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Chinchilla pelt is a rare and expensive fur. Therefore, breeding these animals is a profitable activity. Confirmed acute cases of aflatoxin intoxication have been reported in Argentinean farms. The aims of this study were i) to evaluate mycobiota and AFB1-producing species in chinchilla feeds ii) to investigate their natural AFB1 contamination and iii) to analyze histopathological lesions in chinchilla livers. Feed samples (A: fur chinchillas, B: mother chinchillas, C: lucerne cubes) were collected from a factory and a farm. Livers of sacrificed chinchilla from the farm were macroscopically and microscopically examined. Total fungal counts of feed C exceeded 1x10 4 CFU g-1. Aspergillus, Fusarium and Penicillium were the prevalent genera, while A. flavus, A. fumigatus, F. verticillioides and F. proliferatum were the prevalent species. 50 % of A. flavus strains from factory samples and 69.7 % from farm samples produced 2.78 to 8. 64 µg g-1 and 0.66 to 58.8 µg g-1 AFB1, respectively. Aflatoxin B1was detected only in feeds from the farm, finding the highest incidence in feed C. Toxin levels varied between 1.90 and 97.34 µg kg-1 AFB1. Mean levels in feed A and C exceeded 20 µg kg-1. Macroscopic examination of livers revealed normal appearance, size and color. However, histopathological examination indicated 63.3 % showed slight to moderate lipid degeneration with diffuse cytoplasm vacuolation, 9 % intense lipid cytoplasm vacuolation and 27.3 % hydropic degeneration and nuclear vacuolation in hepatocytes. A periodic monitoring of aflatoxins in feeds and their ingredients can prevent acute outbreaks and economic losses caused by chronic exposure.

Argentina is the fourth global beef producer. Exposure to mycotoxins through contaminated feed is a major hazard for ruminants. In the present study we assess mycobiota, aflatoxin B1 (AFB1), fumonisin B1 (FB1), deoxynivalenol (DON) and zearalenone (ZEA) levels in total mixed rations (TMRs) during two consecutive years. Total fungal counts were evaluated and fungal species were identified. Also, ability of A. flavus isolates to produce AFB1 in vitro was tested. Natural contamination with AFB1 and FB1 was quantified by HPLC. Deoxynivalenol and zearalenone were analysed by immunochromatography and thinlayer chromatogra- phy (TLC), respectively. Fungal counts varied from not detectable (ND) to 2.10 x 108 CFU g-1. The prevalent genera were Aspergillus spp (60 %) and Fusarium spp (66.7 %), respectively The prevalent species was Aspergillus fumigatus. 50 % of A. flavus strains produced 75 to 112.5 μg g-1 AFB1. 46 % of 2007 samples were contaminated with 4 to 10 μg kg-1 AFB1. Deoxynivalenol was detected in 33.3 % of the samples (≥ 1. 25 μg g-1). Fumonisin B1 and ZEA were not detected. This study can be useful to estimate the mycotoxicological risk of cattle TMRs in this region and to compare results with studies from other beef-producing countries.

The harmful effects of fungi in sorghum silage are an important matter in many countries with hot climates. This culture is very susceptible to fungal contamination and both the storage and processing of this substrate are ideal for the development of fungi there are able to produce mycotoxins. The contamination with mycotoxins, secondary products of the metabolism of certain fungi, that promotes risks to both animal and human health, causes numerous losses for the country. The aim of this study was to identify the toxicogenic fungi present in sorghum silage for beef cattle consumption and determine the occurrence of aflatoxin B1 (AFB1) in the same substrate. A total of 60 pre-fermented and 60 post-fermented samples of sorghum silage were analyzed. Total fungal counts and natural incidence of toxigenic Aspergillus, Penicillium y Fusarium species were performed on dichloran rose bengal chloranphenicol agar, dichloran glycerol agar 18 % and Nash-Snyder culture media. Aflatoxin B1 contamination was determined using high pressure liquid chromatography (HPLC). About 30 % of samples from pre-fermented sorghum and 55 % of post-fermented samples were above the recommended limits (1.0 x 104 CFU g-1). The most frequent fungal species in both types of sorghum samples were A. flavus, P. citrinum, P. islandicum and F. verticillioides. An average of 32 % samples, pre and post-fermented, were positive for AFB1. The presence of fungi and AFB1 in the feedstuffs indicates contamination. This toxin could affect animal productivity and health. This fact requires periodic monitoring to prevent the occurrence of mycotoxicosis in animal production.

The objective of the present study was to explore the influence of dietary supplementation with a mixed additive (MA) containing a probiotic and anti-mycotoxin (Saccharomyces cerevisiae RC016 and Lactobacillus rhamnosus RC007) and its interaction on the performance and health (biochemistry and liver/intestine histopathology) of broilers fed diets contaminated with aflatoxin B 1 (AFB 1 ) at 506000±22.1ng/kg. The MA contained S. cerevisiae RC016 (1×10 7 cells/g) and L. rhamnosus RC007 (1×10 8 cells/g) in relation 1:1. A total of sixty-one-day-old Cobb broilers were randomly allocated into four treatment groups with three replicates of 5 birds each for a five-week-old feeding experiment. The experimental diet for each treatment (T) was formulated as follows: T1, a commercial diet (CD); T2, CD+AFB 1 ; T3, CD+0.1% MA; T4, CD+AFB 1 +0.1% MA. The MA improved (p<0.01) production parameters (weight gain, conversion rate, and carcass yield) and reduced (p<0.01) the toxic effect of AFB 1 on the relative weight of the livers. In addition, the macro and microscopic alterations of livers and the possible intestinal injury related to histological damage in the presence of mycotoxin were reduced. The use of probiotic MA based on S. cerevisiae RC016 and L. rhamnosus RC007 in animal feed provides greater protection against mycotoxin contamination and is safe for use as a supplement in animal feed, providing beneficial effects that improve animal health and productivity. This is of great importance at the economic level for the avian production system., (Copyright © 2024 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Fusarium head blight (FHB) is one of the most common diseases in Argentina, affecting the quality and yield of barley grains. Fusarium graminearum sensu stricto (ss) and Fusarium poae are causal agents of FHB and potential sources of mycotoxin contamination in barley. Conventional management strategies do not lead to a complete control of FHB; therefore, biological control emerges as an eco-friendly alternative in the integrated management of the disease. In the present work, Bacillus velezensis, Bacillus inaquosorum, Bacillus nakamurai and Lactobacillus plantarum were evaluated as potential biocontrol agents against F. graminearum ss and F. poae on barley-based media. Bacillus velezensis RC218 was selected to carry out greenhouse and field trials in order to reduce FHB and mycotoxin accumulation. This strain was able to control growth of both Fusarium species and reduced deoxynivalenol (DON) and nivalenol (NIV) production by 66 % and 79 %, respectively. Bacillus inaquosorum and B. nakamurai were more effective in controlling F. poae growth, and the mean levels of reduction in DON accumulation were 50 and 38 %, and 93 and 26 % for NIV, respectively. Lactobacillus plantarum showed variable biocontrol capacity depending on the strain, with no significant mycotoxin reduction. The biocontrol on incidence and severity of FHB in the greenhouse and field trials was effective, being more efficient against F. graminearum ss and DON accumulation than against F. poae and NIV occurrence. This study provides valuable data for the development of an efficient tool based on biocontrol agents to prevent FHB-producing Fusarium species development and mycotoxin occurrence in barley, contributing to food safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A chitosan-glucose derivative (ChG) with lower antimicrobial activity against whey native probiotic yeast K. marxianus VM004 was synthesized by the Maillard reaction. The ChG derivative was characterized by FT-IR, 1 H NMR, and SLS to determine the structure, deacetylation degree (DD), and molecular weight (Mw). In addition, we evaluated the antioxidant, cytotoxic, and antimicrobial activities of ChG. ChG was then used for microencapsulation of K. marxianus VM004 by spray drying. The microcapsules were characterized by evaluating their encapsulation yield, encapsulation efficiency, morphology, tolerance to the gastrointestinal tract, and viability during storage. The results indicated that a non-cytotoxic product with lower MW and DD and higher antioxidant activity than native chitosan was obtained by the Maillard reaction. The yeast ChG microcapsules exhibited an encapsulation efficiency >57 %, improved resistance to gastrointestinal conditions, and enhanced stability during storage. These results demonstrate that ChG may be a promising wall material for the microencapsulation of probiotic yeasts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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.

