Mycobiota and mycotoxins present in finished fish feeds from farms in the Rio de Janeiro State, Brazil

The aim of the present study was to determine species of the fungal genera Aspergillus, Fusarium, and Penicillium and fumonisin B1 (FB1), aflatoxin B1 (AFB1), and ochratoxin A (OTA) contamination from feed intended for fish farms. A total of 60 samples were sampled from tilapia farms in the Rio de Janeiro State, Brazil. The quantitative enumeration of fungi as colony-forming units per gram of feed (CFU/g) was performed using the surface spread method in different culture media. The results were expressed as fungal isolation frequency and relative density. Fungal total counts ranged from <1 × 102 to 4.7 × 104 CFU/g. Fusarium counts were not observed. Among toxigenic genera, Aspergillus (68%) was the most prevalent, followed by Penicillium species (60%). Aspergillus niger aggregate (36%), Aspergillus flavus (35%), and Penicillium citrinum (71%) were the most prevalent species. A high percentage of samples (98%) were contaminated with FB1 levels, while 55% and 3.3% were contaminated with AFB1 and OTA, respectively. The simultaneous occurrence of these mycotoxins emphasizes the need for further research in the area to better assess the risk to the health of fish farms and their implications for the health of consumers of this meat.Fil: Barbosa, Tatiana S.. Universidade Federal Rural do Rio de Janeiro ; 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: Soleiro, Carla A.. Universidade Federal Rural do Rio de Janeiro ; Brasil. Conselho Nacional de Desenvolvimiento Científico y Tecnológico; BrasilFil: Dias, Erica O.. Universidade Federal Rural do Rio de Janeiro ; BrasilFil: Oliveira, Aguida A.. Universidade Federal Rural do Rio de Janeiro ; Brasil. Conselho Nacional de Desenvolvimiento Científico y Tecnológico; BrasilFil: Keller, Kelly M.. Universidade Federal Rural do Rio de Janeiro ; Brasil. Conselho Nacional de Desenvolvimiento Científico y Tecnológico; BrasilFil: Silva, Pedro P. O.. Universidade Federal Rural do 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. Universidade Federal Rural do Rio de Janeiro ; Brasil. Conselho Nacional de Desenvolvimiento Científico y Tecnológico; Brasi

Evaluation of zearalenone in vitro removal by Saccharomyces cerevisiae strains isolated from bovine forage

Zearalenone (ZEN) is a mycotoxin that has relatively low acute toxicity. However, it is a potent oestrogen, interfering with the reproductive tract of animals. Among other effects, ZEN decreases animals fertility, and induces fibrosis in the uterus, breast cancer and endometrial carcinoma (Zinedine et al., 2007). Anti-mycotoxin additives (AMA) are defined as a group of products that, when added to animal feed, are capable of adsorbing, inactivating, or neutralizing mycotoxins in the gastrointestinal tract of animals. One example of these products are adsorbents based on yeast cell walls, a safe and beneficial animal feed additive (Abreu et al., 2008). When based on active cells, yeast based products also act as a probiotic, contributing to improve the general animal health because it stimulates their immune system and promotes the integrity of intestinal mucosa (Albino et al., 2006). Strains of Saccharomyces cerevisiae isolated from silage were tested for their ZEN removal capability. Their effect on - and b-zearalenol (-ZOL and b-ZOL) was also tested. Strains were grown on YPD separately supplemented with ZEN, -ZOL and b-ZOL, and their elimination from culture media was quantified over time by HPLC-FL.This study was carried out with grants from CYTED (Acción 109ac0371), CNPq, CAPES-DS and FAPUR/UFRRJ (Brazil). Luís Abrunhosa was supported by grant SFRH/BPD/43922/2008 from Fundação para a Ciência e Tecnologia – FCT, Portugal.. Authors also acknowledge the support of the Society for Applied Microbiology – SfAM

Binding of Aflatoxin B1 to Lactic Acid Bacteria and Saccharomyces cerevisiae in vitro: A Useful Model to Determine the Most Efficient Microorganism

