First cross-border outbreak of foodborne botulism in the European Union associated with the consumption of commercial dried roach (Rutilus rutilus).

Botulism outbreaks due to commercial products are extremely rare in the European Union. Here we report on the first international outbreak of foodborne botulism caused by commercial salt-cured, dried roach (Rutilus rutilus). Between November and December 2016, an outbreak of six foodborne botulism type E cases from five unrelated households was documented in Germany and Spain. The outbreak involved persons of Russian and Kazakh backgrounds, all consumed unheated salt-cured, dried roach-a snack particularly favored in Easter-European countries. The implicated food batches had been distributed by an international wholesaler and were recalled from Europe-wide outlets of a supermarket chain and other independent retailers. Of interest, and very unlike to other foodborne disease outbreaks which usually involves a single strain or virus variant, different Clostridium botulinum strains and toxin variants could be identified even from a single patient's sample. Foodborne botulism is a rare but potentially life-threatening disease and almost exclusively involves home-made or artisan products and thus, outbreaks are limited to individual or few cases. As a consequence, international outbreaks are the absolute exception and this is the first one within the European Union. Additional cases were likely prevented by a broad product recall, underscoring the importance of timely public health action. Challenges and difficulties on the diagnostic and epidemiological level encountered in the outbreak are highlighted.

Elaboration of a spontaneous gluten-free sourdough with a mixture of amaranth, buckwheat, and quinoa flours analyzing microbial load, acidity, and pH.

Pseudocereals are gluten-free, nutrient-dense raw materials that are being considered for the production of gluten-free products, especially bread. This study proposes a gluten-free sourdough formula based on equal amounts of amaranth, buckwheat, and quinoa with a dough yield of 250, and an elaboration method to obtain ripe sourdough. Sourdough was characterized in terms of microbiology, pH, and total titratable acidity. The established protocol made it possible to obtain a spontaneous ripe sourdough with lactic acid bacteria populations of 9.60 ± 0.02 log CFU/g and total yeasts and non-Saccharomyces yeast populations (lysine positive) of 7.91 ± 0.15 and 7.52 ± 0.10 log CFU/g, respectively. Great pH stability and total titratable acidity were maintained in the ripe sourdough phase, with values of 4.04 ± 0.02 and 18.39 ± 0.56 ml NaOH 0.1 M/10 g, respectively, at the time of the next refreshment. The use of this sourdough could be an interesting alternative for the production of not only gluten-free bread but also other gluten-free products.

Digital Image Analysis of Yeast Single Cells Growing in Two Different Oxygen Concentrations to Analyze the Population Growth and to Assist Individual-Based Modeling.

Nowadays control of the growth of Saccharomyces to obtain biomass or cellular wall components is crucial for specific industrial applications. The general aim of this contribution is to deal with experimental data obtained from yeast cells and from yeast cultures to attempt the integration of the two levels of information, individual and population, to progress in the control of yeast biotechnological processes by means of the overall analysis of this set of experimental data, and to assist in the improvement of an individual-based model, namely, INDISIM-Saccha. Populations of S. cerevisiae growing in liquid batch culture, in aerobic and microaerophilic conditions, were studied. A set of digital images was taken during the population growth, and a protocol for the treatment and analyses of the images obtained was established. The piecewise linear model of Buchanan was adjusted to the temporal evolutions of the yeast populations to determine the kinetic parameters and changes of growth phases. In parallel, for all the yeast cells analyzed, values of direct morphological parameters, such as area, perimeter, major diameter, minor diameter, and derived ones, such as circularity and elongation, were obtained. Graphical and numerical methods from descriptive statistics were applied to these data to characterize the growth phases and the budding state of the yeast cells in both experimental conditions, and inferential statistical methods were used to compare the diverse groups of data achieved. Oxidative metabolism of yeast in a medium with oxygen available and low initial sugar concentration can be taken into account in order to obtain a greater number of cells or larger cells. Morphological parameters were analyzed statistically to identify which were the most useful for the discrimination of the different states, according to budding and/or growth phase, in aerobic and microaerophilic conditions. The use of the experimental data for subsequent modeling work was then discussed and compared to simulation results generated with INDISIM-Saccha, which allowed us to advance in the development of this yeast model, and illustrated the utility of data at different levels of observation and the needs and logic behind the development of a microbial individual-based model.

Evaluation of the relationship between effervescent paracetamol and blood pressure: clinical trial.

BACKGROUND: Paracetamol's solubility is achieved by adding to the excipient sodium salts, either as bicarbonate, carbonate or citrate. As the relationship between salt and hypertension is well known, due to the sodium content it has raised a hypothesis that may interfere with the control of that risk factor. Therefore, the objective of this study is to evaluate the effect on blood pressure of effervescent paracetamol compared to non-effervescent, in hypertensive patients. METHODS/DESIGN: This is the protocol of a phase IV multicenter clinical trial, randomized, controlled, crossover, open, which will compare the effect of two different formulations of paracetamol (effervescent or non-effervescent) in the blood pressure of hypertensive patients, with a seven weeks follow up. 49 controlled hypertensive patients will be included (clinical BP lower than 150 and 95 mmHg, and lower than 135 mmHg and 85 mmHg in patients with diabetes or a history of cardiovascular event, and daytime ambulatory measurements lower than 140 and 90 mmHg) and mild to moderate pain (Visual Analog Scale between 1 and 4). The study was approved by the ethics committee of the Fundació Jordi Gol i Gurina and following standards of good clinical practice. The primary endpoint will be the variations in systolic BP in 24 h Ambulatory Blood Pressure Monitoring, considering significant differences 2 or more mmHg among those treated with non-effervescent and effervescent formulations. Intention-to-treat and per-protocol analysis will be held. DISCUSSION: Despite the broad recommendation not to use effervescent drugs in patients with hypertension, there are relatively little studies that show exactly this pressor effect due to sodium in salt that gives the effervescence of the product. This is the first clinical trial designed to study the effect of effervescence compared to the non-effervescent, in well-controlled hypertensive patients with mild to moderate pain, performed in routine clinical practice. TRIAL REGISTRATION: NCT 02514538.

