Assessment of mycotoxin contamination in food products for human consumption

L'aflatossina B1 (AFB1), l'aflatossina M1 (AFM1), e l'ocratossina A (OTA) sono micotossine prodotte da funghi che possono contaminare diverse colture o prodotti di origine animale. Sono sostanze tossiche che rappresentano un rischio per la salute umana, in quanto dotate di potere cancerogeno e mutageno. Il presente progetto di ricerca nasce con l’obiettivo di valutare presenza e livelli di AFB1, AFM1, e OTA in diversi alimenti destinati al consumo umano. La ricerca di AFB1 e di AFM1 è stata condotta su 58 confezioni di latte e 90 confezioni di farina acquistati in diversi punti vendita distribuiti sul territorio di Bologna e provincia. Il monitoraggio condotto sul latte d’asina è stato incentrato sull’analisi di 63 campioni provenienti da un allevamento situato in Piemonte. La ricerca di OTA è stata effettuata su 100 campioni di salami (50 salami calabresi e 50 salami siciliani) prodotti artigianalmente. Dopo aver messo a punto la metodica estrattiva che prevedeva l’utilizzo di colonnine di immunoaffinità (IAC), le analisi sono state eseguite mediante cromatografia liquida ad alta prestazione con rivelazione a fluorescenza (HPLC-FD). Di tutti i campioni analizzati, solo uno dei campioni analizzati presentava un livello di contaminazione superiore ai limiti definiti per legge. Dai risultati ottenuti si evince come sia importante non solo tenere sotto controllo la filiera produttiva di alimenti destinati all'uomo e agli animali, ma anche effettuare un monitoraggio continuo della contaminazione da micotossine in questi prodotti.Aflatoxin B1 (AFB1), aflatoxin M1 (AFM1), and ochratoxin A (OTA) are mycotoxins produced by fungi that contaminate agricultural commodities or food of animal origin. These substances present a serious health hazards such as carcinogenicity and mutagenicity. The aim of this research was to assess the mycotoxin contamination in different food products for human consumption. To verify the levels of aflatoxin B1 and AFM1 in flour (40 wheat flour and 50 corn flour) and milk from conventional and organic production purchased from local stores in the surrounding of Bologna, 90 samples of flour and 58 samples were analyzed. The presence of AFM1 was also investigated in donkey milk. For this purpose, 63 samples of milk collected in a farm of the North of Italy were analyzed. As regards OTA, 100 handmade salami (50 produced in Calabria and 50 produced in Sicilia) were investigated. For AFB1, a fast and sensitive high performances liquid chromatography with fluorescent detection (HPLC-FD) was developed and validated. For AFM1 and OTA, immunoaffinity-based techniques were used and analyses were performed by HPLC-FD. Thirteen samples of corn flour were positive for AFB1, but only one showed levels of contamination above the limit defined at Community level (2 µg/kg). No significant differences were observed in the concentration of AFB1 by comparing conventional and organic corn flour samples. With regard to AFM1, 35 samples of milk purchased from local stores and 1 sample of donkey milk were contaminated but none showed a level of contamination above the legal limit of 0.05 µg/kg. OTA was detected in 25 samples of salami, but only one exceeded the guideline level of 1 µg/kg established by Italian Ministry of Health. The food analyzed are safe as regards the considered mycotoxins. However, is very important to undertake a continuous monitoring to assure safe products to consumers

Pharmacokinetic profile and partitioning in red blood cells of romifidine after single intravenous administration in the horse

