Detection of fibrous microplastics and natural microfibers in fish species (Engraulis encrasicolus, Mullus barbatus and Merluccius merluccius) for human consumption from the Tyrrhenian sea.

The occurrence of natural/artificial and synthetic microfibers was assessed in three commercial fish species (Engraulis encrasicolus, Mullus barbatus, Merluccius merluccius) from the Tyrrhenian Sea sold for human consumption. The gastrointestinal tracts of n. 150 samples were analyzed, the isolated microfibers were classified applying a morphological approach, based on the analysis of their morphological features, coupled with the identification of the chemical composition of a subsample of microfibers. All the species contained microfibers at levels ranging from 0 to 49 items/individual and the number of ingested microfibers significantly differed between pelagic and demersal fishes. The evaluation of fiber morphologies highlighted that natural/artificial microfibers were the most numerous among the isolated microfibers, while the dominant colors were blue, black, and clear in all the species. Chemical characterization confirmed the morphological identification and indicated cellulose and polyester as the most common polymer types. Considering the analytical issues that may affect the evaluation of microfiber pollution, the results pointed out the importance of an accurate morphological approach that allows the distinction between different fiber types, before the spectroscopic analyses. Moreover, the implementation of fast and accessible methods to identify microfibers in fish species intended for human consumption will be beneficial also to make an adequate risk assessment to consumer health.

A study on textile microfiber contamination in the gastrointestinal tracts of Merluccius merluccius samples from the Tyrrhenian Sea.

The increased demand and consumption of synthetic textiles have contributed to microplastic pollution in the form of microfibers. These particles are widely spread in the aquatic environment, leading to the exposure of marine biota, including edible species. The current study aimed to assess the extent of microfiber contamination in a commercially relevant fish species, Merluccius merluccius, which is considered a small-scale bioindicator for the monitoring of plastic ingestion in the Mediterranean coastal environment. The frequency of ingestion, abundance, and composition of textile microfibers isolated from the fish gut were characterized. Results showed the occurrence of microfibers in 75% of the samples, with a mean number of 10.6 microfibers/individual, of which 70% were classified as natural microfibers. The spectroscopic analyses confirmed both the visual identification of microfibers and the prevalence of cellulosic fibers. The obtained findings provided evidence of both natural/artificial and synthetic microfiber exposure in an important commercial fish species that, considering the consumption of small individuals without being eviscerated, may be a potential route of microfiber exposure in humans. Monitoring programs for fishery products from markets are needed to assess contamination levels and human health risks. In addition, measures to control microfiber pollution need to occur at multiple levels, from textile industries to international governments.

Impact of Fibrous Microplastic Pollution on Commercial Seafood and Consumer Health: A Review.

The omnipresence of microfibers in marine environments has raised concerns about their availability to aquatic biota, including commercial fish species. Due to their tiny size and wide distribution, microfibers may be ingested by wild-captured pelagic or benthic fish and farmed species. Humans are exposed via seafood consumption. Despite the fact that research on the impact of microfibers on marine biota is increasing, knowledge on their role in food security and safety is limited. The present review aims to examine the current knowledge about microfiber contamination in commercially relevant fish species, their impact on the marine food chain, and their probable threat to consumer health. The available information suggests that among the marine biota, edible species are also contaminated, but there is an urgent need to standardize data collection methods to assess the extent of microfiber occurrence in seafood. In this context, natural microfibers should also be investigated. A multidisciplinary approach to the microfiber issue that recognizes the interrelationship and connection of environmental health with that of animals and humans should be used, leading to the application of strategies to reduce microfiber pollution through the control of the sources and the development of remediation technologies.

Research and characterization of fibrous microplastics and natural microfibers in pelagic and benthic fish species of commercial interest.

The ingestion of synthetic microfibers, the most prevalent type of microplastics in marine environments, and natural fibers was assessed in Engraulis engrasicolus and Mullus barbatus, two commercially important fish species in the Mediterranean Sea. Microfibers were isolated from the fish gastrointestinal tract using a 10% potassium hydroxide solution. For the microfiber characterization, the evaluation of specific morphological features using a light microscope, coupled with the Fourier-transform infrared (FTIR) analysis of a subsample of isolated particles, was applied. The preliminary results showed the occurrence of microfibers in 53 and 60% of European anchovy and Red mullet, respectively. A mean of 6.9 microfibers/individual was detected in anchovies, while on average Red mullet samples contained 9.2 microfibers/individual. The most common colors of fibers in both species were black, blue, and transparent. Visual characterization of fibers allowed the classification of 40% of the items as synthetic microfibers. FTIR spectroscopy confirmed the visual classification by fiber morphology. Microfibers were made of different typologies of polymers, represented by cellulose, cotton, and polyester. These findings confirm as the wide distribution of fibrous microplastics, and natural microfibers may impact both pelagic and deep-sea trophic webs. Despite the presence of microfibers in fish species poses a potential risk to human health, the literature is scarce regarding studies on the uptake by commercial marine fish mostly due to methodological issues. The visual characterization, corroborated by spectroscopic techniques, may be useful to differentiate synthetic and natural fibers, representing a fast and easy method to assess fibrous microplastic pollution in commercially important fish species.

