Acute chest syndrome in children with sickle cell disease: Data from a national AIEOP cohort identify priority areas of intervention in a hub-and-spoke system.

Acute chest syndrome (ACS) is a frequent cause of hospitalization in sickle cell disease (SCD). Despite advances in acute care, many settings still lack knowledge about ACS best practices. After the AIEOP Guidelines were published in 2012, suggesting standardized management in Italy, a retrospective study was performed to assess the diagnostic and therapeutic pathways of ACS in children. From 2013 to 2018, 208 ACS episodes were presented by 122/583 kids in 11 centres. 73 were male, mean age 10.9 years, 85% African, 92% HbSS or Sß°. In our hub-and-spoke system, a good adherence to Guidelines was documented, but discrepancies between reference centres and general hospitals were noted. Improvement is needed for timely transfer to reference centres, use of incentive spirometry, oxygen therapy and pain management.

Oxylipins generation in Lactobacillus helveticus in relation to unsaturated fatty acid supplementation.

AIMS: Oxylipins are regarded as unsaturated fatty acids (UFAs) oxidation products, whose accumulation in plants and fungi is associated with stress. The aim of this study was to investigate if a metabolic pathway from UFAs to oxylipins was present also in lactic acid bacteria (LAB). METHODS AND RESULTS: A strain of Lactobacillus helveticus, incubated in the presence of oleic, linoleic and linolenic acids released, after 2 h, fatty acid oxidation products, mainly C6 , C8 , C9 aldehydes and alcohols. An experiment with total carbon labelled linoleic acid, in the presence or not of an oxidative stress, demonstrated that oxylipins, such as hexanal, octanal, nonanal, 2-octenal, 2-octanal, originated mainly from the oxidation of this fatty acid. CONCLUSIONS: Since lipoxygenase, dioxygenase and cytochrome P450 genes have never been found in L. helveticus, a possible pathway for linoleic conversion and oxylipins formation could include, as a first step, the transient formation of hydroxylated linoleic acids by fatty acids hydratases. However, the sequence of steps from the linoleic acid to the C6 and C8 aldehydes needs to be more deeply investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the multiple role of oxylipins which are flavouring agents, antimicrobial compounds and interspecific signalling molecules, the knowledge of the mechanisms involved in their biosynthesis in food related bacteria could have an important biotechnological impact, also allowing the overproduction of selected bioactive molecules.

Effect of capric, lauric and alpha-linolenic acids on the division time distributions of single cells of Staphylococcus aureus.

The effect of non-inhibitory concentrations of capric, lauric and alpha-linolenic acids (C10:0, C12:0 and C18:3 respectively) on the division time distribution of single cells of Staphylococcus aureus was evaluated at pH 7 and pH 5. The effect of the initial cell concentration on the lag time of growing cell populations was also assessed. The statistical properties of the division times (defined as the time interval from birth to next binary fission for a single cell) were studied using the method of Elfwing et al. [Elfwing, A., Le Marc, Y., Baranyi, J., Ballagi, A., 2004. Observing the growth and division of large number of individual bacteria using image analysis. Applied and Environmental Microbiology 70, 675-678]. The division times were significantly longer in the presence of free fatty acids than in the control. Shorter division intervals were detected at pH 7 than at pH 5 in the control experiment and in the presence of C10:0. However, both C12:0 and C18:3 slowed down the growth, regardless of the pH. The observed division time distributions were used to simulate growth curves from different inoculum sizes using the stochastic birth process described by Pin and Baranyi [Pin, C., Baranyi, J., 2006. Kinetics of single cells: observation and modelling of a stochastic process. Applied and Environmental Microbiology 72, 2163-2169]. The output of the simulation results were compared with observed data. The lag times fitted to simulated growth curves were in good agreement with those fitted to growth curves measured by plate counts. The averaged out effect of the population masked the effect of the free fatty acids and pH on the division times of single cells.

Growth and metabolites production by Penicillium brevicompactum in yoghurt.

Penicillium brevicompactum, commonly encountered in the indoor air, is known to produce a mycotoxin, mycophenolic acid (MPA). This mould has been isolated from a wide range of foods; considering that we had previously isolated this species from contaminated yoghurt, in this study we have evaluated its growth in yoghurt sweetened with sucrose, fructose and fructose added with fruit pieces. Fungal growth was evaluated monitoring CO(2) production in the headspace during yoghurt storage at 4+/-1, 8+/-1 and 10+/-1 degrees C throughout 21 days. P. brevicompactum grew well in the samples sweetened with fructose at 8 and 10 degrees C. The addition of sucrose influenced the growth negatively, particularly at 4 degrees C. Volatile Organic Compounds (VOC) and MPA production was determined at 8 degrees C in inoculated and uninoculated yoghurt, as well as in liquid malt extract. Differences in VOC profiles and in MPA production were correlated with the age of the fungus and with the growth medium. This study points out for the first time the early qualitative changes in volatile production patterns of a common indoor mould, grown in yoghurt, as well as the production of MPA during storage at refrigeration temperatures.

