Rapid fluorescence-based measurement of toxicity in anaerobic digestion

Chen, Jian Lin; Ortiz, Raphael; Xiao, Yeyuan; Steele, Terry W. J.; Stuckey, David C.

doi:10.1016/j.watres.2015.02.035

Cited by 21 publications

(23 citation statements)

References 33 publications

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“…In addition to being more convenient, rapid assays reduce the chance of artefacts caused by interactions of the test compound with the chemical compounds in the assay. The resazurin reduction assay is functionally similar to the tetrazolium assay, except it has the advantage of using a fluorescent detection method, which means it has a higher sensitivity to toxicants, for example, detecting the toxicity of 0.05 mg L −1 PCP to E. faecalis in 8 min, and to mixed anaerobic sludge in 10 min without pre‐exposure (Chen, Ortiz, et al, ). Resazurin has been used to continuously monitor living cell proliferation, and the viability of bacterial and eukaryotic cells, for hours or even days due to its non‐toxicity to cells.…”

Section: Resultsmentioning

confidence: 99%

“…Appropriate amounts of subcultured bacteria of E. faecalis (described above) were mixed with fresh autoclaved BHI broth in a 96‐well microplate to result in a final OD 600nm of 0.1, with PCP addition at 10 mg L −1 (IC 50 of PCP to E. faecalis . has been revealed at 10.2 mg L −1 (Chen, Ortiz, Xiao, Steele, & Stuckey, )). The 96‐well microplate was sealed immediately with sterile film with N 2 flushing over the tray to maintain anaerobic conditions.…”

Section: Methodsmentioning

confidence: 99%

“…; k 1 was obtained as a measure of the kinetics of reduction of resazurin to resorufin. This constant, k 1 , was chosen as the “toxicity indicator” (Chen, Ortiz, et al, ), and the difference in toxicity of PCP to different systems was determined using a two‐sample t ‐test.…”

Section: Methodsmentioning

confidence: 99%

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Metabolic reduction of resazurin; location within the cell for cytotoxicity assays

Chen

Steele

Stuckey

2017

Biotech & Bioengineering

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Resazurin is widely used as a metabolic indicator for living cells, however, there has been considerable debate in the literature with regards to the specific location in the cell where the non-fluorescent resazurin is reduced to the strongly fluorescent resorufin. This lack of clarity about the reduction site makes the use of resazurin reduction data in cytotoxicity studies difficult to interpret. In this study, E. faecalis, a Gram-positive and facultative anaerobic bacterial strain, and the most toxic chlorophenol, pentachlorophenol (PCP), were chosen as models for an anaerobe and toxicant, respectively. By studying the kinetics of resazurin reduction by E. faecalis after different treatments (cell disruption, bacterial filtration, and pre-exposure to toxicant), we confirmed that resazurin reduction to resorufin by live Gram-positive and facultative anaerobic bacterial cells can only happen intracellularly under anaerobic conditions, while resorufin reduction to dihydroresorufin can happen both intracellularly and extracellularly. Based on the understanding of these fundamental mechanisms, we suggest that resazurin reduction can be used as a quick bioassay for measuring cytotoxicity.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Metabolic reduction of resazurin; location within the cell for cytotoxicity assays

Chen

Steele

Stuckey

2017

Biotech & Bioengineering

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“…Therefore, a sensitive and comfort system is necessary to analyze the environmental (both ground and surface) and portable water. The existing analytical methods such as fluorescence and Raman spectrophotometry [44][45][46] , photo-electrochemical methods [47][48][49][50] , chromatography 51,52 , chemi-luminescence [53][54][55] , and electrochemical methods 29,[56][57][58][59][60] are used widely to detect the chlorophenol in aqueous medium. Among the chlorophenol detection methods, the electrochemical method has better advantages and the electrochemical method based on current versus potential is becoming more popular to detect directly the toxic chemicals which are environmental unsafe and carcinogenic [61][62][63][64] .…”

mentioning

confidence: 99%

Facile and efficient 3-chlorophenol sensor development based on photolumenescent core-shell CdSe/ZnS quantum dots

