Aflatoxin occurrence in milk and supplied concentrates of goat farms of north‐eastern Italy

BACKGROUND: There is little information about the occurrence of aflatoxin M₁ in goat milk. A survey involving 17 dairy goat farms of north‐eastern Italy was completed during 2005 and 2006, in order to evaluate the prevalence of milk contamination and its relationship with type and level of concentrate supplied. RESULTS: 132 concentrate and 85 milk samples were collected during five farm visits and analysed for aflatoxins. Aflatoxin B₁ (AFB₁) was > 0.1 µg kg^?1 in two‐thirds of the feeds and > 5 µg kg^?1 in nine. Contamination was higher in maize than in other pure feeds (median: 0.8 versus 0.1 µg kg^?1); complementary feeds showed intermediate values. Aflatoxin M₁ (AFM₁) was > 3 ng kg^?1 in one‐third of milks and > 25 ng kg^?1 in three. All the milk samples were below EU statutory limits. The farm ranks for milk AFM₁ level and the peak of concentrate AFB₁ contamination were significantly correlated (0.642). CONCLUSIONS: Risk to human health was generally found to be absent, with only a few cases involving feed contamination to be monitored. The main aflatoxin risk for goat milk could arise from maize and maize‐based concentrates in the more intensive breeding conditions. Copyright © 2008 Society of Chemical Industry     

Screening of aflatoxins in duck feedstuffs in West Java,Indonesia

During the period 1979-1985, incoming feed ingredients (some of imported origin) for duck diets were screened for their aflatoxin level. As a result. records of aflatoxin data from 533 corn, 88 rice, 136 rice bran, 47 wheat pollard, 207 soya bean meal. 55 copra meal and 166 fish meal samples were obtained. Aflatoxin B ₁ was the predominant form, the incidence of samples containing it ranging from 69 to 94%; then accompanied by aflatoxin G₁, 37 to 81%: by B ₂, 33 to 70%; by G₂, 13 to 66%. Levels of total aflatoxin > 20 μg kg ^?1 were most frequently encountered in corn (71%) and copra meal (58%), followed by soya bean meal (20%), wheat pollard (15%), rice bran (12%), fish meal (10%) and rice (4%). Most feed ingredients contained aflatoxins < 100 μg kg^?1 except in corn of which 202 samples (38%) had a higher level. Corn is concluded to be the main source of alfatoxin contamination in corn-based mixed feeds.     

Aflatoxin determination in commonly consumed foods in Tunisia

BACKGROUND: To investigate natural aflatoxin occurrence, a total of 180 samples of different foods widely consumed in Tunisia were analysed by an in‐house‐validated high‐performance liquid chromatography method including affinity column clean‐up and post‐column bromination techniques. RESULTS: The method used appeared to be rapid, selective and reproducible, and its performances were established. Detection limits were 0.05 ng g^?1 for aflatoxin B1 and 0.025 ng g^?1 for aflatoxins B2, G1 and G2. Aflatoxins were detected in all investigated commodities except rice, with an overall contamination frequency of 34.4% and concentrations ranging from 0.1 to 40.6 ng g^?1. Aflatoxin B1 was found in all contaminated samples. Sorghum, spices and nuts were most contaminated. CONCLUSION: This study has provided an effective analytical method for the reliable determination of aflatoxins in food samples. Over one‐third of the samples investigated were contaminated with aflatoxins. Sorghum, spices and nuts were most contaminated, whereas rice showed no contamination. Copyright © 2010 Society of Chemical Industry     

Polychlorinated dibenzo-p-dioxins,dibenzofurans and dioxin-like polychlorinated biphenyls in food and feed in Latvia in 2009–2011

During 2009–2011 a monitoring programme for 17 polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs) and 12 dioxin-like polychlorinated biphenyls (DL-PCBs) was conducted in the Latvian food and feed market. Using ISO 17025-accredited analytical methodology, investigation of 121 food (milk, dairy products, meat, eggs, fish, fish products) and 66 feed samples (fish meal and oil, compound and mineral feed, vegetable and animal fats) was performed. Most samples showed contamination below the European Commission (EC) Regulation No. 1881/2006 and Commission Directive 2006/13/EC limits. Average total toxicological equivalent (total-TEQ₍₁₉₉₈₎) concentrations within the food sample groups, except fish and fish products, ranged between 0.41 and 15.1 pg total-TEQ₍₁₉₉₈₎ g^?1 fat. Fish and fish products showed contamination levels from 0.18 to 46.0 pg total-TEQ₍₁₉₉₈₎ g^?1 fresh weight (f.w.). Fifty-seven per cent of cod liver samples were non-compliant. The most contaminated feed samples were fish meal and fish oil. A comparison with WHO-TEF₍₂₀₀₅₎ data is given.     

