Subsequent Storage Research Articles

Insights into the mechanisms of L. salivarius CECT5713 resistance to freeze-dried storage

Ligilactobacillus salivarius is a lactic acid bacterium exhibiting several health benefits. However, it is sensitive to freeze-drying and storage in the dried state, thus limiting its commercial exploitation. Our objective was to identify markers of cell resistance by applying multiscale characterization to L. salivarius CECT5713 cell populations exhibiting different resistance to freeze-dried storage. Cells were produced under two different sets of production conditions differing in the culture parameters (temperature, neutralizing solution, and harvesting time) and the protective formulation composition. The culturability, membrane integrity, and cell biochemical composition assessed by Fourier transform infrared (FTIR) micro-spectroscopy were evaluated after freezing, freeze-drying, and subsequent storage at 37 °C. Membrane properties (fatty acid composition, membrane fluidity, and phospholipid organization), as well as matrix physical properties (glass transition temperature and water activity), were determined. The most resistant cells to freeze-dried storage exhibited the highest cyclic fatty acid content and the most rigid membrane. Freeze-drying and storage induced damage to membrane integrity, proteins, nucleic acids, and constituents of the peptidoglycan cell wall. From the FTIR spectra analysis, we propose the minimization of the variations of the 1058 and 1714 cm−1 vibration bands (that arise mainly from symmetric C–O–C stretching and CO stretching, respectively) induced by the freeze-drying process as a marker of storage stability. We confirmed that a matrix with a glass transition temperature at least 50 °C higher than the storage temperature is crucial for L. salivarius CECT5713 storage stability. In addition, this work explored promising FTIR methods for a better understanding of the protection mechanisms involved.

Growth Kinetics of Listeria monocytogenes and Salmonella enterica on Dehydrated Vegetables during Rehydration and Subsequent Storage.

Dehydrated vegetables have low water activities and do not support the proliferation of pathogenic bacteria. Once rehydrated, vegetables can be incorporated into other foods or held for later use. The aim of this study was to examine the survival and proliferation of Listeria monocytogenes and Salmonella enterica on dehydrated vegetables during rehydration and subsequent storage. Carrots, corn, onion, bell peppers, and potatoes were heat dehydrated, inoculated at 4 log CFU/g, and rehydrated at either 5 or 25 °C for 24 h. Following rehydration, vegetables were stored at 5, 10, or 25 °C for 7 d. Both L. monocytogenes and S. enterica survived on all vegetables under all conditions examined. After 24 h of rehydration at 5 °C, pathogen populations on the vegetables were generally <1.70 log CFU/g, whereas rehydration at 25 °C resulted in populations of 2.28 to 6.25 log CFU/g. The highest growth rates during storage were observed by L. monocytogenes on potatoes and S. enterica on carrots (2.37 ± 0.61 and 1.63 ± 0.18 log CFU/g/d, respectively) at 25 °C when rehydration occurred at 5 °C. Results indicate that pathogen proliferation on the vegetables is both rehydration temperature and matrix dependent and highlight the importance of holding rehydrated vegetables at refrigeration temperatures to hinder pathogen proliferation. Results from this study inform time and temperature controls for the safety of these food products.

Antibiotic Sensitivity of Escherichia Coli Museum Strains After Long-Term Storage in a Lyophilized State at Moderately Low Temperatures

Sensitivity of microbial cultures to antibiotics was determined using disco-diffusion method, broth serial dilution and agar serial dilution. Restored viability and detection of antibiotic resistance in the museum Escherichia coli strains stored for a lasting time in a lyophilized state indicates that freeze-drying in combination with moderately low temperatures for subsequent long-term storage is an eff ective method of maintaining collection microorganism strains in a functionally stable state, including preservation of their antibiotic resistance. It was found that one of the 21 studied strains was resistant to ampicillin, ampicillin-sulbactam, cephalosporins of the I–III generations, two strains were resistant to chloramphenicol, and four strains were insensitive to tetracycline. Detection of the resistance to cephalosporins and protected penicillins in E. coli strain isolated in 1963, long before the discovery and use of these antibiotics in clinical practice, confirms that antibiotic resistance is a phenomenon associated rather with the general mechanisms of adaptation of pathogens to adverse external environmental factors than with the widespread use of antimicrobial drugs.

