Preferences of group-housed female mice regarding structure of softwood bedding.

Bedding influences various parameters in the housing of laboratory mice, such as health, physiology and behaviour (often considered as being integral parts of welfare). Notwithstanding existent studies about bedding preferences of individually tested mice, data about group-housed mice are still lacking. The aim of this study was to find out the structure preference for softwood bedding of group-housed mice. One hundred and eight 8-week-old female mice (C57BL6/JOlaHsd and BALB/cOlaHsd) were housed in groups of three and were given one-week free access to two different bedding structures at a time. In three test combinations, softwood shaving bedding was tested versus softwood chip bedding products of three different particle sizes (fine/medium/coarse-grained). The preference test was performed in a DoubleCage system composed of two Makrolon type IIL cages, connected by a perspex tunnel. This validated system was able to detect the crossings of each individual animal with correct crossing time and direction. On the basis of these data, dwelling times on the particular bedding structures were statistically analysed as a parameter for bedding preferences. In all three test combinations, a highly significant shaving preference was detected. On average, mice spent 70% of their dwelling time on the shavings. This preference was more explicit during the light period and in C57BL/6J mice. The relative ranking of the bedding structures was: shavings >> coarse-grained chips > medium chips = fine chips. By means of these results, a shaving structure as bedding can be recommended for laboratory mice, whereas fine chip structures should be avoided.

Validation of an automatic system (DoubleCage) for detecting the location of animals during preference tests.

Preference tests have often been performed for collecting information about animals' acceptance of environmental refinement objects. In numerous published studies animals were individually tested during preference experiments, as it is difficult to observe group-housed animals with an automatic system. Thus, videotaping is still the most favoured method for observing preferences of socially-housed animals. To reduce the observation workload and to be able to carry out preference testing of socially-housed animals, an automatic recording system (DoubleCage) was developed for determining the location of group-housed animals in a preference test set-up. This system is able to distinguish the transition of individual animals between two cages and to record up to 16 animals at the same time (four animals per cage). The present study evaluated the reliability of the DoubleCage system. The data recorded by the DoubleCage program and the data obtained by human observation were compared. The measurements of the DoubleCage system and manual observation of the videotapes are comparable and significantly correlated (P < 0.0001) with good agreement. Using the DoubleCage system enables precise and reliable recording of the preferences of group-housed animals and a considerable reduction of animal observation time.

Are the effects of different enrichment designs on the physiology and behaviour of DBA/2 mice consistent?

Environmental enrichment is intended to improve the well-being of laboratory animals. Although many researchers have indicated that environmental enrichment may enhance animal well-being, there is some evidence that enrichment differs in its effects on physiology and behaviour between species and strains. The present study focuses on the effects of different enrichment designs on the physiology and behaviour of male and female DBA/2 mice. A total of 48 DBA/2J mice, 24 males and 24 females were used for this experiment. Upon arrival at about 3 weeks of age, the animals were randomly allotted to three experimental groups: NE, non-enrichment; E1, enriched with nest box, wooden climbing bar and nest material according to Scharmann (1993); E2, enriched with horizontal and vertical dividers, modified from Haemisch and Gärtner (1994). Same-sex groups of four mice were housed for 12 weeks in type III Makrolon cages with (E1 or E2) or without (NE) enrichment objects. Behavioural performance (Open Field, Food Drive and Elevated Plus Maze tests) and physiological traits (haematological variables, body weight and organ weights, corticosterone and thyroxine levels) were measured. This study observed that enrichment had significant effects on the mean values of body weight (females), Open Field and Food Drive tests. The most significant housing differences were found between the E2 and NE/E1 groups. Furthermore, sex differences in the NE, E1 and E2 groups were not consistent for several variables (growth rate, relative weights of spleen, kidney and heart, Food Drive and Elevated Plus Maze behavioural performance). There was often a higher coefficient of variation (CV) in the E1 and E2 groups as compared to the NE group, chiefly in physiological traits and in the Open Field and Food Drive tests. The results of this study indicate, that the effects of enrichment designs used in the present study are not consistent, but vary according to sex and the variable studied.

The effects of different rack systems on the breeding performance of DBA/2 mice.

Housing systems for laboratory animals have been developed over a long time. Micro-environmental systems such as positive, individually ventilated caging systems and forced-air-ventilated systems are increasingly used by many researchers to reduce cross contamination between cages. There have been many investigations of the impact of these systems on the health of animals, the light intensity, the relative humidity and temperature of cages, the concentration of ammonia and CO(2), and other factors in the cages. The aim of the present study was to compare the effects of different rack systems and to understand the influence of environmental enrichment on the breeding performance of mice. Sixty DBA/2 breeding pairs were used for this experiment. Animals were kept in three rack systems: a ventilated cabinet, a normal open rack and an individually ventilated cage rack (IVC rack) with enriched or non-enriched type II elongated Makrolon cages. Reproduction performance was recorded from 10 to 40 weeks of age. In all three rack systems there was a similar breeding index (pups/dam/week) in non-enriched groups during the long-term breeding period, but the coefficients of variation in the IVC rack were higher for most parameters. This type of enrichment seems to lead to a decrease in the number of pups born, especially in the IVC group. However, there was no significant difference in breeding index (young weaned/female/week).