Decontamination of aflatoxin‐contaminated maize by dehulling

Dehulling of maize grains as an aflatoxin decontamination method was investigated. Sixty kilograms of maize (whose average moisture content was 110 g kg^?1) were thoroughly mixed and divided into two samples. The kernel moisture content of one sample was adjusted to 200 g kg^?1 while the other (control) was left at 110 g kg^?1. The two samples were kept at ambient temperature (25–30 °C) for 21 days. Twenty sub‐samples, ten from each, were further divided into two so that one lot was dehulled while the other was not dehulled. These sub‐samples were then milled to pass through a 1 mm screen and the meal was evaluated for aflatoxin contamination. It was found that there was a 92% decrease in aflatoxin levels in dehulled maize meal compared with undehulled maize meal. We therefore suggest that dehulling the grain can be used to reduce aflatoxin levels in maize. Copyright © 2005 Society of Chemical Industry     

Analysis of industry-generated data. Part 2: Risk-based sampling plan for efficient self-control of aflatoxin M1 contamination in raw milk

Aflatoxin M₁ (AFM1) contamination in 21 969 milk samples taken in Italy during 2005–08 and 2010 provided the basis for designing an early warning self-control plan. Additionally, 4148 AFM1 data points from the mycotoxin crisis (2003–04) represented the worst case. No parametric function provided a good fit for the skewed and scattered AFM1 concentrations. The acceptable reference values, reflecting the combined uncertainty of AFM1 measured in consignments consisting of milk from one to six farms, ranged from 40 to 16.7 ng kg^?1, respectively. Asymmetric control charts with these reference values, 40 and 50 ng kg^?1 warning and action limits are recommended to assess immediately the distribution of AFM1 concentration in incoming consignments. The moving window method, presented as a worked example including 5 days with five samples/day, enabled verification of compliance of production with the legal limit in 98% of the consignments at a 94% probability level. The sampling plan developed assumes consecutive analyses of samples taken from individual farms, which makes early detection of contamination possible and also immediate corrective actions if the AFM1 concentration in a consignment exceeds the reference value. In the latter case different control plans with increased sampling frequency should be applied depending on the level and frequency of contamination. As aflatoxin B₁ increases in feed at about the same time, therefore a coordinated sampling programme performed by the milk processing plants operating in a confined geographic area is more effective and economical then the individual ones. The applicability of the sample size calculation based on binomial theorem and the fast response rate resulting from the recommended sampling plan were verified by taking 1000–10 000 random samples with replacement from the experimental databases representing the normal, moderately and highly contaminated periods. The efficiency of the control plan could be substantially enhanced if the dairy farms used feed with a tolerable level of AFB1.     

Analysis of industry-generated data. Part 1: A baseline for the development of a tool to assist the milk industry in designing sampling plans for controlling aflatoxin M1 in milk

The presence of aflatoxin M₁ (AFM1) in milk was assessed in Italy in the framework of designing a monitoring plan actuated by the milk industry in the period 2005–10. Overall, 21 969 samples were taken from tankers collecting milk from 690 dairy farms. The milk samples were representative of the consignments of co-mingled milk received from multiple (two to six) farms. Systematic, biweekly sampling of consignments involved each of the 121 districts (70 in the North, 17 in the Central and 34 in the South regions of Italy). AFM1 concentration was measured using an enzyme-linked immunoassay method (validated within the range of 5–100 ng kg^?1) whereas an HPLC method was used for the quantification of levels in the samples that had concentrations higher than 100 ng kg^?1. Process control charts using data collected in three processing plants illustrate, as an example, the seasonal variation of the contamination. The mean concentration of AFM1 was in the range between 11 and 19 ng kg^?1. The 90th and 99th percentile values were 19–34 and 41–91 ng kg^?1, respectively, and values as high as 280 ng kg^?1 were reached in 2008. The number of non-compliant consignments (those with an AFM1 concentration above the statutory limit of 50 ng kg^?1) varied between 0.3% and 3.1% per year, with peaks in September, after the maize harvest season. The variability between different regions was not significant. The results show that controlling the aflatoxins in feed at farm level was inadequate, consequently screening of raw milk prior to processing was needed. The evaluation of the AFM1 contamination level observed during a long-term period can provide useful data for defining the frequency of sampling.     

