Background The administration of antibiotics to food-producing animals, including bees, is a common practice employed to prevent or treat bacterial diseases. Such practices may result in the presence of veterinary residues in honey, potentially compromising the safety and health of consumers. In the European Union, maximum residue limits (MRLs) have not been established for pharmacological substances in honey. In this context, the development of accurate and robust analytical methodologies is of paramount importance for appropriate risk assessment.Objective This study aimed to develop and optimize a multi-class method for the determination of veterinary drug residues from seven classes (beta-lactams, lincosamides, macrolides, quinolones, sulfonamides, tetracyclines, and diaminopyrimidines) using ultrahigh-performance liquid chromatography coupled to time-of-flight mass spectrometry (UHPLC-ToF-MS).Methods A preliminary assessment was conducted to evaluate the feasibility of solid-liquid extraction, given the complex matrix of compounds present in honey, by analyzing different solvent solutions in acetonitrile, methanol, water, and/or McIlvaine buffer. The subsequent phase of the study involved the optimization of a purification/cleanup method. This was achieved by comparing the efficacy of several solid-phase extraction (SPE) and QuEChERS protocols.Results The results indicated that the most efficacious extraction were acetonitrile-based solutions, specifically acetic acid 1% and formic acid 0.1% in ACN-H2O (80:20, v/v). Finally, the method with the most optimal analytical performance in terms of recovery and matrix effect entailed an initial extraction step with formic acid 0.1% in ACN-H2O (80:20, v/v), followed by a modified QuEChERS protocol.Conclusions Acidified organic solvents in combination with QuEChERS method revealed to be the most effective for the determination of antibiotics in honey, ensuring optimum extraction with minimum matrix interference.Highlights The developed analytical protocol with further detection by high-resolution mass spectrometry will be important to assess health risks and to add value to the consumption of honey and honey-based products.

Background/Objectives: Water scarcity is driving the development of strategies for treating municipal wastewater (MW) to enable its safe reuse. Nonetheless, MW contains contaminants of emerging concern (CECs), such as pharmaceuticals and antimicrobial-resistant (AMR) bacteria, which require innovative treatment technologies. In this context, Corbicula fluminea, an invasive freshwater clam, presents a high biofiltration capacity, and its environmental impact could be mitigated by assigning it a beneficial role in wastewater treatment. Methods: The ability of C. fluminea to remove chemical and biological CECs from real MW secondary-treated effluents was assessed. The effects of real wastewater on the clams’ microbiome and on colony-forming unit (CFU) counts in their soft tissues were also assessed. Results: Under real conditions, the clams achieved over 73% removal for 3 chemical CECs after 24 h, with an average removal of approximately 39%. The clams showed recovery of both CFU counts and microbial community composition, dominated by opportunistic and stress-tolerant groups in the presence of pharmaceuticals. The removal of multidrug-resistant bacteria was evaluated; despite real wastewater reducing clearance rates, the clams significantly reduced these bacteria within 24 h. Conclusions: These results demonstrate that C. fluminea can serve as an effective polishing treatment, improving effluent quality, supporting control of this invasive species.

To target pollution remediation efforts and predict pollutant impacts on fisheries and food security, new tools are needed to reconstruct lifetime exposure histories for endangered and commercially valuable fishes. However, current methods used to quantify chemical burden typically use soft tissues which lack temporal information and can miss pulse exposures. Fish eye lenses are proteinaceous structures that form metabolically inert layers (laminae) through life. Their isotopic chronologies are widely used to track lifetime habitat use and diet, but organic pollutants have not previously been quantified in this tissue for fish. Other metabolically inert archival tissues, such as otoliths, are also popular for isotopic life history reconstruction, but almost entirely comprise calcium carbonate with very low protein or lipid content compared with the eye lens, which we show here contain low but measurable levels of lipids, as much as 3.5% w/w in European eels, and so have a greater affinity to bind organic chemicals. To reconstruct pollutant exposure events, a key first step is to show which, if any, chemicals bind to eye lenses. Hence, the present study developed methods to detect organic pollutants in the eye lenses of common dab (Limanda limanda) and European eel (Anguilla anguilla) using GC-MS/MS. For the first time, multiple polycyclic aromatic hydrocarbons were detected in lens tissue, with total concentrations positively correlated to those measured in paired liver samples. However, eye lenses generally contained fewer analytes, likely reflecting a lower bioaccumulation capacity compared with lipid-rich livers. More targeted analysis is needed to achieve lamina-level (chronological) resolution, and controlled exposure experiments to validate relationships with ambient concentrations and establish the underlying pathway(s). Nevertheless, eye lenses show potential as archives of historical organic pollutant exposure.