Rahman

Karim

Alam

et al. 2020

Sci Rep

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Quantum dots (QDs) are semiconducting inorganic nanoparticles, tiny molecules of 2-10 nm sizes to strength the quantum confinements of electrons. The QDs are good enough to emit light onto electrons for exciting and returning to the ground state. Here, CdSe/ZnS core/shell QDs have been prepared and applied for electrochemical sensor development in this approach. Flat glassy carbon electrode (GCE) was coated with CdSe/ZnS QDs as very thin uniform layer to result of the selective and efficient sensor of 3-CP (3-chlorophenol). The significant analytical parameters were calculated from the calibration plot such as sensitivity (3.6392 µA µM −1 cm −2) and detection limit (26.09 ± 1.30 pM) with CdSe/ZnS/ GCE sensor probe by electrochemical approach. The calibration curve was fitted with the regression coefficient r 2 = 0.9906 in the range of 0.1 nM ∼ 0.1 mM concentration, which denoted as linear dynamic range (LDR). Besides these, it was performed the reproducibility in short response time and successfully validated the fabricated sensor for 3-CP in the real environmental and extracted samples. It is introduced as a noble route to detect the environmental phenolic contaminants using CdSe/ZnS QDs modified sensor by electrochemical method for the safety of healthcare and environmental fields at broad scales. Semiconductor quantum dots (QDs), in particular those of cadmium chalcogenides (CdS, CdSe, CdTe), have received steadily growing attention due to their high potential for various applications, such as sensing 1,2 , biological labeling 3-12 , light-emitting devices (LED) 13-17 , lasers 18,19 , and solar cells 20-22. Recently, the number of interest has been developed in the area of electrochemical sensors and biosensors in which CdSe are employed for the determination of clinical markers 23 and food 24 and environmental pollutants 25,26. CdSe have been using as electrochemical signal tracers to exploit the oxidation potentials of the metal ions forming them. This can also be attributed to some of the characteristics of CdSe, such as a large surface area-to-volume ratio, good interfacial properties with high surface reaction activity, high electron-transfer efficiency, excellent biocompatibility, and feasibility for surface modification 27-30. Moreover, metallic QDs also constitute a feasible option for electrochemical multiplexed assays due to their different stripping peak potentials 31,32. This is particularly true for the development of efficient, reliable, rapid, and cost-affordable analytical procedures for the determination of anthropogenic and natural substances of either organic or mineral nature that have toxic, persistent, and bio-accumulative properties. Due to the prevalent pollution of ground and surface water by chloro-organic chemicals such as chlorophenol derivatives, the public concern has been growing over the several few decades. Because, the chlorophenols (including 3-CP) have high toxicity and able to produce mutagenic, estrogenic and carcinogenic effects in human beings 33-36 and even unsafe for t...

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“…It has been shown in the past that there is a strong correlation between the kinetics of resazurin reduction and the activity of anaerobic sludge, suggesting that the faster the resazurin reduction by anaerobic sludge, the higher the activity of the anaerobic sludge (Chen et al, 2015a(Chen et al, , 2015b. Figure 2 shows the activity of anaerobic sludge with two types of MNPs addition analyzed by the resazurin reduction assay on day 7 and 8 (after refeeding), and k1 value of reaction rate constants for resazurin to resorufin are summarized in Table 2.…”

Section: Effect Of Nanoparticles On the Activity Of Anaerobic Sludgementioning

confidence: 99%

The effect of Fe2NiO4 and Fe4NiO4Zn magnetic nanoparticles on anaerobic digestion activity

Chen

Steele

Stuckey

2018

Science of The Total Environment

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Rapid fluorescence-based measurement of toxicity in anaerobic digestion

Cited by 21 publications

References 33 publications

Metabolic reduction of resazurin; location within the cell for cytotoxicity assays

Metabolic reduction of resazurin; location within the cell for cytotoxicity assays

Facile and efficient 3-chlorophenol sensor development based on photolumenescent core-shell CdSe/ZnS quantum dots

The effect of Fe2NiO4 and Fe4NiO4Zn magnetic nanoparticles on anaerobic digestion activity

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