Household dietary exposure to aflatoxins from maize and maize products in Kenya

Aflatoxicosis has repeatedly affected Kenyans, particularly in the eastern region, due to consumption of contaminated maize. However, save for the cases of acute toxicity, the levels of sub-lethal exposure have not been adequately assessed. It is believed that this type of exposure does exist even during the seasons when acute toxicity does not occur. This study, therefore, was designed to assess the exposure of households to aflatoxins through consumption of maize and maize products. Twenty samples each of maize kernels, muthokoi and maize meal were randomly sampled from households in Kibwezi District of Makueni County in Eastern Kenya and analysed for aflatoxin contamination. The samples were quantitatively analysed for aflatoxin contamination using HPLC. The uncertainty and variability in dietary exposure was quantitatively modelled in Ms Excel using Monte Carlo simulation in @Risk software. Aflatoxins were found in 45% of maize kernels at between 18 and 480 μg kg^–1, 20% of muthokoi at between 12 and 123 μg kg^–1, and 35% of maize meal at between 6 and 30 μg kg^–1. The mean dietary exposure to aflatoxin in maize kernels was 292 ± 1567 ng kg^?1 body weight day^?1, while the mean dietary exposure to aflatoxin in maize meal and muthokoi were 59 ± 62 and 27 ± 154 ng kg^?1 body weight day^?1 respectively. The results showed that the amount and frequency of consumption of the three foods is the more important contributing factor than the mean aflatoxin concentration levels, to the risk of dietary exposure to aflatoxins.     

Aflatoxins and cyclopiazonic acid in feed and milk from dairy farms in São Paulo,Brazil

The occurrence of aflatoxins (AF) B₁, B₂, G₁, G₂ and cyclopiazonic acid (CPA) in feeds, and AFM₁ and CPA in milk was determined in dairy farms located in the northeastern region of São Paulo state, Brazil, between October 2005 and February 2006. AF and CPA determinations were performed by HPLC. AFB₁ was found in 42% of feed at levels of 1.0–26.4 µg kg^?1 (mean: 7.1 ± 7.2 µg kg^?1). The concentrations of AFM₁ in raw milk varied between 0.010 and 0.645 µg l^?1 (mean: 0.104 ± 0.138 µg l^?1). Only one sample was above the tolerance limit adopted in Brazil (0.50 µg l^?1) for AFM₁ in milk. Regarding CPA in feed, six (12%) samples showed concentrations of 12.5–153.3 µg kg^?1 (mean: 57.6 ± 48.7 µg kg^?1). CPA was detected in only three milk samples (6%) at levels of 6.4, 8.8 and 9.1 µg l^?1. Concentrations of aflatoxins and CPA in feed and milk were relatively low, although the high frequency of both mycotoxins indicates the necessity to continuously monitor dairy farms to prevent contamination of feed ingredients.     

Aflatoxins in acid-treated grain in Sweden and occurrence of aflatoxin M1 in milk

A survey of aflatoxins in acid-treated grain and milk from farms using such grain was conducted in Sweden during 1986. Aflatoxins occurred most frequently (40%) in grain treated with a new formula of diluted (700 g litre^?1) aqueous formic acid, but also in 31% of the samples of grain treated with 850 g litre^?1 aqueous formic acid. The lowest incidence was found in grain treated with propionic acid, where aflatoxins were found in only one sample (3%). Aspergillus flavus/A parasiticus occurred in the same manner, but were more frequent than the aflatoxins. When cultivated on aflatoxin-producing agar, positive reactions were more common (56%) among strains originating from grain treated with formic acid than among strains originating from grain treated with propionic acid (4%). Aflatoxin M¹ in concentrations over 50 ng kg^?1 was mainly found in milk from farms using formic acid, and in most of these cases aflatoxins were also detected in the grain samples. In some cases, milk from a single farm was contaminated enough to generate consumption milk from the dairy with aflatoxin M¹ concentrations above or close to 50 ng kg^?1. The risk of aflatoxin formation after inadequate treatment of grain with formic acid is very high and is considerably lower with propionic acid. Formic acid has now been prohibited for use as a preservative of high moisture grain in Sweden.     

