Risk assessment of exposure to multiple mycotoxins in food

Moulds may produce a diversity of toxins such as aflatoxins, ochratoxins, trichothecenes, zearalenone, fumonisins and others. Although toxicological, environmental and epidemiological studies have addressed the problem of these toxins one by one, more than one mycotoxin are found usually in the same contaminated food. Risk assessment for humans potentially exposed to multimycotoxins suffers very much from the lack of adequate food consumption data. Furthermore, for a given mycotoxin, synergism and antagonism with other mycotoxins, found in the same food commodities, are not taken into account. Aflatoxin B1 and ochratoxin A belong to the most frequently occurring mycotoxins. This has repeatedly been demonstrated, however, normally, the risk resulting from their simultaneous occurrence is not considered. A descriptive study was developed to monitor air fungal contamination in one hospital food unit.

Are air-borne mycotoxins a public health concern in Portugal?

Introduction - Microscopic filamentous fungi, under suitable environmental conditions, can lead to the production of highly toxic chemical substances, commonly known as mycotoxins. The most widespread and studied mycotoxins are metabolites of some genera of moulds such as Aspergillus, Penicillium and Fusarium. Quite peculiar conditions may influence mycotoxin biosynthesis, such as climate, geographical location, cultivation practices, storage and type of substrate. Toxicity has been extensively investigated for the most important mycotoxins, such as aflatoxins, ochratoxin A and Fusarium toxins, and much information derived from toxicokinetics in animal models has also been obtained. The adverse effects are mainly related to genotoxicity, carcinogenicity, mutagenicity, teratogenicity and immunotoxicity. Aim of the study - To identify fungal species able to produce important mycotoxins in different Portuguese settings.

Occupational co-exposure to several mycotoxins in the waste management setting

Contrary to fungi, exposure to mycotoxins is not usually identified as a risk factor present in occupational settings. This is probably due to the inexistence of limits regarding concentration of airborne mycotoxins, and also due to the fact that these compounds are rarely monitored in occupational environments. Despite the optimal conditions for fungal growth and, consequently, for mycotoxins production in all the waste management chain, only a few articles were dedicated to study occupational exposure to mycotoxins in this occupational setting. Aim of study: A study was developed in Portugal aiming to assess occupational co-exposure to mycotoxins in the waste management setting.

Fungal contamination in feed production in Portugal: what to expect regarding mycotoxins contamination?

Fungi on crops produce mycotoxins in the field, during handling, and in storage. Exposure of animals and humans are usually through consumption of contaminated feedstuffs or foods. Molds can grow and mycotoxins can be produced either pre-harvest or post-harvest, during storage, transport, processing, or feeding. Worldwide, approximately 25% of crops are affected by mycotoxins annually. Because of this is possible to concluded that mycotoxins occur frequently in a variety of feedstuffs that are given to animals causing several effects: subclinical losses in performance, increases the incidence of disease and reduced reproductive performance. Aim of study: A study was developed intending to know environmental fungal contamination in a Portuguese feed production unit. Corn, wheat and soybeans were the most common cereals used in the feed production.

Environmental mycology in public health: fungi and mycotoxins risk assessment and management

Fungi are essential to the survival of our global ecology, but they might pose a significant threat to the health of occupants when they grow in our buildings. The exposure to fungi in homes is a significant risk factor for a number of respiratory symptoms. Well-known illnesses caused by fungi include allergy and hypersensitivity pneumonitis. Environmental monitoring for fungi and their disease agents are important aspects of exposure assessment, but few guidelines exist for interpreting their health impacts. This book answers the questions: How does one detect and measure the presence of indoor fungi? What is an acceptable level of indoor fungi? How do we relate this information to human health problems?

A review of mycotoxins in food and feed products in Portugal and estimation of probable daily intakes

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that occur naturally in agricultural commodities worldwide. Aflatoxins, ochratoxin A, patulin, fumonisins, zearalenone, trichothecenes and ergot alkaloids are presently the most important for food and feed safety. These compounds are produced by several species that belong to the Aspergillus, Penicillium, Fusarium and Claviceps genera and can be carcinogenic, mutagenic, teratogenic, cytotoxic, neurotoxic, nephrotoxic, estrogenic and immunosuppressant. Human and animal exposure to mycotoxins is generally assessed by taking into account data on the occurrence of mycotoxins in food and feed as well as data on the consumption patterns of the concerned population. This evaluation is crucial to support measures to reduce consumer exposure to mycotoxins. This work reviews the occurrence and levels of mycotoxins in Portuguese food and feed to provide a global overview of this issue in Portugal. With the information collected, the exposure of the Portuguese population to those mycotoxins is assessed, and the estimated dietary intakes are presented.