Effect of aerobic and microaerophilic culture in the growth dynamics of Saccharomyces cerevisiae and in training of quiescent and non-quiescent subpopulations.

Saccharomyces cerevisiae is industrially the most important yeast, and its growth in different concentrations of oxygen can be used to improve various application processes. The aims of this work were to study in aerobic and microaerophilic growth conditions the cell size and tendency of morphological changes in S. cerevisiae in different stages of growth and to assess the effect of the two growth conditions in the differentiation of quiescent and non-quiescent subpopulations in the stationary phase. Dissolved oxygen levels in the culture medium for aerobic and microaerophilic conditions were 6.6 and 5.2 mg L(-1), respectively. In both growth conditions, similar viable cell populations were obtained, although in aerobic conditions the stationary phase was reached and the quiescent and non-quiescent subpopulations were also differentiated. The microaerophilic growth produced a significant reduction in the specific growth rate and consequently also in glucose and oxygen consumption. The most notable changes in cellular size and morphology occurred with the depletion of glucose and oxygen. The concentration of dissolved oxygen in the culture medium significantly modulated the growth kinetics of S. cerevisiae and their development and differentiation to quiescent cells. This could justify the need to readjust small variations in oxygen levels during yeast cultures in biotechnological processes.

Comparison of the microbial dynamics and biochemistry of laboratory sourdoughs prepared with grape, apple and yogurt.

The microbiological culture-dependent characterization and physicochemical characteristics of laboratory sourdough prepared with grape (GS) were evaluated and compared with apple (AS) and yogurt (YS), which are the usual Spanish sourdough ingredients. Ripe GS took longer than AS and YS to reach the appropriate acidity and achieved lower values of lactic acid. In all sourdoughs, the lactic acid bacteria (LAB) increased during processing and were the dominant microorganisms (>1E+8 CFU/g). GS, as well as AS, had high diversity of LAB species. In ripe YS, Pediococcus pentosaceus was the only species identified; in GS and AS, several Lactobacilli were also found, Lb. plantarum, Lb. brevis, and Lb. sakei; in addition, in GS Weisella cibaria also appeared. Regarding the yeast population, non-Saccharomyces yeasts from GS and AS showed a very high specific population (>1E+7 CFU/g), but this was reduced in ripe sourdough (<1E+4 CFU/g). Finally, the Saccharomyces group dominated in all sourdoughs. Starting ingredients or raw material provided microbiological specificity to sourdoughs, and grape could be considered one of them.

Population analysis of a commercial Saccharomyces cerevisiae wine yeast in a batch culture by electric particle analysis, light diffraction and flow cytometry.

Data from electric particle analysis, light diffraction and flow cytometry analysis provide information on changes in cell morphology. Here, we report analyses of Saccharomyces cerevisiae populations growing in a batch culture using these techniques. The size distributions were determined by electric particle analysis and by light diffraction in order to compare their outcomes. Flow cytometry parameters forward (related to cell size) and side (related to cell granularity) scatter were also determined to complement this information. These distributions of yeast properties were analysed statistically and by a complexity index. The cell size of Saccharomyces at the lag phase was smaller than that at the beginning of the exponential phase, whereas during the stationary phase, the cell size converged with the values observed during the lag phase. These experimental techniques, when used together, allow us to distinguish among and characterize the cell size, cell granularity and the structure of the yeast population through the three growth phases. Flow cytometry patterns are better than light diffraction and electric particle analysis in showing the existence of subpopulations during the different phases, especially during the stationary phase. The use of a complexity index in this context helped to differentiate these phases and confirmed the yeast cell heterogeneity.

Patient-to-patient transmission of hepatitis C virus (HCV) during colonoscopy diagnosis.

BACKGROUND: No recognized risk factors can be identified in 10-40% of hepatitis C virus (HCV)-infected patients suggesting that the modes of transmission involved could be underestimated or unidentified. Invasive diagnostic procedures, such as endoscopy, have been considered as a potential HCV transmission route; although the actual extent of transmission in endoscopy procedures remains controversial. Most reported HCV outbreaks related to nosocomial acquisition have been attributed to unsafe injection practices and use of multi-dose vials. Only a few cases of likely patient-to-patient HCV transmission via a contaminated colonoscope have been reported to date. Nosocomial HCV infection may have important medical and legal implications and, therefore, possible transmission routes should be investigated. In this study, a case of nosocomial transmission of HCV from a common source to two patients who underwent colonoscopy in an endoscopy unit is reported. RESULTS: A retrospective epidemiological search after detection of index cases revealed several potentially infective procedures: sample blood collection, use of a peripheral catheter, anesthesia and colonoscopy procedures. The epidemiological investigation showed breaches in colonoscope reprocessing and deficiencies in the recording of valuable tracing data. Direct sequences from the NS5B region were obtained to determine the extent of the outbreak and cloned sequences from the E1-E2 region were used to establish the relationships among intrapatient viral populations. Phylogenetic analyses of individual sequences from viral populations infecting the three patients involved in the outbreak confirmed the patient pointed out by the epidemiological search as the source of the outbreak. Furthermore, the sequential order in which the patients underwent colonoscopy correlates with viral genetic variability estimates. CONCLUSIONS: Patient-to-patient transmission of HCV could be demonstrated although the precise route of transmission remained unclear. Viral genetic variability is proposed as a useful tool for tracing HCV transmission, especially in recent transmissions.