Openness Statement on the Use of Animals in Project Plains Zebra
The researchers involved in ‘Project Plains Zebra’ are fully aware of the spectrum of views regarding the use of animals in scientific research, ranging from the Utilitarian concept that it can be useful to both the animals and humans to use animals in scientific research, through to the Deontological view that it is intrinsically wrong to use animals in scientific experiments.
As we are seeking non-traditional financial support for this project, and from necessity will be using animals in this research project, it is appropriate that we acknowledge this spectrum of views and detail how we address both specific and general concerns that may be raised. By doing this we allow those who are considering supporting to this project, but who may have concerns about the use of animals in research, to have clarity as to why we aim to undertake the work proposed and how animals will be used in this research.
It is our hope that this openness regarding the proposed research will build trust in our endeavour, the individuals undertaking the research, and the intentions of this project.
Aims and Benefits of Project Plains Zebra:
The central aim of this project is to develop a pragmatic data-based paradigm, based on intensive, targeted scientific research, from which appropriate conservation and management strategies for the African Savannah Ecosystem can be developed. There are benefits for the Fauna and Flora, as well as humans, by preserving this ecosystem.
Benefits to Fauna and Flora: Understanding and conserving the African savannah ecosystem will have widespread benefits to all the species, from the micro-organisms through to the complex multicellular flora and fauna that inhabit this ecosystem. Unregulated, or inappropriately managed, human activity would be an existential threat to the flora and fauna of this ecosystem – the hundreds of thousands of species that inhabit this ecosystem would likely be doomed to extinction.
Benefits to Humans: (1) The African savannah ecosystem is a vast carbon sink, storing approximately 4.5% of global terrestrial carbon. If this carbon is released into the atmosphere by unregulated human activities, this will be the equivalent of 15 years of current annual usage of carbon by humans, greatly exacerbating the process and speed of human-induced climate change. By preserving this ecosystem, and potentially increasing the health of this ecosystem, more carbon may be stored, thus helping to mitigate the effects of human-induced climate change. This will also be of benefit to the fauna and flora of this and all other ecosystems on the planet. (2) A healthy ecosystem means that the evolutionary pressures for endemic viruses to mutate and find new hosts is reduced – viruses, like all life forms, need to adapt to survive in a changing ecosystem. However, a healthy ecosystem will ease these evolutionary pressures and may avert the emergence of novel zoonotic diseases that can cause widespread human and animal suffering. (3) The identification of the various migratory pathways of the plains zebras and co-migrating species will open opportunities for new eco-tourism development, which, if planned and developed with due care and consideration to the ecosystem, can benefit the fauna, flora, as well as the peoples of the region.
Ethical Permissions for the Use of Animals in Research:
All the research undertaken using animals in ‘Project Plains Zebra’ will be assessed for approval by the University of the Witwatersrand Animal Research Ethics Committee (Wits AREC). The Wits AREC is composed of academics and representatives from the Society for the Prevention of Cruelty to Animals (SPCA) and lay public. All decisions reached by the Wits AREC are done through consensus and adhere to the principles of Reduction, Replacement, and Refinement. The Wits AREC follows the guidelines set out by the South African National Standards for Research and Teaching, that are based on Australian and European Union Standards. The Wits AREC is also registered and audited by the National Health Research Ethics Committee of South Africa. Additional ethical approvals will be obtained as necessary from the various institutions in which the scientists involved in this project are employed. All ethical committees from which approval for the research will be sought follow the appropriate national standards, laws, and regulations.
Governmental Approvals:
Plains zebras are found in 14 countries within Africa. For each country in which we plan to undertake the proposed research, the appropriate governmental approvals and oversight will be obtained. No research will take place without such approvals.
Animal Use in Project Plains Zebra:
The proposed scientific research is comprised of many sub-projects, several of which require interventions to animals that range from minor through to major. With due transparency, we outline the proposed usage of animals in detail, indicating the severity of the intervention and where refinements in the protocols are employed to reduce the number and potential suffering of animals used, as well as justifying the use of the numbers of animals.
All work undertaken on animals in this project will be supervised by highly experienced wildlife veterinarians, who are either part of the project, or who work for the local authorities/organizations in the regions in which the work will be undertaken. The work will be undertaken according to the highest veterinary standards currently in use and the veterinarians involved have the authority to stop the work if there are any concerns regarding animal health or welfare.