Impact of environmental enrichment in mice. 1: effect of housing conditions on body weight, organ weights and haematology in different strains.

Currently, environmental enrichment is a very common means of improving animal well-being, especially for laboratory animals. Although environmental enrichment seems to be a possible way for improving the well-being of animals, the consideration of housing laboratory animals should not only focus solely on animal well-being, manpower and economics but also on the precision and accuracy of the experimental results. The purpose of the present study was to evaluate the effects of enriched cages (nest box, nesting material, climbing bar) on body weight, haematological data and final organ weights. BALB/c, C57BL/6 and A/J mice, originated from Harlan Winkelmann, were used for the experiments - 16 animals of each strain. Animals at 3 weeks of age were marked and separated randomly to enriched or non-enriched cages, in groups of four, half for each housing condition. Both cages were type III Makrolon cages, only the enriched cages contained a nest box, a wood bar for climbing and nesting material. Animals were kept in a clean animal room under specific pathogen free (SPF) conditions. Body weights were recorded every week. Blood samples were collected at 14 weeks of age (white blood cells (WBC), red blood cells (RBC), haemoglobin (HGB), and haematocrit (HCT) were analysed). At 15 weeks of age, the animals were euthanized by CO(2) in their home cages, and final body weight and organ weights (heart, liver, kidney, adrenal, spleen and uterus) were recorded immediately. Although nearly all the test variables were not affected by environmental enrichment in their mean values, the enriched group showed higher coefficients of variation in many variables, and strain differences of both housing conditions were not consistent. The influences of enrichment were shown to be strain- and test-dependent. Such effects may lead to an increase in the number of animals which is necessary or may change the experimental results, especially when a study, using enriched housing conditions, focuses on strain differences. Since the same enrichment design can result in different influences, a positive or a negative or no adverse effect, due to the strain and the variables studied, researchers need to collect more information before enrichment designs are introduced into experimental plans.

Prescreening Slash Pine and Cronartium Pedigrees for Evaluation of Complementary Gene Action in Fusiform Rust Disease.

Single-urediniospore cultures of the fusiform rust fungus were used to inoculate seedlings from 10 full-sib families of a five-parent slash pine diallel at two different times in 1994. The presence or absence of fusiform rust galls was recorded for each inoculated seedling at 9 months postinoculation, and percent infection levels for each family-inoculum-time combination were used for detecting differences among host families and fungal cultures and for identifying differential interactions. The existence of differential interactions between two or more fungal cultures and two or more host families verifies that complementary gene action does exist in this pathosystem. Some host families may be excluded from more detailed interaction studies on the basis of their redundancy and lack of participation in differential interactions.

Spreading of trypsinized cells: cytoskeletal dynamics and energy requirements.

The spreading of trypsinized XTH-2 cells (a line derived from Xenopus laevis tadpole heart endothelia) on glass was investigated. Three phases can be distinguished: (1) blebbing of rounded cells, first attachment to a solid substratum and formation of a broad smooth contact area; (2) organization of a peripheral zone of actin fibrils and reinforcement of the basal cytoplasm by a stress fibre-like pattern; (3) extension of lamellae. The first phase seems to be independent of a supply of metabolic energy, while the others clearly depend on it. This is concluded from the close relationship between cellular projection area and energization of mitochondria as revealed by (a) the fluorescence intensity of cells vitally stained with the mitochondria-specific fluorochrome DASPMI (2-4-(dimethylamino)-styryl-1-methylpyridinium-iodine); (b) the degree of spreading in the presence of inhibitors of respiration; (c) effective amelioration of spreading (phases (2) and (3] under conditions of high ATP content. In phase (2) the extension of the central part of the cells becomes stabilized, the cell body settles on the basal cytoplasmic layer and further expansion of the projection area is achieved by lamella formation (phase (3]; motile and stabile regions of the cells become separated. This sequence of events is interpreted as a self-organizing process based on the development of internal hydraulic pressure, actin polymerization and contraction of the newly developed actomyosin network. During trypsinization, depolymerization of actin does not occur but rather on addition of Ca2(+)-containing media. Cellular ATP content drops as well on trypsinization, as on addition of Ca2+. Manganese promotes spreading by decreasing F-actin disassembly and maintaining a high level of cytosolic ATP, most probably because it is not accepted by the calcium pumps. Regarding the association of glycolytic enzymes with F-actin and their influence on actin assembly, lactate dehydrogenase has been inhibited with oxamic acid. This treatment improves the correlation between F-actin content and the degree of spreading; however, the total amount of F-actin remains smaller and the cells spread more.