Seasonal variability of trace elements bioaccumulation in Pacific Oysters (Crassostrea gigas) from an experimental pilot farm in the Calich Lagoon (Sardinia, Italy).

BACKGROUND: Metals pollution is a worldwide environmental issue due to their persistence in the ecosystems, non-degradability, and bioaccumulation in marine biota. Pacific Oysters (Crassostrea gigas) are highly nutritious bivalve representing an important dietary constituent but may accumulate metals through feeding on suspended sediments from surrounding water, then represent a suitable tool for biomonitoring. MATERIALS AND METHODS: The occurrence of trace elements (Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Rb, Se, Sn, V, Zn) was investigated in Pacific Oysters (Cassostrea gigas) collected from Calich Lagoon in each season of 2019. Samples were homogenized and subjected to microwave acid digestion before being analyzed by inductively coupled plasma-mass spectrometer (ICP-MS). RESULTS: The results showed a significant seasonal variation for temperature, dissolved oxygen, chlorophyll, and pH. Moreover, high significant seasonal variation in concentrations of Cd, Mn, Ni, and V was recorded. The highest values were found for Fe (128 mg kg⁻1 w.w.), and Al (112 mg kg⁻1 w.w.) in October, for Zn (113 mg kg⁻1 w.w.) in March and May. CONCLUSIONS: Pacific Oysters were confirmed as suitable bioindicators of the health status of coastal lagoons; trace elements concentrations were highly affected by season of collection, and according to literature the highest values were recorded in autumn and summer. The EU legal limits for Cd and Pb were not exceeded, then the farmed oysters were safe to consumers.

Trend of Pathogenic Vibrio parahaemolyticus Occurrences in Bivalve Molluscs Harvested in Sardinian Coastal Environments Between 2011 and 2018.

The aim of the present study was to evaluate Vibrio parahaemolyticus occurrences in bivalve molluscs harvested from Sardinian coastal environments between 2013 and 2015. The prevalence of pathogenic V. parahaemolyticus isolates is based on the detection of the two major virulence genes thermostable direct hemolysin (tdh) and thermolabile hemolysin (trh) To assess changes between 2011 and 2018 in the prevalence of V. parahaemolyticus in bivalve molluscs, we compared our results with those of previous investigations. In total, 2,933 samples were collected: 1,079 in 2013, 1,288 in 2014, and 566 in 2015. The mean prevalence of V. parahaemolyticus in shellfish was 3.5% in 2013, 1.7% in 2014, and 3.5% in 2015. The highest percentage of positive samples in 2013 and 2014 was observed in clams (3.5% and 2.7%, respectively), whereas in 2015, it was reported in oysters (15.1%). By comparing the sampling period of 2011-2014 with that of 2015-2018, an increase in the prevalence of V. parahaemolyticus was observed in shellfish (p < 0.05). In parallel, 208 potentially enteropathogenic V. parahaemolyticus strains were identified through the years 2011-2018 and, in particular, 10 trh+ and six tdh+ isolates. Our present study provides information regarding trends of V. parahaemolyticus occurrences in bivalve molluscs harvested from Sardinian coastal environments between 2011 and 2018 suggesting that the prevalence varies depending on the sampling period and shellfish species.

Effects of Age, Fulton's Condition Index (K) and Muscle Fat on Total Mercury Content in Raw, Pre-Canning and Canned Samples of Atlantic Bluefin Tuna (Thunnus thynnus).

A total of 30 samples of Atlantic bluefin tuna were analysed for total mercury concentration. Relationships between total mercury content and age, Fulton's condition index (K) and fat content were statistically evaluated. The effect of muscle status (raw, pre-canning, canned) on mercury content was also investigated. The average total mercury content was: 1.185 ± 0.662 mg/kg in raw, 1.481 ± 0.893 mg/kg in pre-canning and 1.393 ± 0.882 mg/kg in canned samples, respectively. Canning promotes a statistically significant increase in the concentration of mercury. The weight of fish, K and fat content are useful tools to estimate the mercury accumulation in Atlantic bluefin tuna. The results of the present study represent a contribution to the assessment of the EU mercury levels in Atlantic bluefin tuna.