Probiotic Crescenza cheese containing Lactobacillus casei and Lactobacillus acidophilus manufactured with high-pressure homogenized milk.

High-pressure homogenization (HPH) is one of the most promising alternatives to traditional thermal treatment of food preservation and diversification. Its effectiveness on the deactivation of pathogenic and spoilage microorganisms in model systems and real food is well documented. To evaluate the potential of milk treated by HPH for the production of Crescenza cheese with commercial probiotic lactobacilli added, 4 types of cheeses were made: HPH (from HPH-treated milk), P (from pasteurized milk), HPH-P (HPH-treated milk plus probiotics), and P-P (pasteurized milk plus probiotics) cheeses. A strain of Streptococcus thermophilus was used as starter culture for cheese production. Compositional, microbiological, physicochemical, and organoleptic analyses were carried out at 1, 5, 8, and 12 d of refrigerated storage (4 degrees C). According to results obtained, no significant differences among the 4 cheese types were observed for gross composition (protein, fat, moisture) and pH. Differently, the HPH treatment of milk increased the cheese yield about 1% and positively affected the viability during the refrigerated storage of the probiotic bacteria. In fact, after 12 d of storage, the Lactobacillus paracasei A13 cell loads were 8 log cfu/ g, whereas Lactobacillus acidophilus H5 exhibited, in P-P cheese, a cell load decrease of about 1 log cfu/g with respect to the HPH-P cheese. The hyperbaric treatment had a significant positive effect on free fatty acids release and cheese proteolysis. Also, probiotic cultures affected proteolytic and lipolytic cheese patterns. No significant differences were found for the sensory descriptors salty and creamy among HPH and P cheeses as well as for acid, piquant, sweet, milky, salty, creamy, and overall acceptance among HPH, HPH-P, and P-P Crescenza cheeses.

Effect of high-pressure homogenization, nonfat milk solids, and milkfat on the technological performance of a functional strain for the production of probiotic fermented milks.

The aim of this research was the evaluation of the effects of milkfat content, nonfat milk solids content, and high-pressure homogenization on 1) fermentation rates of the probiotic strain Lactobacillus paracasei BFE 5264 inoculated in milk; 2) viability loss of this strain during refrigerated storage; and 3) texture parameters, volatile compounds, and sensorial properties of the coagula obtained. The data achieved suggested a very strong effect of the independent variables on the measured attributes of fermented milks. In fact, the coagulation times were significantly affected by pressure and added milkfat, and the rheological parameters of the fermented milk increased with the pressure applied to the milk for added nonfat milk solids concentrations lower than 3%. Moreover, the polynomial models and the relative response surfaces obtained permitted us to identify the levels of the 3 independent variables that minimized the viability loss of the probiotic strain used during refrigerated storage.

Use of Yarrowia lipolytica strains for the treatment of olive mill wastewater.

The principal aim of this research was to evaluate the ability of different Yarrowia lipolytica strains, having different origin, to grow in olive mill wastewater (OMW) and reduce its COD level. All the strains were able to grow in undiluted OMW; the comparison between the data obtained in a semi-synthetic medium and in OMW suggests that lipases with different specificity can be produced in relation to the medium composition. Under the adopted conditions, the reduction of the OMW COD values varied from 1.47% and 41.22% of the initial value. Some strains determined a significant reduction of polyphenol content, while other ones caused its apparent increase. Moreover, some Y. lipolytica strains, isolated from chilled foods, produced the highest citric acid concentrations. These results evidenced that some Y. lipolytica strains are good candidates for the reduction of the pollution potential of OMW and for the production of enzymes and metabolites such as lipase and citric acid.

Survey of the physiological properties of the most frequent yeasts associated with commercial chilled foods.