Качество и безопасность продукции в процессе хранения в системе «вино – упаковка, укупорка»

Poor packaging often affects the quality and safety indicators of wine during storage. The need to preserve and prolong wine stability during long-term storage requires a comprehensive study of processes that occur in wines packaged and capped in various ways.&#x0D; This research featured 56 samples of white and red dry wines, which were bottled in four types of packages and stopper for 18 months. The effect of synthetic and natural stopper methods involved a model system that simulated the main composition of wine in line with the EAEU standards. The contents of free SO2 and dissolved oxygen, as well as the microbiology of natural corks, were studied using standard methods. The analysis of volatile substances relied on gas chromatography, mass spectrometry, and solid phase micro-extraction.&#x0D; The study recorded critical changes in the quality of wine products, characterized by increased oxidation markers and low free sulfur dioxide, which fell below 10 mg/dm3. In polyethylene terephthalate bottles, these changes occurred after 12 months of storage, while it took them 18 months to develop in multilayer bags and combined packaging materials. The components of natural and artificial plugs of microbial and chemical origin entered the model systems after 10 days of contact. The microbiological contamination was twice as high as the permissible level. The migration of microbial and chemical substances into the wine samples spoiled their quality and safety.&#x0D; The effect of packaging on the oxidative processes in white and red dry wines during storage resulted in different concentrations of dissolved oxygen and sulfur dioxide. The results can help wine producers to choose the most appropriate packaging and stopper means depending on the wine sort and its subsequent storage conditions. Alternative types of packaging proved to have a limited shelf life.

High-pressure processing applied to sliced dry-cured Iberian loin: Effect of category, company, and storage temperature.

The main purpose of this study was to evaluate the effect of high-pressure processing (HPP) and storage temperature on the microbial counts, the instrumental color, and the oxidation stability of sliced dry-cured Iberian loin from two categories and two leading companies. 600 MPa for 8 min was sufficient to decrease all the microbial counts without affecting the color and the oxidation status, the effect being modulated by the loin category and company, whose effect on those variables was marked. However, the subsequent 90-day storage softened the initial effect of HPP on microorganisms and allowed a significant effect of HPP to develop on color and oxidation. In addition, the coliform counts were higher after storage at 20°C than at 4°C, suggesting that refrigeration may be needed during long-term storage to ensure loin safety.

Mechanistic calendar aging model for lithium-ion batteries

In this work we present a novel mechanistic calendar aging model for a commercial lithium-ion cell with NCA cathode and silicon-graphite anode. The mechanistic calendar aging model is a semi-empirical aging model that is parameterized on component states of health, instead of capacity. Three component states of health are derived from the degradation modes, which are calculated by fitting the electrode potential curves at every check-up measurement. The aging data used for model parameterization spans 672days of storage at 27 different combinations of ambient temperature (Tamb) and state of charge (SOC). To compensate for the influence of the check-up measurements on cell degradation, the aging data is pre-processed in two steps, considering immediate degradation caused by the check-up cycles and accelerated degradation during subsequent storage. The loss of active anode material is negligible during check-up-compensated calendar aging. For loss of lithium inventory and loss of active cathode material, Tafel and Arrhenius terms are successfully used to model Tamb and SOC dependence. The mechanistic calendar aging model predicts the capacity with <1% mean deviation for 7 different storage conditions after 672days without check-ups. The check-up compensation increases predicted lifetime by >150% for exemplary storage at Tamb=60°C and SOC=50%.

Probing Sodium Storage Mechanism in Hollow Carbon Nanospheres Using Liquid Phase Transmission Electron Microscopy.

Carbonaceous materials are promising sodium-ion battery anodes. Improving their performance requires a detailed understanding of the ion transport in these materials, some important aspects of which are still under debate. In this work, nitrogen-doped porous hollow carbon spheres (N-PHCSs) are employed as a model system for operando analysis of sodium storage behavior in a commercial liquid electrolyte at the nanoscale. By combining the ex situ characterization at different states of charge with operando transmission electron microscopy experiments, it is found that a solvated ionic layer forms on the surface of N-PHCSs at the beginning of sodiation, followed by the irreversible shell expansion due to the solid-electrolyte interphase (SEI) formation and subsequent storage of Na(0) within the porous carbon shell. This shows that binding between Na(0) and C creates a Schottky junction making Na deposition inside the spheres more energetically favorable at low current densities. During sodiation, the SEI fills the gap between N-PHCSs, binding spheres together and facilitating the sodium ions' transport toward the current collector and subsequent plating underneath the electrode. The N-PHCSs layer acts as a protective layer between the electrolyte and the current collector, suppressing the possible growth of dendrites at the anode.