Current situation on the occurrence of mycotoxins and toxigenic fungi in Portugal

Mycotoxins are secondary metabolites produced by filamentous fungi that are toxic for humans and animals in small amounts and that are found worldwide in a large number of agricultural commodities. They are usually ingested involuntarily, when contaminated plant products are consumed, and represent a great risk for public health. Therefore, governments throughout the world have imposed strict legal limits for their levels in food and feed products in order to reduce potential health risks for consumers. Despite of its ubiquity, the mycotoxin problem is mainly dependent on regional factors, such as the mycotoxigenic characteristics of the local mycoflora, the local climate conditions, and the local agricultural practices. For this reason, a constant vigilance from local governmental food safety agencies and from the local researcher community is needed. This communication will review the current situation on the occurrence of mycotoxigenic fungi in some Portuguese cultures, such as wine grapes, corn and dried fruits. Particular attention will be given to the incidence of mycotoxigenic Aspergillus strains in those cultures and to the levels of ochratoxin A, aflatoxins, cyclopiazonic acid and fumonisin B2 produced. Data will be discussed taking into account the geographical origin of the isolates and the particular climate conditions of each sampling region. An updated review on the levels of the main mycotoxins found in local products and in imported commodities will also be presented.

Bio-detoxification of mycotoxins by lactic acid bacteria from different food matrices

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP- PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 g/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 m pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Estudio de la micobiota y sus micotoxinas en ingredientes y raciones de animales domésticos. Estrategias orientadas a la prevención de las micotoxicosis. Studies on mycobiota and incidence of mycotoxins in raw material and feedstuff for domestic animals. Different strategies for mycotoxicosis prevention.

Numerosas investigaciones han desarrollado estrategias para la remoción de micotoxinas en alimentos por diferentes métodos, aunque muchos de ellos no han llegado a ser utilizados debido a los elevados costos o a las dificultades prácticas involucradas en el proceso de detoxificación. Estos argumentos estimulan a los investigadores a desarrollar nuevas estrategias de decontaminación que eviten el uso de agentes químicos y que reduzcan las pérdidas en el valor nutritivo y la palatabilidad de los alimentos decontaminados. Una de las alternativas promisorias es la detoxificación biológica. Las levaduras capaces de adsorber micotoxinas y con habilidades probióticas o prebióticas son promisorias para reducir la exposición humana a las micotoxinas. En el tracto gastrointestinal se encuentra normalmente un gran número de especies de bacterias comensales y patógenas; sin embargo, cuando se incrementa la cantidad de microorganismos patógenos se pueden producir alteraciones de la salud y muerte. La industria argentina de alimentos destinados a animales necesita reducir los niveles de micotoxinas presentes en ingredientes o en insumos terminados. Si bien los resultados obtenidos en el mundo en la temática son preliminares y promisorios, en nuestro país aún no se han desarrollado estrategias biológicas de decontaminación de micotoxinas aplicadas a estos alimentos. Estudios de incidencia de micoflora y detección de micotoxinas en alimentos balanceados para aves, llevados a cabo por nuestro grupo de investigación en la región del sur de Córdoba demostraron la presencia de los principales géneros toxicogénicos (Aspergillus, Penicillium y Fusarium) y sus micotoxinas asociadas (aflatoxinas, zearalenona y fumonisinas). En relación a porcinotecnia, la zona sur de la provincia de Córdoba es considerada una de las tres zonas de mayor densidad porcina en Argentina. Sin embargo, la contaminación de los granos con micotoxinas representa un serio problema debido a que producen rechazo del alimento, disminución de la tasa de crecimiento y reducción inmunológica. Si consideramos la evolución en la producción lechera en los últimos años ha seguido una línea de intensificación que ha conllevado un cambio en la utilización de los alimentos, evolucionando del simple pastoreo a los sistemas de alimentación única, basados en la formulación de alimentos balanceados que constituyen la clave de la alimentación de los animales. Diferentes estudios epidemiológicos usando técnicas moleculares han demostrado que con frecuencia la infección por A. fumigatus ocurre como consecuencia de la adquisición exógena del hongo. La magnitud del problema se manifiesta en la continua búsqueda de medidas de prevención y control de estas micotoxicosis. Debido a este impacto negativo que ejercen las toxinas fúngicas lo cual, afecta los parámetros productivos como ganancia de peso y conversión alimenticia con graves pérdidas a la industria animal tanto en el mercado interno como externo.

