Preliminary Investigation towards the Use of Infrared Technology for Raw Milk Treatment.

Infrared (IR) technology offers a promising solution for reducing microbiological loads in various food types while preserving their quality traits, such as flavour. However, research on IR's application in complex matrices is limited. Therefore, our preliminary study aimed to evaluate its effectiveness in sanitizing bovine raw milk. We assessed the bacterial count before and after IR treatment by comparing volatile organic compound profiles via headspace extraction and GC/MS analysis. Our findings showed that higher energy levels led to a greater bacterial reduction. IR85 was the most effective in reducing Coliforms and Enterobacteriaceae in non-homogenised samples, with a reduction ranging from -1.01 to >-2.99 and from -1.66 to -3.09 Log CFU/mL, respectively. IR60 and 70 showed no efficacy, while IR80 had intermediate but still satisfactory effect. IR85 notably affected volatile compounds, particularly increasing hexanal (from 0.08 to 4.21 ng g-1) and dimethyl sulphone (from 10.76 to 26.40 ng g-1), while IR80 better preserved the aroma profile. As a result, only IR80 was tested with homogenised raw milk, demonstrating significant bacterial reduction (from >2.39 to 3.06 Log CFU/mL for Coliforms and from 1.90 to >2.45 Log CFU/mL for Enterobacteriaceae) and maintaining the aroma profile quality.

Evaluation of the Risk from Potentially Toxic Elements (PTEs) in Italy's Most Consumed Processed Fish Products.

In a balanced diet, regular fish consumption provides positive outcomes for human health. On the other hand, the seafood supply chain faces a significant food safety risk due to the presence of potentially toxic elements (PTEs). In the present study, to assess the risk for Italian consumers, the concentrations of five PTEs, namely lead, chromium, cadmium, mercury, nickel, and aluminum, were determined in the three most consumed preserved fish in Italy: tuna (Thunnus albacares, Katsuwonus pelamis), mackerel (Scomber scombrus) and anchovy (Engraulis encrasicolus). Samples were collected from the national market, and the instrumental analysis was performed by inductively coupled plasma mass spectrometry (ICP-MS). The analyzed PTEs were found in all the species that were investigated. However, after considering the target hazard quotient (THQ) and the hazard index (HI), it was observed that the three fish preserves did not pose any risk of chronic toxicity for the average consumer, even at the highest concentrations detected. However, for significant consumers, mercury detected in tuna samples represented almost 90% of the tolerable weekly intake (TWI) reported by the European Food Safety Authority (EFSA), representing a matter of concern for consumers, particularly regarding developmental neurotoxicity, whose HI exceeded 111%. The acute toxicity of nickel was also considered for significant consumers at the highest concentration detected, and the margin of exposure (MOE) calculated was above 7000, much higher than the value of 30 indicated by EFSA. Due to the lack of data on non-professional carcinogenicity or human intake through foods with low cancer risk, this toxicity was not considered in the analysis of PTEs.

Plastic Contamination in Seabass and Seabream from Off-Shore Aquaculture Facilities from the Mediterranean Sea.

We characterized the presence of plastics in different organs of the gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax) from some off-shore aquaculture facilities of the Mediterranean Sea. Plastics were detected in 38% of analyzed fish. Higher contamination was observed in fish from Turkey and Greece with respect to Italy, without significant differences between the geographical areas. Plastics accumulated mostly in the gastrointestinal tract and, to a lower extent, in the muscle, which represents the edible part of fish. Based on the particle detected, a maximum amount of 0.01 plastic/g wet weight (w.w.) can occur in muscles, suggesting a low input for humans through consumption. A large portion of the particles identified was represented by man-made cellulose-based fibers. The characterization of the polymeric composition suggests that plastics taken up by fish can have land-based and pelagic origins, but plastics can be introduced also from different aquaculture practices.

Incidence of Perfluoroalkyl Substances in Commercial Eggs and Their Impact on Consumer's Safety.