CARD6: Warm Ischemic Time, Cold Static Storage and Impairment of Cardiac Endoplasmic Reticular Stress in Human Hearts Donated After Circulatory Death and Brain Death

Background: Endoplasmic reticulum (ER) plays an important role in maintaining normal myocardial function at the cellular level. Overexpression of ER stress-associated proteins during ischemia impact cardiac function. Initial result of heart transplantation using donation after circulatory death (DCD) heart with short warm ischemic time (WIT, less than 30 minutes) was encouraging. However, little is known about the impact of various WIT on donor heart and with subsequent cold storage as used in normothermic regional perfusion(NRP) on the DCD hearts. In this study we investigated expression of myocardial ER stress response proteins in the DCD hearts to evaluate the impact of warm ischemic time (WIT) and subsequent cold storage. We also studied expression of myocardial ER stress response proteins in the brain death donor (DBD) hearts as a control group. Methods: A total of 23 human hearts, DCD group, n=17, with 3 WIT sub-groups [WIT 20 (n=6), 40 (n=6), ≥60 (n=5) minutes] and DBD group, n=6, no warm ischemia (control: WIT:0) were studied. All the DCD hearts received del Nido cardioplegia and all DBD hearts received Celsior solution before they were placed at cold storage for 6 hours. Left ventricular biopsies were performed at hour 0 (T0), 2 (T2), 4 (T4) and 6 (T6). Myocardial ER stress response proteins ATF6 (Activating Transcription Factor 6) and GRP78 (78-kDa glucose-regulated protein) were measured through immunoblotting after normalizing to GAPDH loading control in all cardiac biopsies. Results: Representative western blots showing the protein expressions patterns among the DBD and three different groups of DCD hearts under increasing time points (T0:0hrs, T2:2hrs, T4:4hrs, T6:6hrs) of cold static storage conditions (Fig.1A). At baseline, the proteins expression found to be comparable between DBD, DCD:20 and 40 groups, while DCD:≥60 group remained significantly higher (Fig.1B,C). We noticed differential temporal increase of ER stress proteins after 2 hours of cold storage and became prominent at hour 6 in DCD groups when compared to DBD group; while the DCD:60 group always remained highly significant throughout the cold storage compared to all other groups (Fig.1D,E). Conclusions: Our study concluded that duration of warm ischemia and subsequent cold storage condition in DCD hearts were associated with elevated ER stress response in myocardium. The length of cold storage was also found to be linked with elevated ER stress in DCD hearts. The DCD hearts with more than 60 min warm ischemic has the highest ES stress response suggesting profound alteration in ER homeostasis which may promote cardiac damage. Such findings are important in guiding clinical decision making on selecting DCD donor hearts. On the other hand the DBD hearts with up to 6 hour cold storage exerted minimal ER stress, supporting current DBD heart transplant donor preservation practice.

CARD2: Plasma Homocysteine and Oxidative Stress Markers May Predict the Stress Burden of Human Hearts Donated After Circulatory Death

