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Abstract Background Pharmacogenomics of hydroxyurea is an important aspect in the management of sickle cell disease (SCD), especially in the era of genomic medicine. Genetic variations in loci associated with HbF induction and drug metabolism are prime targets for hydroxyurea (HU) pharmacogenomics, as these can significantly impact the therapeutic efficacy and safety of HU in SCD patients. Methods This study involved designing of a custom panel targeting BCL11A, ARG2, HBB, HBG1, WAC, HBG2, HAO2, MYB, SAR1A, KLF10, CYP2C9, CYP2E1 and NOS1 as potential HU pharmacogenomics targets. These genes were selected based on their known roles in HbF induction and HU metabolism. The panel was designed using the Illumina Design Studio (Illumina, San Diego, CA, USA) and achieved a total coverage of 96% of all genomic targets over a span of 51.6 kilobases (kb). This custom panel was then sequenced using the Illumina MiSeq platform to ensure high coverage and accuracy. Results We are reporting a successfully designed Illumina (MiSeq) HU pharmacogenomics custom panel encompassing 51.6 kilobases. The designed panel achieved greater than 1000x amplicon coverage which is sufficient for genomic analysis. Conclusions This study provides a valuable tool for research in HU pharmacogenomics, especially in Africa where SCD is highly prevalent, and personalized medicine approaches are crucial for improving patient outcomes. The custom-designed Illumina (MiSeq) panel, with its extensive coverage and high sequencing depth, provides a robust platform for studying genetic variations associated with HU response. This panel can contribute to the development of tailored therapeutic strategies, ultimately enhancing the management of SCD through more effective and safer use of hydroxyurea.

Despite native whey (hereafter termed WHEY) being a watery by-product rich in high-value nutritional components, it poses environmental concerns. Recently, the techniques of thermal-mechanical microparticulation to produce microparticulated whey (MPW) and a further process to ferment MPW (FMPW) were suggested, in order to recycle whey into a protein concentrated soft dairy cream that acquires also fat mimicking functionality. Aiming at monitoring the effects of this recycling process on the products’ metabolic profiles, samples (n = 8) of WHEY, MPW and two sub-types of FMPW were analysed by combining direct analysis in real time coupled with high resolution mass spectrometry (DART-HRMS). The most informative ions were used to build a partial least squared discriminant analysis (PLS-DA) model to assess the biochemical compounds that characterize and differentiate WHEY from MPW and FMPW. The DART-HRMS fingerprints acquired in negative mode accurately differentiated samples throughout the steps of microparticulation and fermentation. Post-fermentation, greater relative abundances of lactic acid, glucosamine and histidyl-aspartic acid were detected in both subtypes of FMPW, which were moderately differentiated by DART-HRMS. Moreover, while WHEY and MPW contained high relative abundances of hydroxyglutaric and malic acids, the fermented derivates (FMPW) were characterised by elevated levels of volatile compounds (tetradecanal, hexadecene and tetradecene) and fermentative end-products (diethyltartrate and histidyl-aspartic acid). DART-HRMS successfully captured a pool of informative biomarkers useful to understand the chemical and metabolomic changes occurring in microparticulation and fermentation processes used to recycle whey. This analytical technique is an operative supporting step needed for comprehensive assessment of the nutritional and organoleptic properties of the proposed whey-based soft cream dairy food.

This research aims to monitor the conservation status of the lipid and mineral contents of four shelf-stable insect-based products (yellow mealworm, house cricket, mole cricket, and silkworm) marketed online. A total of 32 single-species packs were purchased from various online commercial suppliers. Moisture, lipids, fatty acids, titratable acidity, mineral elements, and primary and secondary lipid oxidation products were determined. Statistical multivariate approaches were applied to investigate the contribution of each chemical variable to the characterization of edible insects. Titratable acidity (up to 37.3 g oleic acid per 100 g of crickets), as well as primary and secondary lipid oxidation products, showed great variability within and between species. The study revealed a significant occurrence of rancidity (45.5% of the samples exceeded the peroxide limit of 10 mEqO2/kg; 100% of the samples exceeded the indication of 1 mg/kg malondialdehyde), whereas the heavy metal content indicated a relatively safe condition, suggesting the absence of potential risks to human health. Both the chemical and the elemental properties could be regarded as potential characteristics suitable for authenticating this food matrix. This study contributes to the description of several chemical features in commercialized processed insect-based products, aiming to highlight possible safety concerns and identify those unfit for human consumption.

This study aimed to identify critical issues in artisanal bakery and pastry production in Italy that could improve food safety and quality. Fifteen voluntary Italian companies underwent on-site inspections and interviews from 2018 to 2021. The inspection concerned the production site characteristics, processing flows, materials, and personnel to pinpoint potential product contamination and record objective data collection through a 126-question demerit scoring system. The examined areas encompassed various aspects, such as the point of sale, management of raw materials and packaging, hygiene practices, finished product quality, sanitization procedures, external spaces, personnel, and other factors impacting food safety and hygiene. Additionally, assessments of the microbiological air quality were carried out. Two cohorts (six and nine companies in 2018-2019 and in 2020-2021, respectively) revealed critical issues in warehouse/packaging (70% non-compliance), finished product/cooling (50%), and people/products flow (38%). Site visits identified pest management and raw material handling challenges. Significant airborne mold and yeast contamination (30-50 colony-forming units/plate/h) was observed in areas near processing sites or with air turbulence. This study facilitated constructive discussions and proposed solutions with the participating companies.