GPS collars: The vast majority of interventions undertaken for Project Plains Zebra will be those where the fitting of a GPS collar is required. For the sub-projects aimed at revealing novel migratory routes and the temporo-spatial relationships between co-migrating species, approximately 3200 plains zebras, 50 blue wildebeest, 50 Grant’s gazelles, 50 Thomson’s gazelles, 50 white-eared kob, 50 tiang antelopes, and 100 Mongalla gazelles, will be immobilized and GPS collars will be fitted. This is considered a minor intervention. The immobilization and fitting of the collar will only occur once, and the animals will be released back to their natural habitat within 30 minutes. We acknowledge that localizing and darting the animals (either from a helicopter or other vehicle) is stressful, not only to the darted animals but also to those in the vicinity, and that the darting itself will cause short-duration discomfort (similar to the injection of a vaccine in the arm of humans) at the site where the dart delivers the immobilization drugs. For larger animals living in the natural environment, this is currently the most efficient and least stressful way in which an animal can be immobilized (i.e., we cannot acclimatize these animals prior to immobilization due to their size and that they are living in their natural habitat). The attachment of the GPS collar to the immobilized animal will not cause any physical pain, although the animals may experience some discomfort for the first few days before becoming accustomed to the wearing of the collar. The injection of the immobilization reversal drugs into ear veins following the fitting of the collar may cause a minor short-duration discomfort at the injection site. While the fitting of approximately 3550 collars may seem excessive, it must be noted that the area across which collars will be attached is approximately 6.5 million km2 (the size of the continental USA), and we have carefully calculated the proposed number of collars to allow for accurate scientific results to be obtained while minimizing the number of interventions.
Genetic material: From all animals in which interventions will be made throughout this project, approximately 10 ml of blood will be drawn and the full genome of each individual animal sequenced from this material. This is considered a minor intervention, as the blood will be extracted from a vein on the ear while the animals are immobilized, i.e., the animals will be unconscious during this procedure. The drawing of blood may cause minor short-duration discomfort at the extraction site. It should also be noted that animals in the wild are exposed to many small injuries, thus the drawing of blood will not have a significant impact on these animals. Due to the size of the animals used for this research, the extraction of 10 ml of blood should present no complications, and as this is a once-off procedure for each animal, there should be no long-term consequences of this intervention. Detailing the full genomes of all the animals in which interventions will take place is this project has far-reaching importance. In terms of Project Plains Zebra it will allow us to determine, through the study of relatedness, whether the novel migrations observed can be considered single units, or whether they need to be considered as clusters (link to genetics page). This is central to the development of the pragmatic approach to conservation strategies and determining the potential effects of human activities. The data will also be of importance beyond this project, as it will provide data useful for a vast range of analytical studies. To this end, at the conclusion of Project Plains Zebra we plan to make all genetic data freely accessible, thus lowering the need to obtain genetic material from these species in the future, reducing potential future interventions on these animals.
Sleep Recordings in a Captive Semi-Natural Environment: At present, the gold-standard for recording sleep in mammals is through the use of the polysomnographic technique. This technique requires the surgical implantation of electrodes to record from the surface of the brain, to record movements of the eyes, and to record muscle activity levels. As the plains zebras are large mammals, to obtain the best results we need to record sleep in freely-moving animals in an enclosure that mimics their natural habitat. To do this requires the subcutaneous implantation of the recording device (a box approximately 5 x 2 x 8 cm in size) and electrodes (over the brain, one eye and in the deep nuchal musculature). In addition, we need to subcutaneously implant activity meters (approximately 2.5 x 1 x 2.5 cm in size, one under the skin of the neck and one under the skin on the rump) that we can then use to record sleep from in truly wild plains zebras during the migration. This is not a trivial surgery and given that there will be several incisions made in the skin and 3 devices implanted under the skin, we acknowledge this procedure to be a major intervention. In order to obtain scientifically valid results that accurately define what sleep is like in the adult plains zebras, we need to use a minimum of 3 males and 3 females for this work. We acknowledge that the anaesthesia of these 6 animals and the surgical implantation of the sleep recording devices (approximately 5 x 2 x 8 cm) and electrodes will cause localized discomfort in these animals. Our experience in other large mammal species (Davimes et al., 2017; Malungo et al., 2021) indicates that the animals show full recovery within 5 days of surgery. In addition, we acknowledge that maintaining these animals in a captive environment may cause a certain level of stress, but we aim to overcome this by providing an approximately 20 ha semi-natural enclosure for these animals (to which they will be habituated for at least 2 weeks prior to the surgical procedure, and for 4 weeks after the implantation procedure). Due to the nature of this aspect of the research, the plains zebra used (to be sourced from breeding populations at local game farms) will be euthanised at the end of the recording period. The implanted devices will be removed and the data collected, and in addition, biological material for other aspects of the study will be taken (see anatomical studies below) thereby reducing the number of animals required for those studies. It should be noted that the veterinarians and researchers on the project have undertaken similar studies successfully in several mammalian species with no complications. Understanding sleep in the plains zebra is crucial for understanding how the migrations have evolved and how they might change in relation to human activities. At present we know virtually nothing about sleep in plains zebras, and given the potential importance of this aspect of their biology we believe it necessary to undertake this study.