Background: Initial result of heart transplantation using donation after circulatory death (DCD) heart with warm ischemic time (WIT) less than 30 minutes was encouraging. However, little is known about how donor respond to WIT systemically and its impact on the heart with subsequent cold storage as used in normothermic regional perfusion(NRP) on the DCD hearts. Since elevated homocysteine (Hcy) and oxidative stress burden in cardiovascular system have been associated with ischemia, we investigated if circulating Hcy, oxidative stress and antioxidant were elevated in DCD blood and myocardium and if there is temporal relationship with the WIT and subsequent cold storage. Methods: We studied 12 human DCD hearts for research (WIT range:20-110 min). They were assigned to 3 groups of WIT [20 (n=4), 40 (n=5), ≥60 (n=3) min.]. Two time-point donor blood samples were collected from each donor at the baseline (prior to or at extubation) and at the cross-clamp. All hearts received del Nido cardioplegia before they were placed at cold storage for 6 hours. Left ventricular (LV) biopsies were performed at hour 0,2,4 and 6. Circulating Hcy, oxidative stress (oxidized low-density lipoprotein: OxLDL, malondialdehyde: MDA) and total antioxidant in blood were quantified. The MDA and GPx activity (glutathione peroxidase: antioxidant) were quantified in cardiac biopsies. Results: Change in plasma Hcy (Fig.1A), oxidative stress (Fig.1B,C) and antioxidant (Fig.1D) found to be comparable between the 3 groups at baseline; while significant differences were noticed in samples at cross-clamping; especially in DCD≥60 groups. The WIT found to be positively correlated Hcy and oxidative stress at cross-clamping; and negatively correlated with antioxidant (p<0.05,Table:1). The myocardial oxidative stress (MDA) found to be minimal at baseline (DCD20 and 40), and slightly elevated at 2 hrs. and maintained at similar levels at 4 and 6 hrs. of cold storage (Fig.1E); while a preexisting high oxidative stress at baseline in DCD≥60, an exponential increase during the cold storage indicating severity of stress burden. Temporal decrease in cardiac antioxidant (GPx) found to be more pronounced in DCD≥60 group, while the rates of change are comparable in DCD:20 and 40 groups (Fig.1F). The change of blood markers was found to be correlated with change of myocardial markers(Table:1). Conclusions: Systemic warm ischemia promoted hyperhomocystenemia and increased oxidative stress and might be used as initial screening biomarkers for the stress burden on the donor hearts. The correlation between blood and myocardial markers suggested the systemic markers can potentially be used in predicting stress burden of donor hearts. The duration of cold storage was related to progressive increase in myocardial oxidative stress and diminished antioxidant status. We recommend rejecting DCD hearts with WIT≥60 minutes, as both the blood and tissue data indicating maximum oxidative stress burden in this group.

R-MFNet: Analysis of Urban Carbon Stock Change against the Background of Land-Use Change Based on a Residual Multi-Module Fusion Network

Regional land-use change is the leading cause of ecosystem carbon stock change; it is essential to investigate the response of LUCC to carbon stock to achieve the strategic goal of “double carbon” in a region. This paper proposes a residual network algorithm, the Residual Multi-module Fusion Network (R-MFNet), to address the problems of blurred feature boundary information, low classification accuracy, and high noise, which are often encountered in traditional classification methods. The network algorithm uses an R-ASPP module to expand the receptive field of the feature map to extract sufficient and multi-scale target features; it uses the attention mechanism to assign weights to the multi-scale information of each channel and space. It can fully preserve the remote sensing image features extracted by the convolutional layer through the residual connection. Using this classification network method, the classification of three Landsat-TM/OLI images of Zhengzhou City (the capital of Henan Province) from 2001 to 2020 was realized (the years that the three images were taken are 2001, 2009, and 2020). Compared with SVM, 2D-CNN, and deep residual networks (ResNet), the overall accuracy of the test dataset is increased by 10.07%, 3.96%, and 1.33%, respectively. The classification achieved using this method is closer to the real land surface, and its accuracy is higher than that of the finished product data obtained using the traditional classification method, providing high-precision land-use classification data for the subsequent carbon storage estimation research. Based on the land-use classification data and the carbon density data corrected by meteorological data (temperature and precipitation data), the InVEST model is used to analyze the land-use change and its impact on carbon storage in the region. The results showed that, from 2001 to 2020, the carbon stock in the study area showed a downward trend, with a total decrease of 1.48 × 107 t. Over the course of this 19-year period, the farmland area in Zhengzhou decreased by 1101.72 km2, and the built land area increased sharply by 936.16 km2. The area of land transfer accounted for 29.26% of the total area of Zhengzhou City from 2001 to 2009, and 31.20% from 2009 to 2020. The conversion of farmland to built land is the primary type of land transfer and the most important reason for decreasing carbon stock. The research results can provide support, in the form of scientific data, for land-use management decisions and carbon storage function protections in Zhengzhou and other cities around the world undergoing rapid urbanization.

Microbiological profiling and knowledge of food preservation technology to support guidance on a neutropenic diet for immunocompromised patients.