The application of an electrochemical (impedance) tool for monitoring Escherichia coli contamination in shellfish was evaluated after 13 months of observation. The primary aim of the present study was to compare the standard most probable number (MPN) and µ-trac 4200 (log imped/100 g) for the assessment of E. coli contamination (log MPN/100 g) in non-depurated bivalve mollusks (BM) from five sampling areas of the Veneto-Emilian coast (Italy) (118 samples). The secondary aim was to evaluate the correlation between E. coli concentrations in BM and environmental factors on a large data set (690). The methods showed a moderate, positive correlation (0.60 and 0.69 Pearson and Spearman coefficients, respectively; P4,600 MPN/100 g). The results highlighted the suitability of the impedance method for a faster evaluation and routine use especially in clams, while in Mytilus it seemed less effective. Different models built by multivariate permutational variance analysis and multinomial logistic regression selected the suitable environmental features able to predict the E. coli load. Overall, salinity and season affected the E. coli contamination, whereas locally it was mainly influenced by hydrometry and salinity. The application of the impedance method coupled with environmental data analysis could help purification phase management to adhere to legal limits and could represent an advantage for local control authorities to define actions, considering extreme meteorological events' effects as a proactive reaction to climate change.

The genus Vibrio includes bacteria with different morphological and metabolic characteristics responsible for different human and animal diseases. An accurate identification is essential to assess the risks in regard to aquatic organisms and consequently to public health. The Multilocus Sequence Analysis (MLSA) scheme developed on the basis of 4 housekeeping genes (gyrB, pyrH, recA and atpA) was applied to identify 92 Vibrio strains isolated from crustaceans in 2011. Concatenated sequences were used for the phylogenetic and population analyses and the results were compared with those from biochemical identification tests. From the phylogenetic analysis, 10 clusters and 4 singletons emerged, whereas the population analysis highlighted 12 subpopulations that were well supported by phylogeny with few exceptions. The retrospective analysis allowed correct re-attribution of isolated species, indicating how, for some pathogens, there may be an overestimation of phenotypic identification (e.g. V. parahaemolyticus). Use of the PubMLST Vibrio database highlighted a possible genetic link between Sequence Type (ST) 529 and ST195 (V. alginolyticus) isolated from a human case in Norway during 2018. In addition to the identification of major risk groups of V. cholerae, V. vulnificus and V. parahaemolyticus, MLSA could be a valid support for species considered a minor risk, such as V. alginolyticus, V. mimicus and V. fluvialis. Due to the increased incidence of vibriosis in Europe, the application of different tools will also have to be considered to investigate the possible epidemiological links of the various species in the perspective of Open Science to protect the consumer.

Abstract Metabolomics approaches, such as direct analysis in real time-high resolution mass spectrometry (DART-HRMS), allow characterising many polar and non-polar compounds useful as authentication biomarkers of dairy chains. By using both a partial least squares discriminant analysis (PLS-DA) and a linear discriminant analysis (LDA), this study aimed to assess the capability of DART-HRMS, coupled with a low-level data fusion, discriminate among milk samples from lowland (silages vs. hay) and Alpine (grazing; APS) systems and identify the most informative biomarkers associated with the main dietary forage. As confirmed also by the LDA performed against the test set, DART-HRMS analysis provided an accurate discrimination of Alpine samples; meanwhile, there was a limited capacity to correctly recognise silage- vs. hay-milks. Supervised multivariate statistics followed by metabolomics hierarchical cluster analysis allowed extrapolating the most significant metabolites. Lowland milk was characterised by a pool of energetic compounds, ketoacid derivates, amines and organic acids. Seven informative DART-HRMS molecular features, mainly monoacylglycerols, could strongly explain the metabolomic variation of Alpine grazing milk and contributed to its classification. The misclassification between the two lowland groups confirmed that the intensive dairy systems would be characterised by a small variation in milk composition.

Accurate species identification, especially in the fishery sector, is critical for ensuring food safety, consumer protection and to prevent economic losses. In this study, a total of 93 individual frozen–thawed cuttlefish samples from four different species (S. officinalis, S. bertheloti, S. aculeata, and Sepiella inermis) were collected from two wholesale fish plants in Chioggia, Italy. Species identification was carried out by inspection through morphological features using dichotomic keys and then through near-infrared spectroscopy (NIRS) measurements. The NIRS data were collected using a handled-portable spectrophotometer, and the spectral range scanned was from 900–1680 nm. The collected spectra were processed using principal component analysis for unsupervised analysis and a support vector machine for supervised analysis to evaluate the species identification capability. The results showed that NIRS classification had a high overall accuracy of 93% in identifying the cuttlefish species. This finding highlights the robustness and effectiveness of spectral analysis as a tool for species identification, even in complex spatial contexts. The findings emphasize the potential of NIRS as a valuable tool in the field of fishery product authentication, offering a rapid and eco-friendly approach to species identification in the post-processing stages.