Diatom communities respond to water pollution with changes in species and abundances. Some contaminants may cause teratologies in cell walls which may be caused by environmental stress. Among others, pharmaceuticals have become an emerging concern in urban streams, presenting risks to the health of ecosystems and humans. This study evaluates the use of diatom communities and their teratologies as indicators of water contamination, namely by pharmaceuticals. Fifteen urban streams in Coimbra, Portugal, were sampled and diatom communities analysed, alongside in situ parameters, nutrients, and 19 pharmaceuticals. Diatom communities were affected by nutrients and organic carbon, and teratologies were detected in over half of the sites, with deformities ranging from 1 % to more than 20 % of the counted diatoms. Tolerant taxa, including Nitzschia amphibia, Craticula subminuscula, Sellaphora nigri, and Sellaphora saugerresii, exhibited high incidences of deformities, even in sites with good biological quality. In the stream with the highest frequency of diatom teratologies, 18 pharmaceuticals were detected, with 14 reaching the highest concentrations observed among all sites. These compounds included stimulants, anti-inflammatories, antibiotics, anticonvulsants, antidepressants, among others. Findings suggest a link between biological health of aquatic organisms and emerging contaminants, especially in contributing to the segregation of sites between the boundaries of good/moderate from poor biological quality. In addition, the results highlight the importance of using diatom deformities as early warning indicators of environmental degradation and risks for One Health, as they were more sensitive indicators of environmental degradation of streams than biological indices’ classifications and community structure analyses.

The increase in livestock and agricultural production has led to the widespread use of veterinary drugs, including antibiotics, antiparasitic, nonsteroidal anti-inflammatory drugs (NSAIDs), hormones, and pesticides, to accelerate productivity and control disease. After excretion, these compounds remain unmetabolized and can contaminate the surrounding environment through uncontrolled use of manure, wastewater, and direct treatment practices. This review focuses on the multiple environmental exposure pathways through which pharmaceuticals and agrochemicals may contaminate ecosystems, particularly emphasizing the impact that this problem may have on the health of pollinators. For example, honeybees can be exposed to pharmaceutical residues through contaminated soil, water, nectar, and pollen, which often leads to acute toxicity and sublethal effects on reproduction, immune function, and behavior. The persistence of these residues in the environment exacerbates antimicrobial resistance, which poses a global threat. In addition, this study reveals different methods of extraction and detection of pharmaceutical residues from honey, encompassing advanced analytical techniques such as liquid chromatography coupled with mass spectrometry detection. This review highlights the chronic and synergistic effects of pharmaceuticals on bee health and emphasizes risk assessment strategies to mitigate the ecological impact of pharmaceuticals on livestock production.

Plant-based meat alternatives (PBMAs) are increasing their consumption, yet their complex formulations may simultaneously contribute to dietary exposure to mycotoxins and provide bioactive polyphenols. In this study, 100 PBMAs commercialised in Spain and Portugal were analysed for 20 mycotoxins and 37 polyphenols by LCMS/MS, and polyphenol bioaccessibility was assessed using the INFOGEST in vitro digestion model. Twelve mycotoxins were detected, with a predominance of emerging Fusarium toxins, ochratoxin A (OTA) and Alternaria toxins (TEA, TTX). MON was the most frequent (89%), followed by ENNB, TTX and OTA (44%, 44% and 43%, respectively). Co-occurrence was frequent, with 70% of products containing >= 2 mycotoxins. Polyphenol profiles varied by ingredient-based group, and TPC bioaccessibility increased during digestion, reaching its highest values in the intestinal phase. Mycotoxins' risk assessment based on middle-bound concentrations considered two intake scenarios: (i) an apparent per capita intake derived from market consumption data and (ii) a portionbased intake reflecting regular PBMA servings (mean and high consumers). No concern was identified under the market-based scenario. Under the portion-based scenario, aflatoxins (B1 and G2) indicated potential concern for both mean and high consumers, while ochratoxin A suggested concern only for high consumers. Overall, PBMAs generally showed low mycotoxin risk. Simultaneously, PBMAs provided bioaccessible polyphenols, supporting integrated assessment of contaminants and bioactives to better characterise consumer exposure.