Sleep Recordings During Migration: As an extension of the preceding study of sleep in captivity, we aim to determine what sleep is like in free-ranging migrating zebras. To achieve this, we need to subcutaneously implant two activity meters (approximately 2.5 x 1 x 2.5 cm in size, one under the skin of the neck and one under the skin on the rump) in 20 wild plains zebras (10 males, 10 females). We will need to immobilize each animal, implant the two activity meters, and fit a GPS collar (as described above). Following the migration (that will be tracked by the GPS collars), we will relocate the implanted animals, again immobilise the animals, and subsequently remove the activity meters (although we will leave the GPS collars). We acknowledge that all the these procedures will cause potential irritations and localized short-term discomfort, but consider this a moderate intervention. This procedure has been successfully undertaken by the team members in African elephants, Arabian oryx and blue wildebeest (Gravett et al., 2017; Davimes et al., 2016; Malungo et al., 2021), all without complications. We reduce the number of plains zebras that will undergo interventions by having these specific animals form a subset of those used for understanding the dynamics of the known migration in Botswana. We hypothesize that sleep reduction during the migration is essential to the evolution of the migration-centred ecology of the African Savannah Ecosystem, and thus this sub-project is a very important part of the scientific knowledge required to support the development of appropriate management and conservation strategies for the ecosystem.
Movement Paths and Decision Making: In order to understand what causes plains zebras to decide to move towards, away from, or ignore and continue onwards, is central to understanding the process of migration and the identification of landmarks. To achieve this understanding we need to outfit up to 20 plains zebras, located in a game reserve in South Africa, with a GPS collar with an inbuilt triaxial accelerometer tag, plus attach a similar tag to the head of the plains zebras. Using the same processes described above, these animals will need to be immobilised twice, once to attach the collar and tags, and once to remove these. The same intervention concerns regarding immobilisation and the attachment of collars and tags apply to these zebras. In addition, the attachment of the tag to the head (to determine head direction movements) will provide an additional irritation for a few days after attachment, but the plains zebras should become accustomed to this quite rapidly. The head-mounted tag will be attached to a halter that will be fitted on the plains zebras. This halter will be readily removed at the end of the recording period. In this sub-project the skin of the plains zebras will not be opened for implantation, making this a minor intervention. We hypothesize that decision making in terms of the movement paths by the plains zebras will be able to be linked to specific landmarks. The determination of the types of features of the landscape used as landmarks by the plains zebras is of importance, as similar features can be identified across all the migratory paths revealed for plains zebras, and these can be specifically targeted for preservation.