Clay minerals are naturally occurring rock and soil materials primarily composed of fine-grained aluminosilicate minerals, characterized by high hygroscopicity. In animal production, clays are often mixed with feed and, due to their high binding capacity towards organic molecules, used to limit animal absorption of feed contaminants, such as mycotoxins and other toxicants. Binding capacity of clays is not specific and these minerals can form complexes with different compounds, such as nutrients and pharmaceuticals, thus possibly affecting the intestinal absorption of important substances. Indeed, clays cannot be considered a completely inert feed additive, as they can interfere with gastro-intestinal (GI) metabolism, with possible consequences on animal physiology. Moreover, clays may contain impurities, constituted of inorganic micronutrients and/or toxic trace elements, and their ingestion can affect animal health. Furthermore, clays may also have effects on the GI mucosa, possibly modifying nutrient digestibility and animal microbiome. Finally, clays may directly interact with GI cells and, depending on their mineral grain size, shape, superficial charge and hydrophilicity, can elicit an inflammatory response. As in the near future due to climate change the presence of mycotoxins in feedstuffs will probably become a major problem, the use of clays in feedstuff, given their physico-chemical properties, low cost, apparent low toxicity and eco-compatibility, is expected to increase. The present review focuses on the characteristics and properties of clays as feed additives, evidencing pros and cons. Aims of future studies are suggested, evidencing that, in particular, possible interferences of these minerals with animal microbiome, nutrient absorption and drug delivery should be assessed. Finally, the fate of clay particles during their transit within the GI system and their long-term administration/accumulation should be clarified.

Abstract Background Acute chest syndrome (ACS) is the leading cause of mortality in sickle cell disease (SCD). The pathogenesis of ACS is complex and not entirely understood with multiple etiologies likely contributing simultaneously. One particular etiology is pulmonary vascular occlusion due to thrombosis. Thus, anticoagulation is an attractive therapeutic modality. Methods This was a single-center, randomized controlled, open-label, pilot study to determine the feasibility of performing a larger multicenter phase III trial to assess the effects of unfractionated heparin (UFH) in ACS. Subjects were randomized within 24 h of diagnosis of ACS to one of two treatment arms, UFH, and standard of care (SOC), or no UFH and SOC. UFH was given intravenously for 7 days, or until discharge, if discharge was shorter than 7 days. SOC consisted of intravenous fluids, antibiotics, supplemental oxygen, analgesia, red blood cell transfusion, and exchange transfusion. Results From July 2014 to June 2018, a total of 7 patients underwent randomization (four patients received UFH in addition to SOC and 3 patients received SOC only). Two of the prespecified feasibility criteria were not met: the capacity to consent eligible individuals and the timely notification of hospitalized patients with ACS necessary to permit randomization within 24 h of diagnosis; thus, as a result of poor enrollment, the study was terminated early. The duration of hospitalization was 279.43 (SD 267.98) and 127.31 (SD 137.70) h in the UFH and SOC arms, respectively. The duration of hypoxemia, leukocytosis, fever, and moderate to severe pain was 117.52 (SD 60.52), 24.90 (SD 29.69), 117.52 (SD 60.52), and 117.52 (SD 60.52) h, respectively, in the UFH group, and 51.49 (SD 44.79), 0, 53.11 (SD 25.06), and 88.68 (SD 72.77) h, respectively, in the SOC group. No major bleeding was noted in either group. Conclusions Our study did not achieve prespecified feasibility criteria, resulting in poor enrollment and early termination, and serves to highlight some of the pitfalls experienced in clinical research in SCD. It did show the use of UFH without any major adverse events in 7 subjects. No future large-scale study is planned. Trials registration Registered at ClinicalTrials.gov (NCT #02098993) on March 28, 2014.

Food fraud, corresponding to any intentional action to deceive purchasers and gain an undue economical advantage, is estimated to result in a 10 to 65 billion US dollars/year economical cost worldwide. Dairy products, such as cheese, in particular cheeses with protected land- and tradition-related labels, have been listed as among the most impacted as consumers are ready to pay a premium price for traditional and typical products. In this context, efficient food authentication methods are needed to counteract current and emerging frauds. This review reports the available authentication methods, either chemical, physical, or DNA-based methods, currently used for origin authentication, highlighting their principle, reported application to cheese geographical origin authentication, performance, and respective advantages and limits. Isotope and elemental fingerprinting showed consistent accuracy in origin authentication. Other chemical and physical methods, such as near-infrared spectroscopy and nuclear magnetic resonance, require more studies and larger sampling to assess their discriminative power. Emerging DNA-based methods, such as metabarcoding, showed good potential for origin authentication. However, metagenomics, providing a more in-depth view of the cheese microbiota (up to the strain level), but also the combination of methods relying on different targets, can be of interest for this field.

