Tetracycline Residues in Bovine Muscle and Liver Samples from Sicily (Southern Italy) by LC-MS/MS Method: A Six-Year Study.

We examined a total of 369 bovine liver and muscle samples for the detection of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DOX) residues by implementation and validation of a LC-MS/MS method. The method showed good recovery values between 86% and 92% at three levels of concentrations. The linearity tests revealed r² > 0.996 for all the tetracyclines examined. Furthermore, the Youden test revealed that the method was robust. Only 14.4% of the samples showed OTC and TC residues in a concentration range of 10.4⁻40.2 µg kg-1. No CTC and DOX residues were found in all the samples analyzed. Liver samples showed the highest average values (31.5 ± 20.6 and 21.8 ± 18.9 for OTC and TC, respectively). The results showed a low incidence of TCs in all the samples examined, in comparison with other studies reported in the literature. A significant decrease in TC residues frequency was found from 2013 (p < 0.05). This work reports for the first time epidemiological data on the presence of TC residues in liver and muscle samples of cattle farmed in Sicily (Southern Italy). The very low incidence of TC residues indicates a continuous improvement in farming techniques in Southern Italy, which is essential to ensure consumers' protection.

Evaluating the inhibition effect of a novel bio-packaging on histamine content in mackerel (Scomber scombrus): a new strategy with environment respect.

This is the first study in which the impact of a new bio-packaging on histamine formation in mackerel was investigated. To monitor the preservation of fresh fish samples a treatment with innovative polymeric film and a soaking procedure in liquid new biomaterial was adopted. Ultra HPLC diode array detection (UHPLC-DAD) analyses for histamine monitoring were performed on fresh, packaged, and soaked mackerel samples at different time. The histamine content threshold value was up to 7 days, and after this period the results showed that histamine levels were affected by application of biomaterial. Significant increase was detected in sample that was not treated with biofilm. The new biofilm extended the shelf-life and reveals a promising packaging system to prevent histamine biosynthesis.

[Recurrent multifocal cancer lesions in a patient on hemodialysis after kidney pancreas transplant failure]. / Lesioni neoplastiche multifocali e recidivanti in un paziente in emodialisi dopo fallimento di trapianto combinato rene-pancreas.

Cancer is an important cause of mortality in patients on hemodialysis and kidney transplant recipients. Immunodepression and the genotoxic action of uremia are critical pathogenic agents. A 59-year-old man, ex-smoker, who had been on hemodialysis for seven months because of uremic degeneration of diabetic nephropathy, underwent a combined kidney-pancreas transplant in 1991, complicated by slow-resolution CMV infection. In 1993, after kidney graft failure due to chronic rejection, hemodialysis treatment was restarted with good pancreatic function. Steroid therapy was interrupted and azathioprine and cyclosporine immunosuppressive therapy maintained. In September 2007 the patient was diagnosed with two neoplasms of the oral mucosa: a well-differentiated squamous carcinoma and a spinocellular carcinoma associated with field cancerization. The tumors were resected, followed by laser treatment. Histological examination revealed squamous cell carcinoma without lymph node involvement. Azathioprine was interrupted. In January 2008 adjuvant radiotherapy to the surgical areas of the oral mucosa and neck was started. In February a verrucous nevus on the patient's chest turned out to be a spinocellular carcinoma in situ. In May 2008 recurrence of keratinizing squamous carcinoma of the oral mucosa was found, this time with nodal involvement. Cyclosporine administration was interrupted and after consultation with the oncology committee it was decided to continue with supportive therapy only, until the patient's death in August 2008.

Development of a Rapid and Eco-Friendly UHPLC Analytical Method for the Detection of Histamine in Fish Products.