Anatomical Studies of the Brain and Retina: We propose that the neural navigation system of the plains zebra, centred around the size of the hippocampal formation forms the central neural substrate allowing the plains zebras to be the expert navigators of, and primary leaders of, the migrations of the African Savannah. A crucial part of this system is the acquisition of information via the visual and olfactory systems, along with other portions of the brain. Combined, these features of the plains zebra nervous system are likely to form a uniquely evolved cognitive specialisation. However, in order to understand how this system works, we need to examine this system in detail and contextualize the observations in plains zebras in relation to species phylogenetically related to the plains zebras (i.e., other equids, mountain zebra, domestic horse, and domestic donkey), those that migrate with the plains zebras (i.e., blue wildebeest, Grant’s gazelle, Thomson’s gazelle, white-eared kob, tiang antelopes, and Mongalla gazelles), and those that occupy the same habitat but that do not migrate (i.e., impala, waterbuck, nyala, kudu, and eland). At present, the only way to achieve this successfully is by employing a range of modern neuroanatomical techniques, by obtaining actual specimens of brains and eyes from animals that will be euthanised. In order to achieve a proper contextualised understanding of the neural navigation system and the sensory systems involved in navigation in the plains zebra, we aim to euthanise 12 individuals of each of the 15 species listed above and then prepare the tissue in various ways allowing for the studies proposed. We acknowledge that in order to do this essential research we need to euthanize 180 animals, and while we realise this may not be acceptable to all those with an interest in preserving the African Savannah Ecosystem, it is the minimum number of individual animals required to undertake the project successfully. Euthanasia will begin by immobilising the animals with a dart, which can cause some discomfort and stress. After the animals are immobilised will we administer a body mass appropriate intravenous injection of sodium pentobarbital to achieve euthanasia. We consider this process of euthanasia a moderate intervention in terms of the direct effect of suffering and stress to the animals, while acknowledging that this will lead to death. After the heart has ceased beating, the neural tissue will be prepared for scientific research. We aim to acquire the majority of these animals from well-established breeding populations from local game farms/abattoirs (it should be acknowledged that several of these species are farmed for human consumption), with a minimal number of animals being taken from wild populations (those that are not farmed for consumption). In addition, to reduce the numbers used, we will use the same specimens for overlapping or complementary studies. There are no models or other methods currently available that allow this aspect of the research to be undertaken successfully and meaningfully. Having a detailed, contextualised, understanding of how the neural navigation system is specialised in the plains zebra will provide a very solid scientific basis upon which management and conservation strategies can be developed. In addition, these observations will provide significant insights into how the behaviour of the plains zebra is accomplished, and may reveal novel aspects of neural process that will lead to greater insights into plains zebra behaviour.
The 3 Rs of animal experimentation:
The scientific research using animals outlined in Project Plains Zebra will follow the 3 Rs of animal experimentation: Reduction, Replacement, and Refinement. In terms of Reduction of the number of interventions undertaken to achieve the positive outcomes expected from this scientific research, we have carefully assessed the number of interventions required for each aspect of each sub-project and will use the minimum numbers of animals as outlined above to obtain scientifically valid results. At present, there are no alternatives that will allow Replacement of the species to be investigated, we just do not have the data or technological capacity to do this currently. We have carefully chosen the research to be undertaken through a process of Refinement, in that we plan to do only the research that is essential to provide the information needed to successfully develop pragmatic conservation and management strategies for this ecosystem. In addition, we add two more Rs to this list - Responsibility and Respect. The veterinarians and scientists involved in this project take Responsibility for the use of animals in this research and undertake this with the utmost Respect for these animals, their lives, and the minimization/mitigation of any suffering associated with the interventions that will be undertaken.
A Personal Statement on the Use of Animals in Project Plains Zebra by Prof. Paul Manger:
As the individual that conceptualized and leads ‘Project Plains Zebra’, it is appropriate that I share my views regarding the use of animals in this scientific project. In my career I have been fortunate to undertake scientific research on a range of mammal species across the globe, but with a specific focus on African mammals over the last 20 years. My experiences in Africa have led to a deep respect and awe of the animals, plants, fungi, and people that inhabit this amazing continent, and specifically those within the Savannah Ecosystem. This is an ecosystem that is not only worthy of preserving for the lifeforms within, but for the planet more broadly. It would be a great loss for this ecosystem to succumb to the vagaries of unmanaged human expansion when it can be preserved. It would personally be heart-breaking not to be able to show my children a healthy African Savannah Ecosystem, in all its grandeur, when I have the ideas, capacity, and dedicated team of colleagues capable of developing a plan to preserve this ecosystem. Humans are at a crossroad in terms of preserving the planet, and it is my strongly held conviction that ‘Project Plains Zebra’ can contribute significantly to improving our chances of overcoming our self-inflicted climate and biodiversity crises.
To achieve this end, with our current technology and biological understanding, it is very unfortunate that we will have to sacrifice several animals and intervene in the lives of many more, as detailed above; however, at present the only pragmatic solution to preserve the African Savannah Ecosystem is to gather the appropriate scientific data, develop and disseminate conservation plans, and hope that these may be taken notice of and adopted for the benefit of the ecosystem and humanity more broadly. At present we cannot do this without the sacrifice of the few to save the many. While I understand that many may disagree with me, I do hope that those with differing views understand that this project has been designed with the 5Rs of animal experimentation firmly in mind and that I undertake to ensure that all animal experimentation will be done with the utmost respect to the animals and to the best of our capabilities, ensuring that those animals that will be sacrificed, or have their daily lives disrupted to differing degrees, in order to reach the desired end will not have been disrupted in vain.