Determination of Mycotoxins in Plant-Based Meat Alternatives (PBMAs) and Ingredients after Microwave Cooking.

In this study, we investigate the role of microwave cooking in reducing mycotoxin contamination in plant-based food matrices, with a focus on veggie burgers (purchased and home-made) and their ingredients (soybean, potatoes, zucchini, carrots). Two different conditions were studied (Max-Min) that were 800 W for 60 s and 800 W for 90 s, respectively. The degradation patterns of aflatoxins (AFB1, AFB2, AFG1, AFG2), fumonisins (FB1, FB2, FB3), trichothecenes (T2, HT2, ZEA), and ochratoxin A (OTA) were studied. The extraction procedures were conducted with the QuEChERS extraction, and the analyses were conducted with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Principal component analysis (PCA) showed that degradation under microwave cooking varies considerably across different food matrices and cooking conditions. This study provides valuable insights into the degradation of mycotoxins during microwave cooking and underscores the need for more research in this area to ensure food safety.

Essential and non-essential elements in white lupin (Lupinus albus L.) cultivated in Southern Italy.

We assessed the presence of V, Cr, Ni, Cu, Zn, As, Se, Sb, Cd, and Pb in white lupin samples cultivated in Southern Italy by the validation of an Inductively Coupled Plasma Mass Spectrometry (ICP-MS) method. The ICP-MS method validation showed satisfactory values of linearity (r2 > 0.999), recovery (87.4-100.7%), repeatability, and reproducibility values. Zinc was the most abundant element; showing mean concentrations of 0.778 ± 0.09 mg/Kg wet weight (w.w.) and a maximum of 1.013 mg/Kg w.w., followed by copper (0.191 ± 0.05 mg/Kg w.w.). Among the non-essential elements, important levels of cadmium were found (0.017 ± 0.004 mg/Kg w.w.), with 28% exceeding the limits set by the EU Regulation. The results of this work confirm the role of white lupins and other legumes in reducing the pH of the soil, increasing the exchangeable forms of Cd. This work also provides the first data on the nutritional and antinutritional properties of white lupins cultivated in Italy.

Antibiotic Resistance Genes Carried by Commensal Escherichia coli from Shelter Cats in Italy.

Antimicrobial resistance is a widespread global health problem. The presence of resistant bacteria and antibiotic resistance genes has been demonstrated not only in humans but also in animals, including pets. Stray cats share the urban environment with people and pets. This may facilitate transmission of resistant bacteria and resistance genes between stray animals, people and domestic animals. Several studies have investigated the role of stray cats as a fecal carrier of ESBL-producing bacteria. However, there are many genes and resistance mechanisms that can be detected in commensal E. coli, which, because of its genetic plasticity, is considered an indicator for monitoring antibiotic resistance. In this study, rectal swabs were collected from stray cats from colonies and shelters in the city of Monza (Monza Brianza, Italy) to isolate commensal E. coli. Phenotypic tests, such as the minimum inhibitory concentration (MIC) and the double disc test (DDST), and molecular analyses to detect antimicrobial resistance genes (ARGs) were used to study the resistance of these isolates. The results obtained confirm that stray cats can carry ESBL-producing E. coli (6.7%) and genes conferring resistance to other important antibiotic classes such as tetracyclines and sulfonamides.

Polyphenols of white lupin (Lupinus albus L.) seeds cultivated in Southern Italy by a LC-HRMS method.

In this work we examined the contents of 14 polyphenols in white lupin (Lupinus albus L.) samples cultivated in Southern Italy by the optimisation and validation of a LC-HRMS method. The validation of the LC-HRMS method showed linearity results r2 > 0.989 and recovery values between 71 and 119% for a very wide range of concentrations. Ellagic acid was the most abundant polyphenol, with mean concentrations of 16271.86 ± 19798.53 µg/Kg, followed by apigenin (2749.51 ± 889.95 µg/Kg). A significant variability in ellagic acid contents was found between the areas of cultivation examined (p < 0.05). As far as we know, this work provides the first data on the polyphenols contents of white lupins cultivated in Italy. The comparison with other study confirms the role of the cultivation area for the determination of the polyphenol's contents. The study also confirms white lupins as a promising source of antioxidant and anti-inflammatory substances in a balanced diet.

Toxic metals in Loggerhead sea turtles (Caretta caretta) stranded freshly dead along Sicilian coasts.

