DataSheet1_A pilot investigation of the efficacy and safety of magnesium chloride and ethanol as anesthetics in Loligo vulgaris embryos.docx

The inclusion of cephalopods in the legislation related to the use of animals for experimental purposes has been based on the precautionary principle that these animals have the capacity to experience pain, suffering, distress, and lasting harm. Recent studies have expanded this view and supported it. Handling cephalopod mollusks in research is challenging and whenever more invasive procedures are required, sedation and/or anesthesia becomes necessary. Therefore, finding adequate, safe, and effective anesthetics appears mandatory. Several substances have been considered in sedating cephalopods, in some instances applying those utilized for fish. However, species-specific variability requires more detailed studies. Despite long-lasting experience being linked to classic studies on squid giant axons, evidence of action on putative anesthetic substances is scarce for Loligo vulgaris and particularly for their embryos. The aim of the current study was to evaluate effects elicited by immersion of squid embryos in anesthetic solutions and examine whether these forms display a similar reaction to anesthetics as adults do. Different concentrations of ethanol (EtOH; 2, 2.5, and 3%) and magnesium chloride (MgCl2; 1, 1.5, and 1.8%) were tested by adopting a set of indicators aimed at exploring the physiological responses of squid embryos. Forty-two embryos of the common squid Loligo vulgaris (stages 27–28) were assigned to three conditions (EtOH, MgCl2, and controls) and video recorded for 15 min (5 min before, 5 min during, and 5 min after immersion in the anesthetic solutions). In each group, the heart rate, respiratory rate, buoyancy, chromatophore activity, and tentacles/arms responses were assessed to evaluate the embryos' vitality and responsiveness to stimulation. Both substances provoked a decrease in heart and respiratory rates and inhibited buoyancy, chromatophores, and tentacles/arms responses; no adverse effects were observed. EtOH had a faster onset of action and faster recovery than MgCl2, being potentially more adequate as an anesthetic for shorter procedures. Even though MgCl2 caused a longer muscle relaxation, the reversibility was not confirmed for the 1.8% concentration; however, lower concentrations triggered similar results as the ones obtained with the highest EtOH concentrations. We have shown that the late developmental stages of Loligo vulgaris embryos could represent a good model to evaluate anesthetics for cephalopods since they can display similar reactions to anesthetics as adults animals do.</p

data_sheet_1.PDF

Introduction<p>The use of a surgical safety checklist is recommended by the World Health Organization and is associated with advantages: improved communication and reduced complications and mortality. Adapting checklists to the environment in which they are used improves their efficiency, but their implementation can be challenging. The aim of this study was to develop and implement a perianesthetic safety checklist for a small animal hospital.</p>Materials and methods<p>A panel of eight anesthesia diplomates and seven residents and doctoral students were gathered. The Delphi method was used to generate a checklist. The checklist was presented individually to each user by the primary investigator and introduced into the clinical routine over a 5-week period. An interdisciplinary meeting was then held, and the checklist was modified further. Six months after introduction, the use of the checklist was directly observed during 69 anesthetic cases and a survey was sent to the users. A second implementation was organized after formally presenting the checklist to the staff, designating the anesthesia clinical lead as the person responsible for printing and controlling use of the checklist. A second evaluation was performed 3 months later (64 anesthetic cases).</p>Results<p>Using the Delphi process led to the creation of a checklist consisting of three parts: “sign in” (before induction of anesthesia), “time out” (before the beginning of the procedure), “sign out” (at the end of the procedure). At the first assessment, the checklist was printed and used in 32% of cases and not printed in 41% of cases. Response rate of the survey was fair (19/32 surveys): 14/19 users thought the checklist contributed to improving communication; 15/19 reported improved safety and better management of the animals; 9/19 users avoided mistakes (77% would have omitted the administration of antimicrobial prophylaxis); 10/19 thought it was time consuming. At the second assessment, the checklist was used in 45% of cases (printed but not used in 55%). The use of the sign-out section of the checklist was significantly improved.</p>Conclusion and clinical relevance<p>This study illustrates an innovative use of the Delphi method to create a safety checklist. Challenges associated with implementation are reported.</p