The current society consists of an increasing number of people vulnerable to infections. For certain people with severe immunodeficiency, a neutropenic or low-microbial diet is being prescribed, which substitutes high-risk foods that are more likely to contain human (opportunistic) pathogens with lower-risk alternatives. These neutropenic dietary guidelines are typically set up from a clinical and nutritional perspective, rather than from a food processing and food preservation perspective. In this study, the current guidelines in use by the Ghent University Hospital were evaluated based on the current knowledge of food processing and preservation technologies and the scientific evidence on microbiological quality, safety, and hygiene of processed foods. Three criteria are identified to be important: (1) the microbial contamination level and composition; (2) the potential presence of established foodborne pathogens such as Salmonella spp. (to which a zero-tolerance policy is recommended); and (3) an increased vigilance for L. monocytogenes as an opportunistic foodborne pathogen with a high mortality rate in immunocompromised individuals (to which a zero-tolerance policy should apply). A combination of these three criteria was used as a framework for the evaluation of the suitability of foodstuffs to be included in a low-microbial diet. Differences in processing technologies, initial contamination of products, etc., however, lead to a high degree of variability in microbial contamination and make it difficult to unambiguously accept or reject a certain type of foodstuff without prior knowledge of the ingredients and the processing and preservation technologies applied during manufacturing and subsequent storage conditions. A restricted screening on a selection of (minimally processed) plant-based foodstuffs on the retail market in Flanders, Belgium supported decision-making on the inclusion of these food types in a low-microbial diet. Still, when determining the suitability of a foodstuff to be included in a low-microbial diet, not only the microbiological status but also nutritional and sensorial properties should be assessed, which requires multidisciplinary communication and collaboration.

Storage, accumulation and deceleration of secondary beams for nuclear astrophysics

Low-energy investigations on rare ion beams are often limited by the available intensity and purity of the ion species in focus. Here, we present the first application of a technique that combines in-flight production at relativistic energies with subsequent secondary beam storage, accumulation and finally deceleration to the energy of interest. Using the FRS and ESR facilities at GSI, this scheme was pioneered to provide a secondary beam of 118Te52+ for the measurement of nuclear proton-capture at energies of 6 and 7 MeV/u. The technique provided stored beam intensities of about 106 ions at high purity and brilliance, representing a major step towards low-energy nuclear physics studies using rare ion beams.

The Fate of the Chlorophyll Derivatives in Olives Preserved and/or Packaged in Presence of Exogenous Copper.

Chlorophyll pigments are thought to be responsible for the highly appreciated green color of unfermented Castelvetrano-style table olives, but no studies have considered the effects of a controlled addition of copper during storage or packaging at the industrial level. For this purpose, chlorophyll derivatives were analyzed in Nocellara cultivar table olives debittered industrially using the Castelvetrano method, via means of HPLC and MS analyses, following the addition of copper in alkaline brines stored at 4 °C for 3 months in 220 L barrels, and during the subsequent storage in acid brines in commercial 400 g packages at 4 °C for up to 18 months. The presence of copper in storage or in packaging brines both contributed significantly to maintaining the green color of the olives, which was associated with a specific pattern of chlorophyll derivatives, as evidenced by principal component analysis. Notably, re-greening was rapidly achievable also for olives that had yellowed for 18 months at a copper concentration below the limit of EU legislation. Finally, by means of PCA, we also demonstrated that a short-term thermic treatment can work as an accelerated predictive tool in determining the fate of chlorophyll derivatives.

Rosmarinus officinalis L. Essential Oils Impact on the Microbiological and Oxidative Stability of Sarshir (Kaymak).

This study investigated the effect of Rosmarinus officinalis L. essential oil, REO (one, two and three percent) on the microbiological and oxidative stability of Sarshir during 20 days of refrigerated storage (4 °C). Initially, the chemical composition (gas chromatography/mass spectrometry, GC/MS), antimicrobial (paper disc diffusion) and antioxidant (DPPH) properties of REO were evaluated. Then, the microbial safety, oxidative stability (peroxide and anisidine values) and overall acceptability of the product after addition of REO to Sarshir and the subsequent storage period were determined. According to GC/MS analysis, the major components of REO were α-pinene (24.6%), 1,8-cineole (14.1%), camphor (13.5%), camphene (8.1%) and limonene (6.1%), respectively. Moreover, it was also found that Limosilactobacillus fermentum (inhibition zone (IZ) of 23.5 mm) and Salmonella Typhi (IZ of 16.4 mm) were the most sensitive and resistant spoilage and pathogenic bacteria against REO, respectively. In addition, the half-maximal inhibitory concentration (IC50) of the REO was measured at 24.8 mg/mL, while the IC50 value of butylated hydroxytoluene (BHT) was 16.6 mg/mL. The highest and lowest bacterial populations were detected in the control and the sample containing 3% REO, respectively. The control had the highest extent of lipid oxidation, while the lowest peroxide and anisidine values were measured in Sarshir containing 3% REO.

