Aflatoxin M1 in raw milk and aflatoxin B1 in feed from household cows in Singida,Tanzania

Aflatoxin M₁ (AFM₁) contamination in raw milk from household cows fed with sunflower seedcakes or sunflower-based seedcake feeds was determined in 37 milk samples collected randomly from different locations in Singida region, Tanzania. Aflatoxin B₁ (AFB₁) contamination in sunflower-based seedcake feed was determined in 20 feed samples collected from the same household dairy farmers. The samples were analysed by RP-HPLC using fluorescent detection after immunoaffinity column clean-up. Recoveries were 88.0% and 94.5%, while the limits of detection (LOD) were 0.026 ng mL^?1 and 0.364 ng g^?1 for AFM₁ and AFB₁, respectively. Of the analysed cow’s milk samples, 83.8% (31/37) contained AFM₁, with levels ranging from LOD to 2.007 ng mL^?1, exceeding both the European Commission (EC) and Tanzania Food and Drug Authority (TFDA) limit of 0.05 ng mL^?1. Of the contaminated samples, 16.1% exceeded the Codex Alimentarius limit of 0.5 ng mL^?1. AFB₁ was present in 65% (13/20) of the feed samples with levels ranging from LOD to 20.47 ng g^?1, 61.53% exceeding the TFDA and EC maximum limits of 5 ng g^?1 for complete dairy animal feed. The observed AFM₁ and AFB₁ contamination necessitates the need to raise awareness to dairy farmers in Tanzania to safeguard the health of the end-users.     

Aflatoxin M1 in fresh milk collected from local markets of Karachi,Pakistan

During 2016–2017, 156 samples of fresh milk samples were collected from local markets of Karachi, Pakistan and analysed for aflatoxin M₁ (AFM₁) contamination using ELISA technique. AFM₁ was detected in 143 (91.7%) samples, ranged from 20 to 3090 ng L^?1 with a mean level of 346.2 ng L^?1. In 125 (80.1%) samples, the AFM₁ contamination was greater than the maximum limit (ML = 50 ng L^?1) set by EU. However, in 51 (32.7%) samples, the AFM₁ level was higher than the ML of 500 ng L^?1 as assigned by the USA. Statistical analysis showed that the AFM₁ level in milk samples from summer was significantly (p < 0.05) higher than that obtained in winter. It was concluded that the AFM₁ levels in the tested samples appear to be a serious public health problem. Therefore, immediate measures should be taken and re-evaluation done for the procedures for farming, transportation, refrigeration, and storage for the control of AFM₁ level in milk samples.     

Natural occurrence of aflatoxins (B1 and M1) in feed,plasma and raw milk of lactating dairy cows in Beja,Tunisia, using ELISA

Beja is an agricultural area in northwest Tunisia. It contributes to national needs by offering cereals and milk to the market for human and animal consumption. A small number of studies on mycotoxin occurrence in feedstuffs and raw milk from lactating dairy cows in this region are available. Therefore, 226 samples were collected from farms and local markets during November 2008 until April 2010. Samples consisted of 112 raw cow milk, 56 blood from lactating cows and 58 feed destined for dairy cows. Plasma and feed were analysed for aflatoxin B₁ (AFB₁). Milk samples were analysed for aflatoxin M₁ (AFM₁). All samples were treated using a simultaneous methanolic-aqueous extraction, followed by immunoaffinity column clean-ups and were investigated by competitive enzyme-linked immunoabsorbent assay (ELISA). Recoveries were 80%–95% and 81%–92% for AFB₁ and AFM₁, respectively, while the limit of detection (LOD) was 0.01?µg/kg or µg/l for both mycotoxins. Results revealed the presence of AFB₁ in 84.4% of the feed samples (mean 18.7?±?1.4?µg/kg), and 39.2% of the plasma-examined samples (median 7.1?±?1.0?µg/l) were found to be contaminated at levels higher than the Tunisian and the European Union (EU) limit for dairy animals, which are 20 and 5?µg/kg in animal feed, respectively. AFM₁ was detected in 60.7% of the cow raw milk samples examined (median 13.6?±?1.4?µg/l). Contaminated levels were higher than the EU limit of 0.05?µg/l. It was concluded that more precaution should be taken on hygiene controls in order to prevent fungal contamination.     

