Fast hexavalent chromium (Cr(VI)) determination is important for environmental risk and health-related considerations. We used a microbial fuel cell-based biosensor inoculated with a facultatively anaerobic, Cr(VI)-reducing, and exoelectrogenic Ochrobactrum anthropi YC152 to determine the Cr(VI) concentration in water. The results indicated that O. anthropi YC152 exhibited high adaptability to pH, temperature, salinity, and water quality under anaerobic conditions. The stable performance of the microbial fuel cell (MFC)-based biosensor indicated its potential as a reliable biosensor system. The MFC voltage decreased as the Cr(VI) concentration in the MFC increased. Two satisfactory linear relationships were observed between the Cr(VI) concentration and voltage output for various Cr(VI) concentration ranges (0.0125–0.3 mg/L and 0.3–5 mg/L). The MFC biosensor is a simple device that can accurately measure Cr(VI) concentrations in drinking water, groundwater, and electroplating wastewater in 45 min with low deviations (<10%). The use of the biosensor can help in preventing the violation of effluent regulations and the maximum allowable concentration of Cr(VI) in water. Thus, the developed MFC biosensor has potential as an early warning detection device for Cr(VI) determination even if O. anthropi YC152 is a possible opportunistic pathogen.
The extensive use of Cr(VI) in many industries and the disposal of Cr(VI)-containing wastes have resulted in Cr(VI)-induced environmental contamination. Cr(VI) compounds are associated with increased cancer risks; hence, the detection of toxic Cr(VI) compounds is crucial. Various methods have been developed for Cr(VI) measurement, but they are often conducted offsite and cannot provide real-time toxicity monitoring. A microbial fuel cell (MFC) is an eco-friendly and self-sustaining device that has great potential as a biosensor for in situ Cr(VI) measurement, especially for wastewater generated from different electroplating units. In this study, Exiguobacterium aestuarii YC211, a facultatively anaerobic, Cr(VI)-reducing, salt-tolerant, and exoelectrogenic bacterium, was isolated and inoculated into an MFC to evaluate its feasibility as a Cr(VI) biosensor. The Cr(VI) removal efficiency of E. aestuarii YC211 was not affected by the surrounding environment (pH 5–9, 20–35 °C, coexisting ions, and salinity of 0–15 g/L). The maximum power density of the MFC biosensor was 98.3 ± 1.5 mW/m2 at 1500 Ω. A good linear relationship (r2 = 0.997) was observed between the Cr(VI) concentration (2.5–60 mg/L) and the voltage output. The developed MFC biosensor is a simple device that can accurately measure Cr(VI) concentrations in the actual electroplating wastewater that is generated from different electroplating units within 30 min with low deviations (−6.1% to 2.2%). After treating the actual electroplating wastewater with the MFC, the predominant family in the biofilm was found to be Bacillaceae (95.3%) and was further identified as the originally inoculated E. aestuarii YC211 by next generation sequencing (NGS). Thus, the MFC biosensor can measure Cr(VI) concentrations in situ in the effluents from different electroplating units, and it can potentially help in preventing the violation of effluent regulations.
The conventional Biochemical Oxygen Demand (BOD) method takes five days to analyze samples. A microbial fuel cell (MFC) may be an alternate tool for rapid BOD determination in water. However, a MFC biosensor for continuous BOD measurements of water samples is still unavailable. In this study, a MFC biosensor inoculated with known mixed cultures was used to determine the BOD concentration. Effects of important parameters on establishing a calibration curve between the BOD concentration and output signal from the MFC were evaluated. The results indicate monosaccharides were good fuel, and methionine, phenylalanine, and ethanol were poor fuels for electricity generation by the MFC. Ions in the influent did not significantly affect the MFC performance. CN− in the influent could alleviate the effect of antagonistic electron acceptors on the MFC performance. The regression equation for BOD concentration and current density of the biosensor was y = 0.0145x + 0.3317. It was adopted to measure accurately and continuously the BOD concentration in actual water samples at an acceptable error margin. These results clearly show the developed MFC biosensor has great potential as an alternative BOD sensing device for online measurements of wastewater BOD.
