ferrets were housed in a set-up consisting of a home and enrichment chamber (EC) connected by a weighted door. The maximum weights the ferrets pushed for food (MPPfood) and an empty chamber (MPPempty) were determined to evaluate the maximum push capacity of the animals and as a control. Although the ferrets pushed significantly more for food (1325 ± 213 g)than for the empty chamber (1169 ± 193 g), the weight difference was minor (MPPempty was 89 ± 13% of MPPfood). To evaluate the ferrets’ underlying motivation to push for the empty chamber, a second study was performed in which MPPempty was tested in seven alternative set-ups. The first three set-ups included adapted versions of the standard design (set-up A1, A2and A3), intended to determine the functional value of the empty chamber. The four other set-ups (set-up B0, B1, B3, B4) aimed to evaluate the attractiveness of the door elements by allowing the ferrets to choose whether or not to use the weighted door to enter EC. Results demonstrated no significant differences in MPPempty between the A-set-ups, indicating that the value of the empty chamber could not be reduced by adapting the set-up. MPPempty reduced when allowing the ferrets free access to EC, demonstrating that the empty chamber had reinforcing proper-ties. Nevertheless, the ferrets were still motivated to use the weighted door despite being granted free access to EC, indicating that the door also has reinforcing properties. The ferrets decreased the use of the weighted door most when, in a set-up with free access to EC, the nest box in the home cage (53 ± 22% of MPPfood) was replaced by a manipulable plastic bucket (26 ± 13% of MPPfood). These results indicate that availability of items in the home chamber may influence the results, which should be taken into account when designing motivation studies similar to the one performed in this study. The lack of differences between MPPfood and MPPempty furthermore demonstrates that the two-chamber set-up is not suitable for evaluating the ferrets’ motivation for enrichments, thus necessitating other alternatives, such as at three- or multi-chamber consumer demand study, to be explored

Workaholic ferrets: Does a two-chamber consumer demand study give insight in the preferences of laboratory ferrets (Mustela putorius furo)?

Although provision of environmental enrichment is an effective tool to refine laboratory animal experiments, it is currently unknown which enrichments ferrets prefer. This study aimed to assess the suitability of a closed economy, two-chamber consumer demand set-up to determine ferrets’ preferences for selected enrichments. Twelve female ferrets were housed in a set-up consisting of a home and enrichment chamber (EC) connected by a weighted door. The maximum weights the ferrets pushed for food (MPPfood) and an empty chamber (MPPempty) were determined to evaluate the maximum push capacity of the animals and as a control. Although the ferrets pushed significantly more for food (1325 ± 213 g)than for the empty chamber (1169 ± 193 g), the weight difference was minor (MPPempty was 89 ± 13% of MPPfood). To evaluate the ferrets’ underlying motivation to push for the empty chamber, a second study was performed in which MPPempty was tested in seven alternative set-ups. The first three set-ups included adapted versions of the standard design (set-up A1, A2and A3), intended to determine the functional value of the empty chamber. The four other set-ups (set-up B0, B1, B3, B4) aimed to evaluate the attractiveness of the door elements by allowing the ferrets to choose whether or not to use the weighted door to enter EC. Results demonstrated no significant differences in MPPempty between the A-set-ups, indicating that the value of the empty chamber could not be reduced by adapting the set-up. MPPempty reduced when allowing the ferrets free access to EC, demonstrating that the empty chamber had reinforcing proper-ties. Nevertheless, the ferrets were still motivated to use the weighted door despite being granted free access to EC, indicating that the door also has reinforcing properties. The ferrets decreased the use of the weighted door most when, in a set-up with free access to EC, the nest box in the home cage (53 ± 22% of MPPfood) was replaced by a manipulable plastic bucket (26 ± 13% of MPPfood). These results indicate that availability of items in the home chamber may influence the results, which should be taken into account when designing motivation studies similar to the one performed in this study. The lack of differences between MPPfood and MPPempty furthermore demonstrates that the two-chamber set-up is not suitable for evaluating the ferrets’ motivation for enrichments, thus necessitating other alternatives, such as at three- or multi-chamber consumer demand study, to be explored

Overcoming scientific barriers in the transition from in vivo to non-animal batch testing of human and veterinary vaccines

Introduction Before release, vaccine batches are assessed for quality to evaluate whether they meet the product specifications. Vaccine batch tests, in particular of inactivated and toxoid vaccines, still largely rely on in vivo methods. Improved vaccine production processes, ethical concerns, and suboptimal performance of some in vivo tests have led to the development of in vitro alternatives. Areas covered This review describes the scientific constraints that need to be overcome for replacement of in vivo batch tests, as well as potential solutions. Topics include the critical quality attributes of vaccines that require testing, the use of cell-based assays to mimic aspects of in vivo vaccine-induced immune responses, how difficulties with testing adjuvanted vaccines in vitro can be overcome, the use of altered batches to validate new in vitro test methods, and how cooperation between different stakeholders is key to moving the transition forward. Expert opinion For safety testing, many in vitro alternatives are already available or at an advanced level of development. For potency testing, in vitro alternatives largely comprise immunochemical methods that assess several, but not all critical vaccine properties. One-to-one replacement by in vitro alternatives is not always possible and a combination of methods may be required

The Production of Polyclonal Antibodies in Laboratory Animals

This is the report of the thirty-fifth of a series of workshops organised by the European Centre for the Validation of Alternative Methods (ECVAM). ECVAM\u27s main goal, as defined in 1993 by its Scientific Advisory Committee, is to promote the scientific and regulatory acceptance of alternative methods which are of importance to the biosciences and which reduce, refine or replace the use of laboratory animals. One of the first priorities set by ECVAM was the implementation of procedures which would enable it to become well-informed about the state-of-the-art of non-animal test development and validation, and the potential for the possible incorporation of alternative tests into regulatory procedures. It was decided that this would be best achieved by the organisation of ECVAM workshops on specific topics, at which small groups of invited experts would review the current status of various types of in vitro tests and their potential uses, and make recommendations about the best ways forward (1)

Overcoming scientific barriers in the transition from in vivo to non-animal batch testing of human and veterinary vaccines

Introduction Before release, vaccine batches are assessed for quality to evaluate whether they meet the product specifications. Vaccine batch tests, in particular of inactivated and toxoid vaccines, still largely rely on in vivo methods. Improved vaccine production processes, ethical concerns, and suboptimal performance of some in vivo tests have led to the development of in vitro alternatives. Areas covered This review describes the scientific constraints that need to be overcome for replacement of in vivo batch tests, as well as potential solutions. Topics include the critical quality attributes of vaccines that require testing, the use of cell-based assays to mimic aspects of in vivo vaccine-induced immune responses, how difficulties with testing adjuvanted vaccines in vitro can be overcome, the use of altered batches to validate new in vitro test methods, and how cooperation between different stakeholders is key to moving the transition forward. Expert opinion For safety testing, many in vitro alternatives are already available or at an advanced level of development. For potency testing, in vitro alternatives largely comprise immunochemical methods that assess several, but not all critical vaccine properties. One-to-one replacement by in vitro alternatives is not always possible and a combination of methods may be required