Aflatoxin B1 (AFB1) is a major food safety concern, threatening the health of humans and animals. Bentonite (BEN) is an aluminosilicate clay used as a feed additive to reduce AFB1 presence in contaminated feedstuff. So far, few studies have characterized BEN toxicity and efficacy in vitro. In this study, cytotoxicity (WST-1 test), the effects on cell permeability (trans-epithelial electrical resistance and lucifer yellow dye incorporation), and transcriptional changes (RNA-seq) caused by BEN, AFB1 and their combination (AFB1 + BEN) were investigated in Caco-2 cells. Up to 0.1 mg/mL, BEN did not affect cell viability and permeability, but it reduced AFB1 cytotoxicity; however, at higher concentrations, BEN was cytotoxic. As to RNA-seq, 0.1 mg/mL BEN did not show effects on cell transcriptome, confirming that the interaction between BEN and AFB1 occurs in the medium. Data from AFB1 and AFB1 + BEN suggested AFB1 provoked most of the transcriptional changes, whereas BEN was preventive. The most interesting AFB1-targeted pathways for which BEN was effective were cell integrity, xenobiotic metabolism and transporters, basal metabolism, inflammation and immune response, p53 biological network, apoptosis and carcinogenesis. To our knowledge, this is the first study assessing the in vitro toxicity and whole-transcriptomic effects of BEN, alone or in the presence of AFB1.

This study aims at evaluating the effect of a phenol-rich extract obtained from the concentration and purification of olive mill wastewaters (added at a ratio of 87.5 and 175 mg of phenols/kg meat) on the stability and sensory quality of beef hamburgers packed under modified atmosphere and stored under alternating exposure to fluorescent light at 4 ± 2 °C for 9 days. The hamburgers were sampled at different times (0, 6, and 9 days) and grilled at 200 °C. After 9 days, more than 56% of the added phenols in the raw burgers and more than 20% the grilled ones were retained. The results show that both concentrations of phenolic extract proved to effectively reduce primary and secondary lipid oxidation, as well as cholesterol oxidation products (COPs), during the shelf-life of raw hamburgers. Peroxide value, thiobarbituric acid reactive substances, and total COPs were up to 1.4-, 4.5-, and 8.8-fold lower in phenol-enriched raw hamburgers, respectively, than in the control samples; a similar trend was noted also in phenol-enriched cooked hamburgers (1.3-, 5.7-, and 4-fold lower). The sensory analysis also confirmed the effectiveness of the addition of phenolic extract, resulting in a positive effect on the red color intensity (raw product) and thus reducing browning during storage.

Protein kinase CK2 is largely involved in cell proliferation and apoptosis and is generally recognized as an Achilles’ heel of cancer, being overexpressed in several malignancies. The beneficial effects of (−)-epigallocatechin-3-gallate (EGCG) in the prevention and treatment of several diseases, including cancer, have been widely reported. However, poor stability and limited bioavailability hinder the development of EGCG as an effective therapeutic agent. The combination of innovative nanomaterials and bioactive compounds into nanoparticle-based systems demonstrates the synergistic advantages of nanocomplexes as compared to the individual components. In the present study, we developed a self-assembled core-shell nanohybrid (SAMN@EGCG) combining EGCG and intrinsic dual-signal iron oxide nanoparticles (Surface Active Maghemite Nanoparticles). Interestingly, nano-immobilization on SAMNs protects EGCG from degradation, preventing its auto-oxidation. Most importantly, the nanohybrid was able to successfully deliver EGCG into cancer cells, displaying impressive protein kinase CK2 inhibition comparable to that obtained with the most specific CK2 inhibitor, CX-4945 (5.5 vs. 3 µM), thus promoting the phytochemical exploitation as a valuable alternative for cancer therapy. Finally, to assess the advantages offered by nano-immobilization, we tested SAMN@EGCG against Pseudomonas aeruginosa, a Gram-negative bacterium involved in severe lung infections. An improved antimicrobial effect with a drastic drop of MIC from 500 to 32.7 μM was shown.

Olive vegetation water (OVW) is a by-product with a noticeable environmental impact; however, its polyphenols may be reused food and feed manufacture as high-value ingredients with antioxidant/antimicrobial activities. The effect of dietary supplementation with OVW polyphenols on the gut microbiota, carcass and breast quality, shelf life, and lipid oxidation in broiler chickens has been studied. Chicks were fed diets supplemented with crude phenolic concentrate (CPC) obtained from OVW (220 and 440 mg/kg phenols equivalent) until reaching commercial size. Cloacal microbial community (rRNA16S sequencing) was monitored during the growth period. Breasts were submitted to culture-dependent and -independent microbiological analyses during their shelf-life. Composition, fatty acid concentration, and lipid oxidation of raw and cooked thawed breasts were measured. Growth performance and gut microbiota were only slightly affected by the dietary treatments, while animal age influenced the cloacal microbiota. The supplementation was found to reduce the shelf life of breasts due to the growth of spoilers. Chemical composition and lipid oxidation were not affected. The hydroxytyrosol (HT) concentration varied from 178.6 to 292.4 ug/kg in breast muscle at the beginning of the shelf-life period. The identification of HT in meat demonstrates that the absorption and metabolism of these compounds was occurring efficiently in the chickens.

