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The hydromechanical stress is a relevant parameter for mammalian cell cultivations, especially regarding scale-up processes. It describes the mechanical forces exerted on cells in a bioreactor. The maximum local energy dissipation rate is a suitable parameter to characterize hydromechanical stress. In literature, different studies deal with the effects of hydromechanical stress on CHO cells in stirred tank reactors. However, they often focus on lethal effects. Furthermore, systematic examinations in smaller scales like shake flasks are missing. Thus, this study systematically considers the influence of hydromechanical stress on CHO DP12 cells in shake flask cultivations. By utilizing online monitoring of the oxygen transfer rate, the study simplifies and enhances the resolution of examinations. Results indicate that while lethal effects are absent, numerous sub-lethal effects emerge with increasing hydromechanical stress: The process time is prolonged. The time of glucose and glutamine depletion, and the lactate switch correlate positively linear with the logarithmic average energy dissipation rate while the maximum specific growth rate correlates negatively. Strikingly, the final antibody concentration only declines at the highest tested average energy dissipation rate of 3.84 W kg -1 (only tested condition with a turbulent flow regime and therefore a higher maximal local energy dissipation rate) from about 250 mg L -1 to about 180 mg L -1 . This study presents a straightforward method to examine the impact of hydromechanical stress in shake flasks, easily applicable to any other suspension cell line. Additionally, it offers valuable insights for scale-up processes, for example into stirred tank reactors., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This work presents an experimental evaluation of a Venturi and Venturi-Vortex microbubble aeration system, taking as input variables the water-air flow ratio, water renewal time and area-volume ratio of the water tank. The aeration process response variables are defined in terms of oxygen transfer and aeration efficiency through the standard volumetric mass transfer coefficient (KLa20), standard oxygen transfer rate (SOTR), and standard aeration efficiency (SAE). Two methods of air injection were analyzed: 1. Air injection in the throat chamber of the Venturi generator; 2. air supplying in the suction side of the hydraulic pump of the aeration system. Experimental results indicate that the water renewal time variable (RT) is a statistically significant factor with respect to the KLa20, which can be maximized by decreasing RT. The effects of the variable flow ratio (FR) are greater than the effects of renewal time and area-volume ratio (AVR) concerning SOTR and SAE, indicating a maximum response with a minimum flow ratio, using the Venturi-Vortex microbubble generator. When the flow ratio decreases, the air flow increases, generating and transferring a greater amount of microbubbles (MB) into the water. It was found that increasing the air flow produced an increase in the standard oxygen transfer rate SOTR and standard aeration efficiency SAE. Results allow concluding that the injection of the air flow from the suction side of the pump promotes the generation of microbubbles (MB) for a maximum air flow allowed by the system. SOTR and SAE could be maximized whit the flow ratio factor and the Venturi-Vortex generator, supplying air flow from the suction side of the hydraulic pump.

Background: Alginates are polysaccharides used in a wide range of industrial applications, with their functional properties depending on their molecular weight. In this study, alginate production and the expression of genes involved in polymerization and depolymerization in batch cultures of Azotobacter vinelandii were evaluated under controlled and noncontrolled oxygen transfer rate (OTR) conditions. Results: Using an oxygen transfer rate (OTR) control system, a constant OTR (20.3 ± 1.3 mmol L−1h−1) was maintained during cell growth and stationary phases. In cultures subjected to a controlled OTR, alginate concentrations were higher (5.5 ± 0.2 g L−1) than in cultures under noncontrolled OTR. The molecular weight of alginate decreased from 475 to 325 kDa at the beginning of the growth phase and remained constant until the end of the cultivation period. The expression level of alyA1, which encodes an alginate lyase, was more affected by OTR control than those of other genes involved in alginate biosynthesis. The decrease in alginate molecular weight can be explained by a higher relative expression level of alyA1 under the controlled OTR condition. Conclusions: This report describes the first time that alginate production and alginate lyase (alyA1) expression levels have been evaluated in A. vinelandii cultures subjected to a controlled OTR. The results show that automatic control of OTR may be a suitable strategy for improving alginate production while maintaining a constant molecular weight.How to cite: Ponce B, Urtuvia V, Maturana N, et al. Increases in alginate production and transcription levels of alginate lyase (alyA1) by control of the oxygen transfer rate in Azotobacter vinelandii cultures under diazotrophic conditions. Electron J Biotechnol 2021;51. https://doi.org/10.1016/j.ejbt.2021.04.007