Recent Records on Bacterial Opportunistic Infections via the Dietary Route.

This narrative review was aimed at identifying the opportunistic bacterial pathogens that can be transmitted by contaminated food and represent a current threat for patients particularly susceptible to infections because of underlying conditions or predisposing factors. The analysis was focused on recent case or outbreak reports and systematic reviews published in the years 2019 to 2023 and resulted in sorting 24 bacterial groups comprising the genera or species able to cause a variety of systemic or invasive infections if ingested with food or drinking water. These included both bacteria known to cause mild infections in immunocompetent persons and bacteria considered to be innocuous, which are used in food fermentation or as probiotics. No recent cases of infections transmitted through dietary routes were reported for the critical nosocomial pathogens widely found in food products, primarily Acinetobacter baumannii and Klebsiella pneumoniae. However, the very first sources of their introduction into the clinical environment still need to be established. In many instances, risky dietary habits, such as eating raw fish, seafood, raw meat, unpasteurized milk, and their derived products or the lack of control in fermentation processes, has led to the reported illnesses, pointing out the necessity to improve the hygiene of production and consumer awareness of the risks.

A versatile approach to evaluate the occurrence of microfibers in mussels Mytilus galloprovincialis.

Microplastics of fibrous shape are esteemed to be the most abundant micro-debris form present in the environment. Despite the occurrence of microfibers in fish may pose a risk to human health, the literature is scarce regarding studies on the contamination in commercial marine fish mostly due to methodological issues. In this study, a versatile approach, able to discriminate among natural and synthetic microfibers according to the evaluation of specific morphological features, is proposed in farmed mussels (Mytilus galloprovincialis). The approach was useful to determine that microfibers were present in 74% of mussel samples, with a mean number of 14.57 microfibers/individual, corresponding to 3.13 microfibers/g w.w. A negative correlation between the size of analysed mussels and the amount of microfibers/g w.w. was detected, showing that smaller specimens contained more microfibers than the larger ones. This work paves the way to further studies aimed to adequately assess the risk that microfibers may pose to marine biota, also considering the commercial value as seafood items of many species of the Mytilus genus and the potential implication for human exposure.

Microplastics in Fish and Fishery Products and Risks for Human Health: A Review.

In recent years, plastic waste has become a universally significant environmental problem. Ingestion of food and water contaminated with microplastics is the main route of human exposure. Fishery products are an important source of microplastics in the human diet. Once ingested, microplastics reach the gastrointestinal tract and can be absorbed causing oxidative stress, cytotoxicity, and translocation to other tissues. Furthermore, microplastics can release chemical substances (organic and inorganic) present in their matrix or previously absorbed from the environment and act as carriers of microorganisms. Additives present in microplastics such as polybrominated diphenyl ethers (PBDE), bisphenol A (BPA), nonylphenol (NP), octylphenol (OP), and potentially toxic elements can be harmful for humans. However, to date, the data we have are not sufficient to perform a reliable assessment of the risks to human health. Further studies on the toxicokinetics and toxicity of microplastics in humans are needed.

Study on bisphenol F, a bisphenol A analogue, at a dairy company: Health hazard and risk assessment.

The occurrence of analogues of bisphenol A (BPA), including bisphenol F(BPF) in milk is still not well known. BPF may enter the milk chain at the farm and during milk processing at the dairy company. This study identified the main BPF contamination pathways using a monitoring model based on the identification of the hazard at three stages along the dairy chain: raw milk from the storage tank, pasteurized milk from the storage tank, and cardboard packaged milk. Quantitative analysis was performed by high-performance liquid chromatography with fluorometric detection (HPLC/FD) system. BPF was detected in all analysed stages (from

Extraction and identification of microplastics from mussels: Method development and preliminary results.

