Two-methods approach to follow up biomass by impedance spectroscopy: Bacillus thuringiensis fermentations as a study model.

Impedance spectroscopy is used for the characterization of electrochemical systems as well as for the monitoring of bioprocesses. However, the data obtained using this technique allow multiple interpretations, depending on the methodology implemented. Hence, it is necessary to establish a robust methodology to reliably follow-up biomass in fermentations. In the present work, two methodological approaches, mainly used for the characterization of electrochemical systems, were employed to characterize and determine a frequency that allows the monitoring of biomass in Bacillus thuringiensis fermentations by impedance spectroscopy. The first approach, based on a conventional analysis, revealed a single distribution with a characteristic frequency of around 2 kHz. In contrast, the second approach, based on the distribution of relaxation times, gave three distributions (A, B, and C). The C distribution, found near 9 kHz, was more related to the microbial biomass than the distribution at 2 kHz using the equivalent circuits. The time course of the B. thuringiensis fermentation was followed; bacilli, spores, glucose, and acid and base consumption for pH were determined out of line; and capacitance at 9 kHz was monitored. The correlation between the time course data and the capacitance profile indicated that the monitoring of B. thuringiensis at 9 kHz mainly corresponds to extracellular activity and, in a second instance, to the cellular concentration. These results show that it is necessary to establish a robust and reliable methodology to monitor fermentation processes by impedance spectroscopy, and the distribution of relaxation times was more appropriate. KEY POINTS: • Application of impedance spectroscopy for bioprocess monitoring • Low-frequency monitoring of biomass in fermentations • Analysis of impedance data by two methodological approaches.

Synergistic Assembly of 1DZnO and Anti-CYFRA 21-1: A Physicochemical Approach to Optical Biosensing.

Objective: We conducted a comprehensive physicochemical analysis of one-dimensional ZnO nanowires (1DZnO), incorporating anti-CYFRA 21-1 immobilization to promote fast optical biomarker detection up to 10 ng ml-1. Impact Statement: This study highlights the effectiveness of proof-of-concept 1DZnO nanoplatforms for rapid cancer biomarker detection by examining the nanoscale integration of 1DZnO with these bioreceptors to deliver reliable photoluminescent output signals. Introduction: The urgent need for swift and accurate prognoses in healthcare settings drives the rise of sensitive biosensing nanoplatforms for cancer detection, which has benefited from biomarker identification. CYFRA 21-1 is a reliable target for the early prediction of cancer formation that can be perceptible in blood, saliva, and serum. However, 1DZnO nanostructures have been barely applied for CYFRA 21-1 detection. Methods: We assessed the nanoscale interaction between 1DZnO and anti-CYFRA 21-1 antibodies to develop rapid CYFRA 21-1 detection in two distinct matrices: PhosphateBuffered Saline (PBS) buffer and artificial saliva. The chemical modifications were tracked utilizing Fourier transform infrared spectroscopy, while transmission electron microscopy and energy dispersive spectroscopy confirmed antigen-antibody interplay over nanostructures. Results: Our results show high antibody immobilization efficiencies, affirming the effectiveness of 1DZnO nanoplatforms for rapid CYFRA 21-1 testing within a 5-min detection window in both PBS and artificial saliva. Photoluminescence measurements also revealed distinct optical responses across biomarker concentrations ranging from 10 to 1,000 ng ml-1. Conclusion: Discernible PL signal responses obtained after 5 min affirm the potential of 1DZnO nanoplatforms for further advancement in optical biomarker detection for application in early cancer prognosis.

On-line monitoring of industrial interest Bacillus fermentations, using impedance spectroscopy.

Impedance spectroscopy is a technique used to characterize electrochemical systems, increasing its applicability as well to monitor cell cultures. During their growth, Bacillus species have different phases which involve the production and consumption of different metabolites, culminating in the cell differentiation process that allows the generation of bacterial spores. In order to use impedance spectroscopy as a tool to monitor industrial interest Bacillus cultures, we conducted batch fermentations of Bacillus species such as B. subtilis, B. amyloliquefaciens, and B. licheniformis coupled with this technique. Each fermentation was characterized by the scanning of 50 frequencies between 0.5 and 5 MHz every 30 min. Pearson's correlation between impedance and phase angle profiles (obtained from each frequency scanned) with the kinetic profiles of each strain allowed the selection of fixed frequencies of 0.5, 1.143, and 1.878 MHz to follow-up of the fermentations of B. subtilis, B. amyloliquefaciens and B. licheniformis, respectively. Dielectric profiles of impedance, phase angle, reactance, and resistance obtained at the fixed frequency showed consistent changes with exponential, transition, and spore release phases.