Incidence of Mycotoxins (AFB1 and AFM1) in Feeds and Dairy Farms from Rio de Janeiro State, Brazil

Brazil has regions located at the largest dairy production and milk derivate industry concentration, supplying the major consumer markets, represented by São Paulo, Rio de Janeiro and Belo Horizonte Cities. The milk is the most important product of Brazilian agriculture, because it is always presents in daily diet. The aim of this research was evaluating the occurrence of mycotoxins in the feed of dairy cattle and the occurrence of AFM1 in milk under field conditions in farms from Rio de Janeiro State, Brazil. The results revealed that, total fungal counts was found in 67% of feed samples which exceeded the recommended limit recommended (1×104 UFC.g-1) and the incidence of AFM1 in all milk samples was 26.7% at concentrations ranging from 0.010 to 1.500 μg.L-1. The study revealed toxigenic fungi and their mycotoxins were present in feed intended for bovine feeding in Rio de Janeiro farms. Evaluations of mycotoxin levels are important to provide information so that the assessments of risk for animal feed and livestock environment can be done.Fil: Keller, Luiz Antonio Moura. Universidade Federal Fluminense; BrasilFil: Aronovich, Marcos. No especifíca;Fil: Keller, Kelly Moura. Universidade Federal de Minas Gerais; BrasilFil: Castagna, Antonio Airton. No especifíca;Fil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: da Rocha Rosa, Carlos Alberto. Universidade Federal Rural do Rio de Janeiro; Brasi

Gliotoxin contamination in and pre- and postfermented corn, sorghum and wet brewer's grains silage in Sao Paulo and Rio de Janeiro State, Brazil

The aim of this study was to determine total fungal counts and the relative density of Aspergillus fumigatus and related species in silage samples intended for bovines before and after fermentation as well as to monitor the natural occurrence of gliotoxin in silage samples (pre- and postfermentation). Methods and methods: The survey was performed in farms located in São Paulo and Rio de Janeiro States in Brazil. In addition, the ability of A. fumigatus strains and related species strains to produce gliotoxin was also evaluated. A total of 300 samples were taken, immediately after opening of the silo (3-5 months) and during the ensiling period. Fungal counts were done by the surface-spread method. Gliotoxin production ability of isolates and natural contamination were determined by HPLC. Results: All postfermented samples had a total number of moulds exceeding 1 × 10 4 CFU g -1, with Aspergillus sp. as the most prevalent genus. Frequency of strains, among A. fumigatus and related species, was able to produce gliotoxin was similar in pre- and postfermented samples, except for sorghum, which showed differences between both kinds of samples. The highest toxin levels were produced by strains isolated from postfermented samples. More than 50% of the samples showed gliotoxin contamination levels that exceeded concentrations known to induce immunosuppressive and apoptotic effects in cells. Conclusions: The present data suggest that care should be taken because gliotoxin contamination in feedstuffs could affect productivity and also present a health risk for herds. Significance and Impact of the Study: Gliotoxin was found at quite important concentrations levels in pre- and postfermented substrates and its presence could therefore probably affect the productivity and health of herds. Current conservation and management practices do not avoid contamination with A. fumigatus on silage. Therefore, farm workers should be adequately protected during its handling. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.Fil: Keller, Luiz Antonio Moura. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Keller, Kelly Moura. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Monge, Maria del Pilar. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; ArgentinaFil: Pereyra, Carina Maricel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alonso, Veronica Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Chiacchiera, Stella Maris. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; ArgentinaFil: R Rosa, C. A.. Universidade Federal Rural do Rio de Janeiro; Brasi

Fungi and mycotoxins from pre and post storage brewer’s grain intended for bovine intensive-rearing

The aim of the study was to determine the mycobiota and natural levels of mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA), fumonisin B1 (FB1) and deoxinivalenol (DON) present in brewers grains pre and post stored intended for bovine intensive-rearing. Post stored (80%) samples had counts higher than 1 x 104 colony-forming units (CFU/g). Cladosporium spp., and Aspergillus spp. were isolated at high frequencies. Aspergillus flavus was the prevalent isolated species. Pre stored (70%) and post stored (100%) samples showed AFB1 levels over the recommended limits (20 µg/Kg) and OTA levels were below the recommended limits (50 µg/Kg). While pre and post stored samples did not show FB1 and DON natural contamination levels. The presence of mycotoxins in this substrate indicates the existence of contamination. Regular monitoring of feeds is required in order to prevent chronic and acute toxic syndromes related to this kind of contamination.Fil: Keller, Luiz Antonio Moura. Universidade Federal Rural do Rio de Janeiro; Brasil. Conselho Nacional de Desenvolvimento Científico e Tecnológico; BrasilFil: Pereyra, Carina Maricel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Dalcero, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Rosa, C. A. R. Conselho Nacional de Desenvolvimento Científico e Tecnológico; Brasil. Universidade Federal Rural do Rio de Janeiro; Brasi

