Detection of thermal sublethal injury in escherichia coli via the selective medium plating technique: Mechanisms and improvements

In food preservation, the synergistic combination of different technologies aims to maximize the total lethality of the process and minimize the intensity of each hurdle. This is especially the case when at least one of the treatments can cause sublethal (reparable) injury in a great proportion of the population, so that sublethally injured cells can end up being entirely inactivated by the other hurdle(s). The selective medium plating technique (SMPT) is extensively used to enumerate bacterial sublethal injury after inimical treatments, being sodium chloride added to the recovery medium to detect damaged bacterial envelopes. However, little work has been done to explain the reasons for the inability of sublethally injured cells to outgrow in selective agar media, whereas they are able to grow in non-selective agar. In the present paper, the performance of SMPT on Escherichia coli cells after heat treatments is explored by applying different selective agents in the recovery media, using mutants lacking factors involved in osmoregulation, and also by examining the integrity of the cytoplasmic membrane. In view of the results, the possibility of a specific toxic effect of Na+ as the main mechanism under SMPT was discarded, since the same level of sublethal injury was detected using KCl instead of NaCl. The synthesis of the osmoprotectant trehalose determined the maximum osmotolerance of intact cells to the selective agents, but was not crucial in the quantification of sublethal injury. Moreover, for the first time, the extent of sublethal injury detected via SMPT was directly correlated with the physical loss of integrity of the cell membrane in 99.999% of the initial population. This was achieved through statistical analysis of flow cytometry data using propidium iodide-exclusion technique when that dye was added before thermal treatments. The present work confirms the adequacy of SMPT as a tool for detecting the occurrence and quantity of sublethally injured cells after thermal treatments and thus, for efficiently designing the combination of heat with other preservation techniques. We also propose the study of statistical analysis from flow cytometry data for a more rapid quantification of bacterial sublethal injury in a broad detection range

E pluribus unum : impact entrepreneurship as a solution to grand challenges

Insufficiency of research and theory on the relationship between entrepreneurship and grand challenges means that we know little about who engages and what repertoires of actions they take to tackle socioenvironmental challenges that transcend firms, markets, and nations, and what sorts of solutions they create. Drawing on the five articles featured in this symposium-and focusing especially on their protagonists or actors, the actions these actors take, and their achievements-we begin to conceptualize an impact entrepreneurship perspective. Following the tenet of e pluribus unum ("out of many, one") and adhering to the doctrine that diverse, decentralized human effort can improve the world, our impact entrepreneurship perspective refers to the development of solutions to grand challenges, in a financially, socially, and environmentally sustainable fashion. All in all, then, this symposium provides a starting point to discuss, conceptualize, study, interpret, and enrich our understanding of impact entrepreneurship and collective action to address grand challenges

La personalidad jurídica civil de las fundaciones canónicas en España

La personalidad jurídica constituye un instrumento necesario para que una determinada entidad, de naturaleza asociativa o patrimonial, pueda actuar en el tráfico jurídico, como sujeto de derechos y obligaciones y sujeto capaz de relaciones jurídicas. El reconocimiento de dicha personalidad jurídica juega un papel decisivo, especialmente, en el ámbito patrimonial, porque confiere a la entidad capacidad para adquirir, administrar y enajenar bienes con plena eficacia jurídica. El presente trabajo analiza los requisitos necesarios para el reconocimiento de la personalidad jurídica civil de las fundaciones erigidas canónicamente por la Iglesia Católica.Legal personality is a necessary instrument for a particular entity, be it associative or proprietary, enabling it to act within the legal system as a subject with rights and obligations and capable of forming legal relationships. The recognition of legal personality plays a decisive role especially in proprietary contexts, because it confers on the entity the capacity to acquire, administer and dispose of goods with full legal efficacy. The present study analyzes the requirements for recognition of the civil legal personality of foundations created canonically by the Catholic Church

Estratigrafía y estructura de los Andes Centrales Argentinos entre los 30º y 31º de Latitud Sur

