Fish freshness consists of complex endogenous and exogenous processes; therefore, the use of a few parameters to unravel illicit practices could be insufficient. Moreover, the development of strategies for the identification of such practices based on additives known to prevent and/or delay fish spoilage is still limited. The paper deals with the identification of the effect played by a Cafodos solution on the conservation state of sea bass at both short-term (3 h) and long-term (24 h). Controls and treated samples were characterized by a multi-omic approach involving proteomics, lipidomics, metabolomics, and metagenomics. Different parts of the fish samples were studied (muscle, skin, eye, and gills) and sampled through a non-invasive procedure based on EVA strips functionalized by ionic exchange resins. Data fusion methods were then applied to build models able to discriminate between controls and treated samples and identify the possible markers of the applied treatment. The approach was effective in the identification of the effect played by Cafodos that proved to be different in the short- and long-term and complex, involving proteins, lipids, and small molecules to a different extent.

The recombinant bovine somatotropin (rbST) is a synthetic hormone developed to mimic the effects of the endogenous growth hormone, also known as bovine somatotropin (bST). Although rbST use in dairy cows is authorized in several countries, it is currently banned in Europe. Different methods for screening and confirmatory detection of rbST were developed, mainly based on LC-MS/MS and immune-enzymatic assays. However, some commercial forms of rbST have above the same amino acid sequence of bST, making it difficult to produce a reliable differentiation of recombinant from endogenous forms. Complementary strategies for indirect detection of rbST can therefore be considered as alternative biomarker-based tools. Untargeted transcriptomics was applied to characterize the microRNAs (miRNA) isolated from milk extracellular vesicles (EVs) in rbST-treated animals, aiming the identification of non-coding biomarkers related to its administration. Sequencing analysis of 63 archive samples collected during previous animal trial allowed for the identification of 35 differentially expressed (DE) miRNAs. A validation study performed by qPCR on a further 70 milk samples from a field survey confirmed the significant upregulation of bta-miR-10167-3p in milk EV from rbST-treated cows. The results obtained suggest the potential use of bta-miR-10167-3p as a non-invasive biomarker to be considered in novel screening strategies, needed to tackle rbST misuse in dairy cows.

Global fish and seafood consumption is increasing annually, frequently leading to the emergence of food fraud, mainly related to mislabeling and adulteration like, for example, the use of illicit/unauthorized food additives to mask or delay fish spoilage. Among the available diagnostic tools for control purposes, spectroscopic techniques have often been proposed to identify these kinds of illicit practices in fish and seafood products. The presented study aims to test two cheap and portable near infrared (NIR) spectrometers, a handheld MicroNIR and a pocket-sized SCiO, to uncover use of the illicit food additive Cafodos, a mixture of sodium citrate and hydrogen peroxide used to preserve some fish characteristics (like smell, color, na dtexture). The NIR spectroscopy in combination with chemometric approaches, allowed the successfully classification of (81–100%) samples of sea bass (Dicentrarchus labrax) treated with Cafodos. The study highlights the potential of this technique that, by not requiring pre-treatment of samples with further reagents, is cheaper and safer for the environment. In conclusion, the study confirmed the potential of portable devices for rapid NIR spectroscopy analysis to identify food fraud and ensure consumer safety.

The use of anabolic–androgenic steroids (AASs) as growth promoters in farm animals is banned in the European Union, representing both an illicit practice and a risk for consumer health. However, these compounds are still illegally administered, often in the form of synthetic esters. This work aimed to characterize significant coding RNA perturbations related to the illicit administration of testosterone and nandrolone esters in fattening pigs. A total of 27 clinically healthy 90-day-old pigs were randomly assigned to test and control groups. Nine animals were treated with testosterone esters (Sustanon^®) and other nine with nandrolone esters (Myodine^®). At the end of the trial, liver samples were collected and analyzed using RNAseq, allowing the identification of 491 differentially expressed genes (DEGs). The transcriptional signature was further characterized by a smaller sub-cluster of 143 DEGs, from which a selection of 16 genes was made. The qPCR analysis confirmed that the identified cluster could still give good discrimination between untreated gilt and barrows compared to the relative testosterone-treated counterparts. A conclusive field survey on 67 liver samples collected from pigs of different breeds and weight categories confirmed, in agreement with testosterone residue profiles, the specificity of selected transcriptional biomarkers, showing their potential applications for screening purposes when AAS treatment is suspected, allowing to focus further investigations of competent authorities and confirmatory analysis where needed.

The increasing consumption of raw and minimally processed fish products has raised concerns regarding the risk of anisakiasis, the infection caused by ingesting larvae of the Anisakis genus. Freezing is currently the standard control measure; however, alternative non-thermal technologies are being explored to preserve product quality while ensuring safety. Several studies have investigated the impacts of high-pressure processing (HPP) on seafood products, but limited information is available about the minimum effective pressure required to achieve complete inactivation of Anisakis larvae while maintaining fillet quality. Moreover, no studies have evaluated the use of portable near-infrared (NIR) spectroscopy as a rapid tool to authenticate HPP-treated fish products. This study evaluated the efficacy of HPP in inactivating Anisakis spp. larvae in gilthead sea bream (Sparus aurata) fillets and investigated the impact of treatment on physicochemical quality parameters. In addition, the reliability of portable NIR spectroscopy coupled with chemometrics was assessed for rapid discrimination between treated and untreated samples. HPP treatments were applied with different pressure–time combinations, and the treatment at 200 MPa for 5 min was selected as the optimal treatment since it was able to achieve 100% larval inactivation. Quality evaluation showed significant changes in color (increase in L* values) and texture parameters, consistent with pressure-induced denaturation, while lipid oxidation remained within acceptable limits. NIR spectra analysis combined with chemometrics approach allowed discrimination between not treated and HPP-treated fillets with an overall accuracy of 98%. The results demonstrate that HPP at moderate pressure levels represents a promising alternative to freezing for Anisakis larvae inactivation in farmed sea bream, and that portable NIR spectroscopy may serve as a rapid, non-destructive tool for on-site verification of treatment. This combined approach could support the development of innovative control strategies in seafood safety management

Mechanically separated meat (MSM) is a product of animal origin obtained by applying mechanical force to remove residual meat from bones, a process that results in the loss or modification of muscle fiber structure. Its use is regulated in the European Union. Clearly distinguishing MSM from non-MSM is essential not only to maintain high product standards but also to ensure full compliance with food labeling regulations and to meet consumer expectations for transparency. This study aimed to evaluate a standardized histological protocol for differentiating high-pressure MSM, low-pressure MSM, and non-MSM pork meat using light microscopy, combined with image analysis and statistical evaluation. In total 40 MSM samples (20 high-pressure, 20 low-pressure) and 20 non-MSM samples were analyzed. The histological protocol achieved 100% sensitivity and specificity in distinguishing MSM from non-MSM. Image analysis revealed significant differences between high- and low-pressure MSM in calcium aggregate parameters. The method provides a reliable, cost-effective tool for MSM identification in routine food inspection. However, because the study was conducted exclusively on pork samples, the results should be interpreted within the context of this type of meat. Additional validation across other species and production systems is required to confirm the broader applicability of the method.