Bioaccumulation of PCDD/Fs and PCBs in free-range hens: Congener fingerprints and biotransfer factors.

The bioaccumulation of lipophilic environmental contaminants in farm animals is an important issue to control and prevent human exposure to toxic pollutants. Free-range hens were used as a model to assess the transfer and the bioaccumulation of PCDD/Fs, DL-PCBs and NDL-PCBs from the soil to eggs in an area in the Campania region particularly affected by pollution, the "Land of Fires". We determined the following ranges of concentrations in eggs: 0.90-5.51 pg WHO-TEQ g-1 fat for PCDD/Fs; 1.63-4.24 pg WHO-TEQ g-1 fat for DL-PCBs; 2.77-9.75 pg WHO-TEQ g-1 fat for the sum of PCDD/Fs and DL-PCBs and 12.53-30.94 ng g-1 fat for NDL-PCBs. Higher contamination levels were observed in the eggs from free-range hens, compared to those of the control group, that is hens raised indoors; this indicates that soil remains a major source of contamination. Livers showed low levels of contamination for both outdoor and indoor reared hens. Biotransfer factors (BTFs) were calculated for each of the 35 PCDD/F and PCB congeners analysed. The BTFs highlighted that the transfer and bioaccumulation depend on chlorination degree and substitution pattern, in particular they increase with increasing chlorination degree up to the hexa-chlorinated congeners and then decrease. An exception to these finding was observed for 2,3,7,8 TCDD and 2,3,7,8 TCDF, which showed very high BTFs. These results demonstrate the importance of promoting studies on the environmental contaminants bioaccumulation as they provide an effective support for the risk assessment and management of exposure to toxic chemicals.

Dioxins and dioxin-like PCBs in buffalo milk from the Campania region (Italy): Decreasing trend and baseline assessment over 10 years (2008-2018).

Polychlorodibenzo-p-dioxins, polychlorodibenzo-furans (PCDD/Fs) and dioxin-like polychlorobiphenyls (DL-PCBs) enter the food chain from the environment. In this study, we report the 2008-2018 time-trends in the PCDD/F and DL-PCB contamination of milk from buffaloes fed on local forage in rural areas of the Campania region. Validated according to QA/QC criteria, the dataset (N = 808 on a total of 2068 samples, after excluding follow-up results and outliers) was computed on the upper-bound value pg WHO-TEQ2005 g-1fat. We assessed time-trends and assayed baseline contamination levels, which displayed log-normal distribution. A significant decreasing trend (p < 0.01) was observed from 2008 to 2009 and 2010; the P50-P95 range fell from 2.37-8.48 pg WHO-TEQ2005 g-1fat (N = 393) in 2008 to 1.73-4.61 in 2009 (N = 86) and to 0.67-1.46 in 2010 (N = 42). From 2010 to 2018 (N = 329), no significant variation was found among years and the related dataset fitted a log-normal distribution (p < 0.05). Occurrence descriptors indicated that the baseline contamination of dairy products (mean = 0.54; P50-P95 = 0.47-1.24) in the Campania Region was well below the EU regulatory limit in force (5.5 pg WHO-TEQ2005 g-1fat). Given the Tolerable Weekly Intake of 2 pg WHO-TEQ kg-1 body weight (bw) proposed by the EFSA for PCDD/Fs and DL-PCBs, this level of baseline contamination is discussed with regard to the orientation of food safety and food security risk connected with buffalo mozzarella cheese production.

Contamination by Aflatoxins B/G in Food and Commodities Imported in Southern Italy from 2017 to 2020: A Risk-Based Evaluation.