Effect of yeast cell wall on the performance of broiler chickens intoxicated with aflatoxin B1

Micotoxinas são metabólitos secundários produzidos por diversos fungos filamentosos, tóxicos à animais e ao homem por contato, inalação e principalmente ingestão. Aflatoxinas são micotoxinas hepatotóxicas e carcinogênicas, produzidas principalmente pelos fungos Aspergillus flavus e A. parasiticus, e sua presença constitui grande preocupação para a avicultura mundial por problemas como diminuição da produtividade das aves e lesões de carcaça. Adsorventes à base de parede celular da levedura Saccharomyces cerevisiae, possuem glucomananos esterificadas, e são capazes de ligar-se eficientemente a diversas micotoxinas, como aflatoxinas, fumonisinas e zearalenona.   O objetivo do presente estudo foi o de avaliar o desempenho de um aditivo anti-micotoxinas (AAM) à base de parede de leveduras (PCL), em condição de intoxicação experimental por aflatoxina B1 (AFB1) em frangos de corte até os 21 dias de idade.  A adição de 1,012 mg kg-1 (ppm) de AFB1 na dieta dos frangos de corte no presente estudo foi capaz de alterar negativamente o peso vivo, ganho de peso e conversão alimentar a partir dos 14 dias de idade, e nas mesmas condições experimentais a adição do AAM (0,2%) à base de PCL reverteu tais efeitos. Mais estudos devem ser realizados acerca do assunto para melhor esclarecer o mecanismo de ação destes aditivos na produção animal.Mycotoxins are secondary metabolites produced by several filamentous fungi, which are toxic to animals and humans by contact, inhalation and ingestion mainly. Aflatoxins are hepatotoxic and carcinogenic mycotoxins produced mainly by Aspergillus flavus and A. parasiticus. Its presence is of great concern to the poultry industry due to problems such as decreased productivity and damage to the poultry carcass. Adsorbents based on the yeast cell wall of Saccharomyces cerevisiae have esterified glucomannans, and are able to adsorb several mycotoxins such as aflatoxins, fumonisins and zearalenone. The aim of this study was to evaluate the effect of a yeast cell wall (anti-mycotoxin additive) on the performance of broiler chickens intoxicated with aflatoxin B1 (AFB1 ) until 21 days old. The addition of 1.01 mg kg-1 (ppm) of AFB1 in the diet of broilers in this study could affect negatively body weight, weight gain and feed consumption after 7 days old, and under the same experimental conditions the yeast cell wall (0.2%) used as an anti-mycotoxin additive reversed such effects. More studies should be conducted to better clarify the mechanism of action of these additives in animal production.Fil: Keller, Kelly Moura. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Oliveira, A. A. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Almeida, T. X.. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Keller, Luiz Antonio Moura. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Queiroz, B. D.. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Nunes, L. M. T.. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rosa, Carlos Alberto da Rocha. Universidad Federal Rural de Rio de Janeiro; Brasi

Crambe (Crambe hispanica Subsp. abyssinica) Grains Mycobiota and Natural Occurrence of Aflatoxins, Ochratoxin A, Fumonisin B1 and Zearalenone

Crambe grains are a new feed with high concentrations of proteins and fibers. As there is no control during the pre-harvesting or post-harvesting stages of production other grains, crambe may be contaminated by fungi. Fungal overgrowth may lead to mycotoxins production and nutritional properties decrease of the grains. The aim of this study was to analyze the occurrence of fungi and mycotoxins according to pre-harvesting management. Fungal concentration was higher than that recommended by international regulations (3.4×106 to 1.3×104 CFU.g-1), suggesting that management in pre-harvesting stages of crambe grains production may expose the animals that will feed on these grains to the risk of contamination by fungal toxins. More studies are required about quality of crambe grains, because may be strongly affected by the exposition to variable environmental conditions. But, considering low mycotoxin incidence and levels founded, the crambe proves to be a safe food to be exploited for animal nutrition.Fil: Vieira Rosetto, Claudia Antonia. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Oliveira, Águida Aparecida De. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Keller, Luiz Antonio Moura. Universidade Federal Fluminense; BrasilFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Rodrigues, Eliane. No especifíca;Fil: Rosa, Carlos Alberto da Rocha. Universidade Federal Rural do Rio de Janeiro; Brasi

Inhibitory Effect of GRAS Essential Oils and Plant Extracts on the Growth of Aspergillus westerdijkiae and Aspergillus carbonarius Strains

