Abstract

In this study, the effects of agitation, temperature, and pH on biofilm formation by Mycobacterium fortuitum were studied and quantified through response surface modeling. The microtiter plate assay was optimized to achieve conditions favoring maximum mycobacterial biofilm quantification. Optical density (OD) measurement using a crystal violet assay was performed to estimate the amount of biofilm formed. Response surface methodology (RSM) results revealed an R² value of 96.18%, exhibiting a maximum OD of 2.119 (λ_570 nm) at a temperature of 37 °C and pH 7.0, under a static environment. The conditions were experimentally validated. Statistically significant results showed that the maximum biofilm was produced 96 h after mycobacterial inoculation. Thus, the results provide a basis for using RSM as an efficient optimization method for M. fortuitum biofilm assays. This approach can also be incorporated into strategies for screening anti-biofilm compounds, synthetic chemicals, drugs, or inhibitors against pathogenic mycobacteria.

摘要

在这项研究中，通过响应面模型研究和量化了搅拌、温度和 pH 值对偶然分枝杆菌生物膜形成的影响。微量滴定板测定被优化以达到有利于最大分枝杆菌生物膜定量的条件。使用结晶紫测定法进行光密度 (OD) 测量以估计形成的生物膜的量。响应面方法 (RSM) 结果显示R ²值为 96.18%，最大 OD 为 2.119 (λ _{570 nm}) 在 37 °C 的温度和 pH 7.0 下，在静态环境下。这些条件经过实验验证。统计显着性结果表明，最大的生物膜是在分枝杆菌接种后 96 小时产生的。因此，结果为使用 RSM 作为M的有效优化方法提供了基础。偶然生物膜测定。这种方法也可以纳入筛选抗生物膜化合物、合成化学品、药物或病原体分枝杆菌抑制剂的策略中。