In Argentina there are no reports on Aspergillus fumigatus fumagillin-producing strains. In this study we describe the isolation and mycotoxin production capacity of ten A. fumigatus strains isolated from farm and clinical samples. Farm strains were isolated from milk samples taken from dairy cows in Córdoba province, some of which were associated with subclinical mastitis. A culture medium was defined to optimize fumagillin production and a detection method was developed by HPLC chromatography. It is known that in addition to the host immune status, strain virulence is a fundamental characteristic that will determine its pathogenicity and, in this sense, fumagillin is considered to be among the virulence factors. In the present work, all the strains tested for the production of fumagillin were able to synthesize it, highlighting that the strain A. fumigatus RC2243, from a milk sample from a cow with clinical mastitis, was the most productive. The existence of fumagillin-producing strains represents a potential risk of mycotoxins being transferred to raw milk, constituting a public health risk., (Copyright © 2022 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

This study aimed to evaluate the aflatoxin B 1 (AFB 1 ) adsorption capacity of the seaweed Lithothamnium calcareum in vitro and to prevent aflatoxicosis in broiler chickens. In vitro adsorption assays were performed at a single AFB 1 concentration (1 μg/mL) and four seaweed concentrations (0.50, 1, 1.5 and 2 mg/mL) at pH 3 and pH 6. The maximum adsorption was obtained at the lowest seaweed content (0.62 and 0.78 μg/mg). Male broiler chickens (256) were housed in metallic cages. Experimental diets were T1 (control), 18 μg/kg AFB 1 ; T2, 18 μg/kg AFB 1 and 0.2% L. calcareum (2.0 kg/ton); T3, 1018 μg/kg AFB 1 ; and T4, 1018 μg/kg AFB 1 and 0.2% L. calcareum. Performance parameters (live weight, weight gain and feed conversion rate) improved when seaweed was applied. The aspartate-aminotransferase and alanine-aminotransferase levels tend to decrease in birds receiving only seaweed, also the uric acid levels reduced significantly (P ˂ 0.05), while birds receiving only AFB 1 increased the biochemical parameter levels. The livers from animals fed with AFB 1 showed histopathological alterations with disorganization of periportal hepatocytes, necrosis with multifocal coagulation and mild fat degeneration; the livers from T4 had normal appearance. Lithothamnium calcareum was able to prevent aflatoxicosis in broiler chickens and also improved their zootechnical performance.

The objective was to carry out cytotoxicity assays in Vero cells and cytokines analyses in Balb/c mice as safety assessments to evaluate the probiotic mixture (M) Saccharomyces cerevisiae RC016 (Sc) and Lactobacillus rhamnosus RC007 (Lr) for use as feed additive. Vero cells (10 4 cells per well) were exposed to Sc (2·08 × 10 7 , 2·08 × 10 6 ; 2·08 × 10 5 cells per ml), Lr (8·33 × 10 7 ; 8·33 × 10 6 ; 8·33 × 10 5 cells per ml) and their M (1 : 1). Sc concentrations did not affect the Vero cells viability; in contrast, they were lower when exposed to Lr (P ˂ 0·0001). Vero cells showed increasing viability with M decreasing concentrations (91% viability with M2). Control BALB/c mice received only phosphate buffer saline and the others received the M. The IL-10, IL-6 and TNFα concentrations from intestinal fluid were analysed and no significant differences were observed among treatments. The same occurred with the ratio between IL-10/TNF-α. Beneficial effects of probiotics are associated with the regulation of the excessive inflammatory response; it is desirable they can modulate the cytokines production only under pathological conditions. Here, M administration to healthy mice did not induce negative side effects and expands the knowledge about beneficial effects of using probiotic microorganisms in mixture for feed additives development., (© 2020 The Society for Applied Microbiology.)

Whey protein is one of the most relevant co-products manufactured by the dairy industry and it is a powerful environmental pollutant. Therefore, the enzymatic hydrolysis of whey protein concentrate (WPC 35) to produce antioxidant peptides is an innovative approach which can provide added value to whey. The WPC 35 hydrolysis with trypsin was carried out for 4.31 h at 41.1 °C with an enzyme/substrate ratio of 0.017. Under such hydrolysis conditions, the peptides produced have the highest radical scavenging activity and cytoprotector effect. The WPC hydrolysate and a permeate ≤3 kDa were characterized by SDS-page, RP-HPLC and MALDI-TOF-MS. Furthermore, O 2 •- and HO • scavenging activity and the cytoprotective effect against a stress agent in epithelial cells of the rat ileum (IEC-18) were determined. In this study, strong antioxidant and cytoprotective peptides were obtained from a low-cost dairy industry product, which could improve consumers' health when used as functional ingredients., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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.

The aim of this study was to evaluate the effect of feed restriction with or without monensin supplementation, followed by a re-feeding period on cellular apoptosis and proliferation in at term placenta of Anglo-Nubian goats. To evaluate the induction of apoptosis through the intrinsic pathway, proteins Bax and Bcl-2 were determinated. The apoptosis was related with the cell proliferation indices through Ki67 determination. The treatments were applied for 250 days and were (a) ad libitum feeding (control; n = 5); (b) restricted feeding at 70% of control (restricted; n = 7); and (c) restricted with monensin supplementation (monensin; n = 7). After treatments, all the animals were fed to support their requirements. After parturition, 27 placentas were gathered. The placental cellular structure was studied by high-resolution light microscopy and transmission electron microscopy; the cellular proliferation was determined by Ki67 index, and Bax and Bcl-2 proteins were localized by immunohistochemical analysis. Differences in cell proliferation through the Ki67 index were found in monensin group placentas. Monensin supplementation stimulated the placental cell proliferation reversing the effect of feed restriction during the peripuberal period. A significant increase of Bcl-2 in placentas of restricted group was found, and it would provide a protective effect on the placental structure. A lack of the Bcl-2 protective effect was observed in control and monensin group placentas, probably meaning that the observed apoptosis would be induced through the intrinsic signalling pathway. A balance between apoptosis and cell proliferation is necessary to maintain tissue homoeostasis during caprine placental development., (© 2018 Blackwell Verlag GmbH.)