Ecotoxicity of Copper(I) Chloride in Grooved Carpet Shell (Ruditapes decussatus).

Copper (Cu) is a ubiquitous trace element in the aquatic environment, and is usually found at low levels. Copper environmental concentrations can be altered as a result anthropogenic activities. Shellfish are useful bioindicators to ensure adequate environmental monitoring. Thus, the aim of the present study was as follows: (a) determine the LC50 of copper(I) chloride in grooved carpet shell (Ruditapes decussatus) collected in the Santa Gilla lagoon (Sardinia, Italy), and (b) analyze the antioxidant biomarkers in digestive gland and gills of same specimens exposed to different concentrations of the above-mentioned metal (0.045, 0.45, and 0.90 mg/L) for 96 h. A withdrawal period of 96 h was considered for the treated clam, carrying out the same biochemical analyses, superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathione peroxidase (Se-GPx), glutathione S-transferases (GSTs), and total glutathione (GSH+2GSSG) in the two tissues. Different time and dose responses of the antioxidant biomarkers were recorded in the digestive glands and gills. Oxidative stress biomarkers highlighted the ability of Cu to induce oxidative stress in R. decussatus. Clam, following the withdrawal period of 96 h, has not been able to achieve the control levels of all biochemical markers in the digestive gland and gills. R. decussatus can be a suitable model to assess the ecotoxicity of copper in aquatic ecosystems. These findings may advance knowledge on the role and the effects of copper on oxidative stress biomarkers in grooved carpet shell. The metal ecotoxicity response can be useful to perform accurate biomarker-based monitoring programs using this bivalve species.

Survival of Naturally Contaminating Listeria monocytogenes in Commercial Mediterranean-Style Dry Fermented Sausages during Storage.

ABSTRACT: The aim of the present study was the determination of Listeria monocytogenes, competitive microbiota, microbial hygiene indicators, and physicochemical parameters in the typical Mediterranean-style fermented sausages Salsiccia Sarda. A batch of Salsiccia Sarda (25 samples) naturally contaminated by L. monocytogenes and vacuum packaged after 24 days of ripening was included in the study. Fifteen samples stored at 8°C were analyzed after 13 days, after 90 days, and at the end of shelf life (after 180 days from vacuum packaging). Ten vacuum-packaged samples were stored at 12°C in a domestic fridge simulating temperature abuse and were evaluated at the end of the shelf life. Samples were subjected to physicochemical analysis (pH and water activity) and investigated for the presence and enumeration of L. monocytogenes. Competitive microbiota, lactic acid bacteria, coagulase-negative staphylococci, and microbial hygiene indicators (total mesophilic bacterial counts, Enterobacteriaceae, Enterococcuss spp., and Staphylococcus aureus) were determined in all the samples. Although a decreasing trend in L. monocytogenes prevalence was observed through the shelf life, the detection of this pathogen in fermented sausages confirms its ability to overcome hurdles of the manufacturing process. The results highlight the importance of the careful evaluation of the Salsiccia Sarda production process by food business operators to maintain unfavorable conditions for the growth of L. monocytogenes.

Colorimetric Analysis and Determination of Histamine in Samples of Yellowfin Tuna (Thunnus albacares) Marketed in Sardinia (Italy) by a Combination of Rapid Screening Methods and LC-MS/MS.

The consumption of fishery products has been steadily increasing in recent decades. Among the quantitatively more important species, the yellowfin tuna (Thunnus albacares), is one of the main at-risk species as regards the possibility to present important levels of histamine and to be associated with the so-called "Scombroid Fish Poisoning". The main aim of the present study was to evaluate the colorimetric parameters, the occurrence, and the quantification of histamine contamination in yellowfin tuna samples marketed in Sardinia (Italy) by a combination of rapid screening and official control methods. A total of 20 samples of yellowfin tuna loins collected from large retailers, fishmongers and local markets were analyzed for the qualitative and quantitative determination of histamine by the lateral flow test HistaSure™ Fish Rapid Test and LC-MS/MS, respectively. Moreover, all the samples were examined to assess the conformity with the EU rules on labelling and subjected to colorimetric analysis according to the CIE-L*a*b* standard. Visual inspection of yellowfin tuna labels highlighted a 30% of non-compliances. A significant (p < 0.05) difference was reported for brightness (L *), redness (a *), and yellowness (b *). The results of histamine occurrence agreed with the food safety criteria (<100 mg/kg) laid down in EC Regulation 2073/2005 in the 95% and in the 90% of the samples with the rapid screening methods and LC-MS/MS, respectively. A highly significant sessional variation (p < 0.00001) was pointed out. Moreover, the two methods showed an agreement rate of 85%. The results of the present study confirmed the utility of lateral flow tests for the fast qualitative determination of histamine in yellowfin tuna. Rapid screening test should be strengthened by comparison with the official method especially in case of uncertain or positive results.