Eggs play an important role in a balanced diet; however, the European Food Safety Authority (EFSA) recognizes eggs as a major source of poly and per-fluoroalkyl substances (PFASs). In this study, the presence of PFASs was analysed in eggs produced by hens from Northern Italian regions, a PFASs-contaminated area. Sixty-five samples were analysed by high-performance liquid chromatography coupled with high-resolution mass spectrometry. The greatest presence of PFASs was found in eggs from Veneto and Emilia Romagna, and the most detected PFASs were perfluorobutanoic acid (PFBA) and perfluorooctanesulfonic acid (PFOS) (mean concentrations 0.30 ± 0.15 and 0.05 ± 0.00 ng g-1). Considering the most recent updates for the sum of the main four PFASs, the highest concentration found in the analysed samples was 0.05 ng g-1, well below the maximum limit set by the European Union. The PFAS intake evaluation confirmed that egg consumption does not represent a risk for Italian consumers.

Predicting the duration of action of ß2-adrenergic receptor agonists: Ligand and structure-based approaches.

Agonists of the ß2 adrenergic receptor (ADRB2) are an important class of medications used for the treatment of respiratory diseases. They can be classified as short acting (SABA) or long acting (LABA), with each class playing a different role in patient management. In this work we explored both ligand-based and structure-based high-throughput approaches to classify ß2-agonists based on their duration of action. A completely in-silico prediction pipeline using an AlphaFold generated structure was used for structure-based modelling. Our analysis identified the ligands' 3D structure and lipophilicity as the most relevant features for the prediction of the duration of action. Interaction-based methods were also able to select ligands with the desired duration of action, incorporating the bias directly in the structure-based drug discovery pipeline without the need for further processing.

Monitoring Volatile Organic Compounds and Aroma Profile of Robinia pseudoacacia L. Honey at Different Storage Temperatures during Shelf Life.

Bee honey has different volatile organic compound profiles that depend on the botanical origin and the state of conservation and which are mainly responsible for its specific aroma. During honey storage, the profile of these molecules and other indicators, such as 5-hydroxymethylfurfural and the diastatic index, can change depending on temperature and time. This study analyzed the variations that these parameters in acacia honey stored at three different temperatures for a total period of 550 days, using gas chromatography coupled with mass spectrometry and an electronic nose equipped with 10 different sensors. The results confirm that the composition of acacia honey varies over time due to both the reduction in the concentration of volatile molecules (e.g., formic acid, a natural acaricide) and the increase in compounds resulting from heat-dependent degradations (e.g., 5-hydroxymethylfurfural). This study supports the usefulness of the electronic nose for the early detection of aromatic alterations in honey subjected to high-temperature storage.

Incidence of Potentially Toxic Elements and Perfluoroalkyl Substances Present in Canned Anchovies and Their Impact on Food Safety.

Fish plays a key role in a healthy and balanced Italian diet, but it is also subject to the bioaccumulation of different contaminants depending on the geographical or anthropogenic context from which it is derived. In recent years, the European Food Safety Authority (EFSA) has been focusing its attention on consumer toxicological risk, considering emerging contaminants such as perfluoroalkyl substances (PFASs) and potentially toxic elements (PTEs). Regarding fish, anchovies are among the five small pelagic main commercial species in the European Union and the top five fresh species consumed by households in Italy. Considering the lack of data on PFASs and PTEs in this species, our aim was to investigate the mentioned contaminants in salted and canned anchovies collected over 10 months from different fishing areas, even those far apart, to verify possible variations in bioaccumulation and to consider the risk for the consumer. According to our results, the assessed risk was very reassuring also for large consumers. The only concern, related to Ni acute toxicity, also dependent on the different consumers' sensitivity, was related to only one sample.

In Vitro Evaluations and Comparison of the Efficacy of Two Commercial Products Containing Condensed Tannins and of Saifoin (Onobrychis viciifolia Scop.) Hay against Gastrointestinal Nematodes of Goats.