Probiotics represents an alternative to replace antibiotics as growth promoters in animal feed and are able to control enteric bacterial diseases and to improve gut immunity. Saccharomyces cerevisiae RC016 showed previously inhibition/coagregation of pathogens) and mycotoxins adsorbent ability (aflatoxin B 1 , ochratoxin A and zearalenone). The aim of this work was to evaluate beneficial properties of S. cerevisiae RC016 in a non-inflammatory in vivo model in weaned piglets and in an intestinal inflammation ex vivo model induced by the mycotoxin deoxynivalenol (DON). Secretory immunoglobulin A (s-IgA) levels, intestinal cytokines, goblet cells and production parameters were evaluated in a pig model. For the in vivo assays, twelve pigs were weaned at 21 days and assigned to two groups: Control (n=6) and Yeast (n=6). Animals received yeast strain for three weeks. After 22 days the small intestine was recovered for determination of goblet cells and s-IgA. For the ex vivo assay, jejunal explants were obtained from 5 weeks old crossbred piglets and treated as follow: (1) control; (2) treated for 3 h with 10 μM DON used as an inflammatory stressor; (3) incubated with 10 7 cfu/ml yeast strain; (4) pre-incubated 1 h with 10 7 cfu/ml yeast strain and then treated for 3 h with 10 μM DON. CCL20, interleukin (IL)-1β, IL-8 and IL-22 gene expression was determined by qPCR. Oral administration of S. cerevisiae RC016 increased s-IgA, the number of goblet cells in small intestine and all the growth parameters measured. In the ex vivo model, the cytokine profile studied showed a potential anti-inflammatory effect of the administration of the yeast. In conclusion, S. cerevisiae RC016 is a promising candidate for feed additives formulation to improve animal growth and gut immune system. This yeast strain could be able to improve the gut health through counteracting the weaning-associated intestinal inflammation in piglets.

Microbial degradation of aflatoxins (AFs) is an alternative to the use of mycotoxin binders. The lactone ring is a possible target for microbial enzymes and its cleavage reduces AFs toxicity. The aim of this study was to isolate and identify Bacillus strains able to degrade AFB 1 to less toxic metabolites and to identify aiiA genes encoding for N-acyl-homoserine lactone (AHL) lactonase to possibly correlate detoxification with the production of this enzyme. Eleven soilborne Bacillus strains were isolated and identified by MALDI-TOF MS. Ten cultures and eight cell free culture supernatants (CFCS) were able to significantly (P < 0.05) degrade 27.78-79.78% AFB 1 . Cell lysates were also able to degrade AFB 1 (P < 0.05). Exposure to 70 and 80 °C did not affect enzyme activity. Aflatoxin B 1 toxicity towards Artemia salina was reduced after degradation by each of the Bacillus strains. B. subtilis RC1B, B. cereus RC1C and B. mojavensis RC3B, amplified a fragment of 753 pb corresponding to the aiiA gene encoding for AHL lactonase. AFB 1 degradation by the strains tested was due to the extracellular and intracellular enzymes. If demonstrated to be safe, these could be used to detoxify AFB 1 in contaminated food or feed., (Copyright © 2018. Published by Elsevier Ltd.)

Background: A survey on Fusarium species and moniliformin (MON) occurrence in sorghum grains collected from one of the main sorghum-producing areas of Argentina was conducted. Also, growth of F. thapsinum, one of the main sorghum pathogens, and MON production under different water activity (a w ) conditions on a sorghum-based medium were determined., Results: Infection of sorghum grains by Fusarium species ranged from 82.5 to 99%; closely related species F. verticillioides, F. thapsinum and F. andiyazi were the most frequently recovered, followed by F. proliferatum and F. subglutinans. By sequencing a portion of the translation elongation factor-1α (TEF-1α) gene and by maximum parsimony analysis, F. verticillioides and closely related species were identified as F. thapsinum, F. andiyazi and F. verticillioides. Species within the F. graminearum species complex (FGSC) were isolated in high frequency. Maximum growth rates of 12 F. thapsinum strains were obtained at 0.995 a w . All evaluated strains were able to produce MON at all a w values tested, but MON production was higher at 0.995-0.982 a w . MON was detected in 41% of the samples at levels ranging from 363.2 to 914.2 µg kg -1 ., Conclusion: This study provides new data on the occurrence of Fusarium species in sorghum grains destined for animal consumption in Argentina. The production of MON at different a w values showed that the toxin can be produced under field conditions. The risk to livestock exposed to daily low levels of MON associated with the toxin occurrence in the sorghum grains analyzed is unknown. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Aims: (i) To determine the aflatoxin B 1 (AFB 1 ) adsorption and desorption dynamics in the presence of Lactobacillus rhamnosus RC007 under simulated transit of AFB 1 at each gastrointestinal tract (GIT-saliva, stomach and intestine) stage consecutively and then, separately, (ii) to study the ability of L. rhamnosus RC007 to biotransform AFB 1 as a strategy that complements the adsorption process., Methods and Results: The AFB 1 adsorption and desorption assay simulating the GIT passage of AFB 1 (93·89 ng g -1 ) in the presence of L. rhamnosus RC007 (10 8 CFU per ml) was conducted. Moreover, lactic acid production was determined. Results demonstrated that predominant environmental conditions in salivary solution induced a low AFB 1 adsorption, while the transit through the gastric solution and intestinal solution allowed high percentages of adsorption and did not generate significant AFB 1 desorption., Conclusions: The AFB 1 adsorption and desorption dynamics in the presence of L. rhamnosus RC007 was favoured by gastric and intestinal environment., Significance and Impact of the Study: The knowledge of the adsorption dynamics of AFB 1 with a micro-organism of interest will allow predicting its behaviour at each stage of the GIT., (© 2018 The Society for Applied Microbiology.)

The objectives of this study were: to evaluate the use of dry distillery grain soluble extract - DDGse to produce yeast biomass and to obtain cell wall (CW), to use the CW as an aflatoxin B 1 (AFB 1 ) adsorbent, to study the variation in the composition and thickness of the CW under the influence of DDGse to evaluate their implication on the adsorption process using transmission electron microscopy (TEM) and fourier-transform infrared spectroscopy (FITR). The production of biomass and CW were variable. The CW thickness values showed that S. boulardii strain grown in yeast extract peptone dextrose (YPD) or DDGse medium, with no significant differences observed. The thickness of the CW for S. cerevisiae (RC012 and VM014) were increased when the cells were grown in DDGse medium, the thickness was almost double compared to the values obtained in YPD medium. The spectra IR of each CW in the two culture media shown regions corresponding to polysaccharides, proteins and lipids. Cells grown in DDGse medium adsorbed more AFB 1 than those grown in YPD. The CW adsorbed more AFB 1 than the same amount of whole cell. Future studies should be done to determine the type of carbohydrates and the relationship between chitin - beta glucans responsible for mycotoxin adsorption., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aspergillus fumigatus, the major etiological agent of human and animal aspergillosis, is a gliotoxinogenic species into section Fumigati commonly found in contaminated animal environments. In dairy herds, exposed areas of lactating cows, as mammalian glandule, can be easily contaminated by them. This study was aimed to identify A. fumigatus sensu lato strains (identified based on morphology) isolated from raw cow milk at species level, by morphological and molecular techniques, and to estimate their genetic variability. Forty-five A. fumigatus strains showed similar RAPD profiles (generated with PELF and URP1F primers) to each other and to A. fumigatus sensu stricto reference strains; also, they were almost identical to clinical human and feed-borne A. fumigatus strains included in the assay, since their similarity coefficient ranged from 0.7 to 1.00. Therefore, all strains were characterized as belonging to A. fumigatus sensu stricto species. This result was supported by sequencing the benA gene of selected strains and by maximum parsimony analysis. In addition, RAPD fingerprinting demonstrated intra-specific genetic variability into the A. fumigatus sensu stricto cluster. The results found in this study strengthen the fact that A. fumigatus sensu stricto is the predominant species in the Aspergillus section Fumigati found in animal environments such as dairy herd environments, while other species such as A. novofumigatus, A. fumigatiaffinis, A. udagawae and A. lentulus may be rarely isolated. Since no differences between animal and human strains were observed they can become pathogenic also for farm handlers'. Moreover, the presence of A. fumigatus sensu stricto in raw cow milk is probably a very important risk factor since milk and its by-products are generally indented for human consumption, then gliotoxin could be transferred to them., (Copyright © 2018. Published by Elsevier B.V.)