A comparative analysis of the initial and final population of yeasts, lactic acid bacteria and psychrotrophic bacteria, in a number of chilled foods, varying in ingredients, physico-chemical characters and origin, gave evidence that yeasts could play a significant role in the spoilage. The yeast populations appeared to be unexpectedly uniform and comprised principally strains of Yarrowia lipolytica, Debaryomyces hansenii and Pichia membranaefaciens. A survey of 62 isolates, comprising physiological characteristics such as growth temperatures, proteolytic and lipolytic activities, hydrophobicity, aw and preservative tolerance, in addition to organic acid production, indicated that these dominant species have very few common characters and that they are endowed with a spoilage potential probably linked to different physiological properties. The isolates of Y. lipolytica exhibited the strongest proteolytic and lipolytic activities and a pronounced hydrophobicity while D. hansenii isolates were characterized by a high growth rate at low temperature and at intermediate aw. P. membranaefaciens isolates showed a remarkable tolerance to acetic acid as a sole selective factor. A hypothesis of separate growth loci, in multicomponent or polyphasic food systems, was formulated.

Interactions between inhibition factors on microbial stability of fruit-based systems.

Measurement of metabolic CO2 by head space gas chromatography was used in the preparation of shelf-life maps of fruit-based systems. The systems analysed, all of which were peach-based, differed with respect to aw (0.995, 0.934 and 0.890), pH (3.02, 3.60 and 4.09) and benzoic acid concentration (0, 100 and 200 ppm). The test microorganism was a strain of Saccharomyces cerevisiae isolated from spoiled soft-drinks and chosen for its characteristic of multiple resistance. Three different thermal treatments were applied (54 degrees C for 15 min, 58 degrees C for 8 min, 63 degrees C for 3 min) to the systems and their effectiveness was evaluated in relation to the untreated controls. In contrast with observations for osmotolerant species, the relative preservative effectiveness on the test strain decreased with reducing aw and showed only a slight synergistic action with thermal treatment. Besides, the aw of the systems interacted synergistically with the thermal treatment effectiveness in a manner differing from that described for osmotolerant species.

Suitability of log-linear models to evaluate the microbiological quality of baby clams (Chamelea gallina L.) harvested in the Adriatic Sea.

The presence of fecal coliforms or Escherichia coli in baby clams (Chamelea gallina L.) is considered an indicator related to their safety because they can be correlated with the presence of pathogenic bacteria. For this reason the Italian regulation has defined limits for these microorganisms. The presence of these microbial indicators is dependent on various environmental variables. In this work all the variables considered are categorical and, consequently, the traditional approach of predictive microbiology was not applicable. The data were summarized by means of a cross-tabulation and analyzed using the log-linear model technique. This statistical technique is widely used in social and economic studies but only partially developed in food microbiology. The suitability of the log-linear model to analyse microbiological data in relation to environmental variables was evaluated. In particular, the microbiological quality of baby clams harvested in five different areas of the Adriatic Sea coast in Emilia Romagna (Italy) was considered. The influence of the season and geographical origin on microbiological standards was assessed. A logit model was developed to predict the frequencies, depending on geographical origin and season, of samples with concentrations of the indicator organisms below or above the legal standards provided by Italian regulation.

Growth modelling of Listeria monocytogenes and Yersinia enterocolitica in food model systems and dairy products.

The growth potential of Listeria monocytogenes, measured with a gaschromatographic method, was evaluated in model systems simulating dairy products in which NaCl concentration, pH and lipid concentration ranged from values corresponding to matured hard cheeses to some light cheeses and other dairy products. The growth data from the model systems, calculated on the basis of the polynomial models obtained, were compared with values observed for dairy products. Growth parameters of Yersinia enterocolitica in traditional and light cheeses were also available. The comparison between predicted and observed data suggested that individual or interactive effects of chemicophysical variables such as pH, salt and lipid concentration are not sufficient to predict the fate of L. monocytogenes in dairy products. Microstructural factors as well as the pH effect on water binding capacity of proteins could also be implicated.

Interactions between high pressure homogenization and antimicrobial activity of lysozyme and lactoperoxidase.

It was the objective of this work to evaluate the effect of high pressure homogenization on the activity of antimicrobial enzymes such as lysozyme and lactoperoxidase against a selected group of Gram positive and Gram negative species inoculated in skim milk. Lactobacillus helveticus, Lactobacillus plantarum and Listeria monocytogenes were the most pressure resistant species while Bacillus subtilis, Pseudomonas putida, Salmonella typhimurium, Staphylococcus aureus, Proteus vulgaris and Salmonella enteritidis were found to be very sensitive to the hyperbaric treatment. The enzyme addition enhanced the instantaneous pressure efficacy on almost all the considered species as indicated by their instantaneous viability loss following the treatment. Moreover, the combination of the enzyme and high pressure homogenization significantly affected the recovery and growth dynamics of several of the considered species. Although L. monocytogenes was slightly sensitive to pressure, the combination of the two stress factors induced a significant viability loss within 3 h and an extension of lag phases in skim milk during incubation at 37 degrees C. The hypothesis formulated in this work is that the interaction of high pressure homogenization and lysozyme or lactoperoxidase is associated to conformational modifications of the two proteins with a consequent enhancement of their activity. This hypothesis is supported by the experimental results also regarding the increased antimicrobial activity against L. plantarum of the previously pressurised lysozyme with respect to that of the native enzyme.