Determination of phytoplankton in water samples, algal biotoxins, microbiological parameters and microplastics in Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) from an experimental pilot farm in the Calich Lagoon (Sardinia, Italy).

The aims of this paper were to collect and analyse preliminary data of phytoplankton in the water, biotoxins, Escherichia coli, Salmonella spp., Vibrio spp. and microplastic eventually present in farmed mussels, and to acquire information about the production capability from an experimental pilot farm of the Calich Lagoon. Two sampling sessions were carried out, in February and in May 2019, also monitoring the water condition (pH, temperature, salinity, dissolved oxygen, chlorophyll a). No potentially toxic algae were detected, and moreover no biotoxins (Paralytic Shellfish Poison, Diarrheic Shellfish Poison, Amnesic Shellfish Poison) were found in mussels. E.coli was present with the highest concentration in February (16000 MPN/100g e.p.). Salmonella and Vibrio spp. have not been detected. Almost a microplastic per grams was found, mainly fiber of different colours. Further studies, carried out for several months, will allow to better understand the possible problems related to the production of mussels in a lagoon not yet classified as a shellfish production area.

Occurrence of trace elements in Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) from an experimental pilot farm in the Calich Lagoon (Sardinia, Italy).

The present study aimed to determine trace elements in Mediterranean mussels (Mytilus galloprovincialis) from an experimental pilot farm of the Calich Lagoon, a typical Sardinian brackish area (Italy). Two sampling sessions were scheduled in February and May 2019 and the occurrence of 24 metals (Hg, Ag, Al, As, Be, Bi, Cd, Co, Cr, Cu, Fe, Ga, In, Mg, Mn, Mo, Ni, Pb, Rb, Se, Sn, Ti, V, Zn) in bivalves was considered. Environmental conditions of water (temperature, salinity, pH, dissolved oxygen, and chlorophyll a) were also measured in situ. A high significant (P<0.001) difference was reported for temperature, pH, and dissolved oxygen. Our results showed a significant sessional variation of Mo (P<0.001); Cd, V (P<0.01); Ni, Pb and Co (P<0.05) in examined M. galloprovincialis samples; as all values were higher in February than those for May session samples, meanwhile the highest levels were reported for Mg (mean±s.d. 1151±263 mg kg-1 wet weight), Al (mean±s.d. 341±192 mg kg-1 w.w.), and Fe (mean±s.d. 212 ±75 mg kg-1 w.w.) in February samples. The European Union uppermost values (EC Reg. 1881/2006) for Cd, Hg, and Pb were never overpassed. The results confirmed the role of M. galloprovincialis as one of the most appropriate biological indexes to track the presence of trace elements in brackish environments. It could be concluded that the current ecology of the Calich Lagoon suggests that compatibly with the transitional ecosystem, the classification as a bivalves' production area and the implementation of extensive shellfish farming can improve its production capacities. The knowledge of the lagoon ecology is an essential tool for its sustainable exploitation, preserving biodiversity, and mitigating the effects of anthropogenic activities on public health.

Identification and quantification of potential microplastics in shellfish harvested in Sardinia (Italy) by using transillumination stereomicroscopy.

