A synergistic solution for fighting fraudulent practices in squid using light stable isotope ratios and lanthanide tracers.

To identify a novel optimized strategy for preventing fraudulent substitutions of squid species and origins, forty European squids (Loligo vulgaris) and forty flying squids (Todarodes sagittatus) from the Mediterranean Sea and Atlantic Ocean were analyzed for δ13C, δ15N, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, and Lu using isotope ratio mass spectrometry and inductively coupled plasma-mass spectrometry. While δ13C and δ15N variations were mainly species-related, they alone could not reliably distinguish samples. To address this issue, decision rules were developed using Classification and Regression Tree analysis. Threshold values for δ13C (-19.91‰), δ15N (14.87‰), and Pr (0.49 µg kg-1) enabled successful discrimination among Mediterranean European squids, Atlantic European squids, Mediterranean flying squids, and Atlantic flying squids, achieving over 90% accuracy, 81% precision, 80% sensitivity, and 93% specificity. This method holds promise for enhancing traceability and safety in the seafood industry, ensuring product integrity and consumer trust.

Multi-elemental composition of botanical preparations and probabilistic evaluation of toxic metals and metalloids intake upon dietary exposure.

The aim of this study was to evaluate the inorganic elemental composition (49 elements) of 29 botanical preparations obtained from fruits, leaves, peels, seeds, roots, fungi, and spirulina by using inductively coupled-mass spectrometry and a mercury analyzer. Simultaneously, the risk associated with the chronic dietary exposure to 12 toxic metals and metalloids among the European population was evaluated by using a probabilistic approach based on Monte Carlo simulations. The analysis revealed worrying intake levels of Al, As, and Ni, primarily stemming from the consumption of spirulina-, peel-, and leaf-based botanicals by younger age groups. The intake of As from all analyzed botanicals posed a significant risk for infants, yielding margins of exposure (MOEs) below 1, while those deriving from peel-based botanicals raised concerns across all age groups (MOEs = 0.04-2.3). The consumption of peel-based botanicals contributed substantially (13-130%) also to the tolerable daily intake of Ni for infants, toddlers, and children, while that of spirulina-based botanicals raised concerns related to Al intake also among adults, contributing to 11-176% of the tolerable weekly intake of this element. The findings achieved underscore the importance of implementing a monitoring framework to address chemical contamination of botanicals, thus ensuring their safety for regular consumers.

Elemental profiles of swine tissues as descriptors for the traceability of value-added Italian heavy pig production chains.

The increasing demand for reliable traceability tools in the meat supply chain has prompted the exploration of innovative approaches that meet stringent quality standards. In this work, 57 elements were quantified by inductively coupled plasma mass spectrometry and direct mercury analysis in 80 muscle and 80 liver samples of Italian heavy pigs to investigate the potential of new tools based on multi-elemental profiles in supporting value-added meat supply chains. Samples from three groups of animals belonging to the protected designation of origin (PDO) Parma Ham circuit (conventionally raised; raised with genetically modified organism (GMO)-free feeds; raised with GMO-free feeds plus the supplementation of omega-3 polyunsaturated fatty acids (n-3 PUFA)) and a fourth group of samples from animals not compliant with the PDO Parma Ham production process were analyzed. Hierarchical cluster analysis allowed for the identification of three macro-clusters of liver or muscle samples, highlighting some inhomogeneities among the target groups. Following SIMCA analysis, better classification models were obtained by using liver elemental profiles (95% correct classification rate), with the highest classification accuracy observed for GMO-free livers (100%). The elements contributing the most to the separation of livers by class membership were La, Ce, and Pb for conventional, Li, Cr, Fe, As, and Sr for GMO-free + n-3 PUFA, and Lu for non-PDO samples. Given these findings, the analysis of the elemental profiles of pig tissues can be regarded as a promising method to confirm the declared pig meat label attributes, deter potential complex fraud, and support meat traceability systems.

Cadmium, lead, and mercury in two commercial squid species from the north Adriatic Sea (central Mediterranean): contamination levels and health risk assessment.

In this study, lead (Pb), cadmium (Cd), and total mercury (Hg) concentrations in European squids (Loligo vulgaris) and flying squids (Todarodes sagittatus) from the northern Adriatic Sea (Italy) were analyzed. The risk of the Italian population being exposed to potentially hazardous metal concentrations through the consumption of these products was also assessed. Compared to European squids, flying squids showed three times higher total Hg concentrations and one hundred times higher Cd concentrations to the point that more than 6 and 25% of the samples exceeded the maximum Hg and Cd limits established by the current legislation. From the evaluation of dietary exposure levels, it emerged that the consumption of flying squids was associated with the highest Pb intake by children and, consequently, with the lower margin of exposure values in relation to the risk of neurotoxicity (margin of exposure=33). Consumption of flying squids, especially by children, was also associated with higher intakes of Cd, inorganic, and methyl-Hg, which, respectively, accounted for 156, 113, and 23% of the tolerable weekly intakes established for these contaminants at European level. The obtained results raise concern and it may be necessary to provide specific dietary advice on the moderate dietary consumption of some cephalopod species, especially to the youngest and most vulnerable segment of the population. However, besides the highly conservative deterministic method adopted in this study, a refined consumer exposure assessment should be performed through the probabilistic methodology, which is more suitable to represent the real exposure scenario.