Seasonal patterns of aflatoxin M1 contamination in commercial pasteurised milk from different areas in Thailand

Aflatoxin M₁ (AFM₁) levels were determined in pasteurised milk from five commercial trademarks produced in different areas in Thailand. One hundred and twenty milk samples were collected from local markets in Chiang Mai province, Thailand, to evaluate AFM₁ concentrations using immunoaffinity columns and high-performance liquid chromatography with fluorescence detection. The overall median AFM₁ level was 0.023?µg?L^?1 ranging from 0.004 to 0.293?µg?L^?1. All trademarks had average AFM₁ concentrations lower than 0.05?µg?L^?1, with those in Trademarks 3 to 5 being higher than Trademarks 1 and 2 (P?<?0.01). All trademarks had different seasonal patterns of AFM₁, even though operating in the same area. However, only Trademark 3 showed significant differences of AFM₁ levels between seasons. The results suggested that farm management factors, rather than environment factors, were likely to be the main cause of AFM₁ contamination in dairy products.     

Aflatoxin M1 in buffalo and cow milk in Afyonkarahisar,Turkey

Potential hazardous human exposure to aflatoxin M₁ (AFM₁) via consumption of milk and milk products has been demonstrated by many researchers. The aim of this study was to investigate the presence of this mycotoxin in buffalo and cow milk samples in the city of Afyonkarahisar, Turkey. For this purpose, 126 buffalo and 124 cow milk samples were collected from dairy farms in Afyonkarahisar province. AFM₁ levels were determined by high-performance liquid chromatography with tandem mass spectrometric detection. Although AFM₁ was not detected in cow milk samples, AFM₁ was found above the limit of detection (<0.008–0.032 µg/L) in 27% (34 out of 126) of the buffalo milk samples. The results of this study indicated the importance of continuous surveillance of commonly consumed milk or milk product samples for AFM₁ contamination in Turkey.     

Detection of aflatoxin M1 in milk,cheese and sour cream samples from Costa Rica using enzyme-assisted extraction and HPLC

Aflatoxins are toxic fungal metabolites, which can be found in feed. Aflatoxin M₁ (AFM₁) is excreted into milk when ruminants ingest aflatoxin B₁ contaminated feedstuffs. Due to its carcinogenic potential, contamination of milk and dairy products with AFM₁ may pose a risk for consumers. Hence, it is considered a public health concern. In this survey, the level of AFM₁ contamination of dairy products marketed in Costa Rica was determined by enzyme-assisted extraction, immunoaffinity clean-up and high-performance liquid chromatography coupled with a fluorescent detector (HPLC-FLD) in fluid milk (n = 70), fresh cheese (n = 70) and sour cream (n = 70) collected at local convenience stores and supermarkets. AFM₁ concentrations in milk and fresh cheese ranged from 19 to 629 ng/L and from 31 to 276 ng/L, with mean values of 136 ng/L and 74 ng/L, respectively, whereas none of the sour cream samples analysed tested positive for this aflatoxin. In 30 milk samples, and 10 cheese samples, AFM₁ concentrations surpassed threshold concentrations as established by the European Commission. Thus, sour cream and – to a lesser extent – cheese manufacturing seems to reduce the amount of AFM₁ present in milk, possibly due to fraction redistribution or microbiological degradation. The survey results reveal improper quality control procedures in the Costa Rican dairy industry. Therefore, a surveillance programme for dairy products in our country is recommended.     

Aflatoxin M1 in raw milk from different regions of São Paulo state – Brazil

A total of 635 raw milk samples from 45 dairy farms, from three regions of São Paulo state – Brazil, were evaluated during 15 months for aflatoxin M₁ (AFM₁). AFM₁ was determined by high performance liquid chromatograph with fluorescence detection. AFM₁ was detected (>0.003 µg kg^?1) in 72.9%, 56.3% and 27.5% of the samples from Bauru, Araçatuba and Vale do Paraíba regions, respectively. The mean AFM₁ contamination considering all the samples was 0.021 µg kg^?1. Furthermore, the concentration of AFM₁ was quite different among Bauru (0.038 µg kg^?1), Araçatuba (0.017 µg kg^?1) and Vale do Paraíba (<0.01 µg kg^?1) regions. Only three samples (0.5%) had higher contamination than the tolerated limit in Brazil (0.50 µg kg^?1) and 64 samples (10.1%) had a higher contamination than the maximum limit as set by the European Union (0.050 µg kg^?1). The estimated AFM₁ daily intake was 0.358 and 0.120 ng kg^?1 body weight per day for children and adults, respectively.     

Aflatoxin M1 contamination in cow and buffalo milk samples from the North West Frontier Province (NWFP) and Punjab provinces of Pakistan

A total of 178 milk samples (94 of buffalo and 84 of cow) were randomly taken from Punjab and the North West Frontier Province (NWFP) of Pakistan (n?=?89 in each province) and analysed for the presence of aflatoxin M1 (AFM₁) by HPLC-FLD. From Punjab about 46% of buffalo's and 49% of cow's milks were contaminated with AFM₁ as compared with 52% and 51% for milk samples from NWFP, respectively. Overall, the mean AFM₁ concentration was 0.046?µg?kg^?1 with a maximum of 0.350?µg?kg^?1. All samples complied with the Codex Alimentarius limit of 0.50?µg?kg^?1 for AFM₁ in milk, but 16.3% of samples exceeded the European Union maximum level of 0.05?µg?kg^?1. Another set of 415 buffalo's and cow's milk samples (213 morning milks and 202 evening milks) were analysed. Statistical analysis revealed significant differences (p?<?0.05) between mean AFM₁ concentrations in milk during the morning (0.043?µg?kg^?1) and the evening (0.028?µg?kg^?1) lactation times.     