The aims of this study were to assess the plasma concentrations of romifidine in horses after intravenous injection, to evaluate the red blood cell (RBC) partitioning of the anaesthetic drug, and to improve knowledge regarding its sedative effect in horses describing the pharmacokinetic model. Eight adult Standardbred horses received a single bolus of romifidine at a dosage of 100\ua0\u3bcg/kg. Blood samples (5\ua0mL) were collected immediately before romifidine administration (t0), and at 2, 5, 10, 15, 20, 25, 30, 40, 50, 60, 75, 90, 105, 120, 150 and 180\ua0min after injection. A sedation score was recorded at the same time. The romifidine concentrations in plasma and red blood cells were determined by high performance liquid chromatography (HPLC). The plasma and red blood cell concentrations were correlated with the sedation at each time point. Romifidine produced a satisfactory level of sedation in all animals. The sedation was detectable in all horses for up to 105\ua0min. All the animals returned to normal without any behavioural changes at 180\ua0min. The romifidine concentrations in the red blood cells were significantly higher (P\ua0<\ua00.01) at all time points than those in the plasma. The T1/2\u3b2 was 148.67\ua0\ub1\ua061.59\ua0min and body clearance was 22.55\ua0\ub1\ua06.67\ua0mL/kg per min. The results showed that after a single bolus administration of romifidine, a partitioning in the RBCs was detected

Occurrence of aflatoxin M1 in conventional and organic milk offered for sale in Italy

In the present study, 58 samples of milk were analyzed for the presence of aflatoxin M1 (AFM1). The samples were purchased during the period April–May 2013 in a random manner from local stores (supermarkets, small retail shops, small groceries, and specialized suppliers) located in the surrounding of Bologna (Italy). The commercial samples of milk were either organic (n = 22) or conventional (n = 36); fresh milk samples and UHT milk samples, whole milk samples, and partially skim milk samples were present in both the two considered categories. For the quantification of AFM1 in milk, the extraction-purification technique based on the use of immunoaffinity columns was adopted and analyses were performed using HPLC-FD. AFM1 was detected in 35 samples, 11 from organic production and 24 from conventional production. No statistically (P > 0.05) significant differences were observed in the concentration of AFM1 in the two categories of product. The levels of contamination found in the positive samples ranged between 0.009 and 0.026 ng mL−1. No sample exceeded the limit defined at community level for AFM1 in milk (0.05 μg kg−1). This demonstrates the effectiveness of the checks before the placing on the market of these food products. Thus, the Baflatoxins^ problem that characterized the summer of 2012 does not seem to have had effect on the contamination level of the considered milk samples

Ochratoxin A in artisan salami produced in Veneto (Italy)

Fifty samples of artisan salami purchased in Veneto (Italy) were analysed for the determination of ochratoxin A (OTA). The analytical method, based on a sample preparation procedure with immunoaffinity columns (IACs), together with analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD), has guaranteed a high rate of recovery (about 97%), limit of detection (LOD) and limit of quantification (LOQ), respectively, of 0.06\ua0\ub5g\ua0kg 121 and 0.20\ua0\ub5g\ua0kg 121. OTA was detected in five samples, but only one exceeded the guideline value (1\ua0\ub5g\ua0kg 121) established by the Italian Ministry of Health for pork meat and derived products. The results would seem to suggest that salami made with the traditional, non-industrial production method can be considered safe as regards contamination by OTA. However, the very high concentration observed in one sample proves that a high OTA contamination is also possible in this type of product. Thus, the controls of mycotoxin contamination must consider also salami

Occurrence of aflatoxin B1 in conventional and organic flour in Italy and the role of sampling

Aflatoxins (AFs) are mycotoxins produced by certain species of Aspergillus and aflatoxin B1 (AFB1) is the most toxic, consistently carcinogenic and genotoxic. In the present study, a total of 90 different samples of organic and conventional flours (20 conventional wheat flour, 20 organic wheat flour, 42 conventional corn flour, and 8 organic corn flour) were analyzed for the presence of AFB1. The samples were purchased from local stores in the surrounding of Bologna during the period April to May 2013. For the quantification of AFB1 in flours, a fast and sensitive HPLC-FD method was developed and tested for several validation parameters (linearity and range, specificity, accuracy and precision, LOD, LOQ and recovery). AFB1 was found in 13 samples of corn flour, specifically 4 organic and 9 conventional. These results confirm a higher incidence of contamination in corn compared to wheat, as reported in literature. No significant differences were observed in the concentration of AFB1 by comparing conventional corn flour samples and organic corn flour samples (P > 0.05). The levels of contamination found in the positive samples ranged between 0.17-3.75 \u3bcg kg-1 but only in one case the limit defined at Community level (2 \u3bcg kg-1) was exceeded. This demonstrates the effectiveness of the network of monitoring before the placing on the market of these food products. Furthermore, in this study efforts were made to evaluate the minimum quantity of flour related to a commercial package, which is reliable for the assessment of contamination by AFB1. In fact, in sample preparation, the selection of the sample size for the laboratory analysis is a very important step. The minimum amount to get a reliable result is 20 grams and smaller quantities can lead to incorrect results