Aflatoxin B 1 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 B 1 (AFB 1 ) 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 (r 2 >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 AFB 1 in animal samples (LOD=0.017 and LOQ=0.050ng/g). The method was effective for the detection and quantification of AFB 1 residues in broiler liver and could also be potentially used for detecting AFB 1 in other edible animal tissues after natural or experimental AFB 1 exposure with high sensitivity and precision., (Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Probiotics have been explored to stimulate gut health in weaned pigs, when they started to consume solid diet where mycotoxins could be present. The aim of this study was to evaluate the effect of Lactobacillus rhamnosus RC007 on the intestinal toxicity of deoxynivalenol (DON) in an ex vivo model. Jejunal explants, obtained from 5-week-old crossbred castrated male piglets, were kept as control, exposed for 3 h to 10 μM DON, incubated for 4 h with 10 9 CFU/mL L. rhamnosus, or pre-incubated 1 h with 10 9 L. rhamnosus and exposed to DON. Histological lesions were observed, para- and transcellular intestinal permeability was measured in Ussing chambers. The expression levels of mRNA encoding six inflammatory cytokines (CCL20, IL-10, IL-1β, TNFα, IL-8 and IL-22) were determined by RT-PCR. The expressions of the phosphorylated MAP kinases p42/p44 and p38 were assessed by immunoblotting. Exposure to DON induced histological changes, significantly increased the expression of CCL20, IL-1β, TNFα, IL-8, IL-22 and IL-10, increased the intestinal paracellular permeability and activated MAP kinases. Incubation with L. rhamnosus alone did not have any significant effect. By contrast, the pre-incubation with L. rhamnosus reduced all the effects of DON: the histological alterations, the pro-inflammatory response, the paracellular permeability and the phosphorylation of MAP kinases. Of note, L. rhamnosus did not adsorb DON and only slightly degrade the toxin. In conclusion, L. rhamnosus RC007 is a promising probiotic which, included as feed additive, can decrease the intestinal toxicity of DON.

Probiotics are being used in biological control of bacterial pathogens, as an alternative to antibiotics, to improve health and production parameters in fish farming. Fish farming production is severely affected by aflatoxins (AFs), which are a significant problem in aquaculture systems. Aflatoxins exert substantial impact on production, causing disease with high mortality and a gradual decline of reared fish stock quality. Some aspects of aflatoxicosis in fish, particularly its effects on the gastrointestinal tract, have not been well documented. The aim of the present study was to evaluate probiotic properties of lactic acid bacterial (LAB) strains isolated from rainbow trout intestine and feed. Moreover, AFB 1 -binding and/or degrading abilities were also evaluated to assess their use in the formulation of feed additives. Growth at pH 2, the ability to co-aggregate with bacterial pathogens, inhibition of bacterial pathogens, and determination of the inhibitory mechanism were tested. Aflatoxin B 1 (AFB 1 ) adsorption and degradation ability were also tested. All strains were able to maintain viable (10 7 cells ml -1 ) at pH 2. Pediococcus acidilactici RC001 and RC008 showed the strongest antimicrobial activity, inhibiting all the pathogens tested. The strains produced antimicrobial compounds of different nature, being affected by different treatments (catalase, NaOH and heating), which indicated that they could be H 2 O 2, organic acids or proteins. All LAB strains tested showed the ability to coaggregate pathogenic bacteria, showing inhibition percentages above 40%. Pediococcus acidilactici RC003 was the one with the highest adsorption capacity and all LAB strains were able to degrade AFB 1 with percentages higher than 15%, showing significant differences with respect to the control. The ability of some of the LAB strains isolated in the present work to compete with pathogens, together with stability against bile and gastric pH, reduction of bioavailability and degradation of AFB 1 , may indicate the potential of LAB for use in rainbow trout culture.

The aim of this study was to evaluate the efficacy of autochthonous Pichia kudriavzevii as a novel bioadsorbent for aflatoxin B 1 (AFB 1 ). The selection of this yeast was based on the AFB 1 adsorption capacity previously demonstrated in vitro (Magnoli et al. 2016). One-day-old Cobb broilers (n = 160) were randomly assigned to four dietary treatments (T1: basal diet (B); T2: B + 0.1% yeast; T3: B + AFB 1 , 100 μg/kg; T4: B + 0.1% yeast + AFB 1 , 100 μg/kg). Performance parameters (average daily weight gain body, average daily consumption, feed conversion ratio, carcass weight, and dead weight), biochemical parameters (albumin, globulin, and albumin/globulin), liver pathological changes, and AFB 1 residual levels in the liver and excreta were evaluated. Significant differences (P < 0.05) in performance parameters were observed among treatments and controls: T3 group showed the lowest average daily body weight gain value while in T4 group, the value of this parameter increased significantly (P < 0.05). T3 and T4 groups showed the lowest and highest values for average daily feed consumption, respectively. The feed conversion ratio (FC) showed no significant differences among treatments. T3 group showed the lowest dead weight and carcass weight compared with T1 group. The biochemical parameters showed no significant differences among treatments. T3 group showed macroscopic and microscopic liver changes compared to the control. Aflatoxin B 1 levels (μg/g) were detected in broiler livers and showed significant differences among treatments (P < 0.05). In conclusion, native P. kudriavzevii incorporation (0.1%) in broiler diets containing AFB 1 was shown to be effective in ameliorating the adverse effects of AFB 1 on production.

Whey is the main byproduct of the cheese industry. While the composition is variable, it retains up to 55% of milk nutrients. The beneficial features of whey indicates a promising source of new potentially probiotic strains for the development of food additives destined for animal production. The aim of this study was to identify Kluyveromyces spp. isolated from whey, to study some probiotic properties and to select the best strain to be encapsulated using derivatised chitosan. Kluyveromyces marxianus strains (VM003, VM004 and VM005) were isolated from whey and identified by phenotypic and molecular techniques. These three yeast strains were able to survive under gastrointestinal conditions. Moreover, they exhibited weak auto-aggregation and co-aggregation with pathogenic bacteria (Salmonella sp., Serratia sp., Escherichia coli and Salmonella typhimurium). In general the K. marxianus strains had a strong antimicrobial activity against pathogenic bacteria. The potential probiotic K. marxianus VM004 strain was selected for derivatised-chitosan encapsulation. Material treated with native chitosan exhibited a strong antimicrobial activity of K. marxianus, showing a total growth inhibition at 10 min exposure. However, derivatised-chitosan encapsulation showed a reduced antimicrobial activity. This is the first study to show some probiotic properties of whey-native K. marxianus, in vitro. An encapsulation strategy was applied using derivatised chitosan.