Gastrointestinal nematodes (GINs) is a limiting health factor for dairy goats, and the integration of the diet with fodder containing condensed tannins (CT) is becoming increasingly important to control GINs. To preliminary evaluate their potential role as part of GIN control in goat breeding, the in vitro anthelmintic efficacy of the CTs of Silvafeed BYPRO (SBP), Silvafeed Q powder (SQ), and sainfoin hay (SH) was evaluated, and the untargeted metabolomics profiling of the selected formulations was performed. CTs were extracted in water and in ethanol, their concentration was determined, and their chemical characterization was conducted using High-Performance Liquid Chromatography (HPLC) coupled to High-Resolution Mass Spectrometry (HRMS) platform. The in vitro anthelmintic activity of the extracts was evaluated using the Eggs Hatch Test (EHT) and the Larval Migration Inhibition Test (LMIT) using different extract concentrations (150, 300, 600, and 1200 µg/mL). The metabolomic profile of the ethanol extract showed a high number of flavonoids, while the water extract showed higher levels of hydrolysable tannins. The ethanol extracts were effective on both eggs hatching and larvae migration at low concentrations (150 µg/mL) for the three analyzed samples, while the water extracts showed more varied results: SH showed the greatest ovicidal efficacy (concentration 150 µg/mL, %IH = 40.9), while SBP and SQ were more effective against the larvae migration (concentration 600 µg/mL, %LMI = 69.7% and 88%), respectively. The integration of CT-rich fodder into the diet may be considered for the control of GIN infection in goats.

Mining the Protein Data Bank to inspire fragment library design.

The fragment approach has emerged as a method of choice for drug design, as it allows difficult therapeutic targets to be addressed. Success lies in the choice of the screened chemical library and the biophysical screening method, and also in the quality of the selected fragment and structural information used to develop a drug-like ligand. It has recently been proposed that promiscuous compounds, i.e., those that bind to several proteins, present an advantage for the fragment approach because they are likely to give frequent hits in screening. In this study, we searched the Protein Data Bank for fragments with multiple binding modes and targeting different sites. We identified 203 fragments represented by 90 scaffolds, some of which are not or hardly present in commercial fragment libraries. By contrast to other available fragment libraries, the studied set is enriched in fragments with a marked three-dimensional character (download at 10.5281/zenodo.7554649).

One class classification for the detection of ß2 adrenergic receptor agonists using single-ligand dynamic interaction data.

G protein-coupled receptors are involved in many biological processes, relaying the extracellular signal inside the cell. Signaling is regulated by the interactions between receptors and their ligands, it can be stimulated by agonists, or inhibited by antagonists or inverse agonists. The development of a new drug targeting a member of this family requires to take into account the pharmacological profile of the designed ligands in order to elicit the desired response. The structure-based virtual screening of chemical libraries may prioritize a specific class of ligands by combining docking results and ligand binding information provided by crystallographic structures. The performance of the method depends on the relevance of the structural data, in particular the conformation of the targeted site, the binding mode of the reference ligand, and the approach used to compare the interactions formed by the docked ligand with those formed by the reference ligand in the crystallographic structure. Here, we propose a new method based on the conformational dynamics of a single protein-ligand reference complex to improve the biased selection of ligands with specific pharmacological properties in a structure-based virtual screening exercise. Interactions patterns between a reference agonist and the receptor, here exemplified on the ß2 adrenergic receptor, were extracted from molecular dynamics simulations of the agonist/receptor complex and encoded in graphs used to train a one-class machine learning classifier. Different conditions were tested: low to high affinity agonists, varying simulation duration, considering or ignoring hydrophobic contacts, and tuning of the classifier parametrization. The best models applied to post-process raw data from retrospective virtual screening obtained by docking of test libraries effectively filtered out irrelevant poses, discarding inactive and non-agonist ligands while identifying agonists. Taken together, our results suggest that consistency of the binding mode during the simulation is a key to the success of the method.

