María del Pilar Monge

Naturally occurring aflatoxin M1 in raw bulk milk from farm cooling tanks in Argentina

The aim was to carry out a survey of aflatoxin M1 (AFM1) in raw whole milk from bulk tanks. The sample collection was performed in farms located in one the most important milk-production zones in the centre of Argentina. A total of 94 samples of milk from 47 dairy farms were analysed. AFM1 analysis involved the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) with prior purification of the extracts using immunoaffinity columns. AFM1 incidence in raw milk was high as 63.8% and levels were between not detected to 0.07 µg l−1. Several contaminated samples (39%) were over the European Commission limit for infant milk (0.025 µg l−1), although none of samples were above Argentine legislation. Estimates of AFM1 intake were assessed for different age populations. The average AFM1 estimated daily intakes were 1.6, 0.5, 0.17 ng kg−1 body weight day−1 for 4-year-old babies, young children, and adults, respectively. All tested farms used pastures and silages at similar composition. Even though some farms (13) employed high-risk supplementary feeds, such as peanut pod and/or cotton seed, no statistically significant differences were observed between groups. Information from AFM1 levels in milk in Argentina is limited. A systematic AFM1 monitoring programme must be performed by means of accurate and reliable analytical techniques as a strategy for protecting milk consumers.

Natural co-occurrence of fungi and mycotoxins in poultry feeds from Entre Ríos, Argentina

A total of 120 pelleted poultry feed samples from Entre Ríos Province, Argentina, were evaluated. The aims were to investigate (1) the presence of relevant toxigenic fungi, as well as to determine the ability to produce aflatoxins (AFs) by Aspergillus section Flavi isolated strains; and (2) the natural co-occurrence of AFs, fumonisins (FBs), gliotoxin, diacetoxyscirpenol (DAS), HT-2 and T-2 toxin by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS). Total fungal counts were below the established value (1 × 104 CFU g−1). Aspergillus flavus and A. parasiticus were the only aflatoxigenic species isolated. Co-occurrence of fumonisin B1 (FB1), HT-2 and T-2 toxin was detected in 100% of the feeds, with mean levels from 4502 to 5813; 6.7 to 21.6 and 19.6 to 30.3 µg kg−1, respectively. A large number of starter samples were co-contaminated with aflatoxin B1 (AFB1), FB1, HT-2 and T-2 toxins. Gliotoxin and DAS were not found in this survey.

Growth and gliotoxin production by feed-borne Aspergillus fumigatus sensu stricto strains under different interacting environmental conditions

In this study the effects of temperature, oxygen tension, water activity (aW), pH, incubation time and their interactions on (1) the lag phase prior to growth, (2) growth rate and (3) gliotoxin production of two feed-borne Aspergillus fumigatus sensu stricto strains, isolated from fermented maize silage and brewers grains, were evaluated on an agar medium based on these substrates. Regardless of oxygen tension, the growth rate of the two strains decreased significantly as temperature and aW decreased (P<0.05). The optimum conditions for A. fumigatus growth were 37 °C, 0.98 aW for both strains at reduced oxygen tension, regardless of pH level (P<0.05). The studied A. fumigatus strains were able to grow under several incubation conditions, some of them prevalent in stored animal feeds. Some specific interactions that allowed accumulation of gliotoxin at high levels were found. This study showed that gliotoxin production occurred at more restricted conditions than fungal growth. This fact is important, as b...

Negligible effects of tryptophan on the aflatoxin adsorption of sodium bentonite

The main objective of this study was to determine if the competitive adsorption of tryptophan (Trp) and aflatoxin B1 (AFB1) could potentially affect the ability of a sodium bentonite (NaB) to prevent aflatoxicosis in monogastric animals. The adsorption of Trp and AFB1 on this adsorbent is fast and could be operating on the same time-scale making competition feasible. In vitro competitive adsorption experiments under simulated gastrointestinal conditions were performed. A high affinity of the clay for Trp and NaB was observed. The effect of an excess of KCl to mimic the ionic strength of the physiological conditions were also investigated. A six-times decrease in the Trp surface excess at saturation was observed. A similar behaviour was previously found for AFB1 adsorption. Taking into account the amount of Trp adsorbed by the clay and the usual adsorbent supplementation level in diets, a decrease in Trp bioavailability is not expected to occur. Tryptophan adsorption isotherms on NaB were ‘S’-shaped and were adjusted by the Frumkin–Fowler–Guggenheim model. The reversibility of the adsorption processes was investigated in order to check a potential decrease in the ability of NaB to protect birds against chronic aflatoxicoses. Adsorption processes were completely reversible for Trp, while almost irreversible for AFB1. In spite of the high affinity of the NaB for Trp, probably due to the reversible character of Trp adsorption, no changes in the AFB1 adsorption isotherm were observed when an excess of the amino acid was added to the adsorption medium. As a consequence of the preferential and irreversible AFB1 adsorption and the reversible weak binding of Trp to the NaB, no changes in the aflatoxin sorption ability of the clay are expected to occur in the gastrointestinal tract of birds.