Mycotoxins are toxic fungal metabolites found as contaminants in many agricultural products. Feeds contaminated with mycotoxins have a health risk to animals and, as a consequence, may cause big economical losses due to the low efficacy of animal husbandry (Richard, 2007). In addition, directly or indirectly (animal by-products) contaminated foods may also have a health risk to humans (CAST, 2003; Hussein & Brasel, 2001; Wild, 2007). Aflatoxins (AFs), a group of potent mycotoxins with mutagnic, carcinogenic, teratogenic, hepatotoxic and immunosupresive properties, are of particular importance because of their major occurrence and adverse effects on animal and human health, generalized as ?aflatoxicosis? (CAST, 2003; Hussein & Brasel, 2001; Magnoli et al., 2011). The AFs are produced by genus Aspergillus, mainly A. flavus, A. parasiticus and A. nomius, that grow on a variety of raw material during growth, harvest, storage and transportation of for example, the cereal used in the preparation of food and feed commodities (Ito et al., 2001; Kurtzman et al., 1987; Payne, 1998; Pereyra et al., 2010). The investigation of strategies to prevent the presence of AFs in foods, as well as, to eliminate, inactivate or reduce the bio-availability of these mycotoxins in contaminated products include physical, chemical, and biological methods (Bueno et al., 2001; CAST, 2003; Kabak et al., 2006). Limitations such as the loss of nutritional and sensory qualities of the product, the expensive equipment required for these techniques and the impossibility to guarantee the desired results, have allowed us to consider the hipothesis that foods and feeds can always be potentially contaminated with aflatoxins. For instance, in the poultry industry aflatoxin B1 (AFB1) is almost an unavoidable feed contaminant and levels from 0-200 ng/g have been reported (Dalcero et al., 1997). On the other hand, it is known that lactic acid bacteria (LAB) and some yeast, principally Saccharomyces cerevisiae, are capable to bind AFs in liquid media, apparently to cell wall components, polysaccharides and peptidoglycans of LAB (Haskard et al., 2001; Latinen et al., 2004) and glucomannans of yeast (Karaman et al., 2005; Raju & Devegowda 2000) and therefore could be used as potential mycotoxin decontaminating (Armando et al 2011; El-Nezami et al., 1998; Haskard et al., 2000, 2001; Hernandez-Mendoza et al., 2009; Lee et al., 2003; Peltonen et al., 2001; Shetty et al., 2007). The inclusion of appropriate microorganisms in the contaminated diet could prevent the absorption of mycotoxins during their passage in the gastrointestinal tract and eliminated in the faeces (Bueno et al., 2007; El-Nezami et al., 2000; Gratz et al., 2004; Gratz et al., 2007). Moreover, Kankaanpää et al. (2000) showed that the binding of AFB1 to the surface of LAB reduced their adhesive properties, and the accumulation of aflatoxins in the intestine may therefore be reduced via the increased excretion of an aflatoxin-bacteria complex. These considerations encouraged the recent emphasis on biological methods, but mainly focused on preventing AFs absorption in the gastrointestinal tract of the consumers, including these microorganisms in the diet and so prevent the aflatoxicosis effects. The first step in this direction is the selection of the most efficient microorganism for AFB1 removing and while many researchers have assayed LAB and yeast with AFB1 binding abilities (Ciegler et al., 1966; El-Nezami et al., 1998; Gourama & Bullerman, 1995; Haskard et al., 2001; Line et al., 1994; Oatley et al., 2000) no clear mechanism for this effect has been provided. Thus, this selection frequently is performed using a single concentration of AFB1, but we demonstrated that the microorganism efficiency may change when the mycotoxin concentration is modified (Bueno et al., 2007; Pizzolitto, 2011), therefore the microorganism selected could not be the most competent. In this context, we investigated the nature of the interaction between different microorganisms and AFB1 molecule, which led us to develop a model to explain the binding of AFB1 by LAB and Saccharomyces cerevisiae strains. This model allows an estimation of two important parameters related to a microorganism´s capacity for dietary decontamination: the number of binding sites for AFB1 in the surface microorganism (M) and the equilibrium constant of the process involved (Keq), both of them are useful in the selection of the most suitable microorganism in a wide range of AFB1 concentration (Bueno et al., 2007). In adittion, studies of viability of the microorganisms in the salivary and gastrointestinal tract, cell adhesion, autoaggregation, coaggregation and antimicrobial activity against pathogen strains, were also evaluated as a way to research potential beneficial properties on the host (Armando et al., 2011). Thus, in this chapter we describe the development and application of an in vitro methodology to evaluate the aflatoxin B1 binding ability, gastrointestinal tolerance and potential beneficial properties of Saccharomyces cerevisiae strains, useful to select the more appropriated microorganism to be assayed in expensive, complicated but necessary in vivo studies.Fil: Pizzolitto, Romina Paola. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Bueno, Dante Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Armando, María Romina. 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: Dalcero, Ana Maria. 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: Salvano, Mario Armando. Universidad Nacional de Río Cuarto; Argentin

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

Endothelial Wall Thickness, Cardiorespiratory Fitness And Inflammatory Markers In Obese And Non-obese Adolescents.

