Determination of the efficacy of ultrasound combined with essential oils on the decontamination of Salmonella inoculated lettuce leaves

Salmonella is one of main pathogenic bacteria present in fresh produce. Ultrasound has been reported to be effective at inactivating food-borne pathogens. Moreover, ultrasound can be combined with essential oils to enhance its efficacy. This study evaluates the reduction and inactivation of Salmonella enterica Abony inoculated on lettuce leaves by the application of continuous and pulsed ultrasound as well as ultrasound combined with the essential oil of oregano and thyme. The physicochemical properties of these essential oil nanoemulsions are characterised while the structural damage of treated leaves is determined by the electrolyte leakage. Ultrasound combined with essential oils enhanced the microbial reduction on lettuce leaves and inactivation on the treated water, resulting on significant differences at concentrations higher than 0.018% (v/v) compared to control. Particle size, zeta potential and pH varied between 35 and 133 nm, −26 to −36 mV and 5.67 to 5.38, respectively. Electrolyte leakage was similar for both the control and the treated samples, increasing when essential oils were applied.peer-reviewe

Bioproduction of the Recombinant Sweet Protein Thaumatin: Current State of the Art and Perspectives

There is currently a worldwide trend to reduce sugar consumption. This trend is mostly met by the use of artificial non-nutritive sweeteners. However, these sweeteners have also been proven to have adverse health effects such as dizziness, headaches, gastrointestinal issues, and mood changes for aspartame. One of the solutions lies in the commercialization of sweet proteins, which are not associated with adverse health effects. Of these proteins, thaumatin is one of the most studied and most promising alternatives for sugars and artificial sweeteners. Since the natural production of these proteins is often too expensive, biochemical production methods are currently under investigation. With these methods, recombinant DNA technology is used for the production of sweet proteins in a host organism. The most promising host known today is the methylotrophic yeast, Pichia pastoris. This yeast has a tightly regulated methanol-induced promotor, allowing a good control over the recombinant protein production. Great efforts have been undertaken for improving the yields and purities of thaumatin productions, but a further optimization is still desired. This review focuses on (i) the motivation for using and producing sweet proteins, (ii) the properties and history of thaumatin, (iii) the production of recombinant sweet proteins, and (iv) future possibilities for process optimization based on a systems biology approach

Heat adaptation of Escherichia coli K12: Effect of acid and glucose

AbstractThe objective of this work is to investigate the effect of the (possible) acid adaptation during growth in a glucose rich environment on the heat resistance of Escherichia coli K12 MG1655. E. coli cells were grown in TSB and/or TSB dextrose free broth until they reached the stationary phase. Afterwards, the stationary phase cells were added in TSB and/or TSB dextrose free broth and inactivation took place at 54oC and 58oC. It was observed that growth in a glucose rich environment leads to an increased heat resistance, most likely due to a certain level of acid and further heat adaptation via cross protection

Artificial neural networks as a tool for incorporating microbial stress adaptations in the quantification of microbial inactivation

Quantifying microbial adapted responses due to thermal stresses by an accurate methodology is imperative for assessing the efficacy of a heat process. Two different artificial neural network (ANN) models are constructed for studying the increased induction of the heat resistance of Escherichia coli K12 under a treatment of decreasing heating rates. In the first model structure there are two input vectors, namely, time t and temperature rate dT=dt, whereas in the second case is also added a third one, namely, the microbial load delayed with one time unit Nk¡1. For both models a minimal fully-connected feedforward architecture is used consisting of one hidden neuron and one output neuron. Results as based on the prediction capability of the model structures demonstrate the comparative advantage when an ANN architecture with a delay in its inputs is employed. Incorporation of past events seems to be an essential input for taking into account the observed induced microbial heat resistance.peer-reviewe

The role of temperature and carbon dioxide climatic stress factors on the growth kinetics of Escherichia coli

The global level of carbon dioxide and temperature in the atmosphere is expected to increase, which may affect the survival of the stress-adapted bacteria. In this study, the effect of temperature and dissolved carbon dioxide on the growth rate of Escherichia coli was studied, thus assessing its response to induced environmental stress factors. Methods and results A kinetic assay has been performed using a microplate reader with a spectrofluorometer to determine the specific growth rates. Polynomial models were developed to correlate the environmental conditions of temperature and carbon dioxide with E. coli BL21 (DE3) growth in culture media and dairy by-products. At a temperature of 42 °C, as the dissolved CO2 increased, a decrease of the μmax by 0.76 h−1 was observed. In contrast, at 27 °C, this increase led to a rise of the μmax by 0.99 h−1. Moreover, a correction factor was added when applying the model to dairy whey samples. Conclusions The application of this developed model can be considered a useful tool for predicting the growth of E. coli using climate projections.peer-reviewe

Towards Online Model Predictive Control on a Programmable Logic Controller: Practical Considerations