In a cross-section through the Argentine central Andes, two large groups of rocks can be distinguished: a Gondwanic (F'aleozoic) basement, and an Andean cover. The basement is constituted by manne sedimentary units, intruded by Upper Paleozoic granitoid rocks. The most important Gondwanic structures, are East verging thrust and related folds. The Andean cover has a volcanic and volcanoclastic origin with some interbedded continental sedimentary rocks. The lower Permo-Tnassic unit (Choiyoi Group) is linked to an extensional tectonic event, and a Neogene sequence is connected to a compressional tectonic event that produced the inversion of the previous extensional features. The amount of Andean shortening calculated fiom cross sections is of about 10%. Most of the shortening in the Andean Cordillera was transferred to the Precordillera through the detachment fault. The up-lift of the Pampeanas area is linked with a new lower detachment fault

Antimicrobial activity of suspensions and nanoemulsions of citral in combination with heat or pulsed electric fields

The application of essential oils in form of nanoemulsions has been proposed as a method to improve their solubility in aqueous solutions, and hence their antimicrobial activity. The objective of this study was to evaluate the antimicrobial activity of citral, applied directly or in combined treatments with heat or pulsed electric fields (PEF), as a function of the inoculation procedure assayed: (i) a simple, vigorous shaking method by vortex agitation (suspension of citral; s-citral) or (ii) the previous preparation of nanoemulsions by the emulsion phase inversion (EPI) method (nanoemulsion of citral; n-citral). n-Citral was more effective in either inhibiting or inactivating Escherichia coli O157:H7 Sakai than s-citral. However, when combined with heat, a greater synergistic effect was observed with s-citral rather than with n-citral, either in lab media (pH 7·0 and 4·0) or apple juice. For instance, while almost 5 log10 cell cycles were inactivated in apple juice after 15 min at 53°C in the presence of 0·1 µl ml-1 of s-citral, the use of n-citral required 30 min. The use of nanoemulsions did not modify the slight synergism observed when citral and mild PEF were combined (150 µs, 30 kV cm-1). Significance and Impact of the Study: The exploration of different delivery systems of antimicrobial compounds such as citral in aqueous food products aids in the establishment of successful combined treatments for food preservation. While at room temperature, citral in form of a nanoemulsion shows a higher antimicrobial activity; its combination with heat would imply a partial loss of the outstanding synergistic lethal effect achieved when added in suspension form. Therefore, the most suitable procedure to magnify the synergism between heat and citral when processing juices would merely require an intense homogenization step prior to the combined treatment

Efficiency Evaluation of Fully Integrated On-board EV Battery Chargers with Nine-Phase Machines

A fully integrated on-board battery charger for future electric vehicles (EVs) has been recently introduced. It re-utilizes all the propulsion components of an EV in charging/vehicle-to-grid (V2G) modes, it does not require any additional components or hardware reconfiguration, and charging/V2G modes are realized with zero electromagnetic torque production. Both fast (three-phase) and slow (single-phase) charging are possible, with unity power factor operation at the grid side. The solution is based on the use of a triple three-phase machine and a nine-phase inverter/rectifier. This paper reports on the results of efficiency evaluation for the said system. Testing is performed using both a nine-phase induction machine and a nine-phase permanent magnet (PM) machine for a range of operating conditions in charging/V2G modes, with both three-phase and single-phase grid connection. Additionally, the impact of converter interleaving on the losses and efficiency is also studied. Losses are separated for different subsystems, thus providing an insight into the importance of optimization of different EV power train components from the efficiency point of view. Promising efficiencies, in the order of 90%, are achieved although none of the system components have been optimized

Mechanism of Bacterial Inactivation by (+)-Limonene and Its Potential Use in Food Preservation Combined Processes

This work explores the bactericidal effect of (+)-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+)-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+)-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+)-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+)-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS) allowed identification of altered E. coli BJ4 structures after (+)-limonene treatments as a function of the treatment pH: ß-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+)-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered ß-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+)-limonene. The study of mechanism of inactivation by (+)-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+)-limonene in food preservation, either acting alone or in combination with lethal heat treatments