An appropriate seafood origin identification is essential for labelling regulation but also economic and ecological issues. Near infrared (NIRS) reflectance spectroscopy was employed to assess the origins of cuttlefish caught from five fishing FAO areas (Adriatic Sea, northeastern and eastern central Atlantic Oceans, and eastern Indian and western central Pacific Oceans). A total of 727 cuttlefishes of the family Sepiidae (Sepia officinalis and Sepiella inermis) were collected with a portable spectrophotometer (902–1680 nm) in a wholesale fish plant. NIR spectra were treated with standard normal variate, detrending, smoothing, and second derivative before performing chemometric approaches. The random forest feature selection procedure was executed to select the most significative wavelengths. The geographical origin classification models were constructed on the most informative bands, applying support vector machine (SVM) and K nearest neighbors algorithms (KNN). The SVM showed the best performance of geographical classification through the hold-out validation according to the overall accuracy (0.92), balanced accuracy (from 0.83 to 1.00), sensitivity (from 0.67 to 1.00), and specificity (from 0.88 to 1.00). Thus, being one of the first studies on cuttlefish traceability using NIRS, the results suggest that this represents a rapid, green, and non-destructive method to support on-site, practical inspection to authenticate geographical origin and to contrast fraudulent activities of cuttlefish mislabeled as local.

This abstract book contains the abstracts presented at the 28th National Congress of the Italian Association of Veterinary Food Hygienists (AIVI) | Bari, 11-13 September 2019

The experiment studied the effect of two different fat inclusion levels (30% and 40%), NaCl contents (2.4 and 2.6%) and starter cultures (lactic acid bacteria (LAB) 6: L. curvatus/S. xylosus; LAB 8: L. sakei/S. xylosus) on the weight loss and nutritional composition of Italian-type ostrich salami. With this purpose, 8 batches of 9 salami each (n = 72) were prepared. Salami were ripened for 20 weeks: weight loss was monitored throughout the experiment, while salami nutritional composition was evaluated at 10 and 20 weeks of ripening. The lowest fat and highest salt inclusion levels provided the highest cumulative weight loss throughout the trial. At 10 weeks of ripening, salami with 40% fat were the richest in moisture and fat, whereas the leanest ones had the highest protein, ash and cholesterol contents. LAB 6 provided salami with the highest moisture and protein, while LAB 8 increased fat and cholesterol contents. At 20 weeks of ripening the proximate composition of ostrich salami was solely affected by fat inclusion level, with similar findings to those observed at 10 weeks. Overall, fat inclusion level had a great impact on the weight loss and nutritional composition of Italian-style ostrich salami. Reducing the NaCl inclusion from 2.6% to 2.4%, the weight loss of ostrich salami was retarded by approximately 1 week, without affecting the nutritional composition of the final product. Results of the study suggested that it is feasible to produce salami with lower fat and salt contents, while ensuring satisfactory product quality.

Processing of alpine milk in malga farms is carried out under conditions that can favor contamination by coliforms, coagulase-positive staphylococci, or pathogens such as Listeria monocytogenes. With the aim to improve the hygienic characteristics and safety of cheese produced in four malga farms the use of lyophilized Natural Milk Culture prepared with selected strains was tested. Two cheesemaking tests were carried out in the same day always starting from the same milk: in the first case following the malga recipe that uses either Natural Whey Culture or without the addition of a starter, in the second one using a Natural Milk Culture. Cheesemaking were carried out in four malga farms located in the west area of Trentino region within the same week. For hygienic and safety evaluation, aerobic colony count, coagulase-positive staphylococci, Escherichia coli, staphylococcal toxins, Listeria monocytogenes , and Salmonella spp, pH and aw were determined in raw milk from evening and morning milking, curd in vat, curd after extraction and two months-ripened cheese. Pathogens or toxins, high values of coagulase- positive staphylococci and E. coli were not found in cheese samples. However, in the curd coagulase-positive staphylococci reached values almost of 5 Log CFU/g in the two malga without starter cultures. The use of Natural Milk Culture reduced E. coli counts. In addition, DNA was extracted from cheese samples and from Natural Milk Culture and the composition of the microbial community determined by Next Generation Sequencing method. The determination of cheese microbial communities demonstrated that the use of Natural Milk Culture exerted different effects in the different malga, in any case preserving bacterial biodiversity.

Near infrared spectroscopy (NIRs) and ultraviolet visible spectroscopy (UV-vis) have been investigated as rapid techniques to characterize foodborne bacteria through the analysis of the spectra of whole cells or microbial suspensions. The use of spectra collected from broth cultures could be used as a fingerprint for strain classification using a combined polyphasic approach. The aim of this study was to evaluate the feasibility of NIRs and UV-vis for the characterization of blue strains belonging to the Pseudomonas fluorescens group. The bacteria were isolated from different food matrices, including some spoiled samples (blue discoloration). Eightyone strains previously identified at the species level were grown in Minimal Bacterial Medium broth under standard conditions at 22°C. Two biological replicates were centrifuged in order to separate the bacterial cells from the extracellular products. Six aliquots per strain were analyzed on a small ring cup in transflectance mode (680-2500 nm, gap 2 nm). A subset of 39 strains was evaluated by UV-vis to determine changes in the spectral characteristics at 48 and 72 hours. Several chemometric approaches were tested to assess the performance of NIRs and UVvis. According to the variable importance in projection (VIP), the 1892-2020 nm spectral region showed the highest level of discrimination between blue strains and others. Additional information was provided in the 680-886 and 1454-1768 nm regions (aromatic C-H bonds) and in the 2036-2134 nm region (fatty acids). Changes in UV-vis spectral data (at 48 and 72 hours) appear to indicate the presence of phenazine and catecholic compounds in extracellular products.