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.     

A review of the current situation of aflatoxin M1 in cow’s milk in Serbia: risk assessment and regulatory aspects

ABSTRACT

The aim of this systematic review is to provide information regarding the incidence and levels of aflatoxin M₁ (AFM₁) in raw and heat processed cow’s milk in Serbia during 2015–16 and to compare these with collected data on the occurrence of AFM₁ in raw milk and dairy products during the last decade in our region. Estimation of dietary exposure (EDI) and hazard index (HI) calculations for different age groups of the population were also carried out, based on the AFM₁ content of milk samples and on available food consumption data in Serbia. AFM₁ was detected in 69.9% (984/1408) of raw milk samples in 2015 versus 84.9% (3094/3646) in 2016, while in heat-processed milk, AFM₁ was detected in 77.8% (364/468) in 2015 versus 98.5% (753/765) in 2016. On the basis of the obtained results, 450 (9%) of raw and 14 (1.1%) of heat-processed milk samples were contaminated with AFM₁ levels above the maximum permitted level in Serbia (0.25 μg kg^?1). However, a large percentage of raw and heat processed milk in Serbia (30.1% and 17.3%, respectively) was contaminated with AFM₁ levels above the maximum permitted level regulated in the European Union (0.05 μg kg^?1). Therefore, in order to protect consumer health, it is extremely important to further control the level of aflatoxins in milk, and this should be considered as a high priority for risk management actions.     

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 composite spices collected from local markets of Karachi,Pakistan

This survey was carried out to evaluate the occurrence of total aflatoxins (AFs; B₁+B₂+G₁+G₂) in unpacked composite spices. A total of 75 samples of composite spices such as biryani, karhai, tikka, nihari and korma masalas were collected from local markets of Karachi, Pakistan, and analysed using HPLC technique. The results indicated that AFs were detected in 77% (n = 58) samples ranging from 0.68 to 25.74 µg kg^?1 with a mean of 4.63 ± 0.95 µg kg^?1. In 88% (n = 66) samples, AFs level was below the maximum limits (ML = 10 µg kg^?1) as imposed by EU. Furthermore, 61% (n = 46) tested samples contained AFs level between 1 and 10 µg kg^?1, 9% (n = 7) exhibited AFs contamination ranged 10?20 µg kg^?1 and only 3% (n = 2) of the investigated samples contained AFs levels higher than the ML of 20 µg kg^?1 for total aflatoxins as set by the USA. It was concluded that there is need to establish a strict and continuous national monitoring plan to improve safety and quality of spices in Pakistan.     

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     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

Simultaneous determination of aflatoxin B1 and M1 in milk,fresh milk and milk powder by LC-MS/MS utilising online turbulent flow chromatography

A novel, fully automated method based on dual-column switching using online turbulent flow chromatography followed by LC-MS/MS was developed for the determination of aflatoxin B₁ and M₁ in milk, fresh milk and milk powder samples. After ultrasound-assisted extraction, samples were directly injected into the chromatographic system and the analytes were concentrated on the clean-up loading column. Through purge switch, analytes were transferred to the analytical column for subsequent detection by mass spectrometry. Different types of TurboFlow^TM columns, transfer flow rates and transfer times were optimised. Method limits of detection obtained for AFB₁ and AFM₁ were 0.05 μg kg^–1, and limits of quantification were 0.1 μg kg^–1. Recoveries of aflatoxin B₁ and M₁ were in range of 81.1–102.1% for all samples. Matrix effects of aflatoxin B₁ and M₁ were in range of 63.1–94.3%. The developed method was successfully used for the analysis of aflatoxin B₁ and M₁ in real samples.     

Aflatoxin M1 in processed milk and infant formulae and corresponding exposure of adult population in Serbia in 2013–2014

Aflatoxin M₁ (AFM1) occurrence was analysed in 80 samples of milk and 21 samples of infant formulae on the Serbian market, using high-performance liquid chromatography with fluorescence detection. All milk samples collected in 2013 showed AFM1 contamination in the range 0.02–0.32 μg kg^?1, with a mean level of 0.13 μg kg^?1. The EU maximum level for AFM1 in milk (0.05 μg kg^?1) was exceeded in 75% of the samples. In 2014, AFM1 was found in 83%, 70%, 80% and 58% of the samples collected in April, July, September and December, respectively, exceeding the limit in 5% of the samples taken in July. The additional number of liver cancer cases per year associated with exposure to AFM1 was estimated to be 0.004 in the adult population. Regarding infant formulae, AFM1 was found in only one sample.     

