Matilde Cecchini

Histamine poisoning is the most common cause of human foodborne illness due to the consumption of fish products. An enzyme-based amperometric biosensor was developed to be used as a screening tool to detect histamine and histamine-producing bacteria (HPB) in tuna. It was developed by immobilizing histidine decarboxylase and horseradish peroxidase on the surface of screen-printed electrodes through a cross-linking procedure employing glutaraldehyde and bovine serum albumin. The signal generated in presence of histamine at the surface of the electrode was measured by chronoamperometry at in presence of a soluble redox mediator. The sensitivity of the electrode was 1.31–1.59 μA/mM, with a linear range from 2 to 20 μg/ml and detection limit of 0.11 μg/ml. In this study fresh tuna filets purchased in supermarkets in different days (n = 8) were analyzed to detect HPB. Samples with different concentration of histamine were analyzed with culture-based counting methods, biosensor and HPLC and also a challenge test was made. Recovery of histamine from cultures and tuna samples was also assessed. The presence of Morganella psychrotolerans, Photobacterium phosphoreum, P. damselae and Hafnia alvei was detected using culture- and PCR-based methods. At the time of purchase these tuna samples had histamine concentrations from below the limit of detection (LOD) to 60 μg/g. HPLC and biosensor methods provided similar results in the range from zero to 432 μg/g (correlation coefficient, R² = 0.990) and the recovery of histamine from cultures and tuna samples was very high (mean bias −12.69 to 1.63%, with root-mean-square error <12%). These results clearly show that fresh tuna is commonly contaminated with strong HPB. The histamine biosensor can be used by the Food Business Operators as a screening tool to detect their presence and to determine whether their process controls are adequate or not.

Photobacterium damselae subsp. damselae (Pdd) is considered to be an emerging pathogen of marine fish and has also been implicated in cases of histamine food poisoning. In this study, eight strains isolated from mullets of the genera Mugil and Liza captured in the Ligurian Sea were characterized, and a method to detect histamine-producing Pdd from fish samples was developed. The histamine-producing potential of the strains was evaluated in culture media (TSB+) using a histamine biosensor. Subsequently, two strains were used to contaminate mackerel fillets (4 or 40 CFU/g), simulating a cross-contamination on the selling fish stalls. Sample homogenates were enriched in TSB+. The cultures were then inoculated on thiosulfate-citrate-bile salts-sucrose agar (TCBS) and the dark green colonies were cultured on Niven agar. The violet isolates were characterized using specific biochemical and PCR based tests. All Pdd strains were histamine producers, yielding concentration varying from 167 and 8977 µg/mL in TSB+ cultures incubated at 30 °C for 24 h. Pdd colonies were detected from the inoculated mackerel samples and their histidine decarboxylase gene was amplified using species-specific primer pairs designed for this study. The results indicate that mullets can be source of Pdd and the fish retailers needs to evaluate the risk posed by cross-contamination on the selling fish stalls.

Beeswax coating of foods put a barrier to oxygen, light, and vapour that can help to prevent oxidation of fat and pigments and water loss. The amounts of 2-thiobarbituric acid reactive substances (TBARS) and water activity (Aw) were assessed in Italian salami coated with beeswax at 55 days of ripening and compared with controls at 5, 6, and 7 months of shelf life. The results were correlated with sensory quality. TBARS levels were below 0.8 mg kg⁻¹ in the beeswax-coated salami until 6 months of aging (median 0.697, max 0.795) and significantly higher in the uncoated salami (median 1.176, max 1.227). A slight correlation between the amount of TBARS and Aw was observed in beeswax-coated salamis, whereas this effect was masked in controls by the large Aw variability observed at 7 months. Beeswax coating prevents case hardening and facilitated the peeling.

Evaluation and identification of histamine-forming bacteria on fish products of middle Adriatic Sea

A square-wave anodic-stripping voltammetric method for the analysis of lead and cadmium in chicken muscle and liver was developed and validated, and the results of a monitoring study relative to chicken and pigeon meat are reported. The voltammetric method allows the analysis of lead and cadmium at the same time in samples after acid digestion. The use of perchloric acid for digestion and of acetate buffer in the supporting electrolyte are suitable to reduce matrix interferences and obtain limits of quantification which were below 10 ng g⁻¹ for meat and liver samples. The regression between the analytical signal and the concentration of the target analytes in spiked samples and Certified Reference Materials proved to be linear within the 10–100 ng g⁻¹ range for meat and within the 50–500 ng g⁻¹ range for liver. The analytical method was verified using available Certified Reference Materials BCR-184 (cattle meat) and BCR-185R (cattle liver) as well as with spiked chicken samples. Precision (i.e. repeatability and intermediate precision) and accuracy (percentage recovery and bias) were of the order of 0.3–4.5% for both lead and cadmium The level of lead in muscle was in the range between 6.4 and 59.8 ng g⁻¹ in chickens and between 7.9 and 63.6 ng g⁻¹ in farmed pigeons, whereas it was between 8.0 and 84.4 ng g⁻¹ in chicken liver. The cadmium concentration was 0.4–10.4 ng g⁻¹ in chicken muscle, 10.4–90.6 ng g⁻¹ in chicken liver and 2.2–8.0 ng g⁻¹ in farmed pigeons.

The present work provides an overview of the most relevant results concerning a five years monitoring programme of wet and dry deposition (1995-1999) in the city of Bologna. The aim of this research is to get an overall picture of atmospheric pollutants deposition inventory in an urban territory and to allow an assessment of the vulnerability of the area by comparing actual fluxes of acidity and nutrient nitrogen with the respective "critical loads" associated to the territory, in the framework of the UNECE LRTAP (Long Range Transboundary Air Pollution Convention). This comparison, for the Bologna monitoring station, shows "exceedance classes" from 4 to 6 for acidity and from 1 to 5 for nutrient nitrogen, reflecting an urban-industrial context, though a trend in reducing occurs from 1995 to 1999. A water layer surface sampler (DAS-MTX sampler) was employed for "surrogate" dry deposition collection. The contribute of dry fraction to the total deposition fluxes appears to prevail on wet fraction for many pollutants (up to more than 90% for total calcium and alkalinity). A comparison with long term monitoring results from some stations in Italian territory, shows that the differences among chemical species deposition fluxes may be ascribed both to the long distance aerial transport of pollutants and to site characteristics.

¹³⁷ Cs activity in samples from lacustrine areas around the Italian base in Antarctica is reported as an integration of a previous work. Preliminary data of cosmogenic ⁷ Be activity determined in snowfalls, total atmospheric depositions, soil and air particulate collected during the 1990–91 and 1991–92 Italian expeditions in Antarctica is presented. The results obtained point out the efficiency of snow in the processes of air particulate scavenging and provide useful information for the development of research in Antarctica in the study of air/snow transfer processes by means of natural radionuclides.