We developed, validated, and confirmed with proficiency tests a fast ultra-high-performance liquid chromatography with diode array detector (UHPLC-DAD) method to determine histamine in fish and fishery products. The proposed method consists of two successive solid-liquid extractions: one with a dilute solution of perchloric acid (6%) and the second only with water. The instrumental analysis with UHPLC provides a very fast run time (only 6 min) with a retention time of approximately 4 min, a limit of quantification (LOQ) of 7.2 mg kg-1, a limit of detection (LOD) of 2.2 mg kg-1, a recovery around 100%, a relative standard deviation (RSD%) between 0.5 and 1.4, and an r2 of calibration curve equal to 0.9995. The method detected optimal values of the validation parameters and required a limited number of reagents in comparison to other methods reported in the literature. Furthermore, the method could detect histamine in a very short time compared with other methods. This method, in addition to being validated, precise, specific, and accurate, avoids wasting time, money, and resources, and limits the use of organic solvents.

Histamine in Fish Products Randomly Collected in Southern Italy: A 6-Year Study.

ABSTRACT: In total, 4,615 fresh and processed fish samples collected from 2010 to 2015 were analyzed for histamine by ultrahigh-performance liquid chromatography with diode array detection. Histamine levels were detected in 352 (7.6%) samples, with a maximum of 4,110 mg kg-1 and mean values of 908.9 ± 1,226.79 and 344.01 ± 451.18 mg kg-1 for fresh and processed fish samples, respectively. No histamine levels were found in canned tuna and smoked fish samples in contrast to most of the data reported in the literature. A low percentage (2.79%) of noncompliant samples was found. The highest mean values were found during 2011 and 2015 for fresh and processed fish samples, respectively, showing a significant (P < 0.05) difference between the sampling years. The histamine contents found in fresh fish samples were significantly higher (P < 0.05) than those of processed samples. Most of the positive samples came from street vendors, suggesting the need to improve inspection measures in these commercial categories to ensure fish product safety.

Aflatoxin M1 in cow, sheep, and donkey milk produced in Sicily, Southern Italy.

Samples (n = 485) of raw (n = 394) or heat-treated (n = 91) milk of three different species (cow, n = 170; sheep, n = 133; donkey, n = 84), collected 2013-2016 in Western Sicily (Southern Italy), were analyzed for aflatoxin M1 (AFM1) by enzyme-linked immunosorbent assay (ELISA). Positive ELISA results were further analyzed by HPLC with fluorescence detection. Both methods had a detection limit for AFM1 in milk of 7 ng kg-1. ELISA yielded 12.9 and 5% positives in cows and sheep milk, respectively, all samples of donkey milk were negative. Levels of AFM1 were in most cases at 0.007-< 0.05 µg kg-1, only two samples (sheep milk) slightly exceeded the European Union maximum level of 0.05 µg kg-1. Only 6% of the samples were positive for AFM1 in a concentration range of 0.008-0.15 µg kg-1. Only milk samples collected directly from farms were positive. Overall, the levels were much lower than previously reported for Southern Italy cow and sheep milk samples purchased in retail stores. The results of this work indicate a continuous improvement of the feeding techniques on dairy farms of Southern Italy, which is essential to ensure consumers' protection.

Presence of Anisakis pegreffii in farmed sea bass (Dicentrarchus labrax L.) commercialized in Southern Italy: A first report.

We examined 151 European sea bass (Dicentrarchus labrax L.) samples from farms and fish markets of Sicily (Southern Italy) for Anisakidae larvae detection. All the samples were examined by visual inspection and modified chloro-peptic digestion. Two nematode larvae were found in the viscera of only one European sea bass sample from a farm located in Greece (FAO 37.3), giving a total prevalence of infestation of 0.7%. No other parasites were found after chloro-peptic digestion of the samples. The larvae were morphologically ascribed, at genus level, to morphotypes I and molecularly identified as Anisakis pegreffii. To the best if our knowledge, this is the first report on the presence of anisakid parasites in farmed European sea bass of Mediterranean Sea. Our findings suggest that the risk of exposure to Anisakidae nematodes in farmed European sea bass remains very low. However, further data on Mediterranean farms are needed to have a detailed risk analysis.

