Sulfate-reducing bacteria (SRB) are anaerobes that cause human diseases, corrosion, and the bioremediation of metals. This study asserts the methyl tetrazolium assay as an accurate cell activity-based enumeration technique for quantifying SRB. The method offers a quick, easy, and reproducible enumeration technique, simplifying abundant calculations and elimination of the sulfide particles that complicate other counting methods, such as fluorescence-activated cell sorting, acridine orange spectrophotometry, and hemocytometer cell counting, simplifying the quantification of live SRB.
Get full access to this article
View all access options for this article.
References
1.
RowanF, DochertyNG, MurphyM, et al.Desulfovibrio bacterial species are increased in ulcerative colitis. Dis Colon Rectum, 2010; 53(11):1530–1536; doi: 10.1007/DCR.0b013e3181f1e620
2.
MurrosKE, HuynhVA, TakalaTM, et al.Desulfovibrio bacteria are associated with Parkinson’s disease. Front Cell Infect Microbiol, 2021; 11:652617; doi: 10.3389/fcimb.2021.652617
3.
FinegoldSM. Desulfovibrio species are potentially important in regressive autism. Med Hypotheses, 2011; 77(2):270–274; doi: 10.1016/j.mehy.2011.04.032
4.
RitzNL, BurnettBJ, SettyP, et al.Sulfate-reducing bacteria impairs working memory in mice. Physiol Behav, 2016; 157:281–287; doi: 10.1016/j.physbeh.2016.01.023
5.
Ilhan-SungurE, CanseverN, CotukA. Microbial corrosion of galvanized steel by a freshwater strain of sulphate reducing bacteria (Desulfovibrio sp.). Corros Sci, 2007; 49(3):1097–1109.
6.
DeplancheK, MacaskieLE. Biorecovery of gold by Escherichia coli and Desulfovibrio desulfuricans. Biotechnol Bioeng, 2008; 99(5):1055–1064; doi: 10.1002/bit.21688
7.
ZhangZ, ZhangC, YangY, et al.A review of sulfate-reducing bacteria: Metabolism, influencing factors and application in wastewater treatment. J Clean Prod, 2022; 376:134109; doi: 10.1016/j.jclepro.2022.134109
8.
RodriguesICB, LeãoVA. Producing electrical energy in microbial fuel cells based on sulphate reduction: A review. Environ Sci Pollut Res Int, 2020; 27(29):36075–36084; doi: 10.1007/s11356-020-09728-7
9.
EvansTM, WaarvickCE, SeidlerRJ, et al.Failure of the most-probable-number technique to detect coliforms in drinking water and raw water supplies. Appl Environ Microbiol, 1981; 41(1):130–138; doi: 10.1128/aem.41.1.130-138.1981
10.
MooreTC. Counting Cells with Hemocytometer V1. 2018; doi: 10.1750/4/protocols.io.nxsdfne
11.
ParkH-S, SchumacherR, KilbaneJJ. New method to characterize microbial diversity using flow cytometry. J Ind Microbiol Biotechnol, 2005; 32(3):94–102; doi: 10.1007/s10295-005-0208-3
12.
GrelaE, KozłowskaJ, GrabowieckaA. Current methodology of MTT assay in bacteria—A review. Acta Histochem, 2018; 120(4):303–311; doi: 10.1016/j.acthis.2018.03.007
13.
HundieGB, WoldemeskelD, GessesseA. Evaluation of direct colorimetric MTT assay for rapid detection of rifampicin and isoniazid resistance in Mycobacterium tuberculosis. PLoS One, 2016; 11(12):e0169188; doi: 10.1371/journal.pone.0169188
14.
BenovL. Effect of growth media on the MTT colorimetric assay in bacteria. PLoS One, 2019; 14(8):e0219713; doi: 10.1371/journal.pone.0219713
15.
GrelaE, ZąbekA, GrabowieckaA. Interferences in the optimization of the MTT assay for viability estimation of proteus mirabilis. Avicenna J Med Biotechnol, 2015; 7(4):159–167.
16.
TrafnyEA, LewandowskiR, Zawistowska-MarciniakI, et al.Use of MTT assay for determination of the biofilm formation capacity of microorganisms in metalworking fluids. World J Microbiol Biotechnol, 2013; 29(9):1635–1643; doi: 10.1007/s11274-013-1326-0
17.
DasKR, KowshikM, Praveen KumarMK, et al.Native hypersaline sulphate reducing bacteria contributes to iron nanoparticle formation in saltpan sediment: A concern for aquaculture. J Environ Manage, 2018; 206:556–564; doi: 10.1016/j.jenvman.2017.10.078
18.
YasirM, DuttaD, WillcoxMDP. Mode of action of the antimicrobial peptide Mel4 is independent of Staphylococcus aureus cell membrane permeability. PLoS One, 2019; 14(7):e0215703; doi: 10.1371/journal.pone.0215703
19.
GuoR, McGoverinC, SwiftS, et al.A rapid and low-cost estimation of bacteria counts in solution using fluorescence spectroscopy. Anal Bioanal Chem, 2017; 409(16):3959–3967; doi: 10.1007/s00216-017-0347-1
20.
LarssonP, EngqvistH, BiermannJ, et al.Optimization of cell viability assays to improve replicability and reproducibility of cancer drug sensitivity screens. Sci Rep, 2020; 10(1):5798; doi: 10.1038/s41598-020-62848-5
21.
ChenC-H, TsaoY-T, YehP-T, et al.Detection of microorganisms in body fluids via MTT-PMS assay. Diagnostics (Basel), 2021; 12(1):46; doi: 10.3390/diagnostics12010046
LemosRS, GomesCM, SantanaM, et al.The ‘strict’ anaerobe Desulfovibrio gigas contains a membrane‐bound oxygen‐reducing respiratory chain. FEBS Lett, 2001; 496(1):40–43; doi: 10.1016/S0014-5793(01)02399-7
24.
WildschutJD, LangRM, VoordouwJK, et al.Rubredoxin: Oxygen oxidoreductase enhances survival of Desulfovibrio vulgaris Hildenborough under microaerophilic conditions. J Bacteriol., 2006; 188(17):6253–6260; doi: 10.1128/JB.00425-06
25.
SpringS, LapidusA, SchröderM, et al.Complete genome sequence of Desulfotomaculum acetoxidans type strain (5575T). Stand Genomic Sci, 2009; 1(3):242–253; doi: 10.4056/sigs.39508
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