BACKGROUND: The Loggerhead sea turtle (Caretta caretta) is a marine reptile belonging to a monophyletic group of chelonians. As these animals are long-lived, they have the ability to accumulate pollutants. AIM: To collect epidemiological data on toxic metals in marine Loggerhead sea turtles. MATERIALS AND METHODS: Forty Loggerhead sea turtles comprising 25 males and 15 females stranded freshly dead between 2013 and 2018 along the coasts of Sicily, Southern Italy, were examined for arsenic, cadmium, and lead accumulation in muscle and adipose tissues by means of a validated ICP-MS method. A modified K index as a growth condition factor, namely Fulton's K index, was used. Samples were tested in duplicate. A Wilcoxon rank sum test was carried out to evaluate metal contents differences between muscle and adipose tissues and between genders. RESULTS: The Fulton's K index suggested a good body condition of the C. caretta recovered with mean values of 5.34 ± 3.40 (n = 40; ±SD). Detectable concentrations of lead were found in 70% of the samples analysed with mean values of 0.65 ± 1.67 mg/kg wet weight and 0.51 ± 1.29 mg/kg wet weight in muscle and adipose tissues, respectively. No significant differences in arsenic, cadmium, and lead were detected between genders. In addition, no significant correlation was found between modified K index and concentrations of arsenic, cadmium, and lead. CLINICAL RELEVANCE: Findings on muscle and adipose tissues suggest chronic exposure of Caretta caretta to high concentrations of especially lead which might negatively affect health and welfare of these marine turtles although body condition was good.

Toxic metals and total lipids comparison between wild and farmed fish of South mediterranean.

A total of 151 wild and farmed fish samples of three species (S. aurata, D. labrax and U. cirrosa) from south Mediterranean were examined for cadmium, lead and mercury and total lipids detection to verify possible differences between wild and farmed fish. Mercury was detected only in S. aurata samples, with mean values of 0.056 ± 0.128 mg/kg and 0.031 ± 0.033 mg/kg for farmed and wild samples, respectively. The results verified no significant differences in Pb, Cd and Hg levels between wild and farmed fish samples (p < 0.05). The farmed fish samples showed higher total lipids contents than wild (p > 0.05). As far as we know, this work report first findings on the toxic metal presence in farmed Umbrina cirrosa, showing no detectable Hg, Cd and Pb values. The results of this work seems to confirm that the presence of toxic metals could be more related to the pollution the site than the type of production.

First Report on the Presence of Toxic Metals and Metalloids in East Asian Bullfrog (Hoplobatrachus rugulosus) Legs.

We examined the presence of As, Cr, Cd, and Pb in 42 samples of farmed East Asian bullfrog (Hoplobatrachus rugulosus) from Vietnam and Thailand by inductively coupled plasma-mass spectrometry (ICP-MS). An estimation of the dietary intake and exposure to the toxic elements analysed was also carried out. The results showed very high As levels, with mean values of 0.094 ± 0.085 mg/Kg w.w. and a maximum of 0.22 mg/Kg. No significant differences were found for As contents between areas of production (p > 0.05). No detectable Cd contents were found in all the samples examined. The Pb concentrations of the East Asian bullfrog legs samples were below the European Commission's permitted levels. The Cr and Pb contents of the East Asian bullfrog produced in Vietnam were significantly higher than that produced in Thailand (p < 0.05). The target hazard quotient (THQ) ratio for Cr was not exceeded for all the samples analysed. In contrast, the benchmark dose lower confidence limit (BMDL) and THQ ratios for As were exceeded, indicating carcinogenic and non-carcinogenic risks for those who consume this type of food. The results of this work confirm the role of As-contaminated water absorption as an important source of arsenic for these adult organisms.

Fatty Acids Composition of Stomach Oil of Scopoli's Shearwater (Calonectris diomedea) from Linosa's Colony.