Plastics are non-biodegradable polymers made up of different groups of petrochemical materials. Several biotic and abiotic factors can change the density of plastic fragmenting it and originating microplastics (MPs). MPs have been defined as small pieces of plastic less than 5 mm in size. Due to their small size, they are an emerging concern in the marine environment since they can be ingested by aquatic organisms, especially filter-feeding organisms, such as bivalve mollusks. Impacts of MPs exposure have been shown at various levels of biological organization, from cellular to tissue to individual and population levels. For example, oxidative stress and inflammation have been observed in copepods and mussels, obstruction and physical damage of the digestive tract were found in fish and swimming behavior alterations, disruption of foraging and feeding behavior and overall reduced fitness and survival were observed in fish and oysters. In addition, MPs can act as a vector for the transfer of chemicals to marine biota. The aim of the present study was the identification and quantification of potential MPs in shellfish harvested in Sardinia (Italy) by using transillumination stereomicroscopy. Bivalves were collected from 4 of the main production areas located along the Sardinian coast and selected according to the principles of the risk assessment. The results of the present study demonstrated the presence of potential MPs in 70% of the analyzed samples: the presence of MPs in bivalve mollusks may pose a threat to food safety, and there is an urgent need to evaluate the potential risks of MPs to human health.

Qualitative determination of histamine in canned yellowfin tuna (Thunnus albacares) marketed in Sardinia (Italy) by rapid screening methods.

Histamine is produced by the bacterial decarboxylation of histidine, an ammino acid present in large amount especially in scombroid fish such as tuna. Fish containing high levels of histamine have been associated with many instances of "scombroid poisoning". Since histamine is heat resistant, its presence has been used as an indicator of the good manufacturing practice and of the preservation state of canned tuna. In this study we have applied a rapid screening method to determine the presence of histamine in canned tuna marketed in Sardinia (Italy). A total of 165 samples of canned tuna were screened for the qualitative determination of histamine by HistaSure™ Fish Rapid Test. The results were consistently in agreement with the food safety criteria (<100 mg/kg of histamine) laid down in EC Regulation 2073/2005 (as amended). The HistaSure™ kit was confirmed as a rapid screening method for the presence of histamine in canned tuna.

Presence, Seasonal Distribution, and Biomolecular Characterization of Vibrio parahaemolyticus and Vibrio vulnificus in Shellfish Harvested and Marketed in Sardinia (Italy) between 2017 and 2018.

ABSTRACT: In the present study, we investigated the presence, seasonal distribution, and biomolecular characteristics of Vibrio parahaemolyticus and Vibrio vulnificus in samples of bivalve mollusks (Mytilus galloprovincialis, Crassostrea gigas, and Ruditapes decussatus) harvested and marketed in Sardinia (Italy) between 2017 and 2018. A total of 435 samples were submitted for qualitative determination of Vibrio spp., V. parahaemolyticus, and V. vulnificus. Potentially enteropathogenic isolates were detected with biomolecular methods. The overall prevalence of Vibrio spp. was 7.6%. The highest Vibrio prevalence was found in R. decussatus (8.3%). The prevalences of V. parahaemolyticus and V. vulnificus were 2.7 and 4.8%, respectively. Higher prevalences of V. parahaemolyticus and V. vulnificus were found in R. decussatus (4.2%) and C. gigas (6.2%), respectively. Only two pathogenic V. parahaemolyticus strains were recovered (genotypes: tdh- and trh+; tdh+ and trh-), both from M. galloprovincialis. None of the isolates were tdh+ and trh+. Pathogenic Vibrio infections are often underestimated, and human infections are increasing in Europe. European data on the true distribution of Vibrionaceae are scarce, and the results of the present study highlight the need of constant monitoring to update the distribution of pathogenic vibrios.

Seasonal accumulation of trace elements in native Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) collected in the Calich Lagoon (Sardinia, Italy).

The occurrence of 21 trace elements in native Mediterranean mussel (Mytilus galloprovincialis) from the Calich Lagoon, a typical brackish area of the northwest of Sardinia (Italy), was investigated. The seasonal variation of metals in bivalves was considered, and the highest values were found in spring and summer; in particular, a high significant (P < 0.001) temporal variation was reported for silver (Ag) and mercury (Hg). The highest and similar concentrations were registered for aluminium (Al, mean 32 mg kg-1wet weight), iron (Fe, mean 32 mg kg-1 w. w.), and zinc (Zn, mean 25 mg kg-1 w. w.). The maximum limits set by European Regulations for cadmium (Cd), mercury (Hg), and lead (Pb) were never exceeded. Speciation analysis revealed negligible risk related to inorganic arsenic (iAs). Therefore, M. galloprovincialis confirmed its role as suitable bioindicator to monitor the contamination of coastal environments. Although the recommended tolerable weekly intake (TWI) was not exceeded, the levels of aluminium should be carefully evaluated in monitoring plans in the studied lagoon. Graphical abstract.