This study reports the results of aflatoxins B/G monitoring in food of vegetal origin, imported in Southern Italy from extra-European Union countries. From 2017 to 2020, we analyzed 1675 samples using an accredited HPLC method with fluorescence detection. We found out 295 samples (17.6%) were contaminated by aflatoxin B1, 204 by aflatoxins B/G (12.2%), while 75 (4.5%) resulted non-compliant to maximum limits set by the European Union law. Most of the batches tested were unprocessed food; the distribution of contamination levels, incidence of non-compliant samples, inference for different kinds of food are reported. The study focuses on the food more susceptible to contamination by aflatoxins; nuts are the food more controlled, showing the higher number of non-compliant samples. Our study confirms that pistachio nuts, hazelnuts and almonds are the major sources of exposure for consumers. Still, other products, such as chili pepper and Brazil nuts, need to get more information about their contamination levels. The study's findings are discussed in the perspective of the last opinion by EFSA about chronic exposure to aflatoxins. A case study to evaluate not compliance of a composed food to the European Union law is reported.

Polycyclic aromatic hydrocarbons (PAHs), arsenic, chromium and lead in warty crab (Eriphia verrucosa): occurrence and risk assessment.

This study assesses the PAH and heavy metal levels in muscle of warty crabs (Eriphia verrucosa), from the northern coast of the Campania region improving the data on toxic contaminants in this crustacean. The results showed a minimal PAH contamination; the mean concentrations were as follows: 0.2, 1.6 and 1.7 µg kg-1 wet weight (ww) for BaP, PAH4 and PAH6, respectively. Regarding the levels of the two PAHs not included in the European regulations, the BkF mean concentration was 0.1 µg kg-1 ww, while DahA was detected only in 10.7% of samples. Pb and Cr were also detected at low levels with mean values of 0.068 and 0.468 mg kg-1 ww, respectively; instead, high As levels, with a mean value of 5.021 mg kg-1 ww, were found. Considering the EWIs and the ILCRs calculated in this study, the PAH, Pb and Cr contamination levels found in the edible part of the crabs resulted safe for human consumption. Contrariwise, the ILCR calculated for the As exceeded the acceptable level of cancer risk, although the calculation did not refer to the inorganic form which is the only one recognized as carcinogenic. Hence, this study shows that warty crabs can accumulate environmental contaminants in their muscle tissue representing an important route of exposure to these toxics for the local population that regularly consumes them. This finding highlights the importance of monitoring the presence of these pollutants in crabs and in general in all fish and seafood in order to ensure food safety for consumers.

Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese.

Buffalo mozzarella cheese is one of the most appreciated traditional Italian products and it is certified as a Protected Designation of Origin (PDO) product under the European Commission Regulation No. 1151/2012. It is obtained exclusively from buffalo milk. If made from cow milk, or a mixture of buffalo and cow milk, buffalo mozzarella cheese does not qualify as a PDO product. In order to maximize their profits, some producers market buffalo mozzarella that also contains cow milk as a PDO product, thus defrauding consumers. New methods for revealing this fraud are therefore needed. One such method is the droplet digital Polymerase Chain Reaction (ddPCR). Thanks to its high precision and sensitivity, the ddPCR could prove an efficacious means for detecting the presence of cow milk in buffalo mozzarella cheese that is marketed as a PDO product. ddPCR has proved able to detect the DNA of cow and/or buffalo milk in 33 buffalo mozzarella cheeses labelled as PDO products, and experimental evidence could support its application in routine analyses.

Polychlorinated organic pollutants (PCDD/Fs and DL-PCBs) in loggerhead (Caretta caretta) and green (Chelonia mydas) turtles from Central-Southern Tyrrhenian Sea.