The increasing global demand for sustainable and nutritious food sources has led to the rapid growth of plant-based meat alternatives (PBMAs) as eco-friendlier replacements of animal-based products. However, the widespread use of cereals, pulses, and nuts in PBMA production raises concerns about mycotoxin contamination. This study validates an analytical workflow for the extraction and determination of 20 mycotoxins in PBMAs, combining an optimized QuEChERS-based extraction method with low- and high resolution mass spectrometry techniques: UHPLC-TOF-MS and UHPLC-QTRAP-MS/MS, respectively. The method was evaluated according to the performance criteria set by the Commission Implementing Regulation (EU) 2023/2782, covering precision, recovery and limits of quantification (LOQ). The results demonstrated high sensitivity for both techniques, with LOQ values below the regulatory limits, ensuring reliable detection of mycotoxins in complex food matrices. The validated method was applied to 12 PBMA burger samples collected from supermarkets in Spain and Portugal. Several mycotoxins, including aflatoxins (B1 and B2), moniliformin, and tentoxin, were detected, ranging 0.3-103 ng g-1. In conclusion, these results highlight the importance of continuous mycotoxin monitoring in PBMAs.

Background/Objectives: Poultry meat is a popular and nutritious food, valued for its high protein content and healthy fat profile. However, like other animal products, it can contain pharmaceutical residues, including coccidiostats, antimicrobials commonly used to prevent parasitic infections caused by Eimeria species. While most monitoring focuses on raw meat, it is important to understand how these compounds behave during cooking to assess potential health risks better and ensure food safety. Methods: This study examined how five different cooking methods (roasting, grilling, and microwaving, beer and wine marinating) affect the levels of eight coccidiostat residues in 45 samples of poultry muscle collected from a supermarket located in the center of mainland Portugal from May to July 2024. After applying different cooking procedures, ionophore and synthetic coccidiostat residue levels were measured using solid-liquid extraction followed by ultrahigh-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). Results are expressed as percentages of the original concentrations: 100% indicates stability, values above 100% suggest a relative increase (often due to moisture loss), and values below 100% reflect a decrease, likely from heat degradation. Results: Roasting, grilling, and microwaving all increased residue concentrations-up to 198.5%, 180.1%, and 158.4%, respectively. In contrast, marinating meat in wine or beer before cooking reduced residues to 73.1% and 72.0%, suggesting a mitigating effect. The initial concentration also influenced the outcome: samples fortified at the maximum residue limit (MRL) had an overall higher mean concentration after cooking (148.3%,) than those fortified at twice the MRL (2 MRL), which averaged 124.5%. Conclusions: These results show that cooking can significantly alter coccidiostat residue levels depending on the cooking procedures and initial concentration. Ongoing monitoring and further research are essential to better understand how cooking affects these residues and their by-products. This knowledge is key to improving food safety practices and refining consumer health risk assessments.