data_sheet_2.docx

Introduction<p>The use of a surgical safety checklist is recommended by the World Health Organization and is associated with advantages: improved communication and reduced complications and mortality. Adapting checklists to the environment in which they are used improves their efficiency, but their implementation can be challenging. The aim of this study was to develop and implement a perianesthetic safety checklist for a small animal hospital.</p>Materials and methods<p>A panel of eight anesthesia diplomates and seven residents and doctoral students were gathered. The Delphi method was used to generate a checklist. The checklist was presented individually to each user by the primary investigator and introduced into the clinical routine over a 5-week period. An interdisciplinary meeting was then held, and the checklist was modified further. Six months after introduction, the use of the checklist was directly observed during 69 anesthetic cases and a survey was sent to the users. A second implementation was organized after formally presenting the checklist to the staff, designating the anesthesia clinical lead as the person responsible for printing and controlling use of the checklist. A second evaluation was performed 3 months later (64 anesthetic cases).</p>Results<p>Using the Delphi process led to the creation of a checklist consisting of three parts: “sign in” (before induction of anesthesia), “time out” (before the beginning of the procedure), “sign out” (at the end of the procedure). At the first assessment, the checklist was printed and used in 32% of cases and not printed in 41% of cases. Response rate of the survey was fair (19/32 surveys): 14/19 users thought the checklist contributed to improving communication; 15/19 reported improved safety and better management of the animals; 9/19 users avoided mistakes (77% would have omitted the administration of antimicrobial prophylaxis); 10/19 thought it was time consuming. At the second assessment, the checklist was used in 45% of cases (printed but not used in 55%). The use of the sign-out section of the checklist was significantly improved.</p>Conclusion and clinical relevance<p>This study illustrates an innovative use of the Delphi method to create a safety checklist. Challenges associated with implementation are reported.</p

Metadata D-values AvrageCOPp.

IntroductionThe Centre of Pressure (COP) is the single point summarising all forces transferred to the hoof during the stance phase of a stride. COP path (COPp) is the trajectory that COP follows from footstrike to lift-off. Aim of the present study was to characterize the COP and COPp in horses affected by osteoarthritis and chronic lameness.Materials and methodsSeventeen adult horses with a diagnosis of osteoarthritis and single limb chronic lameness were recruited. The COP was recorded using a wireless pressure measuring system (TekScan®) with sensors taped to the hooves (either fore- or hind limb, depending on lameness location). The COPp coordinates were further processed. Procrustes analysis was performed to assess the variability of single strides COPp and average COPp among strides, gaits, and limbs by calculating Procrustes distances (D-values). A linear mixed-effects model was run to analyse D-values differences for lame and sound limbs. Additionally, average COPp D-values and COPp hoofprint shape indices were compared for lame and sound limbs with the Signed Rank Test.ResultsAt walk and trot the single-stride COPp D-values were significantly lower in lame than in sound limbs (marginal effects pConclusionOur findings are in line with previous observations that COPp are highly repetitive and characteristic for each horse and limb. There seems to be a further decrease in COPp variability in the presence of a painful limb pathology.</div

Results of the shape index analysis are shown.

IntroductionThe Centre of Pressure (COP) is the single point summarising all forces transferred to the hoof during the stance phase of a stride. COP path (COPp) is the trajectory that COP follows from footstrike to lift-off. Aim of the present study was to characterize the COP and COPp in horses affected by osteoarthritis and chronic lameness.Materials and methodsSeventeen adult horses with a diagnosis of osteoarthritis and single limb chronic lameness were recruited. The COP was recorded using a wireless pressure measuring system (TekScan®) with sensors taped to the hooves (either fore- or hind limb, depending on lameness location). The COPp coordinates were further processed. Procrustes analysis was performed to assess the variability of single strides COPp and average COPp among strides, gaits, and limbs by calculating Procrustes distances (D-values). A linear mixed-effects model was run to analyse D-values differences for lame and sound limbs. Additionally, average COPp D-values and COPp hoofprint shape indices were compared for lame and sound limbs with the Signed Rank Test.ResultsAt walk and trot the single-stride COPp D-values were significantly lower in lame than in sound limbs (marginal effects pConclusionOur findings are in line with previous observations that COPp are highly repetitive and characteristic for each horse and limb. There seems to be a further decrease in COPp variability in the presence of a painful limb pathology.</div