Proteolytic, lipolytic and molecular characterisation of Yarrowia lipolytica isolated from cheese.

This work studied the qualitative and quantitative proteolytic and lipolytic activities of Yarrowia lipolytica strains isolated from two cheese types. Randomly amplified polymorphic DNA-PCR (RAPD-PCR) analysis was used to compare the cheese strains of Y. lipolytica with strains isolated from other food products and with the type strain of the species in order to investigate the genetic diversity and occurrence of specific environmental groups. Diversity of proteolytic and especially lipolytic activity within Y. lipolytica strains isolated from dairy products was observed. In particular, the degree of specificity for saturated or unsaturated fatty acids as well as for even- or odd-numbered carbon free fatty acids (FFAs) varied among the strains. The RAPD-PCR profiles showed low genetic relatedness between many of the food isolates and the type strain of the species. Such genetic variability needs to be further evaluated. Most of the Y. lipolytica strains appeared to be specific to the particular environment from which they were isolated. However, phenotypic characteristics having technological importance in dairy products and, particularly, lipolytic activities did not correspond to the genetic differences observed by RAPD-PCR analysis.

Variability of the lipolytic activity in Yarrowia lipolytica and its dependence on environmental conditions.

This work was aimed to the evaluation of the variability of lipolytic activity in Yarrowia lipolytica strains, as well as to asses for a selected strain, the response to the changes of physico-chemical variables (such as pH, NaCl and lipid content), in order to obtain predictive models describing their effects on the lipolysis pattern. The strains tested, having different environmental origin, showed different patterns of the free fatty acids (FFA) released. The clustering of the free fatty acids profiles evidenced that the unweighted average distance within the strains of the same species did not exceeded 30%. However, the lipolytic activity of some strains generated FFA profiles that differentiated from the majority of the strains considered. Also, when a single strain was inoculated in model systems in which pH, NaCl and milk fat were modulated according to a Central Composite Design (CCD), chemico-physical characteristics of the system led to marked variations in the lipolytic activity with consequent changes in individual fatty acids released. In most cases, when the same Y. lipolytica strain was used, under the experimental conditions adopted, the modulation of the lactic acid, NaCl and lipid content did not generate differences in the fatty acid release exceeding 20-21%. However, some combinations of factors remarkably affected lipase expression or activity, and generated differences in the fatty acid released higher than those observed among different strains of the same species.

Effects of pH, temperature and NaCl concentration on the growth kinetics, proteolytic activity and biogenic amine production of Enterococcus faecalis.

In this work, the combined effects of temperature, pH and NaCl concentration on the growth dynamics of Enterococcus faecalis EF37, its proteolytic activity and its production of biogenic amines have been studied. The effects of the selected variables have been analysed using a Central Composite Design. The production of biogenic amines, under the adopted conditions, was found to be mainly dependent on the extent of growth of E. faecalis. Its proteolytic activity was not a limiting factor for the final amine production, because in the system studied (skim milk) an excess of precursors was guaranteed. Quantitatively, the most important biogenic amine produced was 2-phenylethylamine but substantial amounts of tyramine were detected in all the samples. This work confirms that the main biological feature influencing the biogenic amine formation is the extent of growth of microorganisms, like E. faecalis, characterised by decarboxylase activity. In the traditional and artisanal cheeses produced using raw milk, enterococci usually reach levels of 10(7) cells/g. With this perspective, it is important that the presence of biogenic amines due to the activities of these microorganisms is maintained within safe levels, without affecting the positive effects of enterococci on the final organoleptic characteristics of the cheese.

A collaborative study on media for the enumeration of yeasts in foods.

