Validation of an Eco-Friendly Automated Method for the Determination of Glucose and Fructose in Wines.

Fermentable sugar dosage helps oenologists to establish a harvest's moment and control the fermentation process of the musts. The official analyses recommended for their determination are long, laborious, and must be carried out by specialized personnel. On the contrary, instrumental analysis automation limits human errors, increases precision, and reduces the time and cost of the analyses. In the food production sector, to use methods other than those recommended by supranational bodies in official reports, it is necessary to validate the analytical processes to establish the conformity of the results between the new methods and the reference ones. This work validated an automated enzymatic apparatus to determine the sum of glucose and fructose levels in wine samples. The validation was carried out on wine samples (dry red wine, dry white wine, moderately sweet wine, and sweet wine) containing different sugar concentrations by comparing data obtained using the OIV-MA-AS311-02 method performed by a specialized operator (reference method) and the same method performed by an automated apparatus. The difference between the results' means obtained with the two procedures was significant. Nevertheless, the automated procedure was considered suitable for the intended use since the differences between the averages were lower than the measurement uncertainty at the same concentration, and the repeatability results were better for the automated procedure than the reference method.

Validation of Rapid Enzymatic Quantification of Acetic Acid in Vinegar on Automated Spectrophotometric System.

Vinegar is produced from the fermentation of agricultural materials and diluted acetic acid (diluted with water to 4-30% by volume) via sequential ethanol and acetic acid fermentation. The concentration of acetic acid must be measured during vinegar production. A Community method for analyzing acetic acid in vinegar is a non-specific method based on the assumption that the total acid concentration of the vinegar is attributable to the acetic acid. It consists of titration with a strong base in the presence of an indicator. This test is laborious and has a time-consuming character. In this work, a highly specific automated enzymatic method was validated, for the first time, to quantify the acetic acid in the wine vinegar, in terms of linearity, precision, repeatability, and uncertainty measurement. The results were compared to the Community method of analysis. Regression coefficient ≅ 1 and the normal distribution of residuals in the ANOVA test confirmed the method's linearity. LLOD (0.946 ppm) and LLOQ (2.00 ppm) defined the method's sensitivity. The results of the tested and the Community methods, linearly distributed in the Shapiro-Wilk test, confirmed the method's repeatability. The few anomalous data in the Huber test were due to random errors. The high selectivity of the enzymatic method, which exclusively measures acetic acid concentration, determined the significant differences between the two tests, examined in the accuracy determination. The enzymatic method can be considered applicable since its precision and uncertainty were lower than the Community method values (relative percentage deviations = 10%). The enzymatic method compared to the Community method reduces the analysis time and the risk of errors due to operators (avoid pipetting errors and wrong calculations), minimizes solvent and the sample consumption and guarantees assay quality through method standardization.

First ecotoxicological assessment of Caretta caretta (Linnaeus, 1758) in the Mediterranean Sea using an integrated nondestructive protocol.

The loggerhead sea turtle (Caretta caretta) is a long-lived carnivorous reptile included in the IUCN Red List. The IUCN assessment for this species underlines the lack of information regarding pollution and pathogens and indicates as a priority effort to investigate and reduce the impacts of these threats. In this contest, the aim of our study was to conduct the first ecotoxicological assessment of this species in the Mediterranean Sea using a nondestructive integrated methodology. We set up and applied a monitoring protocol which includes endpoints, such as CYP1A, LPO, ENA assay, B esterases, never investigated before in this species. Seventy-five loggerhead turtles were sampled in a nondestructive way in Italian Sea Turtle Rescue Centers or free-ranging along the Spanish coasts. Blood, skin and carapace samples were used to test biomarker responses (CYP1A, VTG, LPO, Comet and ENA assay, BChE, GGT) and contaminant (OCs, PAHs, Pb, Cd, Hg) levels. Elaboration of experimental results was carried out taking also into consideration different age classes of the specimens. Among the main results obtained we should underline the statistically significant correlation between carcinogenic PAHs in blood and DNA fragmentation as well as between Cd in carapace and GGT in plasma. We also evidenced as youngest animals showed significantly higher DNA fragmentations, BChE inhibition and increase of GGT. Older specimens showed the highest levels of erythrocyte nuclear abnormalities which may indicate a long term toxicological stress. This study contributed to expand the knowledge about the ecotoxicology of C. caretta in the Mediterranean. The nondestructive protocol could also be applied to other marine ecosystems and other sea turtle species.

Fin whales and microplastics: The Mediterranean Sea and the Sea of Cortez scenarios.