The impact of guar gum (GG), crude algae ethanolic extract (CAEE), and turmeric essential oil (TEO) incorporated edible coating formulations on the quality of cut potatoes was investigated at room temperature (27 ± 3 °C, 70-85 % RH) storage using a rotatable central composite design. Besides, 30 % glycerol, 5 % calcium chloride, and 3 % ascorbic acid (w/w) were added to the coating solution as additives. The surface color, respiration rate, water vapor transmission rate, visible mold growth, and sensory analysis were assessed after seven days of storage. The inclusion of ascorbic acid and TEO in edible coating demonstrated a more effective delay in browning. The coated potatoes had lower OTR, CTR, and WVTR values for GG concentrations of 0.5 to 1 g/100 mL than the control. Compared to additives, higher concentrations of GG improved response parameters. The WVTR value of coated potatoes was significantly impacted by the interaction between CAEE and TEO with GG. Incorporating CAEE and TEO into the formulations of guar gum led to a reduction in the permeability of the coating to oxygen and water vapor. The seven days of extended shelf life compared to two days of control were observed with the optimized coating formulation. Furthermore, the application of the coating treatment proved effective in preventing enzymatic browning and creating a barrier against moisture and gases, contributing to prolonged freshness during extended storage periods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

A fast manometric method has been designed for determining the oxygen transfer rate through cork stoppers. Thanks to this technique, it is possible to measure the effective diffusion coefficient of oxygen through a thin cork wafer in a few days, whereas months are required in the case of a full-length cork stopper. A calculation method has also been developed and validated to extrapolate the effective oxygen diffusion coefficient measured on thin wafers to a full-length stopper, based on the statistical analysis of the experimental data distribution. The question of the material heterogeneity and the representative thickness is addressed as well as the effective length of the stopper beyond which the gain in resistance to mass transfer becomes non-significant.

The Ames test is one of the most applied tools in mutagenicity testing of chemicals ever since its introduction by Ames et al. in the 1970s. Its principle is based on histidine auxotrophic bacteria that regain prototrophy through reverse mutations. In the presence of a mutagen, more reverse mutations occur that become visible as increased bacterial growth on medium without histidine. Many miniaturized formats of the Ames test have emerged to enable the testing of environmental water samples, increase experimental throughput, and lower the required amounts of test substances. However, most of these formats still rely on endpoint determinations. In contrast, the recently introduced Ames RAMOS test determines mutagenicity through online monitoring of the oxygen transfer rate. In this study, the oxygen transfer rate of Salmonella typhimurium TA100 during the Ames plate incorporation test was monitored and compared to the Ames RAMOS test to prove its validity further. Furthermore, the Ames RAMOS test in 96-well scale is newly introduced. For both the Ames plate incorporation and the Ames RAMOS test, the influence of the inoculum cell count on the negative control was highlighted: A lower inoculum cell count led to a higher coefficient of variation. However, a lower inoculum cell count also led to a higher separation efficiency in the Ames RAMOS test and, thus, to better detection of a mutagenic substance at lower concentrations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