A collaborative study was made to evaluate the effectivity of a general purpose medium, tryptone glucose yeast extract (TGY) agar on the detection and enumeration of yeasts from food. Nine laboratories participated in the study and compared five media (four kinds of TGY with different concentrations of glucose, one of them without tryptone, and, for comparison, dichloran rose bengal chloramphenicol (DRBC) agar). Six food samples were investigated by each laboratory and 23 additional food samples were investigated individually by different laboratories. No difference was found in the performance of media with either the samples common to all laboratories or the various samples tested in different ones. A medium consisting of tryptone, glucose and yeast extract, at any concentration of glucose tested, appeared reliable for the detection and enumeration of yeasts from foods, and its performance did not differ from that of DRBC. Omission of tryptone as recommended by ISO provided an even simpler medium of equally good performance. TGY without chloramphenicol may result in higher total counts due to the development of bacteria. DRBC incubated in light results in lower counts compared to that incubated in the dark.

Effect of hexanal on the shelf life of fresh apple slices.

In this work the effects of hexanal, as a component of packaging atmosphere, on the shelf life of and evolution of naturally occurring microbial populations in fresh apple slices during storage at 4 and 15 degrees C were evaluated. Although hexanal had no bactericidal effects, in all conditions considered, this volatile molecule significantly extended the shelf life. In fact, the presence of hexanal in the storage atmosphere (at 4 degrees C) totally inhibited mesophilic bacteria and considerably prolonged the lag phase of psychrotrophic bacteria. Also, at 15 degrees C, hexanal strongly inhibited molds, yeasts, and mesophilic and psychrotrophic bacteria. Moreover, hexanal led to a yeast selection favoring species having a reduced spoilage potential due to their prevalent respiratory activity. When added to a modified atmosphere (70% N(2) and 30% CO(2)), this molecule was also very effective in preventing browning reactions for at least 16 days at 15 degrees C. No changes in hue angle values were observed in samples packaged in modified atmosphere with hexanal, even after 16 days of storage at 4 degrees C.

Effects of hexanal, trans-2-hexenal, and storage temperature on shelf life of fresh sliced apples.

In this paper, the effects of hexanal and trans-2-hexenal, which are both natural molecules characterizing apple aroma, on the microbial population and on color retention of fresh sliced apples were evaluated. In particular, a central composite design (CCD) was developed to assess the individual and interactive effects of the chosen volatile molecules and storage temperatures on (i) the growth of the naturally occurring microflora, (ii) the evolution over time of an inoculated spoilage yeast (Pichia subpelliculosa), and (iii) the enzymatic browning reaction in minimally processed apples. The inclusion of hexanal and trans-2-hexenal in the storage atmosphere of apple slices determined a significant extension of shelf life also when P. subpelliculosa was inoculated at levels of 10(3) colony-forming units/g and abusive storage temperatures were used. In fact, the presence of these molecules in the packaging atmospheres considerably prolonged the lag phases of the inoculated yeast and reduced the growth potential of naturally occurring bacteria. Moreover, the addition to the modified atmosphere of low levels of the hexanal increased the color stability of the products up to 16 days.

Microbiological quality of filled pasta in relation to the nature of heat treatment.

The microbial population present in 49 samples of Italian industrially processed filled pasta was characterized and its changes during refrigerated storage were evaluated. The most frequently isolated species belonged to the genus Bacillus. No pathogenic organisms were isolated from the processed industrial pasta. As a consequence of the diversity of composition and thermal treatment a wide variability was observed (from less than 3 days to more than 1 month) in the shelf life at 4 degrees C of the industrial "fresh filled pasta." However, the results obtained suggested that the shelf life of the processed products depend not only on the number of surviving cells but also on the textural or microstructural changes induced by the heat treatment. Challenge tests using Staphylococcus aureus showed that even pasteurization values (P 70(10), expressed as an equivalent process time, in minutes, necessary to obtain at 70 degrees C the same lethal effect as during the actual process) not exceeding 2 were able to remarkably reduce the cell load of this organism. Subsequent growth of the surviving S. aureus cells occurred only at temperatures > 7 degrees C, particularly when the water activity (aw) values were higher than 0.97.

Analysis of the interaction between autochthonous bacteria and packaging material in PVC-bottled mineral water.

A study with about 10,000 bottles produced by a mineral water company was undertaken in order to identify the causal agent of an off-odour occurrence in the bottled water. Some physiological attributes of the dominant species over an 8-month period, as well as their interaction with packaging material, were investigated. Pseudomonas maltophilia, P. acidovorans, Acinetobacter calcoaceticus var. lowffi, frequently associated with bottles having an off-odour, seemed to play a decisive role in the phenomenon due to their elevated lipolytic activity, their cell hydrophobicity and adhesivity to the PVC walls. Their ability to attack the sodium polysulfide included in the ultramarine blue dye present in PVC, transforming it to H2S was investigated.