Presence of Chlorate and Perchlorate Residues in Raw Bovine Milk from Italian Farms.

Chlorates and perchlorates are endocrine disruptors and emerging environmental contaminants found in various types of foods, including milk and dairy products. The presence of chlorate has been associated with the use of chlorine-based sanitizers to disinfect equipment and water used in food processing. Perchlorate, on the other hand, occurs naturally in the environment but is also released from anthropogenic sources. To protect consumers, the EU set an MRL for chlorate in milk but not for perchlorate. Considering that data on chlorates and perchlorates in this field are limited, the objective of this study was to assess the presence of these two anions in 148 samples of raw bovine milk collected from different farms of Lombardy and grouped in three different geographical zones. Chlorate was detected in 73% of the samples analyzed, at concentrations ranging from

Organohalogenated Substances and Polycyclic Aromatic Hydrocarbons in Fish from Mediterranean Sea and North Italian Lakes: Related Risk for the Italian Consumers.

The primary source of persistent organic pollutant (POP) exposure is food, especially fish. European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata) are among the most eaten sea fish in Italy. Fish from lakes in Northern Italy, such as agone (Alosa agone), represent niche consumption for most people, but possibly constitute a much larger percentage of overall consumption volume for local residents. This study dealt with the presence of POPs in the above-mentioned fish species via GC-MS/MS analysis. None of the analytes for which maximum limits are in place showed concentrations above those limits. Moreover, none of the substances without maximum limits exceeded the provisional tolerable daily intake (PTDI) when given, nor did they exceed the more general values considered safe, even for 99th percentile consumers.

Milk Quality and Safety in a One Health Perspective: Results of a Prevalence Study on Dairy Herds in Lombardy (Italy).

Mastitis is one of the major diseases of dairy cows that affects milk quality and quantity and increases the potential risk for the presence of antimicrobial residues (AR) in milk, which could lead to the development of antimicrobial resistance (AMR) among human pathogens. Even if the presence of AR in milk and milk products is low in many countries, the threat is not negligible and cannot be ignored. These problems may be investigated by applying a One Health approach, and this prevalence study aimed to estimate the risks for human health related to milk production applied to dairy herds in Lombardy. Three hundred thirty-one bulk tank milk samples were randomly collected and analyzed by CombiFoss 7 and MilkoScan 7 (milk quality, bacteria, and somatic cell count), an HPLC system coupled to a Q-Exactive Orbitrap (AR), and qPCR (contagious pathogens). The data were analyzed by a generalized linear model. The results showed a relatively high prevalence of contagious pathogens (S. aureus 28.1%; Str. agalactiae 7.3%; M. bovis 3%), which primarily affect milk nutritional components decreasing mainly milk fat content (range 1%-2.5%), but did not show them to be associated to an increase of the risk of antimicrobial residues. These latter ones were recovered only in 7/331 samples at concentrations far below official MLRs. The results support currently active surveillance programs' efficacy in reducing AR risks, which may be further improved by prioritizing them based on geographical area characteristics.

Trends and potential human health risk of trace elements accumulated in transplanted blue mussels during restoration activities of Flekkefjord fjord (Southern Norway).

The monitoring of contaminants represents a priority to preserve the integrity of marine ecosystems, as well as to plan and to manage restoration activities in order to protect environmental and human health. In the present study, a 6-months active biomonitoring was performed to explore the levels of eighteen trace and toxic elements, including heavy metals (TEs; i.e. Al, As, Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Sr, Ti, and Zn), accumulated in soft tissues of blue mussel (Mytilus edulis Linnaeus, 1758) individuals transplanted at different depths (5- and 15-m depth) in five locations within the Flekkefjord fjord (Southern Norway). As this area suffered a long-lasting contamination due to both organic and inorganic contaminants, a series of restoration activities were activated to tackle and to prevent potential risks for ecosystem and local population. Our results demonstrated that the levels of TEs accumulated in edible tissues of transplanted mussels in the Flekkefjord fjord were generally low before the beginning of the restoration activities. However, location- and time-specific differences in the accumulation of TEs were noted after the implementation of such activities. Interestingly, the levels of Fe and Mn significantly increased after the beginning of the restoration activities, likely because the release of these TEs from the slag used in such operations and/or resuspension of contaminated sediments. However, assuming that native mussels can accumulate the same TEs at levels measured in transplanted individuals, our results suggest a substantial safety for human consumption of native mussels from the Flekkefjord fjord, regardless of restoration activities.