Pomelo (Citrus grandis), an important fruit crop grown in tropical and subtropical areas, is cultivated mainly in Asian countries. The dominant pigment in pomelo leaves, chlorophyll, has been reported to possess many biological activities such as antioxidant, anti-inflammation and anticancer. The objectives of this study were to determine chlorophylls in Pomelo leaves by high-performance liquid chromatography-mass spectrometry (HPLC-MS) and to encapsulate the isolated chlorophylls from preparative column chromatography into a nanoemulsion system for elucidating the inhibition mechanism on the growth of melanoma cells A375. The results showed that chlorophyll a and chlorophyll b could be separated within 25 min by using a C18 column and a gradient ternary mobile phase of acetone, acetonitrile and methanol. Pomelo leaves mainly contained chlorophyll a (2278.3 μg/g) and chlorophyll b (785.8 μg/g). A highly stable chlorophyll nanoemulsion was prepared with the mean particle size being 13.2 nm as determined by a dynamic light scattering (DLS) method. The encapsulation efficiency of chlorophyll nanoemulsion was 99%, while the zeta potential was −64.4 mV. In addition, the chlorophyll nanoemulsion possessed high thermal stability up to 100 °C and remained stable over a 90-day storage period at 4 °C. Western blot analysis revealed that chlorophyll nanoemulsion and extract could upregulate p53, p21, cyclin B and cyclin A as well as downregulate CDK1 and CDK2 in a concentration-dependent manner for inhibition of melanoma cells A375. Furthermore, chlorophyll nanoemulsion and extract could upregulate Bax and cytochrome C and downregulate Bcl-2, leading to activation of caspase-9, caspase-8 and caspase-3 for the induction of cell apoptosis. Compared to chlorophyll extract, chlorophyll nanoemulsion was more effective in inhibiting the growth of melanoma cells A375.
This study aims to determine carotenoids in pomelo leaves (Citrus grandis Osbeck), a rich source of nutrients and phytochemicals, by high-performance liquid chromatography-mass spectrometry and prepare carotenoid nanoemulsions for the study of its inhibitory mechanism on melanoma cells A375. Fourteen carotenoids were separated within 27 min by using a YMC-C30 column and a gradient mobile phase of methanol-acetonitrile-water (84:14:2, v/v/v) and methylene chloride with a flow rate of 1 mL/min and detection wavelength of 450 nm. All-trans-lutein plus its cis-isomers were present in the largest amount (3012.97 μg/g), followed by all-trans-neoxanthin (309.2 μg/g), all-trans-violaxanthin (208.5 μg/g), all-trans-β-carotene plus its cis-isomers (203.17 μg/g), all-trans-α-carotene plus its cis-isomers (152.5 μg/g), all-trans-zeaxanthin (54.67 μg/g), and all-trans-β-cryptoxanthin plus its cis-isomers (24.56 μg/g). A stable carotenoid nanoemulsion was prepared with a mean particle size of 13.3 nm, zeta-potential of −66.6 mV, a polydispersity index of 0.132 and an encapsulation efficiency of 99%. Both the carotenoid extract and nanoemulsion could upregulate p53, p21, cyclin B and cyclin A expressions in melanoma A375 cells and downregulate CDK1 and CDK2 in a concentration-dependent manner. Also, they could upregulate Bax and cytochrome-C and downregulate Bcl-2, leading to cell apoptosis through activation of caspase-9, caspase-8 and caspase-3. Compared to extract, carotenoid nanoemulsion was shown to be more effective in inhibiting the growth of melanoma cells A375. This finding further demonstrated that a carotenoid nanoemulsion prepared from pomelo leaves possessed a great potential to be developed into functional foods or even botanic drugs.
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