Biliary ochratoxin A as a biomarker of ochratoxin exposure in laying hens: An experimental study after administration of contaminated diets

To evaluate the levels of ochratoxin (OTA) in kidney, liver and bile of laying hens, forty-five laying hens were enrolled in this study and divided into three equal groups: a control group D0, and two experimental groups, D1 fed with 10 \u3bcg/kg OTA diet and D2 fed with 200 \u3bcg/kg OTA diet for 6 weeks. Kidneys, livers, and bile from all hens were collected and analyzed by HPLC method for the presence of OTA. Eggs collected 2 days before the start of the experiment and 2 days after its end were also analyzed for the presence of OTA. Results show a relevant biliary excretion of the mycotoxin, with high levels of OTA in the bile after administration of the toxin. OTA level in eggs was below the limit of detection (LOD). These results suggest the suitability of using bile as a matrix for screening measurements of OTA in laying hens

LC–MS/MS Analysis of Five Neonicotinoid Pesticides in Sheep and Cow Milk Samples Collected in Jordan Valley

The purpose of the present study was to evaluate the presence of five neonicotinoid pesticides, acetamiprid, imidacloprid, clothianidin, thiacloprid, and thiamethoxam, in sheep and cow milk samples collected from animals bred in the Jordan Valley. In this area, numerous citrus plantations are present, and these insecticides are commonly used to protect plants from pests and diseases. Thirty-seven sheep milk samples and 31 cow milk samples were analysed. The analytical method, based on a single cleanup extraction step with SPE cartridges packed with diatomaceous earth material, together with analysis by LC–MS/MS, has guaranteed average recoveries between 75.1% and 88.3%, limits of detection (LOD) and quantification (LOQ) of 0.5 and 1&nbsp;µg/kg, respectively, for all the five neonicotinoids. LOQ was much lower than the codex maximum residues limits for these pesticides in milks. No residues of the five neonicotinoids were found in any sample at a concentration level above LOD

Occurrence of imidacloprid, carbendazim, and other biocides in Italian house dust: Potential relevance for intakes in children and pets

The occurrence of pesticides intended for non-agricultural use was investigated in 206 dust samples drawn from vacuum-cleaner bags from residential flats in Italy. The multi-residue analysis targeted on 95 different active principles was performed with UPLC-MS/MS, with a Limit of Quantification (LOQ) of 0.008\ua0\u3bcg/g dry weight. The results indicated the presence of imidacloprid (IMI) and carbendazim (CARB) in 30% and 26% of the samples, with a mean and P95 concentration between 1.6 and 39 and between 0.08 and 4.9\ua0\u3bcg/g, respectively. Combined presence of two biocides was noted in 19.4% samples, of three biocides in 9.2% samples, of four biocides in 3.4% samples, and of five and six biocides in 0.5% and 1% samples, respectively. According to the estimated dust intake in infants/toddlers aged 6-24 months (16-100\ua0mg d(-1)) and cats (200\ua0mg d(-1)), it was possible to obtain risk characterization with respect to the Acceptable Daily Intake (ADI) for IMI of 0.060 mg/kg body weight (bw) proposed by the European Food Safety Authority (EFSA) and the chronic Population Adjusted Dose (cPAD) of 0.019 mg/kg bw d(-1) by US-EPA. Under the worst-case scenario, the presence of IMI in dust indicates potential exceedance of the cPAD in kittens, to be considered as sentinel also accounting for combined exposure. This study highlights the relevance of consumer empowerment about the responsible use of pesticides as biocidal products in indoor environment