This short communication demonstrates for the first time a solely microbial activity driven oxygen influx across a microporous hollow fibre membrane via tracking changes in volume and gas composition of entrapped air supply. A U-shape manometer was used to directly reflect gas influx due to microbial activities. A pressure difference of several hundred pascal was created to draw oxygen while 25 mg-N/L of ammonium was oxidized into nitrite by active biofilm at a hydraulic retention time of 6 h. Calibrated and normalized gas compositions before and after the experiment were processed to unveil the gas exchange and estimate the actual oxygen influx across the membrane. A solely microbial activity driven oxygen influx of 10.7 mg O 2 /m 2 /h was observed. Measuring oxygen transfer from supply side provides a more straight-forward perspective on the role of active biofilm in membrane aerated biofilm reactor. The capability of the microbial activity to uptake oxygen on its own could potentially lead to greater energy savings in some MABR applications when strict aeration control is not needed., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The Ames test is the most commonly used mutagenicity test worldwide. It is based on a microbial system that uses histidine auxotrophic Salmonella typhimurium strains. Due to either spontaneous mutations or mutations induced by a mutagenic compound, the cells can regain their ability to grow without histidine supplementation. The degree of mutagenicity of a sample correlates with the number of cells that are able to grow in media that lack histidine. All test variants published up to now are endpoint determinations providing no information about cell growth and respiration activity during the cultivation time. This study aimed to develop an alternative type of Ames test by characterizing the respiration activity of Salmonella typhimurium over time for dynamic mutagenicity detection. It focuses on elucidating the mechanisms underlying this novel test system, and serves as a general proof of principle. Respiration activity (oxygen transfer and uptake rate) and biomass growth of Salmonella typhimurium TA 100 and TA 98 were mechanistically modeled to understand and predict the behavior of the bacteria during the Ames test. The results simulated by the model were experimentally validated by the online monitoring of respiration activity over cultivation time using a Respiration Activity MOnitoring System (RAMOS). The simulated prediction was observed to fit well to the experimental data. When a mutagenic compound was added, its mutagenicity could be detected online due to the elevated cell number and respiration of histidine prototrophic cells. Laborious manual evaluation of mutagenicity after cultivation is not necessary. Mutagenicity evaluation with the presented alternative Ames RAMOS test fitted well to results from an Ames fluctuation test. In the future, a miniaturized RAMOS device for microtiter plates should allow for a high-throughput Ames RAMOS test., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Ames test is one of the most widely used mutagenicity tests. It employs histidine auxotrophic bacteria, which can mutate back to histidine prototrophy and, thus, grow on a histidine deficient medium. These mutants develop predominantly after adding a mutagenic compound during an initial growth phase on 1 mg/L histidine. In the established test systems, an endpoint determination is performed to determine the relative number of mutants. An alternative Ames test, the Ames RAMOS test, has been developed, which enables the online detection of mutagenicity by monitoring respiration activity. The reproducibility of the newly developed test system was investigated. A strong dependence of the test results on the inoculum volume transferred from the preculture was found. The more inoculum was needed to reach the required initial OD, the more mutagenic a positive control was evaluated. This effect was attributed to the histidine transfer from the preculture to the original Ames RAMOS test. The same problem is evident in the Ames fluctuation test. High reproducibility of the Ames RAMOS test could be achieved by performing the preculture on minimal medium with a defined histidine concentration and termination after histidine depletion. By using 5 mg/L initial histidine within the minimal medium, a higher separation efficiency between negative control and mutagenic samples could be achieved. This separation efficiency could be further increased by lowering the cultivation temperature from 37 to 30 °C, i.e. lowering the maximum growth rate. The optimized Ames RAMOS test was then transferred into a 48-well microtiter plate format (μRAMOS) for obtaining a high throughput test. The online detection of mutagenicity leads to a reduction of working time in the laboratory. Due to the optimization of reproducibility and the increase in separation efficiency, a sound mutagenicity evaluation, even of weak mutagenic compounds, can be achieved., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The oxygen transfer rate (OTR) has a significant impact on the design and operation of vertical flow constructed wetlands (VFCWs) intended for organic matter removal and nitrification. Despite its key role, the information on real oxygen input in VFCWs is limited, being usually estimated by mass balance (stoichiometry), through which is calculated only the oxygen consumption rate (OCR). In this study, for the first time, the gas tracer method was applied to evaluate the oxygen transfer capacity of a real-scale VFCW (24.5 m 2 ) applied to the treatment of domestic wastewater. Propane was used as tracer. The OCR and the OTR were evaluated in VFCW under hydraulic loading rates (HLR) of 60, 90, and 120 mm d -1 corresponding to recirculation rations of 0%, 50%, and 100%. The OTR in standard conditions (20 °C) ranged from 120 to 176 g O 2  m -2  d -1 . The highest OTR was found for the lowest HLR. For the operating conditions tested, the OTR obtained with gas tracer were higher than the OCR calculated by stoichiometry in VFCW, which ranged from 20.6 to 27.8 g O 2  m -2  d -1 . Besides, the OTR were sufficient to satisfy the VFCW oxygen demand for organic matter removal and nitrification. These results show that the gas tracer method for OTR determination may allow advances on the understanding of treatment processes and on the design of new VFCWs since its treatment performance requires aerobic conditions., (Copyright © 2019. Published by Elsevier B.V.)

This study compared the performance of two different gas-permeable membranes, PVDF and PP membrane, in a membrane-aerated biofilm reactor (MABR). The surface characteristics and membrane pore blocking of these two membrane materials were studied utilizing AFM, SEM and CLSM. The PVDF membrane surface was more rough and hydrophilic, and possessed a better microbial affinity compared to PP. Furthermore, the MABR equipped with a PVDF membrane removed more COD (97.06 ± 0.97%) and TN (85.66 ± 0.87%) compared to the MABR with PP membrane (87.13 ± 0.87% and 71.13 ± 0.71%, respectively). As well, the PP membrane exhibited severe membrane pore blocking and had a lower oxygen transfer rate than the PVDF membrane. It is concluded that the PVDF membrane has potential as an aeration membrane material for MABRs., (Copyright © 2018 Elsevier B.V. All rights reserved.)