Food packaging frequently reports the terms natural, 100% natural or similar. Often these indications induce consumers to purchase those products that are considered healthier and fresher. The overall goal of this study was to assess what teen consumers perceive to be genuine and natural foods. A questionnaire was distributed to the students of some high schools (lyceum, technical and professional institutes). It was completed by 349 females and 314 males, with an average age of 17.6 years. Respondents are quite interested in the information on recipes, diet, beauty and food safety; websites were important information retrieval tools. Genuine food is defined mainly as fruits and vegetables, home-made and salubrious, with less or without fat and that is good for health. Meanwhile, natural is demarcated primarily by the absence of additives and manipulation or treatments (negative impact). Also fruits and vegetables and organic production are associated to natural. The existence of a natural food preference is well described and the presence on food label may cause a wrong perception of healthfulness.

Challenge tests are a clear opportunity for manufacturers interested in the evaluation of their management system with the aim to reduce the spread of foodborne pathogens. This is a main concern especially in ready-to-eat food in relation to the risk associated with Listeria monocytogenes. For small and medium-scale food industry the manufacturing practices and products formulation are characterised by a wider variability and poor repeatability. The use of ad hoc challenge test and the comparison among different processing systems are strongly required. This paper reports a preliminary comparison among different challenge tests (n=12) commissioned by three manufacturers of raw-fermented salami during a period of three years (2013-2016). The challenge tests were designed to evaluate the growth potential (δ) of L. monocytogenes during the whole processing period of the salami. The doughs were prepared according to different formulations: the simplest formulation was represented by the use of salt, potassium nitrate, black pepper and starter cultures, while the most composited formulations also included the use of sugars and ascorbic acid in addition to nitrite salt. All the processing steps were conducted within an experimental laboratory dedicated for the processing of meat. After stuffing, the salami were dried and ripened under temperature and relative humidity control. The sugar inclusion can be considered as a protective factor, while the drying step at high temperature (above 20°C) was associated with higher δ values (δ>0.5 log10 cfu/g). The addition of starter cultures, and the subsequent acidification highlighted the importance of pH as the parameter able to affect the L. monocytogenes growth.

Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial proteases, a predominant role in spoilage of dairy products seems to be played by the thermostable metallo-protease AprX, which is produced by various strains of Pseudomonas fluorescens. Differences in AprX enzyme activity among different strains were highlighted, but the most proteolytic strains were not identified. In this study, the presence of the aprX gene was evaluated in 69 strains isolated from food matrices and 18 reference strains belonging to the P. fluorescens group, which had been previously typed by the multi locus sequence typing method. Subsequently, a subset of reference strains was inoculated in ultra-high temperature milk, and the expression of the aprX gene was evaluated at 22 and 6°C. On the same milk samples, the proteolytic activity was then evaluated through Azocasein and trinitrobenzenesulfonic acid solution assays. Finally, to assess the applicability of the former assay directly on dairy products the proteolityc activity was tested on industrial ricotta samples using the Azocasein assay. These results demonstrate the spread of aprX gene in most strains tested and the applicability of Azocasein assay to monitor the proteolytic activity in dairy products.

In the past, milk whey was only a by-product of cheese production, but currently, it has a high commercial value for use in the food industries. However, the regulation of whey management (i.e., storage and hygienic properties) has not been updated, and as a consequence, its microbiological quality is very challenging for food safety. The Next Generation Sequencing (NGS) technique was applied to several whey samples used for Ricotta production to evaluate the microbial community composition in depth using both RNA and DNA as templates for NGS library construction. Whey samples demonstrating a high microbial and aerobic spore load contained mostly Firmicutes; although variable, some samples contained a relevant amount of Gammaproteobacteria. Several lots of whey acquired as raw material for Ricotta production presented defective organoleptic properties. To define the volatile compounds in normal and defective whey samples, a headspace gas chromatography/mass spectrometry (GC/MS) analysis was conducted. The statistical analysis demonstrated that different microbial communities resulted from DNA or cDNA library sequencing, and distinguishable microbiota composed the communities contained in the organoleptic-defective whey samples.

Phenols are plant metabolites characterised by several interesting bioactive properties such as antioxidant and bactericidal activities. In this study the application of a phenols concentrate (PC) from olive vegetation water to two different fresh products – gilt-head seabream (Sparus aurata) and chicken breast – was described. Products were treated in a bath of PC (22 g/L; chicken breast) or sprayed with two different solutions (L1:0.75 and L2:1.5 mg/mL; seabream) and then stored under refrigeration conditions. The shelf life was monitored through microbiological analyses – quality index method for seabream and a specific sensory index for raw breast. The secondary products of lipid-peroxidation of the chicken breast were determined using the thiobarbituric acid reactive substances (TBARs) test on cooked samples. Multivariate statistical techniques were adopted to investigate the impact of phenols and microbiological data were fitted by DMfit software. In seabream, the levels of PC did not highlight any significant difference on microbiological and sensory features. DMfit models suggested an effect only on H2S producing bacteria with an increased lag phase compared to the control samples (C: 87 h vs L2: 136 h). The results on chicken breast showed that the PC bath clearly modified the growth of Pseudomonas and Enterobacteriaceae. The phenol dipping was effective in limiting lipid-peroxidation (TBARs) after cooking. Treated samples disclosed an increase of shelf life of 2 days. These could be considered as preliminary findings suggesting the use of this concentrate as preservative in some fresh products.