Evaluation of total aflatoxin,nitrate and nitrite levels in layer feed samples of companies producing their own feed in Edincik and Bandırma province of Turkey

Feed contamination by fungi can lead to nutrient losses and detrimental effects on animal health and production. The presence of nitrates and nitrites in food can be harmful to both people and animals. The aim of this study was to determine total aflatoxin, nitrate and nitrite levels in layer feed samples from companies producing their own feed in Edincik and Band?rma provinces in Turkey and to discuss the potential risk to animal health. The results of the analyses indicated that mean total aflatoxin (AFT) ranged from 0.4 to 36.8?µg?kg^?1 and from 0.45 to 47.0?µg?kg^?1 in the year 2007 and the year 2008 samples, respectively. It was determined that nitrate levels were 2.4–10 and 1.7–13?µg?kg^?1 and that nitrite levels were 0–2.4?µg?kg^?1 and 0–2.6?µg?kg^?1 in these years, respectively. The levels of total aflatoxin, nitrate and nitrite in the layer samples could not be considered a risk to poultry health and productivity.     

A survey of ochratoxin A contamination in feeds and sera from organic and standard swine farms in northwest Italy

BACKGROUND: A survey was carried out on conventional (n = 11) and organic (n = 4) swine farms in northwest Italy in order to investigate the occurrence of ochratoxin A (OTA) in feed and serum samples collected from September 2006 to March 2009. Each farm was sampled twice and a total of 30 feed samples and 285 serum samples were collected. OTA levels were determined through extraction, immunoaffinity column purification and high‐performance liquid chromatography analysis coupled with fluorimetric detection. RESULTS: All feed samples resulted to be contaminated with OTA at levels ranging from 0.22 to 38.4 µg kg^?1. The OTA concentrations found in organic feed samples were significantly higher (P < 0.05) than those found in conventional feed samples. All serum samples resulted to be contaminated with OTA at levels ranging from 0.03 to 6.24 ng mL^?1. The OTA concentrations found in organic serum samples were significantly higher (P < 0.001) than those found in conventional serum samples. CONCLUSION: None of the feed samples contained more than the maximum level (50 µg OTA kg^?1, considering a feed moisture content of 120 g kg^?1) recommended by the European Commission for OTA in complementary and complete swine feedstuffs. The OTA contamination of organic feed and serum samples was found to be significantly higher than that of conventional feed and serum samples. Copyright © 2010 Society of Chemical Industry     

Doxycycline and sulfadimethoxine transfer from cross-contaminated feed to chicken tissues

During feed preparation at feed mills or during feed mixing in bins at farms, the accidental contamination of feed at trace levels by veterinary drug residues, commonly known as carry-over, can accidentally but frequently occur. To evaluate the concentrations of residual antimicrobials in poultry edible tissues, due to contaminated feed, sulfadimethoxine and doxycycline were administered for 10 days to chickens in poultry feed incurred at the contamination levels frequently found during national feed monitoring programmes (1–5?mg?kg^?1). Sulfadimethoxine and doxycycline residual concentrations detected in muscle (<LOD and 31?µg?kg^?1, respectively), liver (13 and 56?µg?kg^?1, respectively) and kidney (56 and 115?µg?kg^?1, respectively) were compared with their maximum residue limits (MRLs) fixed by EC 470/2009 and EU 37/2010 Regulations for a preliminary risk evaluation.     