Aims: To isolate and characterize native yeast strains from broilers' environment as feedstuff, faeces and gut, and to evaluate their binding capacity for aflatoxin B 1 (AFB 1 )., Methods and Results: A total of nine yeast strains were isolated: three from feedstuff identified as Pichia kudriavzevii (2) and Clavispora lusitaniae (1), two from gut identified as Candida tropicalis and four from faeces identified as Cl. lusitaniae (3) and Cyberlindnera fabianii (1). AFB 1 binding percentages varied among yeast strains and with AFB 1 concentrations. To carry out adsorption studies, one strain from each genus and each origin was selected as follows: Cl. lusitaniae and P. kudriavzevii from feedstuff, Cl. lusitaniae and Cy. fabianii from faeces and Ca. tropicalis from gut. The most appropriate concentrations for cells and toxin were 10 7 cells per ml and 100 ng ml -1 of AFB 1 respectively. All the tested yeast strains showed similar adsorption capacities independently of the origin. The adsorption isotherm studies in all yeasts assayed showed behaviour of L type or Langmuir and a varied affinity for the toxin. The stability of the AFB 1 -yeast complex demonstrated the irreversibility of the binding process., Conclusion: Yeast strains tested in this study constitute potential AFB 1 adsorbents and they possess the advantage to be native from the avian environment., Significance and Impact of the Study: This study makes a contribution to using native yeasts from broilers' environment for controlling chronic aflatoxicosis in avian production., (© 2016 The Society for Applied Microbiology.)

The aim of this work was to study the long-lasting consequences of different weaning age on physiological, immunological and microbiological parameters of weaned piglets. Piglets were weaned at 14 days (14W) or 21 days (21W). Blood samples were taken for IgG and cortisol determination on preweaning day and at 4; 20 and 40 post-weaning days. Three animals of each group were sacrificed. Small intestines for morphometric studies and secretory-IgA determination in fluid were taken. The cecum was obtained for enterobacteria, lactobacilli and total anaerobes enumeration. A significant decrease in piglet's plasma IgG concentrations was observed immediately after weaning and no differences were found between 14W and 21W. An increase in intestinal S-IgA was observed according to piglet's age. This increase was significantly higher in piglets 14W compared to piglets 21W. Animals from 14W group showed a decrease in villus length and in the number of goblet cells and intraepithelial lymphocytes. Other parameters were not affected by the weaning age. A short-term increase in cortisol was observed after weaning in both experimental groups. Enterobacteria decreased significantly after weaning in both groups, reaching values of weaning after 40 days. Lactobacilli counts decreased in both groups after weaning; however their counts were always higher than those obtained for enterobacteria. No differences were observed between 14W and 21W with regards to counts of anaerobes. The shortening of breast feeding time would favor an early synthesis of intestinal S-IgA after weaning. The changes observed in the microbiota could decrease postweaning enteric infections. However, early weaning induced negative effects on the cells of gut innate immunity and villi atrophy. This work provides knowledge about advantages and disadvantages at different weaning and long-lasting consequences on pig health. It is critical that swine producers become aware of the biological impacts of weaning age, so as to be able to decide the appropriate management strategies according to their facilities and rearing environment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Aspergillus section Nigri is a heterogeneous fungal group including some ochratoxin A producer species that usually contaminate raisins. The section contains the Series Carbonaria which includes the toxigenic species Aspergillus carbonarius and nontoxigenic Aspergillus ibericus that are phenotypically undistinguishable. The aim of this study was to examine the diversity of black aspergilli isolated from raisins and to develop a specific genetic marker to distinguish A. ibericus from A. carbonarius. The species most frequently found in raisins in this study were Aspergillus tubingensis (35.4%) and A. carbonarius (32.3%), followed by Aspergillus luchuensis (10.7%), Aspergillus japonicus (7.7%), Aspergillus niger (6.2%), Aspergillus welwitschiae (4.6%) and A. ibericus (3.1%). Based on inter-simple sequence repeat (ISSR) fingerprinting profiles of major Aspergillus section Nigri members, a sequence-characterized amplified region (SCAR) marker was identified. Primers were designed based on the conserved regions of the SCAR marker and were utilized in a PCR for simultaneous identification of A. carbonarius and A. ibericus. The detection level of the SCAR-PCR was found to be 0.01 ng of purified DNA. The present SCAR-PCR is rapid and less cumbersome than conventional identification techniques and could be a supplementary strategy and a reliable tool for high-throughput sample analysis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Aspergillus fumigatus, the major etiological agent of human and animal aspergillosis, is a toxigenic fungus largely regarded as a single species by macroscopic and microscopic features. However, molecular studies have demonstrated that several morphologically identified A. fumigatus strains might be genetically distinct. This work was aimed to apply PCR-restriction length fragment polymorphisms (PCR-RFLP) and random amplification of polymorphic DNA (RAPD) molecular markers to characterize a set of feed-borne and clinical A. fumigatus sensu lato strains isolated from Argentina and Brazil and to determine and compare their genetic variability. All A. fumigatus strains had the same band profile and those typical of A. fumigatus sensu stricto positive controls by PCR-RFLP. Moreover, all Argentinian and Brazilian strains typified by RAPD showed similar band patterns to each other and to A. fumigatus sensu stricto reference strains regardless of their isolation source (animal feeds or human/animal clinical cases) and geographic origin. Genetic similarity coefficients ranged from 0.61 to 1.00, but almost all isolates showed 78% of genetic similarly suggesting that genetic variability was found at intraspecific level. Finally, benA sequencing confirmed its identification as A. fumigatus sensu stricto species. These results suggest that A. fumigatus sensu stricto is a predominant species into Aspergillus section Fumigati found in animal environments as well as in human/animal clinical cases, while other species may be rarely isolated. The strains involved in human and animal aspergillosis could come from the environment where this fungus is frequently found. Rural workers and animals would be constantly exposed., (© The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In this study, gliotoxin production by Aspergillus fumigatus strains from animal environment is studied. Moreover, a rapid, easy and environment-friendly micro-analytical sample treatment procedure coupled with LC-MS/MS was applied for the determination of gliotoxin from A. fumigatus cultures. The ability of gliotoxin production by 143 strains was assayed in yeast extract sucrose agar, and 1 ml of chloroform was used for toxin extraction without further clean-up. Mean recoveries at two spiking levels (2500 and 7000 ng/g; n = 6) were 100.3 ± 6.6 % relative SD (RSD) and 92.4 ± 3.8 % RSD. Repeatability and within-laboratory reproducibility for different concentration levels of gliotoxin (25 to 1000 ng/ml; n = 12) ranged from 0.3 to 5.4 % RSD and from 3.9 to 12.7 % RSD, respectively. The detection limit of the analytical method was 3.5 ng/g. The ability for gliotoxin production by A. fumigatus revealed that 61.5 % of the strains were able to produce the toxin at levels ranging from LOQ to 3430.5 ng/g. However, all the tested samples had similar percentages of producing strains (81.8 to 86.6 %). The micro-analytical sample treatment coupled with LC-MS/MS detection is a precise and useful methodology for determining gliotoxin from fungal extracts of A. fumigatus and allows working both fast and safely and also reducing the effect on the environment. This toxin plays a critical role in the pathobiology of A. fumigatus, and its presence in animal environments could affect animal health and productivity; in addition, there are risks of contamination for rural workers during handling and storage of animal feedstuffs.

