Lead and other heavy metals (cadmium and mercury) accumulation in bivalve mollusks (Mytilus galloprovincialis, Ruditapes spp. and Crassostrea gigas) sampled in Sardinia in 2008-2012

The Sardinian shellfish farming, like the national, is mainly focused on mussels and carpet-shell clams, still less on cupped oyster farming. After Olbia's Gulf, various lagoon areas along the coastal perimeter have been interested to shellfish farming. They are transitional waters, whose state of pollution must be evaluated both as ecosystem's health and as directly/indirectly human risk. This also applies to heavy metals such as lead, cadmium and mercury, arising both from anthropogenic that geological-natural activity. The aim of the present study is to investigate the variability of the concentrations of these metals in different mollusks to make a comparative assessment, detect trends (over the five-years or cyclicrecurring) and identify hot spots. In 2008- 2012, 984 samples have been analysed by inductively coupled plasma mass spectrometry technique. Of them, 599 in pre-marketing (for classification of production areas or their monitoring) and the remaining during marketing. The difference between the average levels of the three metals in the different mollusks species was statistically significant, with Pb>Cd>Hg, and there was evidence of a gradual downward trend, albeit moderate, in the contamination levels, with a significant seasonality in concentrations levels, of lead in particular. Also comparisons between the bio-monitored coastal areas were statistically different. Since the samples were representative of the entire production of bivalve mollusks in Sardinia and the contamination allowable limits have never been exceeded in the products marketed, it can be concluded that these products are safe, pointing out that maintenance of monitoring/surveillance plans provides useful information species-dependent, site-specific and temporal trends

Variability of cadmium accumulation in cephalopods (Octopus vulgaris, Sepia officinalis, Loligo vulgaris and Todarodes sagittatus) collected in Sardinia in 2008-2012

Cephalopods are known to accumulate cadmium and play an important role in its biomagnification. They are an essential link in marine trophic chain and represent an important case in studies on cadmium transfer to man through the food chain. Since cadmium concentration widely varies in different tissues of the cephalopods – mainly accumulating in the hepatopancreas – evisceration represents a recommended preliminary step to reduce cadmium intake in view of cephalopods consumption; yet, the residual concentration in the edible part may still be a risk for public health. This study is intended to assess cadmium levels variability in the muscles of Cephalopoda, considering the different feeding habitats and marine trophic webs. In compliance with EU regulation, a survey on cadmium and other heavy metal levels in various sea food, including cephalopods, was conducted by the Istituto Zooprofilattico of Sardinia in co-operation with local health authorities. During a five-year survey (2008- 2012), 90 samples were collected from the following species: commons octopus (Octopus vulgaris), cuttlefish (Sepia officinalis), European squid (Loligo vulgaris), and flying squid (Todarodes sagittatus), located in different coastal areas and representatives of either benthic or nektonic habitats. Determination of cadmium levels was carried out according to Regulations (EC) No. 882/2004, No. 1881/2006 and No. 333/2007. Analysis of the edible portion (muscle) of fresh homogenised samples was carried out by inductively coupled plasma mass spectrometry (ICP-MS). The results showed a highly skewed distribution of data. No statistically significant differences were observed among four distributions of the natural logs of cadmium levels in the species considered

Analytical Approaches in Official Food Safety Control: An LC-Orbitrap-HRMS Screening Method for the Multiresidue Determination of Antibiotics in Cow, Sheep, and Goat Milk

The presence of unauthorized substances, such as residues of veterinary medicines or chemical contaminants, in food can represent a possible health concern. For this reason, a complete legislative framework has been established in the European Union (EU), which defines the maximum limits allowed in food and carries out surveillance programs to control the presence of these substances. Official food control laboratories, in order to ensure a high level of consumer protection, must respond to the challenge of improving and harmonizing the performance of the analytical methods used for the analysis of residues of authorized, unauthorized, or prohibited pharmacologically active substances. Laboratories must also consider the state of the art of the analytical methodologies and the performance requirements of current legislation. The aim of this work was to develop a multiresidue method for the determination of antibiotics in milk, compliant with the criteria and procedures established by Commission Implementing Regulation (EU) 2021/808. The method uses an LC-Orbitrap-HRMS for the determination of 57 molecules of antibiotic and active antibacterial substances belonging to different chemical classes (beta-lactams, tetracyclines, sulfonamides, quinolones, pleuromutilins, macrolides, and lincosamides) in bovine, ovine, and goat milk samples. It provides a simple and quick sample pretreatment and a subsequent identification phase of analytes, at concentrations equal to or lower than the maximum residual limit (MRL), in compliance with Commission Regulation (EU) 2010/37. The validation parameters: selectivity, stability, applicability, and detection capability (ccβ), are in agreement with the requirements of Commission Implementing Regulation (EU) 2021/808 and demonstrated the effectiveness of the method in detecting veterinary drug residues at the target screening concentration (at the MRL level or below), with a false positive rate of less than 5%. This method represents an effective solution for detecting antibiotics in milk, which can be successfully applied in routine analyses for official food control plans