Enumeration of Escherichia coli and determination of Salmonella spp. and verotoxigenic Escherichia coli in shellfish (Mytilus galloprovincialis and Ruditapes decussatus) harvested in Sardinia, Italy.

The aim of the present study was to evaluate the occurrence of Salmonella spp., Verotoxigenic E. Coli (VTEC) and enumerate E. coli in shellfish (Mytilus galloprovincialis and Ruditapes decussatus) collected before and after depuration from two class B harvesting areas located in Sardinia (Italy). All the samples were analyzed for Salmonella spp. detection according to European Commission Regulation (EC) 2073/2005 and examined using the five tube Most-Probable-Number (MPN) method for enumeration of E. coli in accordance with the European Union reference method ISO 16649-3:2015. E. coli VTEC was investigated following a direct multiplex Polymerase Chain Reaction (PCR) screening test. The enumeration of E. coli met the European law limit for Class A areas of 230 MPN/100g. The averaged enumeration of E. coli in samples of M. galloprovincialis and R. decussatus collected at the harvesting time was 39 and 37 MPN/100 g respectively. The average contamination levels in samples collected after purification were 58 MPN/100g (M. galloprovincialis) and 32 MPN/100 g (R. decussatus). E. coli VTEC was not detected, on the contrary, Salmonella ser. Typhimurium was detected in one sample of M. galloprovincialis and in one sample of R. decussatus collected at the harvesting time. No significant associations were observed between E. coli levels in shellfish and environmental parameters of water or with the detection of Salmonella ser. Typhimurium in M. galloprovincialis and R. decussatus samples. Nevertheless, the occurrence of Salmonella ser. Typhimurium, involved in human infection outbreaks, should be considered a potential risk for consumers.

High-Hydrostatic-Pressure (HHP) Processing Technology as a Novel Control Method for Listeria monocytogenes Occurrence in Mediterranean-Style Dry-Fermented Sausages.

Although conventional microbial control techniques are currently employed and largely successful, their major drawbacks are related to their effects on quality of processed food. In recent years, there has been a growing demand for high-quality foods that are microbially safe and retain most of their natural freshness. Therefore, several modern and innovative methods of microbial control in food processing have been developed. High-hydrostatic-pressure (HHP) processing technology has been mainly used to enhance the food safety of ready-to-eat (RTE) products as a new pre-/post-packaging, non-thermal purification method in the meat industry. Listeria monocytogenes is a pertinent target for microbiological safety and shelf-life; due to its capacity to multiply in a broad range of food environments, is extremely complicated to prevent in fermented-sausage-producing plants. The frequent detection of L. monocytogenes in final products emphasizes the necessity for the producers of fermented sausages to correctly overcome the hurdles of the technological process and to prevent the presence of L. monocytogenes by applying novel control techniques. This review discusses a collection of recent studies describing pressure-induced elimination of L. monocytogenes in fermented sausages produced in the Mediterranean area.

Occurrence, Seasonal Distribution, and Molecular Characterization of Vibrio vulnificus, Vibrio cholerae, and Vibrio parahaemolyticus in Shellfish (Mytilus galloprovincialis and Ruditapes decussatus) Collected in Sardinia (Italy).

In this study, we investigated the occurrence, seasonal distribution, and molecular characterization of pathogenic vibrios in Mediterranean mussels (Mytilus galloprovincialis) and grooved carpet shells (Ruditapes decussatus) from two harvesting areas of Sardinia (Italy). Samples collected before and after depuration were submitted for qualitative and quantitative determination of Vibrio spp. Vibrio spp. isolates were presumptively identified by means of biochemical methods. Identification and virulence profile of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus were performed by molecular methods. The prevalence of Vibrio spp. in M. galloprovincialis and R. decussatus was, respectively, 96 and 77%. The averaged enumeration (mean ± standard deviation) of Vibrio spp. in samples of M. galloprovincialis and R. decussatus collected at the harvesting time was 2.04 ± 0.45 and 2.51 ± 0.65 log CFU/g, respectively. The average contamination levels in samples collected after purification were 2.28 ± 0.58 log CFU/g (M. galloprovincialis) and 2.12 ± 0.67 log CFU/g (R. decussatus). Four potentially pathogenic V. parahaemolyticus isolates (tdh+ or trh+) were recovered from grooved carpet shells samples. No isolate was tdh+/trh+. The presence of potentially pathogenic vibrios in Sardinian waters strengthens the need for rational purification practices under controlled conditions to guarantee the protection of consumers.