Elemental profile identifies metallurgical pollution in epiphytic lichen Xanthoria parietina and (hypo)xanthine correlates with metals.

The accumulation of 55 elements in lichens under the heap of a former nickel smelter (village Dolná Streda, Slovakia) and at eight sites at different distances from the heap plus six sites throughout Slovakia was studied to determine the elemental profile. The major metals in the heap sludge and in the lichens below the heap (Ni, Cr, Fe, Mn, and Co) were surprisingly low in lichens from both the near and far vicinity of the heap (4-25 km), indicating limited airborne spread. However, two different sites with metallurgical activity (another site near the ferroalloy producer in Orava) typically contained the highest amount of individual elements, including rare earth elements, Th, U, Ag, Pd, Bi and Be, and their separation from other sites was confirmed by PCA and HCA analyses. In addition, the amounts of Cd, Ba and Re were highest at sites without a clear source of pollution and further monitoring is needed. It was also an unexpected finding that the enrichment factor calculated using UCC values was increased (often considerably >10) for 12 elements at all 15 sites, indicating eventual anthropogenic contamination with P, Zn, B, As, Sb, Cd, Ag, Bi, Pd, Pt, Te and Re (and other EF values were locally increased). Metabolic analyses showed a negative correlation between some metals and metabolites (ascorbic acid, thiols, phenols and allantoin), but slightly positive (amino acids) or highly positive correlation with purine derivatives hypoxanthine and xanthine. The data suggest that lichens adapt their metabolism to excessive metal loading and that epiphytic lichens are suitable for identifying metal contamination even at apparently clean sites.

Occurrence of Toxic Metals and Metalloids in Muscle and Liver of Italian Heavy Pigs and Potential Health Risk Associated with Dietary Exposure.

Muscle and liver tissues from Italian heavy pigs were analyzed to investigate whether the chronic consumption of these products by local consumers could represent a health risk in relation to the contamination by some toxic metals and metalloids (TMMs). The concentrations of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sn, U, and Zn were measured with an inductively coupled plasma-mass spectrometer, while Hg analysis was performed by using a mercury analyzer. Fe, Zn, and Cu were the most abundant elements in both tissues, while U was detected only at ultra-trace levels. As, Cd, Cu, Fe, Hg, Pb, U, and Zn showed significantly higher concentrations in livers compared to muscles (p ≤ 0.01), with Cd and Cu being 60- and 9-fold more concentrated in the hepatic tissue. Despite this, concentrations of all TMMs were found to be very low in all the samples to the point that the resulting estimated dietary intakes did not suggest any food safety concern. Indeed, intakes were all below the toxicological health-based guidance values or resulted in low margins of exposure. Nevertheless, in the calculation of the worst-case exposure scenario, the children's estimated intake of Cd, Fe, and Zn through the sole consumption of pig liver contributed to more than 23, 38, and 39% of the tolerable weekly intakes of these elements, while the combined consumption of pig liver and pig muscle to more than 24, 46, and 76%. These findings alert about the probability of exceeding the toxicological guidance values of Cd, Fe, and Zn though the whole diet, suggesting long-term negative health effects for the younger population.

Nickel uptake in hydroponics and elemental profile in relation to cultivation reveal variability in three Hypericum species.

The Hypericum species (H. perforatum, H. olympicum, and H. orientale) were cultured in hydroponics with excess nickel (Ni, 1 or 100 µM Ni) to compare the metallic and metabolite content. Identical species were collected outdoor to assess the same parameters (including uranium and lanthanides) with total of 53 elements. The results showed that Ni was less accumulated in shoots in hydroponics (translocation factor of 0.01-0.25) and the highest absolute amount was detected in H. olympicum. Essential elements were typically depleted by Ni excess, but Co and Na increased. Soluble phenols, sum of flavonols and catechin rather increased in response to Ni but quercetin glycosides and free amino acids decreased in the shoots of H. olympicum mainly. Comparison of laboratory and outdoor growing plants showed more phenols in outdoor samples but not in H. olympicum and individual metabolites differed too. Plants cultured in hydroponics contained lower amount of non-essential, toxic and rare earth elements (30-100-fold) and shoot bioaccumulation factor in outdoor samples was low for most elements (<0.01) but not for Cd and Pt. Data reveal that H. olympicum is a potent source of phenolic metabolites whereas H. orientale accumulates many elements (38 out of 53 elements).

Classification of transformed anchovy products based on the use of element patterns and decision trees to assess traceability and country of origin labelling.

Quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS) and direct mercury analysis were used to determine the elemental composition of 180 transformed (salt-ripened) anchovies from three different fishing areas before and after packaging. To this purpose, four decision trees-based algorithms, corresponding to C5.0, classification and regression trees (CART), chi-squareautomatic interaction detection (CHAID), and quick unbiased efficient statistical tree (QUEST) were applied to the elemental datasets to find the most accurate data mining procedure to achieve the ultimate goal of fish origin prediction. Classification rules generated by the trained CHAID model optimally identified unlabelled testing bulk anchovies (93.9% F-score) by using just 6 out of 52 elements (As, K, P, Cd, Li, and Sr). The finished packaged product was better modelled by the QUEST algorithm which recognised the origin of anchovies with F-score of 97.7%, considering the information carried out by 5 elements (B, As, K. Cd, and Pd). Results obtained suggested that the traceability system in the fishery sector may be supported by simplified machine learning techniques applied to a limited but effective number of inorganic predictors of origin.

Multi-element signature of cuttlefish and its potential for the discrimination of different geographical provenances and traceability.

The measurement and analysis of fifty-two elements by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS) and direct mercury analysis were applied to origin discrimination of Italian traditional cuttlefish (Chioggia, Venice lagoon) from Mediterranean and Atlantic samples. A total 68 specimens were analyzed in triplicates to generate 204 mass spectra profiles which were statistically processed by different chemometric techniques. Loading weights from principal component analysis as input for linear discriminant analysis (LW-LDA), stepwise-LDA (S-LDA) and variable influence of projection-partial least square discriminant analysis (VIP-PLS-DA) were used to classify samples while retaining the lowest possible number of key variables. VIP-PLS-DA was found to be the best variable selection-discriminant tool combo since the selected Na-Co-B-K-Cd-V-U-Rb-Ni-Ba-Cu-As-Sr-Mn-Mo-Li-Ca-Mg-Se-Bi-Cs-P-Y elemental pattern allowed the samples to be classified with 100% sensitivity, specificity and accuracy.

Carbon microparticles as a physical carrier for ETV-ICP-MS.

In this study, carbon microparticles (CMs) as a physical carrier to enhance the analyte transport efficiency for inductively coupled plasma mass spectrometry connected with electrothermal vaporization (ETV-ICP-MS) are proposed. Carbon microparticles mixed with samples or calibration standard solutions were dosed as a slurry into the graphite furnace. The optimization of working conditions was done for the standard solution 0.5 µg L-1 of Au and Tl. The pyrolysis and vaporization temperatures were 500 and 2700 °C for Au and 400 and 1900 °C for Tl. The optimized CMs concentration was 1 g L-1 for Au and 2.5 g L-1 for Tl. For the quantification, external calibration standard solutions were used. The result obtained for Au in the digested CRM GBW 07601 (the informatory value 2.1 ±â€¯2 µg kg-1) was 2.2 ±â€¯0.1 µg kg-1. The result for Tl in the digested CRM BCR 679 (the certified value 3 ±â€¯0.3 µg kg-1) was 3.2 ±â€¯0.2 µg kg-1 and in the slurry 2.7 ±â€¯0.1 µg kg-1. The result for Tl in the CRM GBW 10052 (the certified value 57 ±â€¯11 µg kg-1) was 51 ±â€¯3 µg kg-1. The instrumental limits of detection were 0.016 ng L-1 for Au and 0.026 ng L-1 for Tl. The recoveries and repeatabilities measured on calibration standards were in the range 99-100% and 0.2-2.3% for Au and 100-111% and 2.9-6.7% for Tl.

Host specificity of microsporidia pathogenic to the gypsy moth, Lymantria dispar (L.): field studies in Slovakia.

Several species of microsporidia are important chronic pathogens of Lymantria dispar in Europe but have never been recovered from North American gypsy moth populations. The major issue for their introduction into North American L. dispar populations is concern about their safety to native non-target insects. In this study, we evaluated the susceptibility of sympatric non-target Lepidoptera to two species of microsporidia, Nosema lymantriae and Vairimorpha disparis, isolated from European populations of L. dispar and applied in field plots in Slovakia. Application of ultra low volume sprays of the microsporidia maximized coverage of infective spores in a complex natural environment and, thus, exposure of non-target species to the pathogens. Of 653 non-target larvae collected from plots treated with V. disparis in 2002, 18 individual larvae representing nine species in four families were infected. These plots were monitored for two subsequent seasons and V. disparis was not recovered from non-target species. Of 2571 non-target larvae collected in N. lymantriae-treated sites, one larva was found to be infected. Both species of microsporidia, particularly N. lymantriae, appear to have a very narrow host range in the field, even when an inundative technique is used for their introduction. V. disparis infections in L. dispar exceeded 40% of recovered larvae in the treated study sites; infection rates were lower in sites sprayed with N. lymantriae. Several naturally-occurring pathogens were recorded from the non-target species. The most common pathogen, isolated from 21 species in eight families, was a microsporidium in the genus Cystosporogenes.