Pooled D-values for trot (left) and walk (right) trials calculated for the 17 horses included in the study.

At walk, median D-values were 0.106 (0.053–0.190) and 0.089 (0.0436–0.165) for sound and lame limbs respectively, while at trot they were 0.132 (0.052–0.245) and 0.0723 (0.0301–0.154) for sound and lame limbs respectively.</p

Distribution for each quadrant (as in Fig 2) of the footstrike and lift-off coordinates of the average COPp for the left-right comparison and for the lame-sound comparison (Q = Quadrant).

Distribution for each quadrant (as in Fig 2) of the footstrike and lift-off coordinates of the average COPp for the left-right comparison and for the lame-sound comparison (Q = Quadrant).</p

Definition of standard distances and shape indices within the hoof print.

Definition of standard distances and shape indices within the hoof print.</p

Metadata D-values individual horses.

IntroductionThe Centre of Pressure (COP) is the single point summarising all forces transferred to the hoof during the stance phase of a stride. COP path (COPp) is the trajectory that COP follows from footstrike to lift-off. Aim of the present study was to characterize the COP and COPp in horses affected by osteoarthritis and chronic lameness.Materials and methodsSeventeen adult horses with a diagnosis of osteoarthritis and single limb chronic lameness were recruited. The COP was recorded using a wireless pressure measuring system (TekScan®) with sensors taped to the hooves (either fore- or hind limb, depending on lameness location). The COPp coordinates were further processed. Procrustes analysis was performed to assess the variability of single strides COPp and average COPp among strides, gaits, and limbs by calculating Procrustes distances (D-values). A linear mixed-effects model was run to analyse D-values differences for lame and sound limbs. Additionally, average COPp D-values and COPp hoofprint shape indices were compared for lame and sound limbs with the Signed Rank Test.ResultsAt walk and trot the single-stride COPp D-values were significantly lower in lame than in sound limbs (marginal effects pConclusionOur findings are in line with previous observations that COPp are highly repetitive and characteristic for each horse and limb. There seems to be a further decrease in COPp variability in the presence of a painful limb pathology.</div

Metadata mixed model.

IntroductionThe Centre of Pressure (COP) is the single point summarising all forces transferred to the hoof during the stance phase of a stride. COP path (COPp) is the trajectory that COP follows from footstrike to lift-off. Aim of the present study was to characterize the COP and COPp in horses affected by osteoarthritis and chronic lameness.Materials and methodsSeventeen adult horses with a diagnosis of osteoarthritis and single limb chronic lameness were recruited. The COP was recorded using a wireless pressure measuring system (TekScan®) with sensors taped to the hooves (either fore- or hind limb, depending on lameness location). The COPp coordinates were further processed. Procrustes analysis was performed to assess the variability of single strides COPp and average COPp among strides, gaits, and limbs by calculating Procrustes distances (D-values). A linear mixed-effects model was run to analyse D-values differences for lame and sound limbs. Additionally, average COPp D-values and COPp hoofprint shape indices were compared for lame and sound limbs with the Signed Rank Test.ResultsAt walk and trot the single-stride COPp D-values were significantly lower in lame than in sound limbs (marginal effects pConclusionOur findings are in line with previous observations that COPp are highly repetitive and characteristic for each horse and limb. There seems to be a further decrease in COPp variability in the presence of a painful limb pathology.</div