Fumonisins occurrence in maize represents a significant global challenge, impacting economic stability and food safety. This study evaluates the potential of near-infrared (NIR) spectroscopy combined with chemometric algorithms to detect fumonisins in maize. For fumonisin B1 (FB1) and B2 (FB2) levels were developed predictive NIR models using partial least squares (PLS) and artificial neural networks (ANN). PLS models demonstrated strong correlation coefficient (R2) values of 0.90 (FB1), 0.98 (FB2), and 0.91 (FB1 + FB2) for calibration, with ratio of prediction to deviation (RPD) values ranging 2.8-3.6. Similarly, ANN models showed good predictive performance, particularly for FB1 + FB2, with R = 0.99, and the root means square error (RMSE) of 131 μg/kg for calibration; and R = 0.95, RMSE = 656 μg/kg for validation. These findings underscore the efficacy of NIR spectroscopy as a rapid, non-destructive tool for fumonisin screening in maize, with chemometric algorithms enhancing model accuracy, offering a valuable method for ensuring food safety.

Background/Objectives: Antibiotic detection in honey is challenging due to the complexity of this product, the typically low levels of residues, and the absence of Maximum Residue Levels (MRLs) for beehive products. The use of antibiotics in apiculture poses potential risks to human health, including antimicrobial resistance and toxic effects. Reliable, sensitive, and selective analytical methods are essential to ensure food safety and enable accurate monitoring of antibiotic contamination in honey. This study aimed to validate a multi-analyte procedure in accordance with the parameters established in Commission Implementing Regulation (EU) 2021/808 for the identification and quantification of antibiotics, including tetracyclines, lincosamides, quinolones, macrolides, β-lactams, sulfonamides, and diaminopyrimidines. Methods: An extraction protocol was developed using 0.1% formic acid in ACN:H2O (80:20, v/v), followed by a modified QuEChERS with the addition of 1 g NaCl and 2 g MgSO4. The extracts were analyzed by UHPLC-TOF-MS. Results: The method, validated under CIR (EU) 2021/808, demonstrated robust performance, with recoveries ranging from 80.1% to 117.6%, repeatability between 0.5% and 32.2%, reproducibility between 2.3% and 31.6%, and determination coefficients (R2) ranging from 0.9429 to 0.9982. Validation was achieved for 15 antibiotic residues, with CCβ from 3 to 15 μg·kg−1, LODs between 0.09 and 6.19 μg·kg−1, and LOQs between 0.29 and 18.77 μg·kg−1. Application to 10 commercial Portuguese honey revealed no detectable levels of the target antibiotics. Conclusions: The combination of a simplified extraction with UHPLC-TOF-MS provides a reliable approach for the determination of antibiotics in honey. This validated method represents a valuable tool for food safety monitoring and risk assessment of apiculture practices.

Application of antibiotics in beekeeping to treat bacterial diseases can lead to antibiotic residues in honey. This situation can contribute to the broader problem of antibiotic resistance and may cause allergic reactions as well as long-term risks such as carcinogenic effects, gastrointestinal disturbances, aplastic anaemia and reproductive issues. This work aimed at evaluating the safety of honey from the region of Bejaia, in Algeria. A total of ten honey samples sold directly by the producers were analysed. Twenty-four antibiotics were extracted by using a modified QuEChERS methodology and determined by ultra high-performance liquid chromatography coupled with a time-of-flight mass spectrometry (UHPLC-ToF-MS). Method validation yielded satisfactory recovery values, ranging from 82.31 % to 119.54 %. Precision parameters were also within a reasonable range, with minimum values of 0.17 and 3.61 %, and maximum values of 21.78 and 29.87 % for repeatability and reproducibility, respectively. Additionally, detection and quantification limits (LOD and LOQ) ranged from 0.05 to 0.64 mu g kg-1 and 0.14-1.93 mu g kg-1, respectively. Linear regression was excellent with R2 exceeding 0.9664. Finally, 13 out of 24 antibiotic residues were found in the honey samples analysed. Cefazolin, cloxacillin, difloxacin, enoxacin, lincomycin and norfloxacin were detected in all samples. Valnemulin was found in 70 % of the samples. Epi-chlortetracycline, oxolinic acid and sulfadimidine were found in one sample. Ultimately, the findings of the present work demonstrated that the QuEChERS extraction and UHPLC-ToF-MS detection provide a fast, high, and sensitive technique for detecting antibiotics residues in honey.