Calonectris diomedea is a Procellariforms seabird having a very representative colony in Linosa Island (Southern Italy). The adult forms of C. diomedea produce a pasty oil from their proventriculus to feed their chicks during the rearing period. In this work, we examined the fatty acids composition of the stomach oil of C. diomedea from Linosa Island by gas chromatography with flame ionization detection (GC-FID). The samples were collected at 20 and 70 days after hatching. Twenty different fatty acids (FAs) were identified. Saturated fatty acids (SFA) were the most abundant in percentage (41.6%) at day 20 followed by polyunsaturated fatty acids (PUFA, 38.7%) and monounsaturated fatty acids (MUFA, 19.7%). MUFAs were the most abundant in samples collected at day 70 (53.8%), followed by SFAs (36.6%) and PUFAs (9.8%). Oleic acid (C18:1ω9) in the samples on day 70 was 4 times higher than that in the samples on day 20. The Principal Component Analysis (PCA) verified a clear separation of the stomach oil samples in two groups, according to the day of sampling. The results obtained confirm the role of FAs analysis of stomach oil to understand the ecology and breeding behaviour of C. diomedea, highlighting a resemblance with signatures recorded in marine organisms of Linosa Island.

Trace elements in stomach oil of Scopoli's shearwater (Calonectris diomedea) from Linosa's colony.

Calonectris diomedea is a colonial Procellariiform breeding on Mediterranean islands. The stomach oil produced during chick rearing is a peculiar trait of this species. The composition of the stomach oil is likely to reflect the composition of the prey ingested and might reveal the contaminants uptake with prey becoming a possible tool for the marine pollution monitoring. We examined the concentration of 15 trace elements by ICP-MS and direct mercury analyser. The principal component analysis revealed a heterogeneous pattern of metal concentration, showing a significant separation between samples collected 20 and 70 days after hatching. The data obtained in this work give preliminary information on the feeding habits and breeding ecology of Linosa's colony of Scopoli's shearwater. The trace metals variability found suggest that the stomach oil may have a role as trophic markers to understand predator-prey relationships and to have evidence on the accumulation of pollutants in the latter.

QuEChERS LC-MS/MS Screening Method for Mycotoxin Detection in Cereal Products and Spices.

We developed and validated a screening method for mycotoxin analysis in cereal products and spices. Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) was used for the analysis. Dispersive solid-phase extractions (d-SPEs) were used for the extraction of samples. Ochratoxin A (OTA), zearalenone (ZEA), aflatoxins (AFLA; AFB1, AFB2, AFG1, AFG2), deoxynivalenol (DON), fumonisin (FUMO; FB1, FB2, FB3), T2, and HT2 were validated in maize. AFLA and DON were validated in black pepper. The method satisfies the requirements of Commission Regulation (EC) no. 401/2006 and (EC) no. 1881/2006. The screening target concentration (STC) was under maximum permitted levels (MLs) for all mycotoxins validated. The method's performance was assessed by two different proficiencies and tested with 100 real samples.

Acrylamide assessment of wheat bread incorporating chia seeds (Salvia hispanica L.) by LC-MS/MS.

We examined the acrylamide content in samples of wheat bread with chia seeds added at different concentrations (2%, 5%, 7%, 10%) and cooked at predefined conditions (20 min at 200°C) by a validated LC-MS/M method after QuEChERS extraction. The acrylamide contents of the bread samples with added chia seeds were compared with control wheat bread samples. The highest acrylamide values were found in bread with 5% chia seeds, showing a mean value of 156.5 ± 115.4 µg/kg, followed by bread with 10% chia seeds (150.2 ± 103.8 µg/kg). About 6% of the bread samples with added chia seeds reached acrylamide levels above the benchmark level set by the EU Regulation. No significant differences in acrylamide values were found between control samples and bread with different percentages of chia seeds (p > .05). The results obtained provide a first report on the possible contribution of chia to the increase of acrylamide formation in bread.

High hydroxycinnamic acids contents in fennel honey produced in Southern Italy.

A total of 122 honey samples (Apis mellifera ssp. Ligustica) collected from Southern Italy were examined for floral identification by melissopalynological examination and for polyphenols detection by an LC-ESI-OrbitrapTM -MS/MS method. The melissopalynological examination confirmed all the samples examined as fennel (Foeniculum vulgare) unifloral variety. The analytical method carried out for polyphenols detection showed satisfactory linearity and recovery values, achieved during the validation of the method. Very high amounts of flavonols (kaempferol and quercetin) and hydroxycinnamic acids (caffeic acid, chlorogenic acid and ferulic acid), were found in all the samples examined. Among the hydroxycinnamic acids group, caffeic acid showed the highest mean contents (865.90 ± 67.07 µg/kg). The results of this work confirmed the high presence of phenolic acids with strong free radical-scavenging activity in fennel products such as honey, suggesting their use to reduce oxidative stress.

Development of a Rapid and Eco-Friendly UHPLC Analytical Method for the Detection of Histamine in Fish Products.