The sampling activity for this study was performed between September and October 2012. It involved seven shops in Verona, eleven in Vicenza and two in its province (Bassano del Grappa), northern Italy. The scope was to measure the values of energy and nutritional components and to identify the profile of fatty acids in a serving of ready to eat Döner Kebab. The samples were collected according to the usual proportions of this preparation, keeping all the components (bread, meat, vegetables and sauces) separated in different bags. In the laboratory, each component was weighed and, after pooling, processed for the analytical determination of humidity, crude protein, lipid content and fatty acid profile, ashes, sodium (salt), carbohydrate, collagen (measured only in meat) and fibre. The results showed a highly standardized recipe, while the comparison between the two towns showed a significant difference in carbohydrate concentration (mainly due to the quantity of bread used). By observing data on the serving sizes sampled (274 to 618 g) and the nutritional values obtained, Döner Kebab can be seen as a ready to eat dish providing much energy: on average a serving size covers 45 and 36% of the recommended daily intake of energy, 95.7 and 82.1% of protein, 42.5 and 33.4% of saturated fatty acids for females and males, respectively, and 85.5% of salt regardless of gender. Döner Kebab can be considered as an occasional substitute to one of the two main meals of the day.

The aim of the study was to assess the in vitro effect of phenols extracted from oil vegetation water (PEOW) on several food-borne strains. Antibacterial activity of PEOW was based on the minimum bactericidal concentration (MBC) on microtitre assay. The taxa tested were: Staphylococcus (n. 5), Listeria (n. 4), Escherichia (n. 2), Salmonella (n. 1), Pseudomonas (n. 3), Lactobacillus (n. 2) and Pediococcus (n. 1). S. aureus and L. monocytogens showed the lowest level of resistance to PEOW (MBC=1.5-3 mg/mL). In contrast, the Gram negative strains (e.g. S. Typhimurium and Pseudomonas spp.) were in some cases unaffected by the tested doses and the MBCs ranged between 6 to 12 mg/mL. Starter cultures were dramatically reduced on growth (e.g. Staphylococcus xylosus; 0.75 mg/mL MBC). The thresholds for pathogenic strains could be considered for further applications of PEOW in food models (e.g. shelf life or challenge test studies).

The aim of this qualitative survey was to gain an insight into the ways consumers purchase, transport and storage fresh and frozen food. In particular, this paper considered consumers’ behaviour and the knowledge they have about cold chain. An explorative study was held using focus group interviews (n. 4) as the method for data collection. The sampling group was composed of 24 consumers (4 males and 20 females) and the age ranged from 33 to 78. Data revealed that food safety knowledge is at a fairly good level, however consumer practices in certain cases were inappropriate particularly with respect to transport from the store to home, storage and thaw. Consumers were particularly concerned about frozen food that should not be thawed during shopping or transportation. Knowledge about eggs storage seemed to be dodgy as well. Due to the restricted extent of the sample survey the results cannot be generalized to the whole Italian population; still, this method is particularly useful for discovering not only what people think but why they think that way.

A purified extract of phenols compounds (65% of phenolic content of which decarboxymethyl oleuropein aglycone represented 45% of the wet mass) obtained from vegetation water (a by-product of oil mill) was added to a ground meat dough intended for salami manufacture in two concentration levels: 75 and 150 mg/100 g of dough (F1 and F2, respectively). The control batch was composed of lean and fat cuts of pork in 70:30 ratio, 2.7% salt and a mixed starter culture of staphylococci and pediococci. After stuffing into natural casings, salamis were aged until they reached a total weight loss of 30%. The product was then sliced and packaged in a protective atmosphere (nitrogen:carbon dioxide 80:20) and placed in a refrigerator thermostat (2-4°C) with alternating 12 h of artificial light and darkness. The samples were analysed for the measurement of pH, water activity, organic acidity, peroxide number and secondary products of lipid peroxidation at the time of slicing and after 10, 20 and 30 days of storage into the refrigerated thermostat. The pH and water activity were not substantially different between the control and the two enriched batches. The peroxide number and secondary products of lipid peroxidation values in the two batches with phenols were at least substantially lower than the control sample. In conclusion, the phenol compounds obtained from vegetation water have shown no interference with the ripening process while protecting the dough from oxidation.

Apple juice, for its sensory and nutritional qualities, is consumed by people of all ages. Apples are an excellent source of several phenolic compounds and the presence of polyphenols is recognized for their health promoting antioxidant properties. Thermal pasteurization of fruit juices is the conventional method used for their preservation. Therefore, this constitutes the most extensively available methods for the inactivation of microorganisms in fruit juices but it causes side effects on their flavour and nutritional quality. Consumers tend to prefer recently extracted juices with fresh taste and minimal flavor or vitamin losses. To meet consumers’ demand, among the novel technologies that involve non-thermal processes, power ultrasound have been investigated as an alternative to conventional heat treatments. Objective of this study was to evaluate the effectiveness of the use of ultrasound in an attempt to maintain the organoleptic characteristics typical of a natural apple juice. In particular, it was evaluated the action on the microflora residing and shelf life of the product through microbiological and sensory analyses. Juice treated with ultrasound highlighted a reduction of aerobic mesophilic counts and psychrophilic bacteria respectively about 3 and 5 log CFU/mL and an enhanced yeast growth. The general opinion expressed by the panelist was in favour of the sonicated juice. This preliminary study showed that non-thermal methods such as power ultrasound technology may give new opportunities to develop fresh-like apple juice.

