Sampling
Week 1
During the sampling , one of the examiners held the dog , while the other examiner clipped a 3 cm × 2 cm area over the cephalic vein with clippers ; 10 dogs were clipped on the right leg and the other 10 were clipped on the left leg .
The clipped surface was disinfected by rubbing for 10 seconds with 3 ml of alcohol 5 mg / ml ( Fresenius Kabi ) applied to cotton wool using a 5 ml syringe . The time between disinfection and sampling was 30 seconds . Sterile swabs ( AMIES , Copan innovation ) were dipped in a sterile 15 ml tube ( Falcon , Sarstedt ) containing 1 ml of physiological saline ( BVF , SLU ), to soften the swabs before use .
A propylene plastic template was placed on the disinfected leg to standardise the area to be swabbed . The area within the template was swabbed with the soaked swab , using rotating movements . The swab was then placed back in the Falcon tube with physiological saline . The Falcon tube was marked with the identification number given to the dog for the study ( for example , dog # 1 ) and a note of whether the sample was taken from a clipped or unclipped leg .
The plastic template was disinfected before being used on the next leg . The process was repeated by the examiner on the dog ' s other front leg , but the leg was not clipped . The clippers were cleaned with a surface disinfectant ( DAX ) and a toothbrush ( Colgate-Palmolive ) before being used on the next dog . The plastic template was also disinfected with surface disinfection after each use . The process was repeated on all dogs .
One examiner took all the samples one day and the other examiner applied the samples to the blood agar and Sabouraud plates before incubation ; the next day , the roles changed .
Week 2
There was little or no growth of bacteria from the samples taken during the first week so , after sampling the first 10 dogs , the method was adapted to increase the probability of collecting more bacteria in the sampling process . Swabs were replaced with SELEFA cotton-tipped applicators ( OneMed , Finland ). The study was also extended , with the dogs being sampled before disinfection , as the disinfection could have had a greater impact on hygiene than the clipping .
In addition , the time between disinfection and sampling was increased to 60 seconds , when the alcohol was judged to have evaporated . If the alcohol had not evaporated after 60 seconds , the sampling was delayed until it was confirmed it had fully evaporated .
Analysis
Week 1
After sampling , two blood agar plates ( SVA , Uppsala ) and two Sabouraud agar plates ( SVA , Uppsala ) were marked for each dog with the last four digits in the dog ' s microchip number and a note of whether the sample came from the right or left leg .
The physiological saline in the Falcon tube was homogenised using a vortex . From each tube , 0.1 ml of physiological saline was taken and transferred to a blood agar plate and 0.1 ml was transferred to a Sabouraud agar plate . Each sample of saline was spread over the agar with a sterile spreading blade .
The blood agar plates were incubated at 37 ° C for 24 hours and the Sabouraud agar plates were incubated at 30 ° C for either 120 hours or 144 hours . The colonies on the blood agar plates were counted and visually noted after 24 hours and 48 hours . The Sabouraud plates were visually read after 120 hours or 144 hours depending on which day of the week the incubation of the samples began .
Week 2
In the second week , four blood agar plates were used per dog . The blood agar plates were marked with the last four digits of the dog ' s chip number , the leg from which the sample was taken , and whether it was sampled before or after disinfection . The samples were applied to the blood agar and Sabouraud plates before incubation , according to the same method used the first week .
Statistics
The statistics were calculated using Microsoft Excel . p-values were calculated using a paired and onesided Student ' s t-test , which meant that the right leg was compared with the left leg for each dog . Values for clipped legs were compared with values for unclipped legs for 24 hours and 48 hours of incubation , respectively ( blood agar plates ) and 120 or 144 hours , respectively ( Sabouraud agar plates ). The samples taken before disinfection were compared with the samples taken after disinfection . All samples that did not show any growth of bacteria were treated as value 0 when calculated with a one-sided , paired Student ' s t-test , but were included in the analysis .
Results
In the first part of the study , bacteria were detected after 24 hours of incubation in samples from the unclipped sampling sites in two dogs , and from the clipped sampling site in only one dog , following disinfection ( Table 1 , page 33 ).
32 Veterinary Nursing Journal