Monitoring of neonicotinoid pesticides in beekeeping.

The decline of pollinating species is correlated to the extensive use of neonicotinoids against pest insects for crop protection. In this study, the concentrations of neonicotinoid insecticides were determined in honeybees, honeycomb and honey samples, collected in Spring 2015 (blooming period) from different areas in Sicily (IT), to carry out an evaluation of bees products' safety and an overview of neonicotinoid contamination in beekeeping. The results obtained showed only the presence of clothianidin in bee samples and these concentrations don't represent a risk for bees' vitality and safety. The absence of residue in all honey samples, instead, showed the quality of bee products.

Utility of Acute Phase Proteins as Biomarkers of Transport Stress in Ewes and Beef Cattle.

The effect of transport on serum amyloid A (SAA), haptoglobin (Hp), Fibrinogen and white blood cells (WBC) was evaluated in 10 ewes and 10 beef cattle. All animals were transported by road for 6 h over a distance of about 490 km with an average speed of 80 km/h. Blood samples, collected via jugular venepuncture, were obtained before and after transport as well as after 12, 24 and 48 h rest time. One-way repeated measures analysis of variance showed a statistically significant effect of sampling time on SAA, Hp, and WBC in ewes and beef cattle. Based on these results, Hp and SAA levels, together with WBC, may be useful indicators of animal health and welfare and in predicting the risk assessment in meat inspection.

Microbiological and Chemical Analysis of Land Snails Commercialised in Sicily.

In this study 160 samples of snails belonging to the species Helix aspersa maxima and Helix aspersa muller were examined for chemical and microbiological analysis. Samples came from Greece and Poland. Results showed mean concentration of cadmium (0.35±0.036 mg/kg) and lead (0.05±0.013 mg/kg) much higher than the limit of detection. Mercury levels in both species were not detected. Microbiological analysis revealed the absence of Salmonella spp. and Clostridium spp. in both examined species. E. coli and K. oxytoca were observed in Helix aspersa maxima and Helix aspersa muller. Furthermore, one case of fungi positivity in samples of Helix aspersa muller was found. The reported investigations highlight the need to create and adopt a reference legislation to protect the health of consumers.

Utility of Acute Phase Proteins as Biomarkers of Transport Stress in Ewes and Beef Cattle.

The effect of transport on serum amyloid A (SAA), haptoglobin (Hp), Fibrinogen and white blood cells (WBC) was evaluated in 10 ewes and 10 beef cattle. All animals were transported by road for 6 h over a distance of about 490 km with an average speed of 80 km/h. Blood samples, collected via jugular venepuncture, were obtained before and after transport as well as after 12, 24 and 48 h rest time. One-way repeated measures analysis of variance showed a statistically significant effect of sampling time on SAA, Hp, and WBC in ewes and beef cattle. Based on these results, Hp and SAA levels, together with WBC, may be useful indicators of animal health and welfare and in predicting the risk assessment in meat inspection.

Anisakidae in Fishing Products Sold in Sicily.

One of the parasite diseases associated with the consumption of raw fish that occurs with some frequency is the anisakiasis, a human disease caused by the accidental ingestion of larval nematodes of the genus Anisakis, family Anisakidae. At the National Reference Centre for Anisakiasis (C.Re.N.A.) from October 2012 to February 2013, a number of 231 bony fish (Trichiuridae, Engraulidae, Scombridae and Clupeidae) were received from different fishing sites in Sicily. Anisakis pegreffii is the main species detected in fish, as identified by molecular analysis based on polymerase chain reaction-restriction fragment length polymorphism, while Anisakis simplex sensu stricto was found only in Scomber scombrus caught in the Mediterranean Sea (Fishing Areas 37), in the Spanish coast (Fishing Areas 37) and in the Atlantic Ocean (Fishing Areas 34). Larvae of the genus Pseudoterranova were found only in fish caught in the Norwegian Sea.