Minthostachys verticillata (Lamiaceae), popularly known as peperina is largely used in popular medicine for its digestive, carminative, antispasmodic and antirheumatic properties. There are no reports of repeated exposure toxicity to guarantee their safety. The present study investigated the chemical composition, analyzed by GC-FID, and the 90-day toxicity and genotoxicity effect of M. verticillata essential oil (Mv-EO), using Wistar rats as test animals. The rats were divided into four groups (5 rats/sex/group) and Mv-EO was administered on diet at doses of 0, 1, 4 and 7 g/kg feed. The main components of Mv-EO were pulegone (64.65%) and menthone (23.92%). There was no mortality, adverse effects on general conditions or changes in body weight, food consumption and feed conversion efficiency throughout the study in male and female rats. Subchronic administration of Mv-EO did not alter the weights, morphological and histopathological analyses of liver, kidney and intestine. Genotoxicity was tested by micronucleus and comet assays. Mv-EO up to a concentration of 7 g/kg feed for 90 days did not exert a cyto-genotoxic effect on the bone marrow and cells blood of Wistar rats. These results suggest that Mv-EO appears to be safe and could be devoid of any toxic risk., (Copyright © 2014 Elsevier Inc. All rights reserved.)

This study potentiates the adsorbent effect for aflatoxin B1 (AFB1) of a commercial additive (CA) of animal feed, containing inactive lysate of three Saccharomyces cerevisiae strains, active enzymes, adsorbents and a selenium-amino acid complex, when the additive was mixed separately with three S. cerevisiae strains. Levels of AFB1 of 20 and 50 ng g(-1) were used to determine the binding capacity of different concentrations of CA alone and in the presence of yeast strains, as well as toxin desorption, under gastrointestinal conditions. The viability of yeasts in the presence of CA was evaluated. The results show that the CA did not affect the viability of the yeast strains assayed. CA alone showed a low percentage adsorption. At 20 and at 50 ng g(-1), CA was highly efficient in adsorbing AFB1 when combined with RC016 and RC012 strains respectively. Desorption of AFB1 by CA alone and in combination with the yeasts increased with increasing levels of CA. The results demonstrate the improvement of CA in AFB1 adsorption once it is mixed with live yeasts.

Aims: To acquire data on the safety-in-use of the probiotic Saccharomyces cerevisiae RC016 and test its ability to reduce genotoxicity caused by dietary aflatoxins (AFs)., Methods and Results: The probiotic was orally administered to Wistar rats. Six groups (n = 6) were arranged: feed and probiotic controls, two levels of AFs-contaminated feed and two treatments including both the probiotic and the toxin. Genotoxiciy and cytotoxicity were evaluated with the bone marrow micronuclei assay and the comet assay and internal organs were macroscopically and microscopically examined. The tested S. cerevisiae strain did not cause genotoxicity or cytotoxicity in vivo, and it was able to attenuate AFs-caused genotoxicity. Saccharomyces cerevisiae RC016 did not cause any impairment on the rats' health and it showed no negative impact on the weight gain. Moreover, RC016 improved zootechnical parameters in AFs-treated animals. The beneficial effects were likely to be caused by adsorption of AFs to the yeast cell wall in the intestine and the consequent reduction in the toxin's bioavailability., Conclusions: The dietary administration of RC016 does not induce genotoxicity or cytotoxicity to rats., Significance and Impact of the Study: Incorporation of RC016 in the formulation of feed additives increases animal productivity. Similar effects may even occur in human food applications., (© 2014 The Society for Applied Microbiology.)

Unlabelled: Aspergillus fumigatus, a well-known human and animal pathogen causing aspergillosis, has been historically identified by morphological and microscopic features. However, recent studies have shown that species identification on the basis of morphology alone is problematic. The aim of this work was to confirm the taxonomic state at specie level of a set of clinical (human and animal) and animal environment A. fumigatus strains identified by morphological criteria applying a PCR-RFLP assay by an in silico and in situ analysis with three restriction enzymes. The A. fumigatus gliotoxin-producing ability was also determined. Previous to the in situ PCR-RFLP analysis, an in silico assay with BccI, MspI and Sau3AI restriction enzymes was carried out. After that, these enzymes were used for in situ assay. All A. fumigatus strains isolated from corn silage, human aspergillosis and bovine mastitis and high per cent of the strains isolated from cereals, animal feedstuff and sorghum silage were able to produce high gliotoxin levels. Also, all these strains identified by morphological criteria as A. fumigatus, regardless of its isolation source, had band patterns according to A. fumigatus sensu stricto by PCR-RFLP markers., Significance and Impact of the Study: Aspergillus fumigatus is a well-known human and animal pathogen causing aspergillosis. In this study, clinical (human and animal) and animal environment strains were able to produce high gliotoxin levels and had band profiles according to A. fumigatus sensu stricto by PCR-RFLP markers. The results obtained here suggest that strains involved in human and animal aspergillosis could come from the animal environment in which A. fumigatus is frequently found. Its presence in animal environments could affect animal health and productivity; in addition, there are risks of contamination for rural workers during handling and storage of animal feedstuffs., (© 2013 The Society for Applied Microbiology.)

The purposes of this study were to determine the distribution of total mycobiota, to determine the occurrence of Aspergillus spp., Penicillium spp. and Fusarium spp. and to detect and quantify fumonisin B1 and aflatoxin B1 in birds' feedstuffs. Sixty samples from different commercial feeds were collected. Analysis of the total mycobiota was performed and total fungal counts were expressed as CFU g(-1). The isolation frequency (%) and relative density (%) of fungal genera and species were determined. Mycotoxins determination was carried out using commercial ELISA kits. The 48% of standard, 31% of premium and only 9% of super premium feed samples were found above of recommended limit (1 × 10(4) CFU g(-1)). Aspergillus (82%), Cladosporium (50%) and Penicillium (42%) were the most frequently isolated genera. Aspergillus niger aggregate (35%), Aspergillus fumigatus (28%) and Aspergillus flavus (18%) had the highest relative densities. Contamination with fumonisins was detected in 95% of total samples with levels from 0·92 to 6·68 μg g(-1), and the aflatoxins contamination was found in 40% of total samples with levels between 1·2 and 9·02 μg kg(-1). Feed samples contaminated with fumonisins and aflatoxins are potentially toxic to birds., (© 2013 The Society for Applied Microbiology.)

The present revision shows the early and current knowledge in the field of silage fungi and mycotoxins explaining the relevance of fungi and mycotoxins in silage. The problem does not end in animal disease or production losses as mycotoxins in feed can lead to the presence of their metabolic products in dairy products, which will be eventually affecting human health, mainly infants. Silage is green forage preserved by lactic fermentation under anaerobic conditions. This ecosystem maintains its quality and nutritional value depending on interactions among physical, chemical and biological agents. Forages used for ensilage are naturally in contact with yeasts and filamentous fungi, and the contamination often occurs in the field and can also occur during harvesting, transport, storage. Moreover, postharvest poor management can lead to a rapid spoilage. Studies on fungal contamination of dairy cattle feed have shown how corn silage influences the contamination degree of feed supplied to livestock. Increasing knowledge in this area will help elucidate the influence that this microbiota exerts on production and/or degradation of mycotoxins present in silage. Some of these fungi, although opportunist pathogens, are relevant epidemiologically and represent a high risk of contamination to farm workers who handle them improperly., (© 2013 The Society for Applied Microbiology.)