Aflatoxin levels in raw and processed hazelnuts in Turkey

Aflatoxin levels in hazelnut samples obtained from exporter companies were monitored over a 3-year period. A total of 3188 samples of raw and processed hazelnuts were analysed using an HPLC method. The total aflatoxin content of the contaminated samples was in the range of 0.02–78.98?µg?kg^?1 for hazelnut kernels, 0.07–43.59?µg?kg^?1 for roasted hazelnut kernels, 0.02–39.17?µg?kg^?1 for roasted sliced hazelnut kernels, and 0.02–11.20?µg?kg^?1 for hazelnut purees, respectively, showing that the variations of aflatoxin contamination were very high. The results of aflatoxin analysis revealed that the aflatoxin contamination in the hazelnut samples was at a tolerable level. A total of 3147 samples were contaminated with aflatoxins, although below the legal limits. However, the aflatoxin contents of 41 samples exceeded the legal limits. Therefore, aflatoxin contents of hazelnuts should be monitored regularly to minimise the risk of aflatoxin hazard, and pre- and post-harvest strategies should be developed to prevent aflatoxin formation.     

The determination of aflatoxins in spices by immunoaffinity column extraction using HPLC

Seventy‐five samples of different spices marketed in Turkey were purchased from bazaars, herbal shops and supermarkets. Equal amounts of paprika, chilli, black peppers and cumin were purchased and used to test and compare the amount of aflatoxin contamination. Two different analytical methods were examined for their efficacy by adding a known amount of aflatoxin to the blank samples of paprika. Twenty‐seven paprika, all the chilli powder and four ground black pepper samples were contaminated with aflatoxin B₁ in the range of 0.5–116.4, 1.6–80.4 and 0.3–1.2 μg kg^?1 respectively. Twenty‐three (30%) paprika and chilli powder samples were above the regulatory limits used in the European Union. No aflatoxin contamination was detected in the cumin samples at a detection limit of 0.2 μg kg^?1.     

Aflatoxin and its metabolites in tissues from laying hens and broiler chickens fed a contaminated diet

Two groups of 32 hens and broiler chickens were contaminated with 2.5 and 5 mg of aflatoxin (AF) kg^?1 feed for a period of 32 days. During this contamination 16 birds were sacrificed and aflatoxin and its metabolites were detected using thin-layer chrotnatography and fluorescence densisometry. The tissues analysed (liver, muscle, kidney, gizzard and eggs) gave a wide range of concentrations, the lowest was found in ben muscle (0.05 μg kg^?1 of AFB₁) and the highest in gizzards from the 5 mg kg^?1 group of the hens (9.01 μg kg^?1 of AFB₁). Metabolites of AFB₁, AFM, and AFB_2a appeared in the liver but not in other tissues. In broiler's tissues, the following metabolities were isolated: AFM₁ and AFB_2a, in liver, aflatoxicol in muscle and AFM₁ and AFB_2a in kidneys, all having concentrations lower than AFB₁. Aflatoxicol was isolated from one egg sample (0.32 μg kg^?1). For both types of birds, aflatoxin clearance time was only 24 h for muscle and kidneys. In livers from the 5 mg kg^?1 group, AFM₁ and AFB_2a were still found 4 days after removal of the contaminated feed. In eggs and gizzards, aflatoxin residue was still detected on the 8th day of the clearance period although in low quantities. In the broiler's gizzards, clearance time was only 24 h. These results suggest that aflatoxin transfer to edible tissues is very small and the danger of contaminations to humans is also very small, except in the case of gizzards.     

Survey of aflatoxin M1 in cow's milk for human consumption in Kerman Province of Iran

The aim of this study was to assess levels of aflatoxin M₁ (AFM1) in milk samples from Kerman, Iran. AFM1 was detected in 72 samples, ranging in concentration from <0.01 to 0.41?µg?l^?1. The samples were analyzed using immunoaffinity column for clean-up and HPLC for determining AFM1. Milk samples were collected from six dairy farms. AFM1 was found in ～50% of the milk samples. The average level of AFM1 was below the tolerance limit (0.05?µg?l^?1), but 50% of the samples had greater levels than the maximum tolerance limit accepted by EU and the Iranian national standard. The method detection limit and limit of quantification were 0.01 and 0.03?µg?l^?1, respectively, and recovery of the method was 87%. The results showed that AFM1 contamination is a serious problem for public health. To achieve a low level of AFM1 in milk, cattle feed must be monitored regularly for aflatoxin contamination and protected from fungal contamination as much as possible.     