The role of pulmonary hypertension as a cause of mortality in sickle cell disease (SCD) is controversial.We evaluated the relationship between an elevated estimated pulmonary artery systolic pressure and mortality in patients with SCD. We followed patients from the walk-PHaSST screening cohort for a median of 29 months. A tricuspid regurgitation velocity (TRV)≥ 3.0 m/s cuttof, which has a 67-75% positive predictive value for mean pulmonary artery pressure ≥ 25 mm Hg was used. Among 572 subjects, 11.2% had TRV ≥ 3.0 m/sec. Among 582 with a measured NT-proBNP, 24.1% had values ≥ 160 pg/mL. Of 22 deaths during follow-up, 50% had a TRV ≥ 3.0 m/sec. At 24 months the cumulative survival was 83% with TRV ≥ 3.0 m/sec and 98% with TRV < 3.0 m/sec (p < 0.0001). The hazard ratios for death were 11.1 (95% CI 4.1-30.1; p < 0.0001) for TRV ≥ 3.0 m/sec, 4.6 (1.8-11.3; p = 0.001) for NT-proBNP ≥ 160 pg/mL, and 14.9 (5.5-39.9; p < 0.0001) for both TRV ≥ 3.0 m/sec and NT-proBNP ≥ 160 pg/mL. Age > 47 years, male gender, chronic transfusions, WHO class III-IV, increased hemolytic markers, ferritin and creatinine were also associated with increased risk of death.A TRV ≥ 3.0 m/sec occurs in approximately 10% of individuals and has the highest risk for death of any measured variable. The study is registered in ClinicalTrials.gov with identifier: NCT00492531.

Food is exposed to light during processing, packaging, distribution and retail storage, resulting in deterioration of the product quality. Milk and other dairy products are among the most sensitive due to the high content of riboflavin, vitamin B2, which is an efficient photosensitizer for oxidative processes. Photooxidation in cheese induces degradation of nutritional quality such as proteins, lipids, and vitamins. Moreover, it causes formation of off-flavours, off-odours and gradual bleaching. The objective of this study was to evaluate the effect of light exposition on sliced Asiago d’allevo cheese (lowland vs alpine) aerobically packaged stored for 336 h under both fluorescent light and in the dark. Alpine cheese showed higher b* (Pa* indexes (P

The intensity of hemolytic anemia has been proposed as an independent risk factor for the development of certain clinical complications of sickle cell disease, such as pulmonary hypertension, hypoxemia and cutaneous leg ulceration. A composite variable derived from several individual markers of hemolysis could facilitate studies of the underlying mechanisms of hemolysis. In this study, we assessed the association of hemolysis with outcomes in sickle cell anemia. A hemolytic component was calculated by principal component analysis from reticulocyte count, serum lactate dehydrogenase, aspartate aminotransferase and total bilirubin concentrations in 415 hemoglobin SS patients. Association of this component with direct markers of hemolysis and clinical outcomes was assessed. As primary validation, both plasma red blood cell microparticles and cell-free hemoglobin concentration were higher in the highest hemolytic component quartile compared to the lowest quartile (P≤0.0001 for both analyses). The hemolytic component was lower with hydroxyurea therapy, higher hemoglobin F, and alpha-thalassemia (P≤0.0005); it was higher with higher systemic pulse pressure, lower oxygen saturation, and greater values for tricuspid regurgitation velocity, left ventricular diastolic dimension and left ventricular mass (all P

The performance of two-dimensional electrophoresis in conventional gels in Cartesian coordinates (2-DE) vs. polar coordinates (2-PE) is here evaluated. Although 2-DE is performed in much longer Immobiline gels in the first dimension (17 cm) vs. barely 7-cm in 2-PE, an equivalent resolving power is found. Moreover, due to the possibility of running up to seven Immobiline strips in the radial gel format, the reproducibility of spot position is seen to be higher, this resulting in a 20% higher matching efficiency. As an extra bonus, strings of "isobaric" spots (i.e. polypeptides of identical mass with different pI values) are more resolved in the radial gel format, especially in the 10 to 30 kDa region, where the gel area fans out leaving extra space for spot resolution. In conclusion, this novel gel format in the second dimension of 2D gels is seen as an important improvement of this technique, still one of the most popular in proteome analysis.

The research was carried out on Asiago d’Allevo cheese samples produced in a single farm located in the Altopiano dei Sette Comuni (above 1000m a.s.l.). After 6-12-18 and 36 months of ripening, samples were analyzed for quality traits, in order to evaluate the effect of ripening on colour and gross composition. As expected crude protein and fat significantly increased through the considered period. Ripening led to a significant decrease of L*, a* and b* values. Lightness showed a negative relationship with crude protein, meanwhile a* and b* were both negatively related to fat content. Concerning L*, the trend could be explained by water loss and N-soluble compounds concentration, which could alter protein matrix. a* and b* reduction was probably related to degradation processes such as lipolysis, which seemed to be extensive in hard and long ripened cheese. Ripening affected significantly the light reflectance at all λ with green cheese having the higher values.