Supercritical fluid extraction and GC detection of p,p1-DDE and PCB congeners in some samples of dying species from Sardinia

In an environmental pollution research some wild animals, deceased due to accidental causes, were collected from all over Sardinia. In order to detect the possible residues of unknown pollutants the isolated livers of these animals, previously lyophilized, were analyzed. First, by a GC-MS method, we identified the p,p1-DDE and some PCBs belonging to 5, 6, 7, 8 series PCBs. Later we set up a method for quantification of these compounds, using chicken liver as a model, by GC-ECD. The spiked chicken liver, free from p,p1-DDE and PCBs, was lyophilized and extracted by supercritical fluid extraction (SFE), subsequently the solutions obtained were subjected to cleanup by solid phase extraction (SPE). Finally, this method was employed to quantify p,p1-DDE and PCBs in the samples of the specimens collected

Arsenic speciation in fish products and seafood as a prerequisite for proper risk assessment

The Boi Cerbus lagoon, facing a mining and industrial site in Sardinia (Italy), is an important fishing area for the local population. Previous studies showed high concentrations of total arsenic (As_tot) in fish, molluscs and crustaceans sampled in the lagoon, and a possible exceeding of the provisional tolerable weekly intake set by the Joint FAO/WHO Expert Committee on Food Additives by some local consumer groups. However, the percentage of inorganic As (As_inorg) should be known for a correct assessment of potential risk, as its toxicity is much higher than that of the organic forms. Eighty samples of 14 different species of fish, molluscs and crustaceans, sampled in the Boi Cerbus lagoon in 3 different seasons (winter, spring and summer), were analysed for As_tot by inductively coupled plasma mass spectrometry (ICP-MS) and As_inorg by high performance liquid chromatography-ICP-MS. All the data obtained from the analysis were statistically processed to evaluate significant differences based on season, <em>taxon</em> and <em>habitat</em>, in preparation for a subsequent risk assessment

Colorimetric Analysis and Determination of Histamine in Samples of Yellowfin Tuna (<i>Thunnus albacares</i>) 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 L *), redness (a *), and yellowness (b *). The results of histamine occurrence agreed with the food safety criteria (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

Serological and molecular detection of <i>Bartonella</i> spp. in humans, cats and dogs from Northern Sardinia, Italy

Bartonella spp. are aerobic, gram-negative pleomorphic bacteria, belonging to the α2 subgroup of the Proteobacteria. Many Bartonella spp. are pathogens of humans, dogs, and cats. Numerous domestic and wild animals can serve as chronically infected reservoir hosts for Bartonella spp. An increasing number of arthropod vectors have been implicated in the transmission of Bartonella spp. The main source of transmission to humans is cats by means of scratches, whereas infection is less likely to occur by cat bite. Most human cases of infection with Bartonella henselae, named cat scratch disease (CSD), present as an acute febrile lymphadenopathy. The presence of cat fleas is essential for the maintenance of the infection within the cat population. The role of dogs as reservoirs of Bartonella spp. is less clear, because they seem to be only accidental hosts. Nevertheless, dogs are excellent sentinels for human infections, because a similar disease spectrum develops in dogs

Arsenic speciation in fish products and seafood as a prerequisite for proper risk assessment