Pharmaceutical contamination in coastal and marine sediments is an increasing environmental issue. Thousands of biologically active substances continuously enter aquatic ecosystems through wastewater discharge, agricultural runoff, and improper disposal. Due to their persistence and continuous release, these contaminants accumulate in various matrices, including water and sediments, posing long-term ecological risks. Their presence can disrupt microbial communities, bioaccumulate in marine organisms, and heighten the risks of chronic exposure and antimicrobial resistance. To safeguard human, animal, and environmental health, it is crucial to develop accurate methodologies to understand pharmaceutical occurrence profiles and monitor their presence in coastal ecosystems. This study describes the development and validation of a sensitive UHPLC-TOF-MS method for screening and confirming pharmaceuticals in coastal sediments, following Commission Implementing Regulation (EU) 2021/808. Thirty analytes were targeted and evaluated for key performance parameters, including selectivity and sensitivity, linearity, repeatability, reproducibility, trueness, limit of detection (LOD), and limit of quantification (LOQ). Data analysis demonstrated high selectivity and sensitivity, with LODs ranging from 0.02 to 0.23 ng/g and LOQs from 0.06 to 0.69 ng/g. Recovery values at the LOD level ranged from 74.3 % for irbesartan to 115.4 % for indapamide, while repeatability and reproducibility were under 19.5 % and 29.2 %, respectively. All performance criteria met the requirements outlined in European guidelines. To evaluate the method's robustness, it was applied to 12 sediment samples collected from an estuary, with irbesartan detected in 5 samples.

Fungal and mycotoxin control at a primary stage in the food chain is crucial to maintaining the nutritional quality of animal feed. The control of fungal and mycotoxins is one of the essential points that a good biosecurity program must establish to ensure the safe feeding and protection of animal and human health. Acquiring a comprehensive understanding of the role of mycotoxins is vital to identifying breaches of this control and enabling the performance of proper risk assessments and accurate risk management strategies. This study focused on the identification of regulated and emerging mycotoxins in agricultural fields and dairy farms through an analytical methodology by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-QTrap-MS/MS). This allowed us to identify a Portuguese mycotoxin profile in the maize value chain for the first time. Regarding our occurrence data, mycotoxins were identified in all samples, namely seeds, flowering plants, grain and forage at harvest, maize silage, and animal feed. FBs and ENNB were highly present in all stages of the production cycle. This work offers an initial insight into a full screening of regulated and emerging mycotoxins within an important agricultural commodity. The high occurrence of these compounds points to the need to perform occurrence surveys from an integrative perspective to protect consumers’ health, especially within food chains that provide various important staple foods worldwide.

Consumption of misidentified foraged mushrooms containing bicyclic amanitin octapeptides is a worldwide public health and veterinary problem, being considered one of the deadliest accidental human and canine food ingestion due to acute liver failure (ALF). Reversal of advanced ALF and complete clinical recovery can be achieved following definitive removal of accumulated amatoxin laden bile from the gallbladder. An accurate means of quantifying amanitin content in aspirated bile is, therefore, urgently needed. Building on our prior work validating a method to detect and quantify amanitin in hepatic autopsy tissue, the development of an accurate method of measuring α- and β-amanitin in aspirated gallbladder bile was performed to evaluate the efficiency of this emergency procedure applied as a clinical treatment for intoxicated patients. A solid-phase extraction (SPE) procedure was optimized followed by detection based on ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). Low resolution mass spectrometry (LRMS) was compared with high resolution (HRMS) by the validation of UHPLC-MS/MS (triple quadrupole MS) and UHPLC-ToF-MS (time-of-flight MS). Both methods were able to detect amatoxins in bile with limits of detection and quantification ranging from 2.71 to 3.46 µg.kg-1, and 8.36-9.03 µg.kg-1 for α-amanitin and, 0.32-1.69 µg.kg-1 and 0.55-5.62 µg.kg-1 for β-amanitin, respectively. Validation was completed with the evaluation of linearity, specificity, robustness, recovery, and precision following the ICH guidelines and CIR 808/2021. The validated methods were finally applied to bile samples obtained 48-96 hours + post-ingestion from 4 amatoxin poisoning patients who underwent gallbladder drainage procedures in Vietnam, Canada, and California. Gallbladder bile from patients with amatoxin mushroom poisoning contained significant amanitin content, even when aspirated several days post-ingestion, thus confirming the important role of enterohepatic circulation in amatoxin hepatotoxicity. This work represents a high and unique analytical throughput in amanitin poisoning allowing to efficiently respond to this fatal health problem.