Clonal relationship among Vibrio parahaemolyticus isolated from Mediterranean mussels (Mytilus galloprovincialis) and grooved carpet shells (Ruditapes decussatus) harvested in Sardinia (Italy).

The aim of the present study was to investigate the genetic variability of Vibrio parahaemolyticus strains isolated from naturally contaminated Mediterranean mussels (Mytilus galloprovincialis) and Grooved carpet shells (Ruditapes decussatus) from three harvesting areas of Sardinia (Italy) using a combination of different typing methods: traditional phenotypic systems and molecular techniques. Ninety-nine putative V. parahaemolyticus strains isolated from shellfish collected before and after purification were included in the study. Seventy-two isolates were confirmed as V. parahaemolyticus and were submitted to REP, ERIC and BOX PCRs. The combined dendrogram showed the similarity of the data set of the three typing methods and demonstrates how the different techniques grouped the strains in two clusters in accordance with each singular dendrogram. Several strains rendered a unique pattern regardless of the typing method, which indicates the high discriminatory power of the methods. Moreover, the use of multiple typing methods allowed a more accurate characterization of the genetic profiles of isolates and the identification of clones hardly revealed through the common techniques. The intraspecific typing of environmental V. parahaemolyticus can be of great interest in order to recognize clonal relationships between environmental contamination, foodborne disease, and geographical/temporal distribution of this pathogen. The comparative analysis focusing on the obtained genetic profiles supports the possibility for typing methods to discriminate strains with similar phenotypic profile, identifying the level of genetic correlation among the strains and the presence of genetic clones.

Bacterial and Viral Investigations Combined with Determination of Phytoplankton and Algal Biotoxins in Mussels and Water from a Mediterranean Coastal Lagoon (Sardinia, Italy).

Calich Lagoon is a Mediterranean coastal lagoon located along the northwestern coast of Sardinia (Italy). The connection to marine and fresh water determines the high productivity of this coastal lagoon. Despite its great potential and the presence of natural beds of bivalve mollusks (Mytilus galloprovincialis), the lagoon has not yet been classified for shellfish production. In this study, through a multidisciplinary approach, the presence of several bacterial pathogens (Escherichia coli, Salmonella spp., and Vibrio spp.) and viral pathogens (hepatitis A virus and norovirus genogroups I and II) was evaluated from March 2017 to February 2018. In addition, phytoplankton composition in lagoon waters and associated algal biotoxins (paralytic and diarrhetic shellfish poisoning) in mussels were also monitored. The aim of this study was to provide useful data to improve knowledge about their seasonal presence and to assess the potential risk for public health, as well as to provide input for future conservation and management strategies. In mussels, Salmonella spp. were found in spring, along with E. coli, but Salmonella spp. were not found in autumn or winter, even though E. coli was detected in these seasons. Vibrio parahaemolyticus was found in autumn and winter, but not in spring. Norovirus genogroups I and II were found in winter samples. None of the bacteria were found in summer. Algal biotoxins have never been detected in mussel samples. Among potentially harmful phytoplankton, only Pseudo-nitzschia spp. were present, mainly in summer. The results showed that a possible bacterial and viral contamination, together with the presence of potentially toxic microalgae, is a real problem. Therefore, the development of natural resource management strategies is necessary to ensure the good quality of waters and guarantee the protection of consumers.

The bivalve Ruditapes decussatus: A biomonitor of trace elements pollution in Sardinian coastal lagoons (Italy).