The effect of essential oils (obtained using hydrodistillation) and plant extracts (ethanolic, aqueous, and hexanic extractions) of 10 different plants cultivated in Brazil were tested using the diffusion agar method, with the objective of evaluating the inhibitory effect of the oils and extracts on the mycelial growth of Aspergillus westerdijkiae NRRL 3174 and A. carbonarius RC 2054 (UNRC). Of the 40 essential oils and plant extracts analyzed, oregano essential oil and plant extract, rosemary essential oil, and the clove ethanolic extract were the best choice to obtain the growth parameters (radial growth rates (mm day−1) and lag phase (h)) due the good results presented and the volume of oil/extract obtained. Comparing all the essential oils and plant extracts that were tested for growth parameters, the best results were obtained for the clove ethanolic extract for both strains assayed. These results demonstrated an outstanding potential use of some of these products in prevention of fungal contamination in food. However, further studies need to be conducted to determine the ability of these oils and extracts to inhibit or reduce ochratoxin A production.Fil: Rodrigues, Mariana Paiva. Universidade Federal de Minas Gerais; BrasilFil: Oliveira, Águida Aparecida De. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Lago Biscoto, Gabriela. Universidade Federal de Minas Gerais; BrasilFil: Pinto, Priscila Natália. Universidade Federal de Minas Gerais; BrasilFil: de Souza Dias, Raul Roque. Universidade Federal de Minas Gerais; BrasilFil: Salvato, Lauranne Alves. Universidade Federal de Minas Gerais; BrasilFil: Keller, Luiz Antonio Moura. Universidade Federal Fluminense; BrasilFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Rosa, Carlos Alberto da Rocha. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Keller, Kelly Moura. Universidade Federal de Minas Gerais; Brasi

Estudo sobre a micobiota e as micotoxinas em rações comerciais destinadas à alimentação de eqüinos no rio de Janeiro, Brasil

Colonization of feedstuffs and agricultural commodities by mycotoxins producer moulds is a significant problem worldwide. Contamination of feed ingredients, increases the risk of contamination of the finished products such as manufactured commercial feeds. Studies evaluating the risk of mycotoxins in the equine environment are scarce. The aims of this study were to evaluate the mycobiota present in commercial equine feeds and to determine total aflatoxins(AFs) and fumonisins (FBs). Thirty samples of 21 commercial equine feeds were collected at random from 5 different equestrian centers located in Rio de Janeiro, Brazil. Total fungal counts (CFU g-1) of equine feeds ranged from not detectable (ND) to 1.3 x 106 CFU/g onto DRBC medium. Penicillium spp. (52%) was the prevalent genera, followed by Aspergillus spp. (32.5%). Aspergillus niger aggregate (60%) and Aspergillus flavus (22%) showed the highest relative densities. Thirteen Penicillium spp. were isolated. All samples (100%) were contaminated with AFs at levels that varied between 1 and 44 μg/Kg. The 65% of these samples were positive to FBs contamination at levels ranging from 0.2 to 8.5 μg/g. Monitoring mycological and mycotoxicological quality of feed ingredients and commercial feedstuffs used in equine husbandry are critical for improving animal production and performance.A colonização de matérias-primas e produtos agrícolas por fungos produtores de micotoxinas é um problema significativo em todo o mundo. A contaminação dos ingredientes das rações aumenta o risco do produto acabado como as rações comerciais manufaturadas. Estudos avaliando o risco de micotoxinas no ambiente eqüino são escassos. Os objetivos deste estudo foram avaliar a micobiota presente em rações comerciais para eqüinos e determinar aflatoxinas (AFs) e fumonisinas (FBs) totais. Trinta amostras de 21 marcas comerciais foram coletadas, ao azar, de 5 diferentes centros hípicos localizados no Rio de Janeiro, Brasil. As contagens totais de fungos (UFC g-1) das rações de equinos apresentaram intervalos entre não detectado (ND) a 1,3 x 106 UFC/g no meio DRBC. Penicillium spp. (52%) foi o gênero prevalente, seguido por Aspergillus spp. (32,5%). Aspergillus niger agregados (60%) e Aspergillus flavus (22%) apresentaram as maiores densidades relativas. Treze espécies de Penicillium foram isoladas. Todas as amostras (100%) estavam contaminadas com AFs com níveis que variaram entre 1 e 44 μg/Kg. Um total de 65% das amostras estavam positivas para a contaminação por FBs com níveis que variaram entre 0,2 a 8,5 μg/g. O monitoramento micológico e micotoxicológico da qualidade dos ingredientes e rações comerciais usados na alimentação eqüina são críticos para incrementar a produção e performance animal.Fil: Keller, Kelly Moura. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Keller, Luiz Antonio Moura. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Queiroz, Beatriz Dias. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Oliveira, Águida Aparecida De. Universidade Federal Rural do Rio de Janeiro; BrasilFil: de Almeida, Tatiana Xavier. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Marassi, Ana Cláudia. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Gonzalez Pereyra, Maria Laura. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Cavaglieri, Lilia Reneé. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto; ArgentinaFil: Dalcero, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto; ArgentinaFil: Rosa, Carlos Alberto da Rocha. Universidade Federal Rural do Rio de Janeiro; Brasi

