Applying the 3Rs at Pirbright
The principles of the 3Rs, Reduction (in numbers), Refinement (of procedures) and Replacement (with laboratory procedures) were developed over 50 years ago as a framework for humane animal research.
Improvements in veterinary medicines and diagnostics are the result of years of research and much of the time is spent doing research in laboratories (in-vitro). However, the sheer complexity of viral diseases and the hosts’ immune responses to them means that research with animals (in-vivo) is essential to better understand the disease and to develop new methods of control.
At The Pirbright Institute this usually entails working with the species for whose improved health the research is directed (cattle, poultry, sheep and pigs). We also use small numbers of mice and rabbits in our research.
Whilst the number of animals used at the Institute is tiny compared to the millions of animals that benefit from the Institute’s research, we strive to apply the principles of the 3Rs for the benefit of animals used in research and for the quality of the data that they yield.
- ASF research
- FMD vaccine matching
- Feeding our insect colonies
- Tick Cell Biobank
- Statistical review of experiments
Before beginning any research involving animals, experimental design is carried out in collaboration with statisticians to ensure statistically relevant results are obtained. Vaccine candidates are extensively tested in cell cultures before advancing to the stage of animal experiments to reduce the number of animals needed.
The use of standardised clinical and pathology scoring facilitates comparisons between experiments and thus reduces the numbers of groups which have to be used in each experiment. Continued re-evaluation of results from experiments mean the data collected can be refined and the outcomes of experiments can be predicted based on cell culture and analysis of host responses that correlate with protection.
Cells taken from immune animals are also used to identify those ASFV proteins that are recognised by the host immune system which may be capable of inducing protection. This greatly reduces the requirement for animal experiments to obtain this information.
Together with mathematical modellers at Glasgow University, researchers are working towards a robust and challenge-free computational based model for the assessment of FMD vaccine efficacy.
Traditionally, laboratory study of tick-borne pathogens and development of methods to control ticks and the diseases they transmit requires maintenance of ticks by feeding them for up to four weeks on suitable animal hosts (for example rabbits, gerbils, sheep and cattle).
However, continuous tick cell lines derived from the tick vectors of many of these pathogens are increasingly being used as alternatives to whole ticks in experiments to increase fundamental understanding of the relationships between ticks and pathogens, and the mechanisms of pathogen acquisition, survival and transmission by the tick to the vertebrate host. In addition, tick cell lines are now being explored as model systems to understand how acaricides (insecticides specifically targeted at ticks) work, how ticks develop resistance to acaricides, and to find novel targets for new acaricide development. In both cases, most of the preliminary and developmental experiments can be carried out in vitro in tick cell lines, before moving to whole tick feeding on vertebrate hosts in the final stages, thereby greatly reducing experimental animal usage and enabling many more laboratories that lack tick feeding facilities to contribute to this research field.
The Tick Cell Biobank was created at the University of Edinburgh in 2009, using funding from the Wellcome Trust, to house almost all of the world’s surviving tick cell lines. At that time these numbered around 50; the majority were established from the 1970s onwards by just three scientists: Tim Kurtti and Ulrike Munderloh at the University of Minnesota and Lesley Bell-Sakyi in Edinburgh. The Biobank moved to The Pirbright Institute in September 2012 with Dr Lesley Bell-Sakyi.
The Tick Cell Biobank now holds nearly 60 cell lines derived from most of the vectors of the important tick-borne diseases of domestic animals and humans (anaplasmosis, babesiosis, cowdriosis, theileriosis, borreliosis, ehrlichiosis, rickettsiosis, African swine fever, Nairobi sheep disease, tick-borne encephalitis, Crimean-Congo haemorrhagic fever etc). New cell lines are currently being developed from additional tick species of medical and/or veterinary importance.
Tick cell lines are supplied to recipient laboratories under suitable MTAs and at affordable cost (taking into consideration ability to pay). To increase the uptake and improve the rate of successful transfer to the new laboratory, free training is provided on request to the recipient scientists. Since September 2012, 175 tick cell lines have been distributed to over 40 research laboratories in UK, Europe, Asia, Africa, North and South America and 41 recipient scientists have been trained.
Formal statistical review of animal experiments is part of the ethical review process at The Pirbright Institute to ensure that all studies are designed appropriately and have sufficient statistical power to detect effects that are biologically meaningful.
Pirbright's in-house statistican, Dr Simon Gubbins assists researchers with their grant submissions to optimise animal numbers and experimental design.
Experiments are designed based on the magnitude of effect that needs to be detected for a study to be informative (e.g. difference in levels of viraemia; proportion of animals protected by a vaccine), the expected level of variation amongst animals and the power (typically 80%) and confidence (typically 95%) required. In most cases, the required number of animals can be determined by standard methods. For more complex designs, however, group sizes may need to be determined by simulating the statistical model underlying the experiments in computer packages.