This study assesses for the first time the levels of PCDD/Fs and DL-PCBs in sea turtles coming from Tyrrhenian Sea. The concentrations measured in liver of the 24 specimens analysed were 6.90 vs 5.65 pg g-1 wet weight (ww) for PCDD/Fs and 10.95 vs 0.79 ng g-1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The DL-PCB levels resulted very higher in Caretta caretta than Chelonia mydas probably due to the different eating habits between the two species investigated. Furthermore, the highest levels of DL-PCBs were determined in livers of the adult Caretta caretta turtles of male sex. Positive correlations were found out between PCB-81 and the body mass (BM) of turtles (r2 = 0.6561; p = 0.001) and between PCB-81 and the curved carapace length (CCL) (r2 = 0.6250; p = 0.006) suggesting that the body burden of contaminants is related to the body size. The mean TEQ values, as a matter of risk assessment for turtles, were 3.64 vs 1.62 pg TEQ g-1 ww for PCDD/Fs and 8.72 vs 2.16 pg TEQ g-1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The results reported in this study increase the data available about the consequences of the Mediterranean Sea contamination by organochlorine pollutants and highlight an evident PCDD/F and PCB bioaccumulation in sea turtle tissues that threatens the survival of these marine organisms.

A relationship between environmental pollutants and enteric viruses in mussels(Mytilus galloprovincialis).

Mussels can be affected by environmental contaminants, as non-dioxin-like polychlorinated biphenyls (NDL-PCBs), polycyclic aromatic hydrocarbons (PAHs) and cadmium (Cd). Moreover, mussels may concentrate human enteric viruses, like noroviruses (NoVGI/GII), astrovirus (AsV) and rotavirus (RV). Herein, to establish a relationship between environmental and viral contamination, with the aim to ensure human food safety, both chemical and microbiological analysis were carried out in mussels Mytilus galloprovincialis, farmed in Campania region (Italy). Chemical analysis revealed ranges below the European maximum limits, and were: ∑6 NDL-PCBs (28, 52, 101, 138, 153, 180) 0.579-16.857 ng g-1 wet weight (ww); BaP LOQ (<0.2 µg kg-1)- 2.9 µg kg-1 ww, and ∑4 PAHs (BaA, CHR, BbFA, BaP) 0.7-23.7 µg kg-1 ww; Cd LOQ (<0.005 mg kg-1)- 0.078 mg kg-1 ww. In addition, mussels exhibited a prevalence of NoVGI (18%), NoVGII (43%), AsV (40%), RV (30%) and the presence of more than one viruses resulted in about 35%. Overall, a simultaneous contamination, both chemical and viral, was found in 62% of samples. Interestingly, virus-positive mussels showed levels of chemicals higher than negative ones.

Polychlorobiphenyl levels in the serum of cats from residential flats in Italy: Role of the indoor environment.

The domestic cat (Felis catus) was used as a sentinel of exposure to polychlorobiphenyls (PCBs) in indoor urban environments y. Sera from 120 cats were pooled to form 30 different groups selected by age (<2 years; > 2 ≤ 8 years; > 8 years), sex, municipality (Bologna and Turin) and environment (indoor vs. outdoor). Test portions of 1 mL were analyzed by means of gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS) for six selected indicators non-dioxin-like PCBs (∑6 PCBs: congeners #28, #52, #101, #132, #153 and #180) and the results were computed in the upper-bound mode. The internal dose of PCBs attributable to the cats' alimentary lipid intake ranged from 32.4 to 1,446 ng/g (P50 165; mean 258). The Wilcoxon test revealed significantly lower PCB burden in "outdoor" groups than in "indoor" groups. Age correlated well with the heptachlorinated and most bio-accumulative congener, PCB #180, and slightly with hexachlorinated PCBs #138 and #153. Contamination attributable to house dust collected in 15 living-rooms ranged from 10.0 to 279 ng/g dry weight (P50 97.4; mean 94.4). Exposure estimates indicated a 0.6-16 ng/kg bw range of daily ∑6 PCB intake from a default value of 200 mg/cat of dust ingestion. The intake of PCBs due to dust ingestion fell within the same order of magnitude as that computed from a 60 g daily intake of commercial dry pet foods, while inhalation accounted for 0.21-8.2 ng/kg bw/day, on setting the nominal ∑6 PCB contamination in outdoor and indoor air at 0.37 and 15 ng/m3, respectively. Italian indoor cats could be exposed to higher levels of ∑6 PCBs than the Reference Dose (RfD) of 10 ng/kg/bw/day; this supports the World Health Organization's statement that the quality of the indoor environment is a major determinant of health.