The Boi Cerbus lagoon, facing a mining and industrial site in Sardinia (Italy), is an important fishing area for the local population. Previous studies showed high concentrations of total arsenic (As_tot) in fish, molluscs and crustaceans sampled in the lagoon, and a possible exceeding of the provisional tolerable weekly intake set by the Joint FAO/WHO Expert Committee on Food Additives by some local consumer groups. However, the percentage of inorganic As (As_inorg) should be known for a correct assessment of potential risk, as its toxicity is much higher than that of the organic forms. Eighty samples of 14 different species of fish, molluscs and crustaceans, sampled in the Boi Cerbus lagoon in 3 different seasons (winter, spring and summer), were analysed for As_tot by inductively coupled plasma mass spectrometry (ICP-MS) and As_inorg by high performance liquid chromatography-ICP-MS. All the data obtained from the analysis were statistically processed to evaluate significant differences based on season, <em>taxon</em> and <em>habitat</em>, in preparation for a subsequent risk assessment

Microbiological and chemical evaluation of Helix spp. snails from local and non-EU markets, utilised as food in Sardinia

With this study, 28 pools of snails of the genus Helix, respectively Helix aspersa (n=24) and Helix vermiculata (n=4) were analysed. They were taken from snail farming and stores. The snails were from Sardinia, other regions of Italy, and from abroad. All the samples were examined as pool looking for these microbiological target: Salmonella spp., Listeria monocytogenes, Escherichia coli O157, Clostridium perfringens, Norovirus and Hepatitis A Virus (HAV). In the same pools, the concentration of cadmium and lead by inductively coupled plasma mass spectrometry was also determined. The levels of these heavy metals were quite high, especially for cadmium. Two samples were positive for Salmonella spp., while no sample was positive for Escherichia coli O157, HAV and Norovirus. Two samples were positive for Clostridium perfringens and 8 for Listeria monocytogenes. The microrganisms related to Listeria monocytogenes were identified using biochemical techniques, then serotyped and gene sequenced by multiple loci sequence typing technique. Furthermore, antimicrobial restistence was tested on the same samples

Effect of amalgam fillings on the mercury concentration in human amniotic fluid

Background: Methyl mercury (MeHg) and metallic Hg are well known as neurotoxic agents. Dental amalgam contributes significantly to elemental Hg vapour exposure in the general population. There is little information about Hg concentration in human amniotic fluid (AF) of pregnant women and its potential toxic effect on the fetuses. Objective: Primary to assess the relationship between the presence of detectable mercury (Hg) concentration in human AF, number and surface areas of amalgam fillings of pregnant women; secondary to analyse their obstetric history and perinatal complications. Methods: Seventy-two pregnant women took part in this prospective study. One dentist recorded the dental status, presence, number and surface areas of amalgam fillings. Total Hg concentration in AF was determined in digested samples using automatic cold vapour atomic absorption equipment. The detection limit of Hg in AF, determined from blank readings, was 0.08 ng/ml. To estimate the dependence of the explanatory variables (such as number and surface areas of amalgam fillings, fish consumption, presence of liver or neurological diseases and smoking habits) on mercury concentration several linear regression models were built up. Stepwise logistic regression procedures were running on total sample and on patients with at least one amalgam filling (Positive Filling group = PF). Principal component analysis (PCA) provided two factors, which explained for more the 60% of the variance among the variables. Results: The overall mean Hg concentration in AF among all patients was 0.37&#177;0.49 ng/ml. Nineteen (26.4%) women had a Hg concentration &#60;0.08 ng/ml (Hg negative group). In 53 (73.6%) patients, with a concentration ≥ 0.08 ng/ml (Hg positive group), the mean value of Hg was 0.49&#177;0.52 ng/ml. The average number of amalgam fillings was 2.26&#177;3.19 in the Hg negative group and 5.32&#177;3.03 in the Hg positive group (ANOVA one-way p=0.04). A dependence of mercury concentration on number of amalgam fillings (p=0.03), surface area of the amalgam fillings (p=0.04) and fish consumption (p=0.04) was observed but not at a significant level. In stepwise logistic procedure the number of amalgam fillings gave a contribution to the model (p=0.04), although null value was included in the confidence intervals. We observed no statistically significant differences (X &#178;test) among the patients with a Hg concentration &#60;0.08 ng/ml (n=19) and those with a concentration ≥ 0.08 (n=53) with regard to obstetric history and perinatal complications. Conclusions: Number and surface areas of amalgam fillings influenced positively Hg concentration in AF but not at a significant level. Moreover Hg levels detected in AF were low and no adverse outcomes were observed through pregnancies and in the newborns