Protective effects of enzymatic hydrolysis products of pomelo peel cellulose on Lactobacillus plantarum during freeze-drying process

The successful application of probiotics in food industry depends on the use of good cryoprotectants. This paper was aimed to evaluate protective effects of enzymatic hydrolysis products of different degrees of pomelo peel cellulose (EHPC) on Lactobacillus plantarum during freeze-drying process. The data showed that in contrast to trehalose, the survival rate of L. plantarum was increased obviously under the optimal composite cryoprotectants, which consisted of trehalose and EHPC for 2 h with the mass ratio of 8:2. In addition, the morphology, water-related indices, glass transition temperature (Tg) and cell membrane integrity of lyophilized L. plantarum powder were also evaluated. The results showed that the water content of L. plantarum prepared by the optimal composite cryoprotectants was lower, which was in a reasonable range and conducive to subsequent storage; the Tg and cell membrane integrity were significantly improved. Furthermore, the macromolecular fibers of composite cryoprotectants provided a good habitat and attachment space for L. plantarum, and the sheet fibers with large area acted as a physical barrier to prevent the damage to cells from adverse external environment. The composite cryoprotectants from this study could be considered for industrial development of probiotics.

What the Lactate Shuttle Means for Sports Nutrition

The discovery of the lactate shuttle (LS) mechanism may have two opposite perceptions, It may mean very little, because the body normally and inexorably uses the LS mechanism. On the contrary, one may support the viewpoint that understanding the LS mechanism offers immense opportunities for understanding nutrition and metabolism in general, as well as in a sports nutrition supplementation setting. In fact, regardless of the specific form of the carbohydrate (CHO) nutrient taken, the bodily CHO energy flux is from a hexose sugar glucose or glucose polymer (glycogen and starches) to lactate with subsequent somatic tissue oxidation or storage as liver glycogen. In fact, because oxygen and lactate flow together through the circulation to sites of utilization, the bodily carbon energy flow is essentially the lactate disposal rate. Consequently, one can consume glucose or glucose polymers in various forms (glycogen, maltodextrin, potato, corn starch, and fructose or high-fructose corn syrup), and the intestinal wall, liver, integument, and active and inactive muscles make lactate which is the chief energy fuel for red skeletal muscle, heart, brain, erythrocytes, and kidneys. Therefore, if one wants to hasten the delivery of CHO energy delivery, instead of providing CHO foods, supplementation with lactate nutrient compounds can augment body energy flow.

Effects of Perfluorodecanoic Acid on the Photostability and Self-Recovery of Cesium Lead Bromide Nanocrystals

Photoinduced desorption of surface ligands causes photodegradation of nanophosphors due to enhanced non-radiative relaxation, which is attributed to generated surface defects. We previously reported the self-recovery of photodegraded CsPbBr3 nanocrystals (NCs) and revealed that its mechanism can be attributed to readsorption of the desorbed surface ligands. Thus, using different surface ligands would change the photodegradation and subsequent self-recovery behaviors. Accordingly, in the present work, perfluorodecanoic acid (PFDA) was added to pre-synthesized CsPbBr3 NCs modified with oleic acid and oleylamine to evaluate its effects on photodegradation and subsequent self-recovery. The as-prepared NCs (OA-NCs) and PFDA-modified NCs (PFDA-NCs) were dispersed in nanocomposite ethylene-vinyl acetate resin films to suppress concentration quenching. OA-NCs showed monotonical photoluminescence (PL) decrease under blue excitation light and self-recovery during subsequent dark storage. In contrast, the PFDA-NCs exhibited temporary PL enhancement under excitation irradiation. The photo-activation is attributed to optimization of the adsorption states of the surface ligands in the early stage, while the influence of photodegradation due to ligand desorption becomes dominant later. However, increased PFDA modification prevented self-recovery, indicating that desorbed PFDA may be resistant to readsorption on the NC surface.