Microplastics (MPs) are an emerging threat to marine ecosystems. One of the primary environmental risks is their bioavailability for aquatic organisms. Some fish and bivalves are of particular interest because their feeding strategies expose them to particles present in the water column. The aim of the study was to assess an extraction method in order to isolate and quantify MPs from fish gastrointestinal tract (n.8) and muscle (n.4), and bivalves (n.8) samples. The accuracy of the method was assessed through the calculation of the recovery percentage in samples spiked with a known number of MPs using microscopic observation. Successively, the extraction was preliminarily applied on n.20 mussels collected from mariculture plants of the Tyrrhenian and the Adriatic Sea. The results of the digestion protocol showed an average extraction yield of 80% in fish gastrointestinal tracts, 90% in fish muscle samples, and 95% in mussels. Preliminary analysis carried out on farmed mussels showed an average abundance of 3.8 items/individual, and 0.5 items/g of tissue, among those black, was the most represented color.

Occurrence of bisphenol A in the milk chain: A monitoring model for risk assessment at a dairy company.

Bisphenol A (BPA) as a chemical hazard may enter the milk chain during primary production at the farm and, successively, during milk processing at a dairy company. To identify the potential pathways that play a role in the occurrence of BPA, a monitoring model for risk assessment based on the identification of the hazards at each stage of milk processing was investigated. Milk samples were analyzed using liquid chromatography with fluorescence detection. Quantifiable levels were detected in samples obtained from the raw milk storage tank, pasteurized milk from the storage tank, and packaged milk. The highest BPA contamination levels were detected in raw milk from the storage tank (mean 0.265 µg/L). Despite the fact that dietary exposure levels were below the temporary daily intake, BPA may have adverse effects, particularly for vulnerable population groups. New monitoring programs involving each stage of milk processing should therefore be applied.

Occurrence of potential plastic microfibers in mussels and anchovies sold for human consumption: Preliminary results.

There is a global concern over the impact of microplastics on marine species and trophic webs. Microfibers commonly represent the greater portion of microplastics in the aquatic environment, but little is known about fiber uptake and accumulation by marine biota. The aim of the study was to investigate the potential plastic microfiber contamination in mussels (Mytilus galloprovincialis) and anchovies (Engraulis encrasicolus) from the Tyrrhenian Sea sold for human consumption. Anthropogenic debris was extracted from the digestive tracts of fish and the whole shellfish using a 10% KOH solution and quantified under a light microscope. The preliminary results showed the occurrence of potential plastic and natural microfibers in 73% of the samples. On average mussels contained 1.33 microfibers/g w.w. and 7.66 items/individual, while anchovies contained 9.06 microfibers/individual. Considering that mussels are consumed as a whole, and small pelagic fish, as anchovy, may be eaten without removing the gastrointestinal tract, microfiber contamination may lead to human exposure. More research is required to adequately assess the risk that microplastics, including microfibers, may pose for food safety and human health.

Development of a high-performance liquid chromatography method to assess bisphenol F levels in milk.

Bisphenol F (BPF) is a bisphenol A (BPA) analogue. As an endocrine disruptor, BPF shows a similar BPA hormonal activity and greater endocrine effects. To assess BPF levels in milk a selective method based on solvent extraction with acetonitrile, solid-phase extraction (SPE), high-performance liquid chromatography with fluorescence detection (HPLC-FD) system, was developed. The method showed high recovery values (from 97.60 to 107.16%), and good detection and quantification limits (LOD=0.03 µg/L; LOQ=0.1 µg/L). To validate the analytical method, quantitative analyses of n.20 milk samples of whole milk were preliminarily carried out applying a monitoring system based on the control of different stages of pasteurized whole milk processing at a dairy company. The proposed method is simple, sensitive, and might be suitable to detect BPF residues in milk processing. At the dairy company, the occurrence of BPF levels ranging from

Detection and quantification of Campylobacter in foods: New analytic approaches to detect and quantify Campylobacter spp. in food samples.