Cereal grains are widely cultivated agricultural commodities, and they constitute a key source of energy. However, many of them are affected by fungal contamination, and consequently, by the presence of mycotoxin. The development of monitoring techniques to evaluate exposure, is therefore of interest. Immunoassays have recently emerged as potential alternatives for the study of mycotoxins. This study focuses on a validation of a biochip array technology (BAT) for multi-mycotoxins screening in oat, barley, rye, and wheat grains. The Evidence Investigator Myco 7 (RANDOX Food Diagnostic), based in a competitive chemiluminescent immunoassay, was used for the simultaneous semi-quantitative detection of the mycotoxin's immunoassays. According to validation results, spiked rice samples showed low false results (5 % of false negatives and 5 % of false positives for fumonisins, 5 % of false negatives for Zearalenone, Aflatoxin B1, T2HT2 and Deoxynivalenol and 5 % of false positives were found for ochratoxin A). In the spiked samples of other cereals, were found 5 % of false positives for fumonisin B1+fumonisin B2, zearalenone, ochratoxin A and aflatoxin B1 and 5 % of false negatives were found for Fumonisin 1 + Fumonisin 2, Ochratoxin A, Aflatoxin B1 and T2+HT2 in agreement with European Union legislation performance criteria. This BAT immunoassay provides important benefits for the rapid and efficient screening of several mycotoxins in cereal grain samples at various levels.

Honey is a food of great nutritional importance and has always been used for human consumption. The production of honey and other beekeeping products depends on the proper functioning of this extremely important sector, as it has a direct impact on other sectors such as agriculture. The decline in bee colony numbers has been linked, among other factors, to bacterial diseases affecting bees, including American and European foulbrood, and Nosema spp. disease. In this matter, prophylactic or therapeutic use of veterinary drugs in apiculture is common but can lead to their accumulation in bees and in honey. Consumption of contaminated honey can have adverse effects such as allergic or hypersensitivity reactions, carcinogenicity, reproductive effects, and teratogenicity. Commission Regulation (EU) N⍛ 37/2010 sets MRLs for antibiotics in various foods, but these limits are not set for api-products. The lack of harmonized rules has led some countries to set recommended concentrations and minimum performance limits. Nonetheless, to achieve this goal, development of accurate and precise analytical methodologies is crucial. In recent years, the analysis of antibiotics in honey has led to the development of methods in an extensive range of families, including aminoglycosides, amphenicols, lincosamides, macrolides, nitroimidazoles, quinolones, sulfonamides, tetracyclines and nitrofurans. This review work entails an in-depth exploration of occurrence studies, extraction methodologies, and analytical techniques for the determination of antibiotics in apiculture products. It was found that the most used extraction methods include solid-phase extraction, dispersed solid or liquid phase extraction and QuEChERS. Due to the complexity of the honey matrix, samples are often diluted or acidified using McIlvaine buffer, H2O, MeOH, acidified ACN and TCA solution. This is usually followed by a purification step using SPE cartridges or PSA. Golden analytical methodologies include high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (MS/MS) with Orbitrap or Q-ToF detectors.

Common variable immunodeficiency enteropathy is a sprue-like disease, which may manifest as a severe malabsorption syndrome with nutritional deficits and cachexia. The authors report a case of a 33-year-old Afghan man, who presented to the emergency department due to chronic watery diarrhea and severe malnourishment. He had been previously misdiagnosed with celiac disease in his early adulthood; however, this was based on inconclusive findings. After a thorough diagnostic workup, the final diagnosis of common variable immunodeficiency enteropathy with symptomatic norovirus infection of the gut was obtained during his prolonged hospitalization. A slow but progressive improvement was observed with immunoglobulin replacement therapy, corticotherapy, and ribavirin treatment. This is a noteworthy case of a rare malabsorption disorder, and it reviews important aspects concerning the differential diagnosis of small bowel villous atrophy of unknown etiology, as well as gastrointestinal manifestations of common variable immunodeficiency disorder.