Accumulation of Polychlorinated Biphenyls in Mussels: A Proteomic Study.

Polychlorinated biphenyls (PCBs) are environmental pollutants of industrial origin that can contaminate food, mainly food of animal origin. Although production of PCBs has been banned in many countries since the 1980s, they are still present in the environment and are considered dangerous pollutants for human health. In fact, they can bioaccumulate in living organisms such as marine organisms because of their chemical and physical properties. New analytical approaches are useful to monitor the presence of such contaminants in seafood products and in the environment. In this work, we evaluate changes in protein expression of Mytilus galloprovincialis (Lam.) experimentally exposed to a PCB mixture and identify chemically specific protein expression signatures by using a proteomic approach. In particular, we identify 21 proteins whose levels of expression are sensibly modified after 3 weeks of exposure. The present work shows that a proteomic approach can be a useful tool to study alterations of protein expression in mussels exposed to PCBs and represents a first step toward the development of screening protocols to be used for biomonitoring surveys of fishery products.

Occurrence of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in eggs from free-range hens in Campania (southern Italy) and risk evaluation.

Hens' eggs collected in home-producing farms located in different areas of the Campania region of southern Italy were analysed for six NDL-polychlorobiphenyls (PCB indicators), 17 polychlorodibenzo-p-dioxins (PCDDs) and polychlorodibenzofurans (PCDFs), and 12 congeners of dioxin-like PCBs (DL-PCBs). In the investigated area, waste from civil, industrial and hospital activities was illegally dumped and in many cases burned, so the area is called the 'Land of fire'. This illicit practice has been considered the main source of pollution in this territory and the cause of the persistent organic pollutants' (POPs) contamination in the neighbouring lands intended for agricultural use and livestock. The results, which were obtained from the analysis of 56 samples of eggs, showed levels of contamination by dioxins and PCBs that have often exceeded the action levels set by European Union Recommendation 711/2013 and sometimes even exceeded the maximum levels set by Commission Regulation (EU) 1259/2011. The median PCDD/Fs was 0.41 pg TEQ g-1 fat (range = 0.01-6.18 pg TEQ g-1 fat) and the median concentration of DL-PCBs was 0.70 pg TEQ g-1 fat (range = 0.01-14.60 pg TEQ g-1 fat), whereas NDL-PCBs median was 7.35 ng g-1 fat (range = 0.48-67.55 ng g-1 fat). Nevertheless, the estimated weekly intake (EWI) calculated for consumers of eggs home produced in Campania is lower than the tolerable weekly intake (TWI) established by the European Union Scientific Committee on Food (SCF).

Non-dioxin-like PCB and PBDE deposition on Zea mays L. leaves: modelled contamination in milk from dairy animals fed on silage.

Maize (Zea mays L.) can intercept airborne pollutants before their deposition on soil. Selected non-dioxin-like polychlorinated biphenyls (Σ6NDL-PCBs) and polybrominated diphenyl ethers (Σ8PBDEs) with feed and food safety relevance were measured on maize leaves harvested for silage in dairy animals from 28 fields in Italy. Analyses were carried out by gas chromatography coupled to high- and low-resolution mass spectrometry. Contamination ranged from 0.65 to 5.3 ng g(-1) with 12% moisture for Σ6NDL-PCBs, and from 2.7 to 6.2 for Σ8 PBDEs. Modelled contamination in cow's milk was estimated to fall within the range 0.27-16 ng g(-1) for PCBs, 0.17-1.9 for PBDE number 47, and 0.22-2.1 for PBDE number 99 on a lipid basis. The results indicate that maize silage alone may raise Σ6NDL-PCB contamination in dairy milk up to the 95th percentile in the European Union. Results are discussed in terms of air quality standards able to support food safety.

Serum levels of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and polychlorinated biphenyls in a population living in the Naples area, southern Italy.

