An efficient, cost-effective method for determining the growth rate of sulfate-reducing bacteria using spectrophotometry

• Main Authors: J.L. Wood, A. Osman, S.A. Wade
• Format: Article
• Language: English
• Published: Elsevier 2019-01-01
• Series: MethodsX
• Online Access: http://www.sciencedirect.com/science/article/pii/S2215016119302602

The use of sulfate reducing bacteria (SRBs) in laboratory studies is a common approach for investigating microbially influenced corrosion (MIC). The characteristic formation of black iron sulfide precipitates during SRB growth, however, preclude the use of traditional spectrophotometric approaches for capturing growth data instead necessitating labour-intensive or technically specialized approaches. As such, an understanding of SRB growth responses to experimental conditions is often missing from MIC studies. Bernardez and de Andrade Lima (2015) have outlined a spectrophotometric approach for estimating SRB cell mass via the addition of HCl. This method has potential for the study SRB growth however its applicability is currently limited by the use of large aliquot volumes (45 mL), which restrict the number of timepoints that can sampled from one culture, and the extensive time devoted to cell preparation prior to OD readings. • We demonstrate an improved method for capturing SRB growth data via spectrophotometry following acidification. We incorporate lower sample volumes and adapt the method described in Bernardez and de Andrade Lima (2015) to a high throughput microtiter plate approach that increases the efficiency of this method and its applicability to growth rate studies. • Our results allay theoretical concerns that acidification may distort growth rate analysis by impacting cells differently depending on their metabolic state. • We further demonstrate that this method (acid-amended OD measurements) is more accurate and far more cost efficient than traditional methods (dilution spread-plate counting) and popular molecular methods (quantitative PCR) currently in use in SRB growth research. Method name: Spectrophotometric growth analysis of sulfate-reducing bacteria using acid-amendments, Keywords: Optical density, Sulfate-reducing bacteria, SRB, Growth rate, Microbial corrosion, Biocorrosion