The 3Rs work at Lund University

Lund University has a total of five distinct facilities across the region of Skåne, dedicated to animals in research. The Medical Faculty, which conducts the majority of animal research, predominantly utilizes rodents, zebrafish, and larger animals. This includes the life science hub, Medicon Village, with its own animal facility. There is also a specialized facility for studies involving wild animals.

We have interviewed Malin Fex and Marie Sydoff, researchers at the university. Malin serves as the 3Rs coordinator and is part of the university's animal welfare committee, which oversees all animal-related activities. Marie is a radiation physicist at Lund University Bioimaging Centre, that also performs animal experiments.

Consultation and the 3Rs checklist

Before each new ethical application, a consultation takes place involving the principal investigator, approved animal facility managers, veterinarians, and other stakeholders in the animal research. The goal is to ensure meticulous planning of experiments, feasibility of all components, and an experimental design that yields optimal outcomes. A dedicated 3Rs checklist guides these discussions.

Malin explains that the 3Rs checklist is entirely new for this year. It was collaboratively developed by the facility managers, veterinarians, and the animal welfare committee in order to create a comprehensive and inclusive checklist. The welfare committee initiated this checklist, which is meant to be used as a tool during consultations, to ensure a thorough review and discussion with the principal investigator.

— We actively work on refining all procedures, such as anaesthesia, added warmth during awakening and sedation, says Malin. It's vital for everyone working closely with the animals to understand this. We bring up these aspects during our consultation meetings, thanks to the 3Rs checklist.

Education and training

Malin shares an example from a few years ago when they trained all research groups in stress-reduced handling of mice. They learned to lift the animals by using cupped hands and also by using the enrichment ladder as a type of scoop to gently lift the mice. Lifting them by their tails is prohibited. Malin is surprised by how smoothly it went; everyone embraced the change and recognized its benefits.

All new employees involved in animal research need training in laboratory animal science. Afterward, they must "earn a license" to handle the animals and perform specific procedures like injections.

Malin points out that the 3Rs principles are applied right from when the animals arrive. They focus on acclimating animals, getting them comfortable with human interaction, their environment, and preparing them for upcoming procedures. Every animal's unique characteristics are considered — lineage, needs, and specific conditions.

The animals are trained for the procedures they'll undergo. If needed depending on the research study's nature, a plan is developed in collaboration with the research group for training the animals before experiments. This process involves productive dialogue between animal caretakers and researchers. The importance of healthy and trained animals in studies is well understood. Nobody wants to use animals unnecessarily or risk compromised results due to stressed research animals.

Planned breeding of research animals

Another strategy to minimize animal use involves planning breeding and the number of animals required for an experiment. During consultations, calculations regarding animal numbers are discussed, whether it is reasonable and what type of breeding is needed. This approach helps manage breeding and avoid unnecessary surplus animals.

Malin emphasizes that the focus is on optimizing breeding rather than maximizing it. They also strive to share animals and tissues among different research groups to make the most out of each animal and its contributions.

Imaging technology reduces the need for research animals

Lund University Bioimaging Centre, a part of the university, offers research and imaging services. Prior to submitting applications for animal studies, they hold consultation meetings with the research groups and the principal investigators in charge. The purpose of these meetings is to plan and refine experiments, ensuring the best possible results while eliminating unnecessary use of experimental animals.

Marie highlights the growing role of advanced imaging technology. With these tools, researchers can conduct longitudinal studies, monitoring the same animal over an extended period. For instance, during cancer tumour treatment, images captured at various time points reveal the effectiveness of the treatment through tumour shrinkage or the presence of live cells within the tumour.

This technology eliminates the need to use multiple groups of animals for different time points, a practice that required euthanizing animals at each stage. Now, a single group of animals is sufficient, and images are captured at corresponding time points.

Marie also mentions that there of course are limitations to these experiments as well. Animals have a threshold for the number of imaging sessions they can undergo. The animals are anaesthetised before each imaging, which must be factored in. Ethical permits usually set limits on the number of injections each animal can receive.

Refined methods

Technological advancements are constant. Currently, the university is reviewing its CT, PET, and SPECT scan equipment. Upgraded machines promise even better opportunities to study organs with finer detail, such as the brain.

Marie explains that these machines can reveal smaller tumours and finer structures within organs. PET and SPECT cameras highlights the function of a specific organ, a tumour or an organ-system, while CT images depict anatomy. The images are three-dimensional, which enables precise tracking of active substances within animals.

Their advanced microscopy techniques, like light microscopy and electron microscopy, allow tissues to be studied in more detail after imaging, providing additional insights on a smaller scale.

This approach maximizes the utility of each animal, reducing the total number needed. Sharing tissues among research groups is another practice they're working on.

Proximity matters for the 3Rs

Having the Bioimaging Centre physically situated within the university offers significant advantages. The close proximity to the animal welfare body and veterinarians facilitates work within the 3Rs. The understanding of animal welfare and its impact on results is widespread. Researchers realize that consolidating all aspects of a study in one location reduces stress on animals, due to eliminating the need to move them between different procedures.

Marie states that the animals need rest before and after imaging sessions. This principle is to them non-negotiable. The animals' stress levels must be minimized, as it can impact their physiology and consequently study results. For researchers, cost-effectiveness is important, which is achieved by using the same animal over an extended period and getting it right the first time.

Marie recalls a recent doctorate course held on location in Lund and how they presented their various techniques. They showed the PhDs the importance of planning all aspects of a study and proper animal handling. Real-life examples were shared, highlighting good and bad examples in relation to results.

Another current 3Rs initiative involves minimizing radiation exposure during CT scans of the lungs. Marie explains that while higher doses improve image quality to a certain extent, maintaining the balance is vital for the results in longitudinal studies. Using the right dose to achieve perfect image quality relative to the study's objectives is key.

A strong 3Rs culture

Marie and Malin both agree that there is a strong 3Rs culture in Lund University. Everyone is committed to ensuring animals' well-being and understand its crucial role in research.

Malin notes that everyone is instilled with this culture from the outset. They find it easy to pause before beginning a study, ensuring each experiment is meticulously planned and uses the exact number of animals necessary, neither too many nor too few.