An effective self-control strategy for the reduction of aflatoxin M1 content in milk and to decrease the exposure of consumers

The study reports the results of testing the sensitivity of an early warning sampling plan for detecting milk batches with high aflatoxin AFM₁ concentration. The effectiveness of the method was investigated by the analysis of 9017 milk samples collected in Italian milk processing plants that applied control plans with different action limits (AL). For those milk processing plants where 30 ng kg^?1 AL has been applied, the AFM₁ contamination was significantly lower at or above the 95th percentile of the milk samples when compared with plants that used 40 ng kg^?1 AL. The results show that the control plan can be used effectively for early warning of occurrence of high AFM₁ contamination of milk and to carry out pro-active measures to limit the level of contamination. Estimation of dietary exposure was also carried out, based on the aflatoxin M₁ content of the milk samples and on Italian food consumption data. Estimated Daily Intakes (EDI) and Hazard Indices (HI) were calculated for different age groups of the population. HIs show that no adverse effects are expected for the adult population, but in the case of children under age three, the approximate HI values were considerably higher. This underlines the importance of the careful monitoring and control of aflatoxin M₁ in milk and dairy products.     

Natural occurrence of aflatoxin B1 in sorghum grown in different geographical regions of India

BACKGROUND: Sorghum (Sorghum bicolor (L.) Moench) is an important coarse cereal crop grown for grain and fodder in the semi‐arid Tropics, mainly in Asian and African countries. In India sorghum is consumed as human food and poultry feed. Sorghum grain grown in the rainy season (kharif) is becoming severely affected by grain moulds, the major fungi involved being Aspergillus, Fusarium and Curvularia. If the extent of mould is severe, the grain is unsafe for consumption owing to contamination by mycotoxins. RESULTS: This paper presents a multi‐centre study conducted in sorghum to evaluate natural contamination of aflatoxin B₁ in India. A total of 1606 grain sorghum samples were collected during the rainy (kharif) season across 4 years from seven states of India, representing different geographical regions of the country. Aflatoxin B₁ contamination during 2007–08 was the highest (13.1%), followed by samples from the year 2004–05 (2.85%). The samples collected in years 2005–06 and 2006–07 showed contamination below 1%. The number of samples (35) showing aflatoxin B₁ contamination above the safety limit was also highest during 2007–08 as compared to samples from the other years. CONCLUSION: This study, conducted for 4 years, showed that natural contamination of aflatoxin B₁ in sorghum grown in India is within safety limits (20 µg kg^?1) recommended by the Codex Alimentarius Committee and 73% of samples were positive for toxin. However, 0.75% (12) of total samples contained aflatoxin above the safety limit. The overall occurrence of toxin from Madhya Pradesh and Rajasthan was below 5 µg kg^?1. Copyright © 2012 Society of Chemical Industry     

Aspergillus Section Flavi and aflatoxins in dusts generated by agricultural processing facilities in the Philippines

Relatively few data exist regarding concentrations of aflatoxins and their causative organisms in dusts within occupational environments. Here, we examined Aspergillus Section Flavi populations and aflatoxin levels in 54 samples of dusts generated by agricultural processing facilities as possible indicators of aflatoxin exposure in the Philippines. The average incidence of Aspergillus Section Flavi expressed as a percentage of total mould populations in rice dust, corn dust, feed dust and copra dust were 8, 4, 31 and 10%, respectively. Predominant aflatoxigenic fungi isolated were Aspergillus flavus and A. parasiticus with ratios of 31:1, 40:5, 16:4 and 1:1 in rice dust, corn dust, feed dust and copra dust, respectively. Aflatoxins produced by selected isolates in in vitro rice culture ranged from 100 µg kg^?1 to 100.5 mg kg^?1. Toxigenicity of isolates based on the average aflatoxin concentrations produced by positive isolates were in the order of copra dust > corn dust > rice dust > feed dust. Average natural concentrations of aflatoxins in rice dust, corn dust, feed dust, and copra dust were 25, 6, 15 and 10 µg kg^?1, respectively. Estimates of the amount of inhaled aflatoxins by workers in an 8‐h work shift ranged from 0.06 to 114 ng, the average of which is higher than the amount of aflatoxins ingested by Filipinos due to the consumption of polished rice. The presence of highly toxigenic Aspergillus Section Flavi and aflatoxins in agricultural dust is a critical health risk for workers, considering the frequency of exposure and the possibility of inhalation and subsequent absorption of aflatoxins in the respiratory tract. Copyright © 2006 Society of Chemical Industry