A Practical Assessment of the Disinfectant Efficacy of UV Light with and without Ozone Using a Novel Transfer Hatch in a Research Animal Facility.

Most in vivo animal research and breeding using mice and rats in China takes place in facilities under barrier conditions. Items being moved across the barrier are typically disinfected using UV radiation in a transfer hatch. However, the time periods necessary for this disinfection technique are inefficient, and disinfection is frequently incomplete, especially if concealed surfaces are present. The current study used a newly developed transfer hatch incorporating both UV and ozone disinfection to examine disinfection efficacy against 4 bacteria species (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii). Disinfection trials used UV and ozone, applied separately and in combination, for up to 30 min. Separate and combined treatments were also tested with a UV barrier. We found that if UV radiation has direct contact with surfaces, it is an efficient disinfection method. However, where surfaces are concealed by a UV barrier, UV radiation performs relatively poorly. The results of this study indicate that a combination of UV and ozone produces the most effective disinfection and is markedly quicker than current disinfection times for UV applied on its own. This novel transfer hatch design therefore allows more complete and efficient disinfection, improves workflow, and reduces barrier breaches by pathogens that may affect animal health and welfare and compromise research outcomes.

Effectiveness of treatment of bedding and feces of laboratory animal with ozone.

BACKGROUND: The incineration and burying of the soiled bedding of laboratory animals, as well as using detergents to treat their feces, is hazardous to the environment. This highlights the need for an alternative, environmentally friendly solution for the treatment of the waste of laboratory animal facilities. This study aims to evaluate the efficacy of ozone disinfection of the soiled bedding and feces of laboratory animals. METHODS: Two grams of soiled beddings were randomly sampled from the cages of mice and rats. These samples were mixed in a beaker with 40ml saline. Ozone was piped into the beaker at a concentration of 500mg/h. Samples were taken from the beaker at time 0min, 30min, 45min and 60min after ozone treatment for microbiological culturing in an incubator for 48h. Colony form unit of each plate (CFU/plate) at each time point were counted, the mean CFU/plate at each time point after ozone treatment were compared with that present at time zero. Feces of rabbits and dogs were treated and pathogens were counted the similar way as that of bedding of the mice and rats; samples being taken at 0min, 15min, 30min, 45min and 60min. RESULTS: Pathogens were observed in beddings of both mice and rats as well as in feces of rabbits and dogs. Ozone treatment for 30min killed more than 93% of pathogens in the bedding of the two rodent species and 60min of treatment killed over 99% of pathogens. Treatment of rabbit and dog feces for 30min killed over 96% pathogens present, and 60min's treatment killed nearly all the pathogens. Both Gram positive and Gram negative pathogens were sensitive to ozone treatment. CONCLUSION: Ozone treatment of bedding and feces is an effective and environment friendly way to deal with the waste of animal facilities, saving energy and potentially enabling their reuse as fertilizer.

Effective chair training methods for neuroscience research involving rhesus macaques (Macaca mulatta).

BACKGROUND: Neuroscience research using macaques remains fundamental in our endeavours to understand how the human brain functions. Applying the refinement principle of the 3Rs is essential to optimise the monkeys' welfare and still produces high quality science. NEW METHOD: Here we document refinements in our training methods for acclimation to transport devices (i.e. primate chair or transport box) while working with 46 male rhesus macaques. Our training methods always used positive reinforcement training (PRT). However, PRT was sometimes combined with negative reinforcement training (NRT), but not pole and collar techniques, to successfully transfer each monkey from its home enclosure to its transport device. RESULTS AND COMPARISON WITH EXISTING METHODS: Training monkeys in pairs or groups, and starting their PRT training upon arrival within the unit reduced the days required to acclimate them. While the use of PRT is essential to establish a positive relationship with monkeys, NRT techniques are sometimes necessary, and are most effective when withdrawn immediately once the monkey makes the desired response, to reduce the days of acclimation. Once acclimatised to their chair, monkeys succeeded within 10 days to present their head voluntarily for neck-plating using PRT. Space reducers inside the chairs also facilitated head presentations for some monkeys. CONCLUSIONS: Acclimating (shaping) the monkeys to transport devices can be a stressful experience for monkeys and trainers. The adaptations to our training substantially reduced the days spent on this stage. We view this reduction in days as an effective implementation of the 3Rs (refinement) in monkey neuroscience research.

Reference intervals for biochemical and haematological analytes of juvenile captive-bred long-tailed macaques (Macaca fascicularis) from Mauritius with examination of the effects of individual characteristics using principal component analysis.

BACKGROUND: The Mauritian cynomolgus macaque (Macaca fascicularis) is widely used in biomedical research. Determining reference intervals for biochemical and haematological analytes provides an important tool for clinical diagnosis and pre-clinical research. METHODS: Blood samples from 736 Mauritian long-tailed macaques were analysed to determine reference intervals of 13 biochemical and 10 haematological analytes. The need for partitioning the reference interval between males and females was determined. To examine the variation associated with age, body weight and sex on the analytes, our correlating, multivariate data set was first reduced using principal component analysis, and then the effect of these characteristics on factor scores was examined using GLM analysis. RESULTS: Partitioning of reference intervals, based on sex, was recommended for albumin, alkaline phosphatase and mean corpuscular haemoglobin. Sex significantly influenced the concentration of lymphocytes, granulocytes and white blood cells. CONCLUSION: These findings provide useful reference data for research involving Mauritian long-tailed macaques.