The aim of the present study was to develop rapid qualitative and quantitative methods based on the use of Real-Time PCR and Droplet Digital PCR (ddPCR), in order to have reliable techniques to detect and quantify Campylobacter spp. in food samples. The gene 16S-rRNA was used as specific target for Campylobacter spp. Real- Time PCR evaluation assay and a not competitive internal control was ushered in it. To investigate the selectivity of the method, 26 Campylobacter strains and 40 non-Campylobacter strains were tested and in order to verify the application of Real- Time PCR method, 5 pork meat samples were experimentally inoculated with a Campylobacter jejuni strain. Subsequently, dilutions with a bacterial load of Campylobacter jejuni within 10-106 CFU/mL were chosen for the optimization of the ddPCR assay. Lastly, a total of 54 naturally contaminated foods samples were analyzed through molecular (Real-Time PCR and ddPCR) and traditional methods. The Real-Time PCR protocol demonstrated to amplify only the Campylobacter spp. strains and when Campylobacter jejuni was experimentally inoculated in meat samples the pathogen was always detected. The ddPCRs assay allowed to quantify a level of contamination of 10 CFU/mL, but it was unable to quantify levels of 105 - 106 CFU/mL. Lastly, Campylobacter spp. was never detected in the 54 samples tested. In conclusion, the novel analytic approach proposed, based on an initial screening of the samples with Real-Time PCR and then on quantification of Campylobacter spp. with a ddPCR on those positive, represents a quick monitoring tool and, if used correctly, it would allow the implementation of food safety.

Occurrence of Microplastics in Commercial Seafood under the Perspective of the Human Food Chain. A Review.

The occurrence of microplastics in the marine ecosystem and aquatic organisms, their trophic transfer along the food web, and the identification of seafood species as suitable indicators have become a research priority. Despite the high quantity of research in this field, a comparison between the available data and an appropriate risk assessment remains difficult. In this perspective, as an innovative approach, the association of the feeding strategies of commercial seafood and the microplastic level was considered. Further research to assess the occurrence of microplastics in the marine food web, the long-term effects on animals and humans, and the health implications is needed.

Hot topic: Bisphenol A in cow milk and dietary exposure at the farm level.

Chemical hazards may enter the milk chain during primary production. The study, for the first time, investigated the occurrence of bisphenol A (BPA) levels in cow milk samples collected on the farm following manual or mechanical milking and from the cooling tank. We applied a new monitoring model based on the identification of the hazards at each stage of the milk chain to identify potential pathways for contamination along the milk chain. We evaluated exposure to BPA through milk consumption based on detected contamination levels and the temporary tolerable daily intake established by the European Food Safety Authority (EFSA). Milk samples (n = 72) were analyzed using liquid chromatography with fluorescence detection. The mean BPA concentrations were 0.757 µg/L in manually milked samples, 0.580 µg/L in mechanically milked samples, and 0.797 µg/L in milk from the cooling tank. Bisphenol A occurred in the milk chain as a result of different stages of milking, and reached the highest levels at the end of the milk chain. Although the dietary intake of BPA was below the EFSA's temporary tolerable daily intake, exposure to BPA, even at low doses, through milk consumption represents a public health concern. Therefore, to ensure milk safety, new monitoring plans should be applied based on the identification of hazards at each stage of the milk chain.

Study on endocrine disruptors levels in raw milk from cow's farms: Risk assessment.

Diet represents the primary route for human exposure to bisphenol A (BPA). As endocrine disruptor (ED), BPA has raised concerns about its adverse effects on human health. Therefore, EFSA recommended a tolerable daily intake (t-TDI) of 4 µg/kg bw/day and the EU Regulation n. 2018/213 fixed a specific migration limit (SML) of 0.05 mg/kg for BPA in food from plastic materials intended to come in contact with food. BPA could be present in milk due to environmental contamination, and also as a result of the migration from contact materials used during milking and storage. Considering the widespread consumption of milk and milk products, the contamination of dairy products is a matter of public health concern. The aim of the study was to investigate the BPA contamination levels of raw cow's milk from two farms located in Campania region, Italy. The milk samples (n=22), weekly collected from the cooling tank, were analyzed using liquid chromatography with fluorescence detection. In raw milk from both farms, preliminary results showed the occurrence of BPA levels lower than the SML limit, ranging from not detected to 2.34 µg/L. The consumer exposure, calculated considering a hypothetical raw milk consumption and three possible scenarios, was below the t-TDI. Despite the low levels of exposure through milk consumption, low doses can have lasting effects during human development. Thus, new approaches, methods, and plans should be applied to monitor ED contamination, such as BPA and other pollutants, and to assure milk safety.

Investigation on bisphenol A levels in human milk and dairy supply chain: A review.