The anthropogenic contamination of water bodies with metals via fertilizer, industrial sewage and urban wastewater has resulted in widespread problems in aquatic organisms, but also poses a risk to consumer health. Shellfish from coastal and estuarine environments bioaccumulate toxic metals in their tissues due to their ability to concentrate inorganic contaminants several orders of magnitude above ambient levels. The aim of this study was to evaluate the presence of trace elements in adult specimens of indigenous clams (Ruditapes decussatus). To this end, wild clams were collected from four different brackish areas of Sardinia (Western Mediterranean Sea, Italy) devoted to extensive aquacultural practices. The concentration of 16 trace elements (Al, Ag, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, Sn, Tl and Zn) in the tissues of the clams was quantified. The legal limits set by European Regulations for cadmium; mercury and lead were never exceeded. However, unexpectedly high values for aluminium and iron (mean 207 mg kg⁻1 and 113 mg kg⁻1 wet weight) were found in Santa Gilla lagoon, which is close to industrial settlements and had the highest values for the majority of chemical elements investigated. The highest values of the metalloid arsenic (As) were reported in the Porto Pozzo and San Teodoro lagoons (9.6 and 6.8 mg kg⁻1 w.w., respectively). The clam R. decussatus confirmed the capacity of bivalves as suitable bioindicators of trace elements pollution. Further investigations are necessary for constant monitoring of clams, 32 which constitute an important traditional resource for the fishing activity in Sardinia.

Retail Stores Policies for Marketing of Lobsters in Sardinia (Italy) as Influenced by Different Practices Related to Animal Welfare and Product Quality.

The aim of the present study was to evaluate the marketing policies of lobsters as influenced by different practices related to product quality in seven supermarkets located in Italy. Retailers were divided in two categories: large scale and medium scale. The two groups were compared to screen for differences and to assess differences in score distribution attributed to different practices related to product quality. Our results showed no statistical differences (p > 0.05) between the two categories. Lobsters were often marketed alive on ice and/or stocked for long periods in supermarket aquariums, highlighting the need to improve the specific European regulations on health, welfare, and quality at the market stage. Retail shop managers should be encouraged to develop better practices and policies in terms of marketing of lobsters. This will help in keeping the animals in good health and improve product quality at the marketing stages.

Evaluation of short purification cycles in naturally contaminated Mediterranean mussels (Mytilus galloprovincialis) harvested in Sardinia (Italy).

The aim of the present study was to investigate the effect of short purification cycles on the safety of naturally contaminated Mytilus galloprovincialis from harvesting areas of the Gulf of Olbia (Sardinia, Italy). Samples from ten batches of mussels were collected before, during and after purification treatment at two purification centres (A-B). All the samples were analysed for Escherichia coli and Salmonella spp according to Council Regulation (EC) 2285/2015. Detection and enumeration of Vibrio spp were performed according to previously published methods. Presumptive identification of Vibrio spp isolates were performed by means of conventional biochemical tests and polymerase chain reaction. The presence of Hepatitis A virus was detected by nested reverse transcriptase-polymerase chain reaction. Environmental parameters (water temperature and salinity) were also recorded. The results of Escherichia coli counts showed the overall efficacy of the short purification cycles; a purification cycle of 8 h led to a rapid decline in the concentration. The decrease in Escherichia coli counts does not correlate with the presence of naturally occurring vibrios, the decline of which occurs at an even slower rate. The average contamination levels for Vibrio spp before purification were 8.20 ±â€¯0.47 and 7.99 ±â€¯0.62 Log10 CFU/g in samples collected at purification plants A and B, respectively. After purification, the average contamination levels were 8.10 ±â€¯0.60 Log10 CFU/g at purification plant A and 7.85 ±â€¯0.57 Log10 CFU/g at purification plant B. The contaminated samples revealed the presence of Vibrio alginolyticus (n=21), Vibrio fluvialis (n=12), Vibrio cholerae (n=4), Vibrio parahaemolyticus (n=2) and Vibrio vulnificus (n=1). The Vibrio parahaemolyticus isolates carried the tdh or the trh genes. None of the isolates was tdh+/trh+. Salmonella spp and Hepatitis A virus were not detected. The adoption of short purification cycles for Mytilus galloprovincialis in the presence of pathogenic vibrios might not be sufficient to guarantee the safety of consumers.