The impact that microplastics have on baleen whales is a question that remains largely unexplored. This study examined the interaction between free-ranging fin whales (Balaenoptera physalus) and microplastics by comparing populations living in two semi-enclosed basins, the Mediterranean Sea and the Sea of Cortez (Gulf of California, Mexico). The results indicate that a considerable abundance of microplastics and plastic additives exists in the neustonic samples from Pelagos Sanctuary of the Mediterranean Sea, and that pelagic areas containing high densities of microplastics overlap with whale feeding grounds, suggesting that whales are exposed to microplastics during foraging; this was confirmed by the observation of a temporal increase in toxicological stress in whales. Given the abundance of microplastics in the Mediterranean environment, along with the high concentrations of Persistent Bioaccumulative and Toxic (PBT) chemicals, plastic additives and biomarker responses detected in the biopsies of Mediterranean whales as compared to those in whales inhabiting the Sea of Cortez, we believe that exposure to microplastics because of direct ingestion and consumption of contaminated prey poses a major threat to the health of fin whales in the Mediterranean Sea.

Amount and distribution of neustonic micro-plastic off the western Sardinian coast (Central-Western Mediterranean Sea).

A plethora of different sampling methodologies has been used to document the presence of micro-plastic fragments in sea water. European Marine Strategy suggests to improve standard techniques to make future data comparable. We use Manta Trawl sampling technique to quantify abundance and distribution of micro-plastic fragments in Sardinian Sea (Western Mediterranean), and their relation with phthalates and organoclorine in the neustonic habitat. Our results highlight a quite high average plastic abundance value (0.15 items/m(3)), comparable to the levels detected in other areas of the Mediterranean. "Site" is the only factor that significantly explains the differences observed in micro-plastic densities. Contaminant levels show high spatial and temporal variation. In every station, HCB is the contaminant with the lowest concentration while PCBs shows the highest levels. This work, in line with Marine Strategy directives, represents a preliminary study for the analysis of plastic impact on marine environment of Sardinia.

Large filter feeding marine organisms as indicators of microplastic in the pelagic environment: the case studies of the Mediterranean basking shark (Cetorhinus maximus) and fin whale (Balaenoptera physalus).

The impact of microplastics (plastic fragments smaller than 5 mm) on large filter feeding marine organisms such as baleen whales and sharks are largely unknown. These species potentially are ingesting micro-litter by filter feeding activity. Here we present the case studies of the Mediterranean fin whale (Balaenoptera physalus) and basking shark (Cetorhinus maximus) exploring the toxicological effects of microplastics in these species measuring the levels of phthalates in both species. The results show higher concentration of MEHP in the muscle of basking shark in comparison to fin whale blubber. These species can be proposed as indicators of microplastics in the pelagic environment in the implementation of Descriptor 8 and 10 of the EU Marine Strategy Framework Directive (MSFD).

Anthropogenic contaminants in Indo-Pacific humpback and Australian snubfin dolphins from the central and southern Great Barrier Reef.

We present the first evidence of accumulation of organochlorine compounds (DDTs, PCBs, HCB) and polycyclic aromatic hydrocarbons (PAHs) in Indo-Pacific humpback and Australian snubfin dolphins from the central and southern Great Barrier Reef. These dolphins are considered by the Great Barrier Marine Park Authority to be high priority species for management. Analyses of biopsy samples, collected from free ranging individuals, showed PAHs levels comparable to those reported from highly industrialized countries. DDTs and HCB were found at low levels, while in some individuals, PCBs were above thresholds over which immunosuppression and reproductive anomalies occur. These results highlight the need for ongoing monitoring of these and other contaminants, and their potential adverse effects on dolphins and other marine fauna. This is particularly important given the current strategic assessment of the Great Barrier Reef World Heritage Area being undertaken by the Australian Government and the Queensland Government.

Are baleen whales exposed to the threat of microplastics? A case study of the Mediterranean fin whale (Balaenoptera physalus).

Baleen whales are potentially exposed to micro-litter ingestion as a result of their filter-feeding activity. However, the impacts of microplastics on baleen whales are largely unknown. In this case study of the Mediterranean fin whale (Balaenoptera physalus), we explore the toxicological effects of microplastics on mysticetes. The study included the following three steps: (1) the collection/count of microplastics in the Pelagos Sanctuary (Mediterranean Sea), (2) the detection of phthalates in surface neustonic/planktonic samples, and (3) the detection of phthalates in stranded fin whales. A total of 56% of the surface neustonic/planktonic samples contained microplastic particles. The highest abundance of microplastics (9.63 items/m(3)) was found in the Portofino MPA (Ligurian Sea). High concentrations of phthalates (DEHP and MEHP) were detected in the neustonic/planktonic samples. The concentrations of MEHP found in the blubber of stranded fin whales suggested that phthalates could serve as a tracer of the intake of microplastics. The results of this study represent the first warning of this emerging threat to baleen whales.