Comparative Medicine Program, Texas A&M University, College Station, Texas.
Department of Epidemiology and Biostatistics, School of Public Health, Texas A&M University, College Station, Texas.
J Am Assoc Lab Anim Sci. 2024 Jul 1;63(4):368-376. doi: 10.30802/AALAS-JAALAS-23-000122. Epub 2024 Mar 15.
Validated glass bead sterilization protocols to effectively sterilize rodent surgical instruments after bacterial exposure (for example, cecal contamination) are lacking. To refine current approaches, we added either a multienzyme detergent, neutral pH detergent, or chlorhexidine scrub step before glass bead sterilization of forceps or needle drivers exposed to cecal contents. We exposed sets of forceps and needle drivers to cecal contents, which were then air dried for 3 min. Immediately after, the instruments were wiped several times with a clean, dry paper towel. The contaminated tips were soaked in either a multienzyme or neutral pH detergent ( = 5 min), chlorhexidine scrub ( = 2 min), or no pretreatment solution. To further increase debris removal, instruments (from all groups) were brushed using a clean toothbrush. The nonpretreatment instruments were briefly soaked in saline before brushing. After being rinsed with sterile water, all instruments were exposed to a glass bead sterilizer for 60 s at 500 °F (260 °C). Sets were then swabbed for bacterial culturing. Swabs were plated onto either sheep blood agar ( = 23) or chocolate agar ( = 20) for aerobic culturing or agar ( = 20) for anaerobic culturing. A subset of instruments was sampled to determine organic material presence after treatment using an ATP luminometer ( = 21). Multiple agar types and bioluminescence were used to more deeply evaluate tool sterility and to differentiate the relative effectiveness of each protocol. From the saline group, only one pair of forceps yielded growth on agar, and 2 pairs yielded growth on chocolate agar. No other bacterial growth was observed. The use of a pretreatment agent also lowered overall organic contamination levels in needle drivers compared with using only saline. These results indicate that brushing instruments to mechanically remove debris from instruments is paramount to ensure sterility. However, a best practice would be to also use one of the pretreatment options used in this study.
缺乏经过验证的玻璃珠灭菌方案来有效灭菌经细菌暴露（例如盲肠污染）后的啮齿动物手术器械。为了改进当前的方法，我们在将镊子或持针器暴露于盲肠内容物后，用玻璃珠灭菌之前，添加了多酶清洁剂、中性 pH 清洁剂或洗必泰擦洗步骤。我们将一组镊子和持针器暴露于盲肠内容物中，然后将其风干 3 分钟。之后，立即用干净的干纸巾擦拭器械数次。受污染的尖端浸泡在多酶或中性 pH 清洁剂中（= 5 分钟）、洗必泰擦洗（= 2 分钟）或无预处理溶液中（= 5 分钟）。为了进一步去除碎屑，使用干净的牙刷清洁所有器械（来自所有组）。非预处理器械在刷之前短暂浸泡在盐水中。用无菌水冲洗后，将所有器械在 500°F（260°C）下用玻璃珠灭菌器处理 60 秒。然后对器械进行拭子取样以进行细菌培养。将拭子接种到绵羊血琼脂（= 23）或巧克力琼脂（= 20）上进行需氧培养，或接种到 琼脂（= 20）上进行厌氧培养。用 ATP 发光计（= 21）对器械处理后的有机物质存在情况进行抽样。使用多种琼脂类型和生物发光来更深入地评估工具的无菌性，并区分每种方案的相对效果。从盐水组中，只有一对镊子在 琼脂上产生了生长，两对在巧克力琼脂上产生了生长。未观察到其他细菌生长。与仅使用盐水相比，预处理剂的使用还降低了持针器的总体有机污染水平。这些结果表明，用刷子清除器械上的碎屑对于确保无菌性至关重要。但是，最佳实践是还要使用本研究中使用的预处理方案之一。