Aim: To evaluate the ability of probiotic Saccharomyces cerevisiae RC016 strain to reduce fumonisin B(1) (FB(1)) in vitro and to optimize the culture conditions for the growth of the yeast employing surface response methodology., Methods and Results: Using Plackett-Burman screening designs (PBSD) and central composite designs (CCD), an optimized culture medium containing (g l(-1)) fermentable sugars provided by sugar cane molasses (CMs), yeast extract (YE) and (NH(4))(2) HPO(4) (DAP) was formulated. The S. cerevisiae RC016 strain showed the greatest binding at all assayed FB1 concentration. The CMs, YE, DAP concentrations and incubation time influenced significantly the biomass of S. cerevisiae RC016., Conclusion: A combination of CMs 17%; YE 4·61 g l(-1) and incubation time 60 h was optimum for maximum biomass of S. cerevisiae RC016., Significance and Impact of the Study: The importance of this work lies in the search for live strains with both probiotic and fumonisin B1 decontamination properties that could be sustainably produced in a medium just containing cheap carbon, nitrogen and phosphorus sources and would be included in a novel product to animal feed., (© 2013 The Society for Applied Microbiology.)

Aims: To in vitro evaluate the influence of the corn on the adsorption levels of aflatoxin B1 (AFB1) and zearalenone (ZEA) by yeast cell walls (YCWs)., Methods and Results: Two commercial YCWs were studied. The YCWs contain different percentages of polysaccharides. YCW1 and 2 contain 5.9 and 21% of mannans and 17.4 and 23% of β-glucans, respectively. Each YCW was resuspended in pH 2 and pH 6 buffer solutions. Corn was used to study the matrix influence. An aliquot of 500 μl YCW suspension was added to each microtube containing 500 μl of 0.1, 0.25, 0.5, 1, 2.5 and 5 μg ml(-1) AFB(1) or 0.5, 5, 10, 20 and 50 μg ml(-1) ZEA. Microtubes were kept with mechanical agitation at 37 °C for 30 min and then centrifuged for 10 min at 16,873 g and; the supernatants were quantified by high-pressure liquid chromatography. The amount of bound toxin was plotted as a function of the amount of added toxin according to mathematical expressions proposed by three theoretical models. Both YCWs were capable of adsorbing AFB(1) and ZEA in amounts from 0.061 to 0.40 and from 0.10 and 0.26 g g(-1), respectively. In the presence of the matrix, both adsorbents were not able to adsorb AFB(1) . However, they could adsorb ZEA at levels from 0.03 to 0.23 g g(-1)., Conclusions: Both YCWs adsorbed ZEA in the presence of corn and also under simulated gastrointestinal pH conditions. These results suggest that the studied YCWs are potential candidates for ZEA adsorption., Significance and Impact of the Study: Several in vitro assays have informed the ability of different substrates including yeast walls to adsorb AFB(1) and ZEA; none of them have evaluated their ability to adsorb AFB(1) and ZEA in the presence of the corn. The corn matrix can influence the adsorption phenomena of these mycotoxins., (© 2012 The Society for Applied Microbiology.)

The effect of Saccharomyces cerevisiae RC008 and RC016 strains, previously selected based on their aflatoxin B₁ mycotoxin binding ability and beneficial properties, against Aspergillus carbonarius and Fusarium graminearum under different interacting environmental conditions was evaluated. In vitro studies on the lag phase, growth rate and ochratoxin A/zearalenone and DON production were carried out under different regimens of a(w) (0.95 and 0.99); pH (4 and 6); temperature (25 and 37 °C) and oxygen availability (normal and reduced). Both yeast strains showed antagonistic activity and decreasing growth rate compared to the control. In general, the RC016 strain showed the greatest inhibitory activity. Except at the interacting condition 0.95 a(W), normal oxygen availability and 37 °C, at both pH values, A. carbonarius and F. graminearum were able to produce large amounts of mycotoxins in vitro. In general, a significant decrease in levels of mycotoxins in comparison with the control was observed. S. cerevisiae RC008 and RC016 could be considered as effective agents to reduce growth and OTA, ZEA and DON production at different interacting environmental conditions, related to those found in stored feedstuff. The beneficial and biocontrol properties of these strains are important in their use as novel additives for the control of mycotoxigenic fungi in stored feedstuffs., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The main objective of this study was to evaluate the interference of environment components on the in vitro evaluation of aflatoxin B₁ adsorption capacity of sodium bentonite under simulated gastrointestinal conditions of monogastric and ruminant animals. Sodium bentonite showed a high aflatoxin B₁ affinity with all of the assays. Langmuir or sigmoid isotherms were found in different assays. Both the affinities and the surface excesses at monolayer saturation were affected by the buffer components. The specific influence of ions in each buffer solution was investigated. A significant decrease in the surface excess at monolayer saturation was observed under ionic strength control. A change in the isotherm shape from sigmoidal to Langmuir was observed with the increase in the sodium chloride concentration. This was attributed to the decrease in the importance of lateral interaction between adsorbed toxin molecules compared with surface-molecules interactions under a high salt coverage. The presence of rumen fluid components in the adsorption environment decreased the aflatoxin B₁ maximum adsorption capacity of sodium bentonite. Despite the high affinity of this adsorbent to capture aflatoxin B₁, different substances present in the environment could affect the adsorption capacity, at least at low toxin concentrations that mimic chronic exposure. The environment of the gastrointestinal tract, in either monogastric or ruminant animals, affect in vivo aflatoxin B₁ adsorption by sodium bentonite and should be taken into account when an in vitro performance evaluation is done.

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 deoxynivalenol (DON) present in brewers grains pre- and poststored intended for bovine intensive rearing. Poststored (80%) samples had counts higher than 1 × 10(4) colony-forming units (CFU/g). Cladosporium spp. and Aspergillus spp. were isolated at high frequencies. Aspergillus flavus was the prevalent isolated species. Prestored (70%) and poststored (100%) samples showed AFB1 levels over the recommended limits (20  μ g/Kg), and OTA levels were below the recommended limits (50  μ g/Kg) while pre- and poststored 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.

Aims:  To examine Saccharomyces cerevisae strains with previously reported beneficial properties and aflatoxin B₁ binding capacity, for their ability to remove ochratoxin A (OTA) and zearalenone (ZEA) and to study the relation between cell wall thickness and detoxificant ability of yeast strains., Methods and Results:  A mycotoxin binding assay at different toxin concentrations and the effect of gastrointestinal conditions on mycotoxin binding were evaluated. Ultrastructural studies of yeast cells were carried out with transmission electronic microscopy. All tested strains were capable of removing OTA and ZEA. Saccharomyces cerevisiae RC012 and RC016 showed the highest OTA removal percentage, whereas RC009 and RC012 strains showed the highest ZEA removal percentages. The cell diameter/cell wall thickness relation showed a correlation between cell wall amount and mycotoxin removal ability. After exposure to gastrointestinal conditions, a significant increase in mycotoxin binding was observed., Conclusions:  All tested Saccharomyces cerevisiae strains were able to remove OTA and ZEA, and physical adsorption would be the main mechanism involved in ochratoxin A and ZEA removal. Gastrointestinal conditions would enhance adsorption and not decrease mycotoxin-adsorbent interactions., Significance and Impact of the Study:  Live strains with mycotoxin binding ability and beneficial properties are potential probiotics that could be included in animal feed. Previous and present results suggest that the RC008 and RC016 strains are very promising candidates for functional feed product development., (© 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.)

Aims: 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.)

