Composition and flow rate of the anaesthetic mixtures used for anaesthesia induction – pilot study.

<p>The Isoflurane % was read on the dial of the precision vaporizer, the O2 and N2O % referred to the Fi of the gases.</p

The mean frequency of abnormal walking behaviour (+1SE) in C57 and C3H mice in each control and surgery group.

<p>High dose buprenorphine significantly increased abnormal behaviour frequency in controls (group ABupH) making it impossible to determine any post-surgical analgesic effects. (n = 8 per group; 4 of each strain). Symbols indicate individual group comparisons: * Group VSa cf. Asa, ABupL (p = 0.031, 0.004, respectively); § Group ABupL cf. VSa, VBupL, VBupH (p = 0.004, <0.001, <0.001, respectively).</p

The mean frequency of grooming behaviour (+1SE) following automated behaviour analysis with HCS.

<p>Grooming increased in response to surgery but C3H mice groomed less overall. The C57 mice showed a greater increase following surgery which was significantly reduced by high dose buprenorphine. However, this effect was again confounded by a similar response in high dose C57 controls (n = 8 per group; 4 of each strain). Symbols show the results of individual comparisons: * Group ASa cf. VSa (p = 0.001); ∞ ABupL cf. VBupL (p = 0.002); ¥ ASa cf. VBupL (p = 0.002); $ ABupL cf. VSa (p = 0.001); § C57 mice in group VBupH cf. C57 mice in groups VBupL, VSa (p = 0.043, both comparisons).</p

The mean frequency of the Composite normal behaviour measure ‘Score 2’ (+1SE) obtained from manual behaviour analysis.

<p>This was computed from the occurrences of bipedal high rearing, head and face washing (‘Lick Head’), digging, normal posturing, head scratching or of other body regions (e.g. flank). Note the significant reduction in normal behaviour caused by buprenorphine in both strains (ABupH), which was significant in C3H mice in this group compared to both other control groups. (n = 8 per group; 4 of each strain). Symbols indicate individual group and strain results whereby: * Group ABupL, Score 2 significantly greater frequency than all other groups except Asa (p≤0.001); § C3H mice in group ABupH vs. C3H groups Asa, ABupL (p = 0.039, 0.008, respectively).</p

Geometric mean frequency of the composite normal activity measure (G^behave) from HCS (automated) analysis (with 95% confidence intervals).

<p>The text gives details of how this was computed from the Low, Medium and High activity scores. Results were similar to those obtained manually; surgery reduced normal behaviour incidence and C3H mice were more susceptible to the confounding effects of high dose buprenorphine; i.e. normal behaviour was significantly depressed in this strain (group ABupH). (n = 8 per group; 4 of each strain). Symbols indicate results of individual group comparisons: * Groups Asa, ABupL vs. VSa, VBupL (p<0.001 for respective comparisons); ¥ Group ABupL vs. VBupH, p = 0.002); § C3H mice in group ABupH cf. Asa, ABupL (p = 0.01, p = 0.001, respectively); ∞ C57 mice in group VBupH vs. C57 mice in groups VSa, VBupL (p = 0.012, 0.011, respectively); $ C57 mice in group VBupH cf. C57 mice in group ABupL (p = 0.014).</p

Values of pH, P_aO₂ (mmHg) and lactate (mmol.l⁻¹) measured in arterial blood at the time of loss of righting reflex.

<p>Composition of the gas mixtures for the groups 1–3: see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032290#pone-0032290-t003" target="_blank">table 3</a>.</p

Time to loss of righting reflex (LORR) in seconds within the 3 treatment groups and the 2 strains of mice (C57Bl/6 and CD1).

<p>Error bars: +1 SD. Composition of the gas mixtures for the groups 1–3: see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032290#pone-0032290-t003" target="_blank">table 3</a>.</p

The mean frequency of inactive periods ‘Stop’ (+1SE) in each control and surgery group in data collected manually.

<p>Buprenorphine resulted in a decrease in periods of inactivity both in the control and surgery groups given the high dose of drug. A similar decrease in inactivity also occurred in the other two surgery groups (VSa and VLBup). (n = 8 per group; 4 of each strain). Asterisks indicate groups Asa, ABupL were significantly more frequently ‘Stopped’ than all other groups (p≤0.031 for all individual comparisons).</p

Plot of canonical discriminant scores for individual mice in each group according to Low, Medium and High frequency normal behaviour data from analysis with HCS (Automated).

<p>Groups are identified as in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075948#pone-0075948-g001" target="_blank">Figure 1</a> legend; the axis labels show the significance of each Function and its respective percentage contribution to between-group variation. Although activity changes showed significant separation of the surgery and control groups (Function 1; P<0.001), compared with the results of pain scoring there was a lack of differentiation according to whether mice were treated with saline or buprenorphine. Low frequency measures were: ‘Jumping’ and ‘Come Down’; Medium frequency: ‘Rear Up’, ‘Walk Left’ and ‘Walk Right’; High frequency: ‘ComeDown from and to Partially Reared’ (2 behaviours), ‘RearUp from and to Partially Reared, ‘Remain Low’, Remain Partially Reared’, ‘Sniff’ and ‘Walk Slow’. (n = 8 per group).</p

Diagram representing the induction chamber.

<p>The fresh gas inlet (bottom arrow) of the chosen mixture was delivered underneath a multi-perforated floor. A total of 4 mini fans were ensuring that the mixture was optimally blended. The sampling end of the sidestream gas monitoring (<b>X</b>) was positioned at mouse level, 8 cm above the floor. The gas outlet (upper arrow) was connected to an active scavenging system.</p