An efficient, cost-effective method for determining the growth rate of sulfate-reducing bacteria using spectrophotometry
| Autor: | A. Osman, Scott A Wade, J. L. Wood |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Growth data
Clinical Biochemistry Optical density Improved method 010501 environmental sciences 01 natural sciences 03 medical and health sciences Microtiter plate Microbial corrosion Spectrophotometry medicine Growth rate Sulfate-reducing bacteria lcsh:Science 030304 developmental biology 0105 earth and related environmental sciences ComputingMethodologies_COMPUTERGRAPHICS Immunology and Microbiology 0303 health sciences medicine.diagnostic_test Spectrophotometric growth analysis of sulfate-reducing bacteria using acid-amendments Biocorrosion Dilution Medical Laboratory Technology Environmental science lcsh:Q Biochemical engineering SRB |
| Zdroj: | MethodsX MethodsX, Vol 6, Iss, Pp 2248-2257 (2019) |
| ISSN: | 2215-0161 |
| Popis: | Graphical abstract 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. |
| Databáze: | OpenAIRE |
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