Animal feed may be contaminated with different mycotoxins, with aflatoxin B(1) (AFB(1)) being a very common and toxic compound. Considering that birds normally have to cope with different stressful situations at the same time, the present study aims to evaluate the effects of feed contamination with AFB(1) in combination with corticosterone treatment in drinking water (a model to induce physiological stress in birds) on selected performance indices: BW, feed conversion, egg production, and macroscopic and microscopic liver alterations. At 5 wk of age, quails were randomly assigned to 1 of 6 dietary treatment groups that resulted from the combination of the presence or absence of corticosterone in drinking water (5 mg/L) with the presence or absence of AFB(1) contamination (0, 100, or 500 μg/kg). The animals remained in these treatments from 5 to 11 wk of age. There were 6 replicates per treatment, each containing 2 males and 2 females. Contamination with 100 μg of AFB(1) per kilogram of feed induced no changes in BW, feed conversion, and egg production parameters. Quail fed with 500 μg of AFB(1) per kilogram of feed showed significant decreases in BW and feed consumption compared with their control counterparts. Corticosterone in combination with 500 μg of AFB(1) per kilogram of feed intensified the negative effects observed on BW and feed consumption and also had negative effects on feed conversion rate and egg production parameters, suggesting that the adverse effects of contamination with AFB(1) are intensified in situations of chronic stress. Quail treated with 500 µg of AFB(1) per kilogram showed hepatocytes with degree 1 and 2 lesions, and all quail treated with 500 µg of AFB(1) per kilogram of feed in combination with corticosterone showed degree 2 liver lesions (i.e., hepatocytes with fatty macro and microvacuoles and necrosis). This result is also consistent with the hypothesis that chronic stress exacerbates the effect of AFB(1) contamination. In conclusion, this study suggests that the negative effects of AFB(1) contamination are increased when overlapped with chronic stressful stimulation.

In this study the aflatoxin B₁ (AFB₁) removal capacity, the tolerance to salivary and gastrointestinal conditions, autoaggregation and coaggregation with pathogenic bacteria of Saccharomyces cerevisiae strains isolated from broiler feces, were evaluated. Only four of twelve isolated strains were identified as Saccharomyces cerevisiae using molecular techniques. The results obtained in AFB₁ binding studies indicated that the amount of AFB₁ removed was both strain and mycotoxin-concentration dependent. Therefore, a theoretical model was applied in order to select the most efficient strain to remove AFB₁ in a wide range of mycotoxin concentration. The results indicated that S. cerevisiae 08 and S. cerevisiae 01 strains were the most efficient microorganisms in the mycotoxin removal. Viability on simulated salivary and gastrointestinal conditions was investigated and S. cerevisiae 08 strain showed the best results, achieving 98% of total survival whereas S. cerevisiae 01 reached only 75%. Autoaggregation and coaggregation assays showed S. cerevisiae 08 as the most appropriate strain, mainly because it was the unique strain able to coaggregate with the four bacterial pathogens assayed. Consequently, S. cerevisiae 08 is the best candidate for future in vivo studies useful to prevent aflatoxicosis. Further quantitative in vitro and in vivo studies are required to evaluate the real impact of yeast-binding activity on the bioavailability of AFB₁ in poultry. However, this study could be useful in selecting efficient strains in terms of AFB₁ binding and provide an important contribution to research into microorganisms with potential probiotic effects on the host.

The effect of Saccharomyces cerevisiae RC008 and RC016, previously selected based on their aflatoxin B(1) binding ability and beneficial properties, against Aspergillus parasiticus under different interacting environmental conditions was evaluated. Studies concerning the lag phase, growth rate and aflatoxin B(1) production were carried out in vitro under different regimes of a (w) (0.95 and 0.99), pH (4 and 6), temperature (25 and 37°C), and oxygen availability (normal and reduced). Both yeast strains showed great antagonistic activity at pH 4, decreasing growth rate compared with the control. The RC008 strain showed the greatest inhibitory activity at all assayed conditions. A. parasiticus produced large amounts of AFB(1) in vitro. A significant decrease of AFB(1) levels in comparison with the control were observed with yeast interaction. Differences between control and treatment values ranged from 130 to 5400 ng ml(-1). S. cerevisiae RC008 and RC016 could be considered as effective agents in reducing growth and AFB(1) production at different interacting environmental conditions, related to that found in stored feedstuff. The importance of the present work lies in the search for live strains with both probiotic and biocontrol properties able to prolong the safe storage of feedstuff and exert beneficial properties after animal consumption and which could be included in a novel product for animal feed.

Aims: To evaluate mycobiota and aflatoxins B(1) (AFB(1)), B(2) (AFB(2)), G(1) (AFG(1)), G(2) (AFG(2)) and fumonisin B(1) (FB(1)) contamination in different malted barley types and brands and brewer's grain collected from a major Argentinean brewery., Methods and Results: Total fungal counts were performed using the plate count method. Aflatoxin B(1), AFB(2), AFG(1), AFG(2) and Zearalenone (ZEA) analyses were performed by thin-layer chromatography (TLC). Fumonisin B(1) was determined by HPLC. Eighty-three percentage of the malted barley (100% M1, 50% M2 and 100% M3) and 61% of brewer's grain samples had a count >1 × 10(4) CFU g(-1). Yeasts were isolated from all malt and brewer's grain samples. Genera containing some of the most important mycotoxin producer species--Fusarium ssp., Aspergillus ssp., Penicillium ssp. and Alternaria ssp.--were isolated from the analysed samples, along with other environmental saprophytic fungi such as Geotrichum ssp., Mucorales and Cladosporium ssp. All samples were contaminated with 104-145 μg kg(-1) FB(1). Eighteen per cent of brewer's grain samples were contaminated with 19-44.52 μg kg(-1) AFB(1). Aflatoxin B(2), AFG(1), AFG(2) and ZEA were not detected in any of the analysed samples., Conclusions: Fungal and mycotoxin contamination in malt and brewer's grain is an actual risk for animal and human health., Significance and Impact of the Study: This study may be useful for assessing the risk of mycotoxins in Argentinean beers and especially in animal feeds., (© 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.)

The aim of the present study was to determine the mycobiota and natural levels of mycotoxins such as zearalenone, fumonisin B(1), aflatoxin B(1) and ochratoxin A present in raw materials and finished fattening pig feed. Samples were examined for total fungi and genera distribution. Zearalenone, FB(1), AFB(1) and OTA contamination were determined using high pressure liquid chromatography and thin layer chromatography. Milled maize and finished feed samples showed fungal contamination over than 1 × 10(4) CFU/g. All samples contained at least one of the main mycotoxigenic genera Aspergillus, Fusarium and Penicillium. A. flavus and F. verticillioides were the most prevalent species. Only some Aspergillus section Nigri strains from suckling pig to growing pig samples were able to produce OTA. A. flavus strains from milled maize, wheat bran, suckling pig to growing pig samples were able to produce AFB(1). All samples were positive for FB(1). Sucking pig, piglet, growing and boar feed samples showed ZEA natural contamination. AFB(1) and OTA contamination were not detected. There was a 100% correlation between FB(1) and ZEA contamination in sucking pig, piglet, growing and boar feed samples; 50% piglet samples and 67% suckling pig samples showed ZEA levels over the recommended limits. The present study has shown the occurrence of two mycotoxins, FB(1) and ZEA in feed intended for fattening pig consumption. In animal production, the simultaneous presence of toxicogenic fungi and low dietary levels of mycotoxins in field conditions can cause possible health impacts and lost performance in pigs from feeding spoiled feeds.

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 B(1) production, micoflora, and potential aflatoxin B(1) 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 B(1) 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 × 10(5) to 4.4 × 10(9) CFU g(-1). Aspergillus spp. prevailed over other fungal genera. Aspergillus flavus was the prevalent species followed by A. fumigatus. Aflatoxin B(1) 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 B(1)  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.