Bisphenol A (BPA), widely used as additive in food packaging, is an environmental and food contaminant that shows a weak estrogenic activity in general population and toxicity in the infant population. A temporary tolerable daily intake (t-TDI) of 4 µg/kg bw/day and a migration limit of 0.6 mg/kg in food from plastic materials, intended to come in contact with food, were fixed. Dietary milk is important in the human diet. The review investigated the contamination levels in human milk and along the dairy supply chain. Despite the reported levels are generally below the fixed limits, breast milk is considered a continuous low-level exposure to endocrine-active compounds for infants. In addition, BPA residues are detected in milk and dairy products posing a risk to human health. BPA enters into milk chain via multiple pathways at various points during milk production (e.g., PVC tubing used during the milking process, transfer from bulk milk to storage tanks, during milk processing). To prevent or mitigate this hazard, a specific TDI for infants is recommended and evaluation of risk factors at each phase of the dairy supply chain, in the quality systems, is recommended.

Screening of Oxalate Degrading Lactic Acid Bacteria of Food Origin.

A screening for oxalate degrading abilities was initially carried on within Lactic Acid Bacteria cultures of different food origin. Seventy-nine strains were drop-inoculated onto MRS agar plates containing calcium oxalate. By comparing colonies diameters, 31 strains were used to inoculate, in parallel, MRS and MRS modified by sodium oxalate addition. Differences in the strains' growth were assessed by colony forming unit counts. For two strains, the growth in oxalate enriched medium was significantly higher; while, for eleven strains an opposite behaviour was recorded. Two strains - probiotic Lactobacillus rhamnosus LbGG and Enterococcus faecalis 59 - were chosen. The first strain appeared to be able to metabolize oxalate more efficiently than the other tested cultures, while strain 59 appeared unable to gather advantage by oxalates and, indeed, appeared to be inhibited by the salt presence in the medium. Outcomes revealed that higher glucose concentrations may favour oxalates utilization. In MRS with oxalate, but without glucose, citrate was completely metabolized. Evaluation along time confirmed that the oxalate degradation is more significant in presence of glucose. Outcomes may represent a good start for the development of a safe and even probiotic culture able to lower the oxalates content of food.

Preliminary Study on Physicochemical and Biochemical Stress Markers at Poultry Slaughterhouse.

Pre-slaughter stress can result in variations in the glycogen storage and metabolic changes of muscle, responsible for quality poultry meat. Aim of this study was to investigate, as pre-slaughter stress markers and quality meat, physicochemical (pH), biochemical (muscle glycogen content), and chemical (super oxides free radicals) parameters. The carcass quality, as incidence of individual carcass defects, was also evaluated. Twenty broilers were processed with two different electrical stunning: high (250 Hz; 640 mA; 60V) (Lot C or control) and low (150 Hz; 360 mA; 60 V) (Lot A) frequency and intensity, using sinusoidal alternating current. As preliminary results, the use of low frequency and intensity induced faster pH decline post mortem and adequate acidification of pH at 3 hours (6.49 Lot C; 6.37 Lot A), better muscle glycogen reserve (0.770 µL/50 mL Lot C; 1.497 µL/50mL Lot A), and lightly more rapid muscle oxidation (IDF: 0.109 Lot C; 0.122 Lot A), (FOX: 0.131 MeqO2/kg Lot C; 0.140 MeqO2/kg Lot A). The incidence of individual carcass defects sufficient to cause downgrading or rejection, both in Lot C and Lot A, was generally low. In a multidisciplinary approach, to assess animal welfare and quality poultry meat, additional and feasible parameters should be implemented. Monitoring of pH, muscle glycogen reserve and superoxide free radical production measurements might be markers easier to use, routinely, in practice at abattoir. Further studies are needed to evaluate the usefulness of these parameters.

Study on the occurrence of polycyclic aromatic hydrocarbons in milk and meat/fish based baby food available in Italy.

The study compared the profile of 14 polycyclic aromatic hydrocarbons, frequently occurred in food, in milk (N = 22) and meat/fish based (N = 18) baby foods available on the Italian market. PAH total levels, markers (Regulation EC/835/2011) and carcinogenic PAHs were determined by high- performance liquid chromatography with fluorescence detector (HPLC-FD). The average of total PAHs was 52.25 µg kg-1 in milk and 11.82 µg kg-1 in meat/fish based baby foods. The levels of PAH markers were higher than the permissible EU limits of 1 µg kg-1 in 18.2% and 77.7% milk, and 5.6% and 44.4% meat/fish based baby foods. Milk based samples showed significant higher values (P < 0.05) of carcinogenic and possible carcinogenic hydrocarbons than meat/fish based products. The Margins of Exposure (MOE) value of milk based baby food samples indicated a potential concern for consumer health. Monitoring programs, and good agriculture and manufacture practices should be recommended.