We developed, validated, and confirmed with proficiency tests a fast ultra-high-performance liquid chromatography with diode array detector (UHPLC-DAD) method to determine histamine in fish and fishery products. The proposed method consists of two successive solid-liquid extractions: one with a dilute solution of perchloric acid (6%) and the second only with water. The instrumental analysis with UHPLC provides a very fast run time (only 6 min) with a retention time of approximately 4 min, a limit of quantification (LOQ) of 7.2 mg kg-1, a limit of detection (LOD) of 2.2 mg kg-1, a recovery around 100%, a relative standard deviation (RSD%) between 0.5 and 1.4, and an r2 of calibration curve equal to 0.9995. The method detected optimal values of the validation parameters and required a limited number of reagents in comparison to other methods reported in the literature. Furthermore, the method could detect histamine in a very short time compared with other methods. This method, in addition to being validated, precise, specific, and accurate, avoids wasting time, money, and resources, and limits the use of organic solvents.

Histamine in Fish Products Randomly Collected in Southern Italy: A 6-Year Study.

ABSTRACT: In total, 4,615 fresh and processed fish samples collected from 2010 to 2015 were analyzed for histamine by ultrahigh-performance liquid chromatography with diode array detection. Histamine levels were detected in 352 (7.6%) samples, with a maximum of 4,110 mg kg-1 and mean values of 908.9 ± 1,226.79 and 344.01 ± 451.18 mg kg-1 for fresh and processed fish samples, respectively. No histamine levels were found in canned tuna and smoked fish samples in contrast to most of the data reported in the literature. A low percentage (2.79%) of noncompliant samples was found. The highest mean values were found during 2011 and 2015 for fresh and processed fish samples, respectively, showing a significant (P < 0.05) difference between the sampling years. The histamine contents found in fresh fish samples were significantly higher (P < 0.05) than those of processed samples. Most of the positive samples came from street vendors, suggesting the need to improve inspection measures in these commercial categories to ensure fish product safety.

Land Snails as a Valuable Source of Fatty Acids: A Multivariate Statistical Approach.

The fatty acid (FA) profile of wild Theba pisana, Cornu aspersum, and Eobania vermiculata land snail samples, collected in Sicily (Southern Italy), before and after heat treatment at +100 °C were examined by gas chromatography with a flame ionization detector (GC-FID). The results show a higher content of polyunsaturated fatty acids (PUFAs) in all of the examined raw snails samples, representing up to 48.10% of the total fatty acids contents, followed by monounsaturated fatty acids (MUFAs). The thermal processing of the snail samples examined determined an overall reduction of PUFA levels (8.13%, 7.75%, and 4.62% for T. pisana, C. aspersum and E. vermiculata samples, respectively) and a species-specific variation of saturated fatty acid (SFA) contents. Oleic acid remained the most abundant FA of all of the snails species examined, accounting for up to 29.95% of the total FA content. A relevant decrease of É·3/É·6 ratio was found only for T. pisana samples. The principal component analysis (PCA) showed a separation of the snail samples in terms of species and heat treatment. The results of this work suggest land snails as a valuable source of MUFA and PUFA contents and boiling as appropriate treatment, according to the maintenance of healthy properties.

Mercury Detection in Benthic and Pelagic Fish Collected from Western Sicily (Southern Italy).

In highly polluted water, fish can accumulate mercury up to a concentration of 10 mgKg-1. This has occurred on the eastern coasts of Sicily (Southern Italy), probably due to the intense industrial activity of this area. However, little is known about Hg accumulation in fish of the western Sicilian coasts. In this work, we examined the Hg accumulation of 108 fish samples belonging to 14 species collected from western Sicilian coasts using a direct mercury analyzer. The samples showed a mean mercury concentration of 0.165 ± 0.22 mg kg-1 with a maximum in Lepidopus caudatus (1.72 mgKg-1), exceeding the limits provided by EC Reg. 1881/2006. The lowest Hg levels were found in Sparus aurata samples (0.001 mgKg-1). A significant difference was found between the fish species examined (p < 0.05). The comparison between benthic and pelagic species did not show statistical differences (p < 0.05). Fish food constitutes the main route of Hg uptake for humans. Only four of the 130 samples examined reached a mercury concentration over the European limits. The comparative analysis of Hg pollution for benthic and pelagic species did not confirm a different trend in metal contamination.