The extended welfare assessment grid: a matrix for the assessment of welfare and cumulative suffering in experimental animals.

Combining a range of assessment parameters into one usable entity has been identified as an important goal in providing a practical, objective and robust assessment of welfare, particularly in laboratory animals. This paper refines and extends one such previously published method. The proposed Extended Welfare Assessment Grid provides for the incorporation of changes in the state of an animal over time, allowing for predictive, retrospective, scheduled, or event monitoring. It enables the numeric, as well as visual, representation of the animal's welfare, placing this in the context of the careful and realistic justification for experimental use of the animal. This assessment method represents a valuable tool for those tasked with ensuring ethical oversight, as well as for those planning the use, or monitoring, of animals in research. It is particularly applicable to animals used in long-term studies, especially non-human primates. It is believed that this system will draw attention to the temporal component of suffering that is often overlooked in the planning of research schedules and allow an assessment of cumulative suffering imposed by the events that occur.

Bovine tuberculosis (Mycobacterium bovis) in British farmland wildlife: the importance to agriculture.

Bovine tuberculosis (bTB) is an important disease of cattle and an emerging infectious disease of humans. Cow- and badger-based control strategies have failed to eradicate bTB from the British cattle herd, and the incidence is rising by about 18%per year. The annual cost to taxpayers in Britain is currently 74 million UK pounds. Research has focused on the badger as a potential bTB reservoir, with little attention being paid to other mammals common on farmland. We have conducted a systematic survey of wild mammals (n=4393 individuals) present on dairy farms to explore the role of species other than badgers in the epidemiology of bTB. Cultures were prepared from 10397 samples (primarily faeces, urine and tracheal aspirates). One of the 1307 bank voles (Clethrionomys glareolus) live-sampled, and three of the 43 badgers (Meles meles), yielded positive isolates of Mycobacterium bovis. This is the first time the bacterium has been isolated from the bank vole. The strain type was the same as that found in cattle and badgers on the same farm. However, our work indicates that the mean prevalence of infectious individuals among common farmland wildlife is extremely low (the upper 95% confidence interval is < or =2.0 for all of the abundant species). Mathematical models illustrate that it is highly unlikely the disease could be maintained at such low levels. Our results suggest that these animals are relatively unimportant as reservoirs of bTB, having insufficient within-species (or within-group) transmission to sustain the infection, though occasional spill-overs from cattle or badgers may occur.

Alopecia scoring: the quantitative assessment of hair loss in captive macaques.

Many captive animals show forms of pelage loss that are absent in wild or free-living conspecifics, which result from grooming or plucking behaviours directed at themselves or at other individuals. For instance, dorsal hair loss in primates such as rhesus macaques (Macaca mulatta) in research facilities, results from excessive hair-pulling or over-grooming by cage-mates. This behaviour appears to be associated with stress, and is controllable to some extent with environmental enrichment. Quantifying alopecia in primates (as in many species) is therefore potentially useful for welfare assessment. A simple system for scoring alopecia was developed and its reliability was tested. Study 1 showed high interobserver reliability between two independent scorers in assessing the state of monkeys coats from photographs. Study 2 showed that there were no significant differences between the scores derived from photographs and from direct observations. Thus, where hair loss due to hair pulling exists in captive primates, this scoring system provides an easy, rapid, and validated quantitative method, for use in assessing the success of attempts to reduce it via improved husbandry. In the future, such scoring systems might also prove useful for quantifying barbering in laboratory rodents.

Evidence from rhesus macaques suggests that male coloration plays a role in female primate mate choice.

Male animals of many species use conspicuous coloration to attract mates. Among mammals, primates possess the most brilliant secondary sexual coloration. However, whether colour plays a part in primate female mate choice remains unknown. Adult male rhesus macaques undergo a hormonally regulated increased reddening of facial and anogenital skin during their mating season. We experimentally investigated whether red male facial coloration is preferred by simultaneously presenting female rhesus macaques (n = 6) with computer-manipulated pale and red versions of 24 different male faces. The duration and direction of gaze were measured to discern visual preferences. Females exhibited preferences for the red versions of male faces. It is proposed that male coloration might provide a cue to male quality.

Species-typical patterns of infant contact, sleeping site use and social cohesion among nocturnal primates in Africa.

This paper provides a checklist and summary of what is currently known of the variation in infant contact, sleeping site preference and aspects of social cohesion in the nocturnal primates of Africa. Genera and species are compared, based on previously unpublished field observations and a review of the literature. There is a clear pattern of similarity between the species within each genus and distinct differences between genera. Species in the same genus tend to be ecologically equivalent and replace each other allopatrically, whereas species in different genera are more likely to be sympatric, with up to 6 species living together. Maximum sympatry within genera is found in Otolemur and Galagoides, where species are ecologically divergent. This may reflect an ancient origin of species within these genera or suggest that further taxonomic revision is required at the generic level. Some data are recorded for the first time for species that have only recently been separated (cryptic species), but some taxa remain very poorly known. It is concluded that field studies are still at an elementary stage and further research with radio tracking is urgently needed in the face of rapidly declining habitats.