In vitro activity of neem (Azadirachta indica) oil on growth and ochratoxin A production by Aspergillus carbonarius isolates

Aspergillus carbonarius is a saprobic filamentous fungus, food spoiling fungus and a producer of ochratoxin A (OTA) mycotoxin. In this study, the in vitro antifungal activity of neem oil (0.12% p/p of azadirachtin) was evaluated against the growth of six strains of A. carbonarius and the production of OTA. Four different concentrations of neem oil were tested in addition to three incubation times. Only the concentration of 0.3% of neem oil inhibited more than 95% of the strain’s growth (97.6% ± 0.5%), while the use of 0.5% and 1.0% of neem oil showed lower antifungal activity, 40.2% ± 3.1 and 64.7% ± 1.1, respectively. There was a complete inhibition of OTA production with 0.1% and 0.3% neem oil in the four strains isolated in the laboratory from grapes. The present study shows that neem essential oil can be further evaluated as an auxiliary method for the reduction of mycelial growth and OTA production.Fil: Rodrigues, Mariana Paiva. Universidade Federal de Minas Gerais; BrasilFil: Astoreca, Andrea Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: De Oliveira, Águida Aparecida. Universidad Federal Rural de Rio de Janeiro; BrasilFil: Salvato, Lauranne Alves. Universidade Federal de Minas Gerais; BrasilFil: Biscoto, Gabriela Lago. Universidade Federal de Minas Gerais; BrasilFil: Keller, Luiz Antonio Moura. Universidade Federal Fluminense; BrasilFil: Rosa, Carlos Alberto da Rocha. Universidade Federal Do Rio de Janeiro. Instituto de Biología; BrasilFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Quimica. Area Fisicoquimica Organica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: De Azevedo, Maria Isabel. Universidade Federal de Minas Gerais; BrasilFil: Keller, Kelly Moura. Universidade Federal de Minas Gerais; Brasi

In Vitro Activity of Neem (<i>Azadirachta indica</i>) Oil on Growth and Ochratoxin A Production by <i>Aspergillus carbonarius</i> Isolates

Aspergillus carbonarius is a saprobic filamentous fungus, food spoiling fungus and a producer of ochratoxin A (OTA) mycotoxin. In this study, the in vitro antifungal activity of neem oil (0.12% p/p of azadirachtin) was evaluated against the growth of six strains of A. carbonarius and the production of OTA. Four different concentrations of neem oil were tested in addition to three incubation times. Only the concentration of 0.3% of neem oil inhibited more than 95% of the strain’s growth (97.6% ± 0.5%), while the use of 0.5% and 1.0% of neem oil showed lower antifungal activity, 40.2% ± 3.1 and 64.7% ± 1.1, respectively. There was a complete inhibition of OTA production with 0.1% and 0.3% neem oil in the four strains isolated in the laboratory from grapes. The present study shows that neem essential oil can be further evaluated as an auxiliary method for the reduction of mycelial growth and OTA production.Facultad de Ciencias ExactasCentro de Investigación y Desarrollo en Fermentaciones Industriale

In Vitro Activity of Neem (<i>Azadirachta indica</i>) Oil on Growth and Ochratoxin A Production by <i>Aspergillus carbonarius</i> Isolates

Aspergillus carbonarius is a saprobic filamentous fungus, food spoiling fungus and a producer of ochratoxin A (OTA) mycotoxin. In this study, the in vitro antifungal activity of neem oil (0.12% p/p of azadirachtin) was evaluated against the growth of six strains of A. carbonarius and the production of OTA. Four different concentrations of neem oil were tested in addition to three incubation times. Only the concentration of 0.3% of neem oil inhibited more than 95% of the strain’s growth (97.6% ± 0.5%), while the use of 0.5% and 1.0% of neem oil showed lower antifungal activity, 40.2% ± 3.1 and 64.7% ± 1.1, respectively. There was a complete inhibition of OTA production with 0.1% and 0.3% neem oil in the four strains isolated in the laboratory from grapes. The present study shows that neem essential oil can be further evaluated as an auxiliary method for the reduction of mycelial growth and OTA production.Facultad de Ciencias ExactasCentro de Investigación y Desarrollo en Fermentaciones Industriale