SCRAPING AND SONICATION OF IMPLANTS FOR DISLODGING BACTERIA IN A RAT EXPERIMENTAL BIOFILM MODEL

Abstract

Soft tissue biopsies may prove culture negative in biofilm prosthetic infections. Identification of the causative bacteria could be achieved by either scraping of the prosthetic surface or by sonication of the entire implant. These techniques have not been thoroughly studied in experimental models where the biofilm is developed in vivo.

In a novel rat biofilm model we compared scraping and sonication as methods for dislodging biofilm bacteria.

Twenty plates of steel alloy (5×7×1mm), with a surface roughness (Ra) of 0.35 (0.19–0.51) μm, were inserted into 20 standardised pieces of sheep costae, weight: 1.2 (1.0–1.5) gram. To each bone graft was added 50 μL of a Staphylococcus epidermidis suspension containing 1.4 (1.1–1.7)×104 CFU. Ten Sprague Dawley rats were operated with implantation of the bone graft subfascially on each side of the interscapular region. After two weeks the grafts were excised. The plates were removed from the grafts and rinsed twice in saline. Aliquots of 50 μL were cultured. 10 plates were scraped, followed by vortex mixing of the knife blade; and 10 plates were sonicated at 30 kHz for five minutes. 50 μL of the saline used for a) vortex mixing of the knife blade, and b) sonication, was seeded on agar. After overnight incubation the number of CFU was counted.

The total number of CFU recovered after scraping and sonication were 2(0–13) × 102 and 298(8–878) × 102, respectively (p< 0, 01). Compared to the number of CFU in the rinsing fluid, no increase was observed after scraping. For each plate that was sonicated there was a 38 (3–300) fold increase in the number of CFU.

First, sonication is a superior technique for dislodging biofilm bacteria in an in vivo model, compared to scraping. Secondly, the present experimental model is a promising method for developing biofilm in vivo.