Figure 1.
Blood glucose measurements during the study.
For both groups measurements were taken at baseline, on day −5, +2, +9 of grid floor exposure for the test group and at the end of the study (mmol/L, mean and SD).
Table 1.
Cytokine, blood glucose and HbA1c measurements.
Figure 2.
(Blue: control mice, Red: grid floor housed mice). A. DGGE-profile cluster analysis similarity tree from end fecal samples. The mice clustered strongly after gel (green, yellow and purple lines) and after cage allocation (rounded squares). There was no clear clustering according to housing. Overall similarity was 59.61%±4.68. B. DGGE-profile cluster analysis similarity tree from end cecal samples. The mice clustered according to housing (circles) independent of gel (green and yellow) and cage allocation (rounded squares). The overall similarity was 64.07%±5.64. C. Scatter plot of the y-component of similarity-based cecal PCA-plot (mean and SD). Difference between control mice (−107563±108957) and grid floor housed mice (107563±84155) is significant (p<0.01).
Table 2.
Significantly different taxa between control mice and grid floor housed mice.
Figure 3.
16S rRNA gene 454/FLX based pyrosequencing of cecal content.
(Blue: control mice, Red: grid floor housed mice). A. Jackknifed replicate PCA plot. The plot is based on the phylogenetic distance matrix showing clustering of mice according to the floor type. B. Whisker plot of the 2nd principal component values. The significant difference between the two groups observed in the DGGE-based PCA-plot is confirmed (p<0.01***).
Figure 4.
Time spent in each of the three Tripletest compartments and risk assessments.
No significant differences were observed between control mice and grid floor housed mice in the overall time spent in the three compartments. Some significant changes were observed within each group. ”IZ”: time spent in IZ of total test time, “DC”: time spent in DC of total test time, “Entries CA”: number of entries into CA from OF, LDB or CA, “RAOA”: risk assessment OA, “RAOF”: risk assessment OF. “OF/total test time”: time spent in Open Field of total test time, “EPM”/total test time”: time spent in Elevated Plus Maze of total test time, “LDC/total test time”: time spent in Light (LC) or Dark Compartment (DC) of total test time, “RAOA, RAL, RAOF”: time spent risk assessing Open Arms, Light Compartment and Open Field, respectively.
Figure 5.
Scatter plots of amount for control mice (A) and mice housed on grid floor (B) (g, mean and SD). No significant difference was found between the groups. Within the groups, there was a significant difference from baseline to end, which differed between the groups with a mean of 10.77 g removed wood chips for the control mice (p<0.05) compared to 19.64 g for grid floor housed mice (p<0.01).
Figure 6.
End results from three mice were not obtained, as they learned to master tail climbing. A. End results for immobility duration for control mice and grid floor housed mice. There was a significant difference between the groups in total duration of immobility (p<0.05) (s, mean and SD). B. Within-group comparison from baseline to end of grid floor housed mice. There was a significant increase in the number of immobility episodes (p<0.05) (number of episodes, mean and SD).
Table 3.
Bacterial levels of correlation between pyrosequencing data from cecal flora and parameters.
Table 4.
Timeframe with number of days, age of the mice and tests performed.
Table 5.
Construction details of the Tripletest.
Figure 7.
Overview of Tripletest construction.
The Open Field is connected to the closed arm of the Elevated Plus Maze with a 7×7 cm opening and a similar passage connects the other closed arm of the Elevated Plus Maze to the Light/Dark Box. The whole system is elevated 45 cm from the floor.