Lipidomics profile of irradiated ground meat to support food safety.

Meat irradiation is considered as an effective treatment that expose the advantageous effects on meat preservation. This research, based on untargeted LC-HR orbitrap MS-based lipidomics strategy was meant to estimate the alterations in lipid profile of irradiated chicken, turkey and mixed (chicken, turkey and pork) ground meat in order to evaluate if exists any food safety issue concerning the lipidome alteration. Special attention was paid on oxidation triggered by irradiation. All three matrices exhibited a characteristic lipidome profile which was affected differently by five levels of irradiation intensity. Overall, 345 lipids categorized into 14 subclasses were identified. Remarkably, the oxidized glycerophosphoethanolamines and oxidized glycerophosphoserines were identified in irradiated turkey meat, while for all three categories a characteristic diacylglycerols profile was recognised. Our analytical approach highlighted that the estimation of qualitative variations in lipid portion might be valuable in food inspection purposes, especially when the samples from animal origin are suspected on irradiation treatment.

Undeclared (Poly)phosphates Detection in Food of Animal Origin as a Potential Tool toward Fraud Prevention.

(Poly)phosphates are approved as water-preserving and emulsifying agents that improve the appearance and consistency of many food products. The labelling of added (poly)phosphates is essential for protecting vulnerable population groups and to prevent unfair trade practices resulting in economic fraud. The problems with (poly)phosphates' utilisation concerns both analytical and legislative issues, such as: (1) their straightforward detection; (2) excessive addition altering freshness perception and misleading consumers; (3) uncontrolled usage increasing foodstuff weight; (4) application in products where they are not permitted; and (5) no indication on the label. Bearing all these issues in mind, the main purpose of this study was the quantification and screening of the (poly)phosphates profile in meat, marine and dairy products (160 samples), of which 43 were without declared (poly)phosphate treatment. Analysis was completed by high-performance ion-exchange chromatography either with conductometric detection or coupled to Q-Exactive Orbitrap high-resolution mass spectrometry. Although the (poly)phosphates profiles varied greatly according to species and processing type, the following criteria for detection of illicit treatment were established: high orthophosphate level, quantified short-chain (poly)phosphate anions and the presence of long-chain forms. In conclusion, the instrumental platforms used in this study can be recommended to inspection bodies as reliable methods for the detection of food adulteration with (poly)phosphates.

Effect of High-Pressure Processing on Physico-Chemical, Microbiological and Sensory Traits in Fresh Fish Fillets (Salmo salar and Pleuronectes platessa).

High-pressure (HP) treatment could lead to several advantages when applied to fish and seafood since it would affect the extension of the shelf life of this highly perishable food. In this regard, this study aimed to evaluate the effect of high-pressure treatment (500 MPa for 2 min at a temperature of 4 °C) on changes in quality on two different kinds of fresh fish fillets (Salmo salar and Pleuronectes platessa). Specifically, physico-chemical (VOCs, untargeted metabolomics spectra, pH and color), microbiological (Enterobacteriaceae, Pseudomonas spp., mesophilic and psychrotrophic bacteria) and sensory traits were evaluated at different days of refrigerated storage. From the results obtained, it is possible to state that the high pressure significantly (p ≤ 0.05) reduced microbial growth for each investigated microorganism. Regarding the colorimetric coordinates, no remarkable effects on a* and b* indices were found, while a significant effect (p = 0.01) was observed on the colorimetric index L*, making the HP-treated samples lighter than their respective controls. The sensory analysis showed that for the odor attribute, the HP treatment seems to have had a stabilizing action during shelf-life. Moreover, the treated samples obtained a better score than the respective controls (p ≤ 0.05). With regards to texture and appearance attributes, the treatment seems to have had a significant (p ≤ 0.05) effect, making the treated samples more compact and opaque than controls, therefore resulting in the loss of the characteristics of raw fish for the treated samples. Moreover, from a chemical point of view, HP treatment prevents the development of volatile sulfides and delays the formation of histamine (p ≤ 0.05). Very interestingly, the metabolomic approach revealed novel dipeptide markers for the HP procedure.