Given the growing computational power of embedded controllers, the use of model predictive control (MPC) strategies on this type of devices becomes more and more attractive. This paper investigates the use of online MPC, in which at each step, an optimization problem is solved, on both a programmable automation controller (PAC) and a programmable logic controller (PLC). Three different optimization routines to solve the quadratic program were investigated with respect to their applicability on these devices. To this end, an air heating setup was built and selected as a small-scale multi-input single-output system. It turns out that the code generator (CVXGEN) is not suited for the PLC as the required programming language is not available and the programming concept with preallocated memory consumes too much memory. The Hildreth and qpOASES algorithms successfully controlled the setup running on the PLC hardware. Both algorithms perform similarly, although it takes more time to calculate a solution for qpOASES. However, if the problem size increases, it is expected that the high number of required iterations when the constraints are hit will cause the Hildreth algorithm to exceed the necessary time to present a solution. For this small heating problem under test, the Hildreth algorithm is selected as most useful on a PLC

Sublethal Injury Caused to Listeria monocytogenes by Natural Plant Extracts: Case Study on Grape Seed Extract and Garlic Extract

Natural antimicrobials, such as grape seed extract (GSE) and garlic extract (GE), are often used as (a part of) novel food preservation technologies, especially due to their antilisterial effect. However, little is known on the extent of sublethal injury (SI) these extracts cause to Listeria monocytogenes, possibly leading to overestimated efficacies for such novel technologies. The influence of both extracts on the sublethal injury of L. monocytogenes strain LMG23775 was investigated, also using propidium iodide staining to investigate the nature of the injury. Minimum inhibitory concentrations were 500 mg gallic acid equivalents (GAE)/L and 7.5 μg allicin/mL for GSE and GE, respectively. These concentrations slowed down the growth of L. monocytogenes, while only causing a significant amount of SI for GSE. Pure extracts caused inactivation of the cells, with GSE being the most effective. Sublethal injury from pure GSE was mainly caused by membrane damage. In pure GE, a significant amount of SI, not caused by membrane damage, was also present, albeit less pronounced than in pure GSE. In conclusion, both extracts cause a significant amount of sublethal injury to L. monocytogenes, which is not taken into account in relevant studies investigating novel food preservation applications involving natural plant extracts

Recent trends in non-invasive in situ techniques to monitor bacterial colonies in solid (model) food

Planktonic cells typically found in liquid systems, are routinely used for building predictive models or assessing the efficacy of food preserving technologies. However, freely suspended cells often show different susceptibility to environmental hurdles than colony cells in solid matrices. Limited oxygen, water and nutrient availability, metabolite accumulation and physical constraints due to cell immobilization in the matrix, are main factors affecting cell growth. Moreover, intra- and inter-colony interactions, as a consequence of the initial microbial load in solid systems, may affect microbial physiology. Predictive food microbiology approaches are moving towards a more realistic resemblance to food products, performing studies in structured solid systems instead of liquids. Since structured systems promote microbial cells to become immobilized and grow as colonies, it is essential to study the colony behaviour, not only for food safety assurance systems, but also for understanding cell physiology and optimizing food production processes in solid matrices. Traditionally, microbial dynamics in solid systems have been assessed with a macroscopic approach by applying invasive analytical techniques; for instance, viable plate counting, which yield information about overall population. In the last years, this approach is being substituted by more mechanistically-inspired ones at mesoscopic (colony) and microscopic (cell) levels. Therefore, non-invasive and in-situ monitoring is mandatory for a deeper insight into bacterial colony dynamics. Several methodologies that enable high-throughput data collection have been developed, such as microscopy-based techniques coupled with image analysis and OD–based measurements in microplate readers. This research paper provides an overview of non-invasive in-situ techniques to monitor bacterial colonies in solid (model) food and emphasizes their advantages and inconveniences in terms of accuracy, performance and output informatio

Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris

The sweet protein thaumatin is emerging as a promising sugar replacer in the market today, especially in the food and beverage sector. Rising demand for its production necessitates the large-scale extraction of this protein from its natural plant source, which can be limited in terms of raw material availability and production costs. Using a recombinant production technique via a yeast platform, specifically, Pichia pastoris, is more promising to achieve the product economically while maintaining batch-to-batch consistency. However, the bioproduction of recombinant proteins requires the identification of optimal process variables, constituting the maximal yield of the product of interest. These variables have a direct effect on the growth of the host organism and the secretion levels of the recombinant protein. In this study, two important environmental factors, pH, and temperature were assessed by cultivating P. pastoris in shake flasks to understand their influence on growth and the production levels of thaumatin II protein. The results from the pH study indicate that P. pastoris attained a higher viable cell density and secretion of protein at pH 6.0 compared to 5.0 when grown at 30 °C. Furthermore, within the three levels of temperatures investigated when grown at pH 6.0, the protein levels were the highest at 30 °C compared to 20 and 25 °C, whereas 25 °C exhibited the highest viable cell density. Interestingly, the trend observed from the qualitative effects of temperature and pH occurred in all the media that was investigated. These results broaden our understanding of how pH and temperature adjustment during P. pastoris cultivation aid in enhancing the production yields of thaumatin II prior to optimising the fed batch bioreactor operation