Assessment of the Culture of Care working with laboratory animals by using a comprehensive survey tool.

The term Culture of Care, within the scientific community using laboratory animals, is being used more and more frequently after it was introduced in the EU Directive 2010/63/EU, where it is phrased as a 'climate of care', which became effective in national legislation from January 2013. However, there is a risk that the term could become a meaningless phrase if no agreed local definition of the term exists at the animal facility (called establishment in the EU Directive). This paper presents a comprehensive survey tool that provides a means to describe what the Culture of Care in an establishment looks like. The tool is one of the elements that can contribute to the overall picture of the culture; however, it cannot stand alone. Together with an evaluation of the effectiveness of the Culture of Care (e.g. key performance indicators) and a description of the outcomes and achievements in terms of animal welfare and the 3Rs (Replace, Reduce, Refine), the survey tool will constitute a comprehensive picture. The survey tool offers a multilevel and comprehensive view of different subcultures, presenting details on mindset and behaviour of the employees and the different relations within the culture, thus enabling the initiation of improvement projects if required. The tool addresses essential elements of a co-operative culture in terms of what we think, what we do and how we work together.

A new group housing approach for non-human primate metabolism studies.

Understanding the absorption, distribution, metabolism and excretion (ADME) of candidate drugs in preclinical species is an integral part of the safety and efficacy evaluation in drug development. For this purpose, the housing of single animals in metabolism cages has historically been common practice for ADME studies. Whilst mini-pigs and dogs are selected wherever possible, non-human primates (NHPs) are used where there is no suitable scientific alternative. Having undergone only minimal revisions over the past 30 years, the traditional single-housing metabolism cage design for NHPs significantly limits normal vertical movement and social behaviours in primates. Minimising animal suffering and improving welfare is an important aspect of working with animals in research and Novo Nordisk A/S, together with collaborators, has focused on this area for many years. A novel metabolism cage for group housing of NHPs has been designed in a joint collaboration between Novo Nordisk A/S and Covance Inc. The advantages of this novel cage are extensive, including a significantly increased cage volume and ability for socialisation, as well as improvements to alleviate stress and boredom. The excretion balance data from six male NHPs housed in single or group metabolism cages were compared using the radiolabelled test compound [14C]-quetiapine. Welfare, in terms of stress and behaviour, when animals were single or group housed was also assessed. Mean recoveries of radioactivity were shown to be comparable irrespective of housing design (83.2% for group-housed animals vs. 87.1% for single-housed animals), supporting the potential suitability of NHP group housing for future metabolism ADME studies.

A novel welfare and scientific approach to conducting dog metabolism studies allowing dogs to be pair housed.

Metabolism cages are designed to conduct absorption, distribution, metabolism and excretion (ADME) studies, enabling an 'excretion balance' scientific objective to be met. Historically, the design of dog metabolism cages has involved single housing. This type of housing has limitations for normal social behaviours and has been largely unchanged for 25-30 years. Improving animal welfare is a focus area for the authorities as well as the industry throughout the European Union. A collaboration was developed between Novo Nordisk and Covance to enhance the design of metabolism cages, allowing dogs to be pair housed. The purpose of the study was to compare excretion balance data from pair-housed and singly housed dogs in order to demonstrate that conducting excretion balance studies with a pair-housing design improves animal welfare without compromising the scientific integrity of the study. A radiolabelled test compound, [14C]-Quetiapine, was selected for this investigation based on its excretion profile. The assessment of the dogs' stress levels was investigated by measuring the levels of serum cortisol as an indicative biomarker. Results were inconclusive due to large variations in cortisol levels. However, dogs appeared calmer in the pair-housing setting. The overall mean recovery (±standard deviation) for pair-housed animals (94.0 ± 0.66% of the dose) was equivalent to that from singly housed dogs (93.0 ± 2.29%). Based on these data, we conclude that pair housing of dogs for future metabolism ADME studies does not compromise the scientific integrity, and therefore is a major progression in the design of these studies, enhancing welfare.

Effect of local anaesthesia on neuronal c-fos expression in the spinal dorsal horn and hypothalamic paraventricular nucleus after surgery in rats.

The surgical stress response is the neurophysiologic reflex response to surgery, which involves activation of the hypothalamic-pituitary-adrenal axis and is regulated by the hypothalamic paraventricular nucleus. The effect of pre-operative use of local anaesthetics on activation of neurones in the paraventricular nucleus during surgery was studied by quantification of the neuronal expression of the c-fos-gene after a standardized plantar incision in rats. Furthermore, c-fos expression in the spinal dorsal horn was used as a measure of spinal nociception. Six halothane-anaesthetized animals underwent surgery following infiltration with lidocaine and bupivacaine, six animals were operated without local anaesthetics, and six control animals were subjected to the anaesthetic procedures. After two hours, the animals were perfused with 4% formaldehyde and the spinal cords and brains were collected and processed by immunohistochemistry for stereological quantification of the number of neurones with Fos-like immunoreactivity. Furthermore, brain and spinal cord were sampled from nine control animals right after induction of halothane anaesthesia. Surgery without local anaesthetics caused a significant increased number of neurones with Fos-like immunoreactivity in the spinal cord (4258+/-1710; mean+/-S.D.; P<0.01) compared to the anaesthesia control group (1204+/-436). Local anaesthetics reduced this number to 2029+/-919 (P<0.05), which was not significantly different from the anaesthesia control group. After surgery, the number of neurones with Fos-like immunoreactivity in paraventricular nucleus increased from 2948+/-1365 in the anaesthetized control group to 5550+/-3875 and 5191+/-1558 in the surgery and local anaesthetics plus surgery group, respectively, although significance was only reached for the group receiving local anaesthetics (P<0.05). In conclusion, preoperative local anaesthetic infiltration did not reduce the surgery-induced c-fos expression in paraventricular nucleus after paw surgery in rats, although spinal nociception was reduced.