Determination of Carbohydrates in Lactose-Free Dairy Products to Support Food Labelling.

Milk and its derivatives are the basis of human nutrition since childhood. Given their importance within a balanced diet, the determination of carbohydrates in milk and its derivatives is fundamental for two reasons: one of alimentary origin related to intolerances and the other one of technological origin, especially for PDO (Protected Designation of Origin) products. The dietetic approach, including lactose-free products, has a crucial role in the management of lactose intolerance, but labelling and compositional rules indicating the absence or reduced presence of lactose in food are currently not harmonised at Union level. Considering the above-mentioned issues and the absence of official methods for the determination of low sugar concentrations, we propose a new and simple IC-HRMS method to detect carbohydrates in milk and different lactose-free derivatives, which can allow the definition of concentration limits useful to characterise products suitable for specific dietary regimes.

Histamine Control in Raw and Processed Tuna: A Rapid Tool Based on NIR Spectroscopy.

The present study was designed to investigate whether near infrared (NIR) spectroscopy with minimal sample processing could be a suitable technique to rapidly measure histamine levels in raw and processed tuna fish. Calibration models based on orthogonal partial least square regression (OPLSR) were built to predict histamine in the range 10-1000 mg kg-1 using the 1000-2500 nm NIR spectra of artificially-contaminated fish. The two models were then validated using a new set of naturally contaminated samples in which histamine content was determined by conventional high-performance liquid chromatography (HPLC) analysis. As for calibration results, coefficient of determination (r2) > 0.98, root mean square of estimation (RMSEE) ≤ 5 mg kg-1 and root mean square of cross-validation (RMSECV) ≤ 6 mg kg-1 were achieved. Both models were optimal also in the validation stage, showing r2 values > 0.97, root mean square errors of prediction (RMSEP) ≤ 10 mg kg-1 and relative range error (RER) ≥ 25, with better results showed by the model for processed fish. The promising results achieved suggest NIR spectroscopy as an implemental analytical solution in fish industries and markets to effectively determine histamine amounts.

Antibiotics and Non-Targeted Metabolite Residues Detection as a Comprehensive Approach toward Food Safety in Raw Milk.

Antibiotic residues in milk are a serious health and technological problem in dairy processes. This study aims to verify the absence of administered antimicrobials after therapeutic treatments, taking into consideration the withdrawal period, and to evaluate the reliability of screening tests under field conditions after confirmatory HPLC-HRMS (High Performance Liquid Chromatography-High-Resolution Mass Spectrometry) Orbitrap analysis. Moreover, the presence of expected or non-targeted metabolites was investigated using the new Compound Discoverer approach. The presence of antimicrobial drugs was shown in 29% of the samples, and also sometimes their metabolites (for enrofloxacin and lincomycin), despite the fact that samples were collected at the seventh milking. Moreover, in 9% of the samples, undeclared treatments were revealed due to the presence of both parent drugs and metabolites. Lastly, the putative identification of two new enrofloxacin metabolites, ENRO-N-methylacetamide and ENRO-ornithine, was proposed. In the light of this evidence, it must be borne in mind that metabolites, some of which are pharmacologically active, may also pose a risk to consumers and for the entire processing of milk in the cheese industries.