The objective of this study was to estimate the levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (DL-PCBs and NDL-PCBs) in blood serum obtained from non-occupationally exposed volunteers living in the Naples area (Campania Region, southern Italy). The samples were taken from two geographical zones: one was an urban area of Naples and its surroundings and the other was located in an area deemed to be at high environmental risk. Total mean concentrations of these persistent pollutants proved to be in the range 1.43-17.38 pg WHO-TEQ1998 g(-1) lipid for PCDD/Fs, and 0.98-25.45 pg WHO-TEQ1998 g(-1) lipid for DL-PCBs. NDL-PCBs were in the range 316.57-482.90 ng g(-1) lipid. No significant differences were observed between women and men, nor between donors living in the two different areas. The mean levels of PCDD/Fs and PCBs in the population living in the Naples area were lower than those observed in some studies of populations living in exposed areas (near incineration plants or industrial sites) and urban or rural areas.

Predicting dioxin-like PCBs soil contamination levels using milk of grazing animal as indicator.

Dioxin-like PCBs (dl-PCBs) are ubiquitous persistent organic pollutants of recognized negative effects on human health. Assessing highly polluted areas should be an important public health issue. This study proposes to use the milk of grazing animals as a bioindicator of dl-PCB contamination in the environment. The hypothesis is that milk concentration of dl-PCBs are related to soil concentrations of these compounds, and that soils are generally reflective of a larger environmental issue of dl-PCB contamination. In this study, we evaluate the possibility of predicting soil concentrations using milk of sheep, cows and buffalos in a spatial model. For this purpose, samples of soil and milk collected in Campania (Italy) were introduced in a GIS platform to perform geostatistical analysis for building a simple predictive model. The ordinary least squares regressions (OLS) showed a statistically significant correlation (p<0.05) between soil and milk contamination. However, this relationship was spatially variable. Thus, a geographically weighted regression (GWR) was performed, obtaining R(2) values of 0.91, 0.77 and 0.66 for sheep's, buffalo's and cow's milk respectively. Assessed the mathematical relationships between the variables, new data was introduced to evaluate the performance of the model. Predictions of soil contamination with dl-PCBs using sheep's, cow's and buffalo's milk showed a mean error of 23%, 25% and 36% respectively. According to these results the sheep's milk can be considered the best bioindicator of dl-PCBs contamination among the three species. The results of this project evidence the potentialities of the proposed approach to assess bioindicator performance in a spatial predictive model.

Contamination levels and congener distribution of PCDDs, PCDFs and dioxin-like PCBs in buffalo's milk from Caserta province (Italy).

An extraordinary plan of official control was carried out in 2008 in Campania (Italy) with the aim to monitor polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) levels in buffalo milk and to detect the contaminated farms, most of which are located in Caserta province. For these companies has been ordered seizure and execution of additional analyses has been requested in farms falling in the nearness, within a distance of 3km, for a total of 304 farms examined. Moreover, all non-compliant farms were subjected to a periodic sampling in order to monitor trends in the levels of contamination. In this paper the distribution and the concentrations of 17 PCDD/Fs and 12 dioxin-like PCBs in 460 samples of buffalo milk collected in the province of Caserta (Italy) are presented. The range of WHO-TEQ values for the PCDD/Fs in milk was 0.17pgTEQg(-1)fat and 87.0pgTEQg(-1)fat with a mean value 3.63pgTEQg(-1)fat and medium value 2.25pgTEQg(-1)fat. The concentrations of dioxin-like PCBs in the analysed samples ranged from 0.21pgTEQg(-1)fat to 15.9pgTEQg(-1)fat and the WHO-TEQ values of sum of PCDDs, PCDFs and dl-PCBs ranged from 0.45pgTEQg(-1)fat to 103.0pgTEQg(-1)fat. The geo-referencing analysis allowed to individuate a restricted area of the region object of the present study where is located the majority of the non-compliant farms. The study of the congeners distribution has finally suggested that the likely cause of contamination is to be attributed to the illegal burning of waste.