Soybean pod blight and root rot caused by lineages of the Fusarium graminearum and the production of mycotoxins

Surveys of soybean (Glycine max) seed grown in South Brazil revealed infection with Fusarium graminearum. To determine if members of this complex were pathogenic to soybean, six strains derived from soybean were added to soil at a rate of 10³ macroconidia/ ml or individual pods were inoculated with 10(4) macroconidia/ml. Seedlings grown in infested soil developed small necrotic lesions in the crown and upper roots. Pods inoculated with conidia developed large (>1 cm), dark brown, necrotic lesions. Younger pods inoculated with the fungus blighted and dropped from the plant. Strains of the F. graminearum complex recovered from lesions on the crown, roots and pods of soybean plants were identified as lineage 1, 2 or 8 by obtaining the DNA sequence from the EF1-alpha gene and comparing it to strains of the known lineage. Two strains of F. graminearum lineage 7 from the U.S. caused similar symptoms of the disease on soybean. Mycotoxin tests on soybean and wheat (Triticum aestivum) indicate that most Brazilian strains produce nivalenol as the major trichothecene mycotoxin rather than deoxynivalenol. In addition, strains from lineages 2 and 8 produce the novel trichothecene, 3-acetylnivalenol.

Parboiled rice: chemical composition and the occurrence of mycotoxins

The objective of this work was to evaluate the incidence of aflatoxin B1 (AFB B1), deoxynivalenol (DON), ochratoxin A (OTA), and zearalenone (ZEA) in parboiled rice with respect to its chemical composition. Eight lots from five different brands of parboiled rice were collected in four samplings, at different seasons, until the amount of 32 lots. It was observed that: DON was present in 22% of the samples (from 180 to 400 ppb); ZEA in 19% (from 317 to 396 ppb); OTA in 12.5% (from 13 and 26 ppb); and AFB B1 in 9% (from 11 to 74 ppb). The results of the chemical composition were not different from those previously mentioned in the literature concerning parboiled rice. The ash and phenol levels in the contaminated parboiled rice samples suggested that those compounds had a relation to the occurrence of OTA, DON and ZEA mycotoxins.

Importance of mycotoxins in food and feed in India

Various food and feed samples including groundnut seed, maize, sorghum, soyabean cake, groundnut cake, cotton cake, poultry feed, buffalo milk, cow milk and milk powders were collected from farmers' fields, farmer's stores, oil millers storage, traders' storage, retail shops and supermarkets. More than 2000 samples were analysed by ELISA and most of the commodities, with the exception of sorghum seed, contained high levels of aflatoxin. Groundnut cake was one of the major cattle feed ingredients in the peri-urban area of Hyderabad (Andhra Pradesh, India) and >75% of the samples contained >100 µg/kg aflatoxin, leading to a high level of aflatoxin M1, in milk samples. Strategies to reduce aflatoxin levels (especially in groundnut) by management interventions at preharvest, harvest and storage, are discussed.

Fungi, mycotoxins and phytoalexin in peanut varieties, during plant growth in the field

The aim of the present study was to analyze the mycobiota, occurrence of mycotoxins (aflatoxins and cyclopiazonic acid), and production of phytoalexin (trans-resveratrol) in two peanut varieties (Runner IAC 886 and Caiapo) during plant growth in the field. Climatic factors (rainfall, relative humidity and temperature) and water activity were also evaluated. The results showed a predominance of Fusarium spp. in kernels and pods, followed by Penicillium spp. and Aspergillus flavus. Aflatoxins were detected in 20% and 10% of samples of the IAC 886 and Caiapo varieties, respectively. Analysis showed that 65% of kernel samples of the IAC 886 variety and 25% of the Caiapo variety were contaminated with cyclopiazonic acid. trans-Resveratrol was detected in 6.7% of kernel samples of the IAC 886 variety and in 20% of the Caiapo variety. However, trans-resveratrol was found in 73.3% of leaf samples in the two varieties studied. (C) 2011 Published by Elsevier Ltd.

Evaluation of yeast cell wall on the performance of broiles fed diets with or without mycotoxins

This experiment aimed at evaluating the effects of the interactions between aflatoxin (500 or 250 ppb) and ochratoxin (500 or 250 ppb), and the possible benefits of adding yeast cell wall to prevent the effects of these mycotoxins in broiler chickens. Relative organ weight gain and live performance were evaluated at 21 and 42 days of age. Results indicated that at the levels of mycotoxins included in the experimental diets, ochratoxin reduced feed intake and body weight gain, and aflatoxin only affect feed intake of 21-day-old birds. No interaction was observed between aflatoxin and ochratoxin at the levels used in experimental study. Yeast cell wall did not significantly reduced the deleterious effects of ochratoxins. No significant differences were observed in relative organ weight gain. Yeast cell wall improved feed conversion ratio when birds were fed either contaminated or non-contaminated feeds.