Aflatoxins in animal feed in Iran

One hundred and forty-six samples of animal feed (barley, n = 60; wheat bran, n = 22; wheat dry pulp, n = 29; and canola meal, n = 35) were collected in 2011 from Mashhad (Khorasan, Iran). Aflatoxins (AFs) were determined in these samples after immunoaffinity column clean-up by high-performance liquid chromatography (HPLC) with fluorescence detection. Aflatoxin B₁ (AFB₁) contamination was found in 28 samples: in five of the barley samples (8.3%) at a mean level of 0.48 µg·kg^?1, in two wheat bran samples (9.0%) at a mean level of 0.88 µg·kg^?1, in 10 wheat dry pulp samples (34.5%) at a mean level of 0.30 µg·kg^?1 and in 11 canola meal samples (31.4%) at a mean level of 0.92 µg·kg^?1. AFB₁ levels were below the maximum levels of Iran regulations (5 µg·kg^?1) and the EU maximum limit (5 µg·kg^?1).     

Fumonisin B1 in cereal mixtures marketed in Brazil

The aim of this study was to detect and quantify fumonisin B₁ (FB₁) in cereal mixtures marketed in Brazil. Fifteen samples from different lots were acquired each month by internet from supermarkets during seven months, adding up to 105 analysed samples. The unit sample constituted of an original package with a minimum of 250 g. Extraction and clean-up of samples for FB₁ determination were carried out using immunoaffinity columns. Identification and quantification of FB₁ were performed by high performance liquid chromatography. Eighty-eight (83.8%) samples were contaminated with FB₁ and four (3.8%) presented levels above 500 µg kg^?1 (634, 703, 1269 and 1876 µg kg^?1). Maximum FB₁ + FB₂ levels allowed by Brazilian regulations will reach 1500 µg kg^?1 for corn flour in 2016 and 1000 µg kg^?1 for others corn products. This study showed that even at levels below the legislative limits, human exposure to this toxin can occur constantly.     

Urinary aflatoxin exposure monitoring in rural and semi-urban populations in Ogun state,Nigeria

Aflatoxins are a major class of fungal toxins that have food safety importance due to their economic and health impacts. This pilot aflatoxin exposure biomonitoring study on 84 individuals was conducted in a rural (Ilumafon) and a semi-urban community (Ilishan Remo) of Ogun state, Nigeria, to compare aflatoxin exposures among the two population cohorts. First morning urine samples were obtained from the participants, and the urinary aflatoxin M₁ (AFM₁) levels were measured by a quantitative Helica Biosystems Inc. ELISA kit assay. About 99% (83 out of 84) of the urine samples had detectable AFM₁ levels in the range of 0.06 to 0.51 ng mL^?1 (median: 0.27 ng mL^?1). The mean urinary AFM₁ levels were significantly (p = 0.001) higher in the semi-urban population (0.31 ± 0.09 ng mL^?1) compared to the rural population (0.24 ± 0.07 ng mL^?1). There were, however, no significant differences in mean urinary AFM₁ levels of males and females, and among children, adolescents and adults. This study indicates high aflatoxin exposure to the extent of public health concerns in the studied populations. Thus, more efforts are required for aflatoxin exposure monitoring and control in high-risk regions.     

Multiresidue method for simultaneous analysis of aflatoxin M1, avermectins,organophosphate pesticides and milbemycin in milk by ultra-performance liquid chromatography coupled to tandem mass spectrometry

A method developed for the simultaneous analysis of aflatoxin M₁, abamectin, doramectin, eprinomectin, ivermectin, moxidectin, acephate, azinphos-ethyl, azinphos-methyl, diazinon, methamidophos, methidathion, mevinphos, pirimiphos-ethyl and pirimiphos-methyl in whole raw milk, based on the QuEChERS method for extraction and clean-up, with detection and quantification by ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) is described. The method was validated according to parameters of the Analytical Quality Assurance Manual from the Brazilian Ministry of Agriculture and Commission Decision 2002/657/EC, and proved suitable for analysis of these analytes within the proposed working range, with recovery values between 77% and 110%, a standard deviation lower than 20%, limits of detection between 0.05 and 0.99 µg l^?1, and limits of quantification between 0.15 and 1.98 µg l^?1. Samples from animals treated with abamectin, doramectin, ivermectin and diazinon were analysed by the validated method. Residues of aflatoxin M₁ were also found in field samples at levels below the established maximum residue limit.