Increased carotid intima-media thickness (c-IMT) is considered a marker of early-onset atherosclerosis and it has been found in obese children and adolescents, but the risk factors associated with this population remain to be elucidated. To compare and verify the relationship between c-IMT, metabolic profile, inflammatory markers, and cardiorespiratory fitness in obese and non-obese children and adolescents. Thirty-five obese subjects (19 boys) and 18 non-obese subjects (9 boys), aged 10-16 years, were included. Anthropometry, body composition, blood pressure, maximal oxygen consumption (VO2max), and basal metabolic rate were evaluated. Serum glucose, insulin, homeostasis model assessment of insulin resistance (HOMA-IR), blood lipids, C-reactive protein (CRP), and adiponectin were assessed. c-IMT was measured by ultrasound. The results showed that c-IMT, triglycerides, insulin, HOMA-IR, and CRP values were significantly higher in the obese group than in the non-obese group, and high-density lipoprotein cholesterol (HDL-c), adiponectin, and VO2max values were significantly lower in the obese group than in the non-obese group. The c-IMT was directly correlated with body weight, waist circumference, % body fat, and HOMA-IR and inversely correlated with % free fat mass, HDL-c, and VO2max. Our findings show that c-IMT correlates not only with body composition, lipids, insulin resistance, and inflammation but also with low VO2max values in children and adolescents.1847-5

Surveillance of Aflatoxin and Microbiota Related to Brewer's Grain Destined for Swine Feed in Argentina

Córdoba province in the center of Argentina is an important area of swine production. The use of industry by-product (brewer's grain) as feedstuff for swine is a regular practice and increases animal performance on these animals production. The occurrence of aflatoxin contamination is global, causing severe problems especially in developing countries. No reports on aflatoxin B1 production, micoflora, and potential aflatoxin B1 producing microorganism from brewer's grain are available. The aims of this study were (1) to isolate the microbiota species from brewer's grain, (2) to determine aflatoxin B1 natural contamination levels, and (3) to determine the ability of Aspergillus section Flavi isolates to produce aflatoxins in vitro. Physical properties, total fungal counts, lactic acid bacteria, and fungal genera distribution were determined on this substrate. In 65% of the samples, fungal counts were higher than recommended by GMP, and lactic bacterium counts ranged from 1.9 × 105 to 4.4 × 109 CFU g−1. Aspergillus spp. prevailed over other fungal genera. Aspergillus flavus was the prevalent species followed by A. fumigatus. Aflatoxin B1 levels in the samples were higher than the recommended limits (20 ng g−1) for complementary feedstuffs. Several Aspergillus section Flavi strains were able to produce aflatoxin B1  in vitro. Inadequate storage conditions promote the proliferation of mycotoxin-producing fungal species. Regular monitoring of feeds is required in order to prevent chronic and acute toxic syndromes related to this kind of contamination

Effect Of Concurrent Training With Blood Flow Restriction In The Elderly.

The aim of this present study was to investigate on the effects of concurrent training with blood flow restriction (BFR-CT) and concurrent training (CT) on the aerobic fitness, muscle mass and muscle strength in a cohort of older individuals. 25 healthy older adults (64.7±4.1 years; 69.33±10.8 kg; 1.6±0.1 m) were randomly assigned to experimental groups: CT (n=8, endurance training (ET), 2 days/week for 30-40 min, 50-80% VO2peak and RT, 2 days/week, leg press with 4 sets of 10 reps at 70-80% of 1-RM with 60 s rest), BFR-CT (n=10, ET, similar to CT, but resistance training with blood flow restriction: 2 days/week, leg press with 1 set of 30 and 3 sets of 15 reps at 20-30% 1-RM with 60 s rest) or control group (n=7). Quadriceps cross-sectional area (CSAq), 1-RM and VO2peak were assessed pre- and post-examination (12 wk). The CT and BFR-CT showed similar increases in CSAq post-test (7.3%, P<0.001; 7.6%, P<0.0001, respectively), 1-RM (38.1%, P<0.001; 35.4%, P=0.001, respectively) and VO2peak (9.5%, P=0.04; 10.3%, P=0.02, respectively). The BFR-CT promotes similar neuromuscular and cardiorespiratory adaptations as CT

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