Impact of the calcium concentration on the efficacy of phage phiIPLA-RODI, LysRODIΔAmi and nisin for the elimination of Staphylococcus aureus during lab-scale cheese production

Staphylococcus aureus is a Gram-positive human opportunistic pathogen that may also cause food poisoning because of the ability of some strains to produce heat stable enterotoxins that can persist in food even after the pathogen is successfully eliminated. In this context, biopreservation may be a forward-looking strategy to help eliminate staphylococcal contamination in dairy products by using natural compounds. However, these antimicrobials exhibit individual limitations that may be overcome by combining them. This work investigates the combination of a virulent bacteriophage, phiIPLA-RODI, a phage-derived engineered lytic protein, LysRODIΔAmi, and the bacteriocin nisin for the elimination of S. aureus during lab-scale cheese production at two CaCl2 concentrations (0.2 % and 0.02 %), and subsequent storage at two different temperatures (4 °C and 12 °C). In most of the assayed conditions, our results demonstrate that the combined action of the antimicrobials led to a greater reduction of the pathogen population than the compounds individually, albeit this effect was additive and not synergistic. However, our results did show synergy between the three antimicrobials for reducing the bacterial load after 14 days of storage at 12 °C, temperature at which there is growth of the S. aureus population. Additionally, we tested the impact of the calcium concentration on the activity of the combination treatment and observed that higher CaCl2 levels led to a notable increase in endolysin activity that allowed the utilization of approximately 10-times less protein to attain the same efficacy. Overall, our data show that the combination of LysRODIΔAmi with nisin and/or phage phiIPLA-RODI, and an increase in the calcium concentration are successful strategies to decrease the amount of protein required for the control of S. aureus contamination in the dairy sector with a low potential for resistance selection, thereby reducing costs.

Synthesis of Pneumatic Systems in the Control of the Transport Line of Rolling Elements

Abstract This paper presents the synthesis of a pneumatic control system for a selected configuration of the transport path for the delivery of rolling elements to spiral storage in inter-operational transport. The sequential control system sets the state of the manifolds to ensure a flow of workpieces to serve the subsequent storage. The essential module of the control system is the memory block. It is developed based on a storage filling sequence graph. The filling level of the storages can be monitored in one or two points using sensors. The rolling element displacement control sensors work together with appropriately designed systems to execute the delay of the rising and falling edge input signal. By using a two-level control of the filling level of the storages, it is possible to control the emptying status of the storages as a function of the technological time of removal of the items from the storage between the two control points. Control systems were synthesised and verified using Festo’s FluidSim computer programme.

Applying Material Science Principles to Chemical Stability: Modelling Solid State Autoxidation in Mifepristone Containing Different Degrees of Crystal Disorder

Ball-milling and harsh manufacturing processes often generate crystal disorder which have practical implications on the physical and chemical stabilities of solid drugs during subsequent storage, transport, and handling. The impact of the physical state of solid drugs, containing different degrees/levels of crystal disorder, on their autoxidative stability under storage has not been widely investigated. This study investigates the impact of differing degrees of crystal disorder on the autoxidation of Mifepristone (MFP) to develop a predictive (semi-empirical) stability model. Crystalline MFP was subjected to different durations of ambient ball milling, and the resulting disorder/ amorphous content was quantified using a partial least square (PLS) regression model based on Raman spectroscopy data. Samples of MFP milled to generate varying levels of disorder were subjected to a range of (accelerated) stability conditions, and periodically sampled to examine their recrystallization and degradation extents. Crystallinity was monitored by Raman spectroscopy, and the degradation was evaluated by liquid chromatography. The analyses of milled samples demonstrated a competition between recrystallization and degradation via autoxidation of MFP, to different extents depending on stability conditions/exposure time. The degradation kinetics were analyzed by accounting for the preceding amorphous content, and fitted with a diffusion model. An extended Arrhenius equation was used to predict the degradation of stored samples under long-term (25°C/60% RH) and accelerated (40°C/75% RH, 50°C/75% RH) stability conditions. This study highlights the utility of such a predictive stability model for identifying the autoxidative instability in non-crystalline/partially crystalline MFP, owing to the degradation of the amorphous phases. This study is particularly useful for identifying drug-product instability by leveraging the concept of material sciences.