Influence of water activity and temperature on growth and fumonisin production by Fusarium proliferatum strains on irradiated wheat grains

Wheat is the most important cereal consumed by the Argentine population. In previous studies performed in durum and common wheat grains in this country it has been observed fumonisin contamination as well as high incidence of Fusarium proliferatum. Fumonisins are toxic fungal metabolites, and consumption of fumonisin-contaminated maize has been epidemiologically associated with oesophageal cancer and neural tube defects in some human populations. Using irradiated wheat-grains, the effects of abiotic factors, temperature (15, 25, and 30°C) and water activity (aW; 0.995, 0.98, 0.96, 0.94, 0.92, and 0.88), on mycelial growth and fumonisin biosynthesis were compared for three F. proliferatum strains isolated from wheat grains in Argentina. Although all isolates showed similar profiles of growth, the fumonisin production profiles were slightly different. Maximum growth rates were obtained at the highest aW (0.995) and 25°C, with growth decreasing as the aW of the medium was reduced. Maximum amounts of total fumonisins (FB1, FB2 and FB3) were produced at 0.995 aW and 15°C for 2 strains, and at 25°C and 0.995 aW for the third one. Fumonisins concentrations varied considerably depending on the aW and temperature interactions assayed. Studied strains showed different fumonisin production profiles. Two-dimensional profiles of aW by temperature interactions were developed from these data to identify areas where conditions indicate a significant risk of fumonisins accumulation on wheat. As a result, temperature and aW conditions that resulted in fumonisins production are those found during wheat grain development (especially milk and dough stages) in the field. This is the first study made using irradiated wheat grains and provides useful baseline data on conditions representing a low or a high risk for fumonisins contamination of wheat grains which is of concern because this cereal is destined mainly for human consumption.

Validation of a liquid chromatography/tandem mass spectrometry method for the detection of aflatoxin B 1 residues in broiler liver

Aflatoxin B1 is a carcinogenic and mutagenic mycotoxin produced mainly by Aspergillus flavus and Aspergillus parasiticus. It is the predominant mycotoxin found in raw materials used for the manufacture of broiler feeds. The aim of the present study was to develop a new and optimized method for the detection and quantification of aflatoxin B1 (AFB1) residues in broiler liver using solid phase extraction (SPE) clean-up and liquid chromatography-electrospray ionization/tandem mass spectrometry (LC-ESI-MS/MS) detection. The method was validated for linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ). The validation parameters indicated satisfactory linearity (r2>0.99), accuracy and precision (4.57% intra-day RSD; 14.65% inter-day RSD) a very high recovery (99±13%) and high sensitivity achieved for AFB1 in animal samples (LOD=0.017 and LOQ=0.050ng/g). The method was effective for the detection and quantification of AFB1 residues in broiler liver and could also be potentially used for detecting AFB1 in other edible animal tissues after natural or experimental AFB1 exposure with high sensitivity and precision.

Activated carbons as potentially useful non-nutritive additives to prevent the effect of fumonisin B1 on sodium bentonite activity against chronic aflatoxicosis

ABSTRACT Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are mycotoxins that often co-occur in feedstuffs. The ingestion of AFB1 causes aflatoxicosis in humans and animals. Sodium bentonite (NaB), a cheap non-nutritive unselective sequestering agent incorporated in animal diets, can effectively prevent aflatoxicosis. Fumonisins are responsible for equine leukoencephalomalacia and porcine pulmonary oedema, and often have subclinical toxic effects in poultries. Fumonisin B1 and aflatoxin B1 are both strongly adsorbed in vitro on sodium bentonite. Co-adsorption studies, carried out with a weight ratio of FB1 to AFB1 that mimics the natural occurrence (200:1), showed that FB1 greatly decreases the in vitro ability of NaB to adsorb AFB1. The ability of two activated carbons to adsorb FB1 was also investigated. Both carbons showed high affinity for FB1. A complex behaviour of the FB1 adsorption isotherms with pH was observed. In vitro results suggest that under natural contamination levels of AFB1 and FB1, a mixture of activated carbon and sodium bentonite might be potentially useful for prevention of sub-acute aflatoxicosis.

Fumonisin occurrence in wheat-based products from Argentina

ABSTRACT In Argentina, wheat is the most consumed cereal by the human population. Since fumonisins occurence in wheat grains and wheat-based products have been reported worldwide, a survey was conducted in order to determine fumonisin contamination in 91 wheat-based products (white wheat flour samples, wheat flour used at bakery products and whole-wheat flour samples) collected from different retail stores of Rio Cuarto city in Argentina using HPLC-MS/MS. Sixty-seven samples (74%) showed contamination by fumonisins. From these samples, 16 showed fumonisin levels between LOD and LOQ (between 0.01 to 0.05 ng/g), while fumonisins (FB1 + FB2) in quantifiable samples ranged from 0.05 ng/g to 18.9 ng/g. Although FB1 was more prevalent, FB2 was foun3d in higher levels than FB1. Overall, fumonisin prevalence was high, but concentrations were far below EU or USA limits set for maize and maize-based products.