Zinc and copper have been used as growth promotors in alternative to antibiotics in pig's diet. The aim was the ascertainment of the Zn and Cu concentrations in piglets' liver and kidney and their impact in the reduced susceptibility to Zn, Cu, and antibiotics in enterococci, used as microbiota biomarker. Zn and Cu were determined in the livers and kidneys of 43 piglets slaughtered in Portugal, by flame atomic absorption spectrometry. Enterococci were isolated from feces for determining the identification of species (E. faecalis, E. faecium, and Enterococcus spp.); susceptibility to vancomycin, ciprofloxacin, linezolid, tigecycline, ampicillin, imipenem, and metals; and Cu tolerance genes. In piglets with Zn and Cu high or toxic levels, enterococci had reduced susceptibility to ions, reinforced by the presence of Cu tolerance genes and by resistance to antibiotics. The study relevance is to show the relationship between these metals' levels and decreased susceptibility to Cu, Zn, and antibiotics by enterococci. From the results, it could be supposed that the piglets were being fed with high doses of Zn and Cu which could select more resistant bacteria to both antibiotics and metals that could spread to environment and humans.

Residues of antibiotics applied in chicken husbandry can be found in eggs for human consumption. To prevent this issue and avoid adverse effects on consumers' health, such as allergies and antimicrobial resistance (AMR), maximum residue limits (LMR) have been established by the European Commission. The aim of this study was to develop a method to extract, detect and quantify antibiotics in eggs, using UHPLC-TOF-MS for semi-quantitative screening and UHPLC-QTRAP-MS/MS for quantitative confirmation, following the performance criteria established by Commission Regulation (EU) N degrees 2021/808 for analytical method validation. Different extraction methods were analyzed, with liquid-liquid extraction (LLE) based on acetonitrile (ACN) and ethylenediaminetetraacetic acid (EDTA), proving to be the most efficient method for extraction of antibiotic residues from eggs. UHPLC-TOF-MS and UHPLC-QTRAP-MS/MS were validated for 46 compounds, including penicillin, cephalosporins, tetracyclines, quinolones, macrolides, sulfonamides, and other antibiotics. After validation, robustness of both methods was evaluated by analyzing ten egg samples from backyard production and supermarkets. Erythromycin residues were detected in two eggs from backyard production; residues of up to seven antibiotics were detected in two samples of supermarket eggs. Nonetheless, all positive samples were below the established MRL. The developed methods will be a useful tool to monitor possible contamination in eggs, as they allow the simultaneous detection of several molecules. Furthermore, this methodology can be optimized to reduce the relative matrix effect, extended to a larger group of molecules, include other types of drugs, and be validated for other matrices of animal origin.

Ulva, a genus of green macroalgae commonly known as sea lettuce, has long been recognized for its nutritional benefits for food and feed. As the demand for sustainable food and feed sources continues to grow, so does the interest in alternative, plant-based protein sources. With its abundance along coastal waters and high protein content, Ulva spp. have emerged as promising candidates. While the use of Ulva in food and feed has its challenges, the utilization of Ulva in other industries, including in biomaterials, biostimulants, and biorefineries, has been growing. This review aims to provide a comprehensive overview of the current status, challenges and opportunities associated with using Ulva in food, feed, and beyond. Drawing on the expertise of leading researchers and industry professionals, it explores the latest knowledge on Ulva's nutritional value, processing methods, and potential benefits for human nutrition, aquaculture feeds, terrestrial feeds, biomaterials, biostimulants and biorefineries. In addition, it examines the economic feasibility of incorporating Ulva into aquafeed. Through its comprehensive and insightful analysis, including a critical review of the challenges and future research needs, this review will be a valuable resource for anyone interested in sustainable aquaculture and Ulva's role in food, feed, biomaterials, biostimulants and beyond.