RESUMO
AIMS: To isolate and characterize native yeast strains from broilers' environment as feedstuff, faeces and gut, and to evaluate their binding capacity for aflatoxin B1 (AFB1 ). 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). AFB1 binding percentages varied among yeast strains and with AFB1 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 107 cells per ml and 100 ng ml-1 of AFB1 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 AFB1 -yeast complex demonstrated the irreversibility of the binding process. CONCLUSION: Yeast strains tested in this study constitute potential AFB1 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.
Assuntos
Aflatoxina B1/metabolismo , Galinhas/microbiologia , Leveduras/metabolismo , Adsorção , Ração Animal/microbiologia , Animais , Fezes/microbiologia , Intestinos/microbiologia , Leveduras/isolamento & purificaçãoRESUMO
AIM: This study evaluated the binding capacity of aflatoxin B1 (AFB1 ) by two Enterococcus faecium strains (MF4 and GJ40) isolated from faeces from healthy dogs. MATERIALS AND METHODS: The binding assay was performed using 50 and 100 ppb of AFB1 analysing the effects of the viability, incubation time and pH on AFB1 binding. Binding stability was determined by washing three times the bacteria-AFB1 complexes with phosphate buffer saline. RESULTS: Both GJ40 and MF4 strains have the ability to remove AFB1 from aqueous solution. Viable cells were slightly more effective in AFB1 binding than nonviable ones for both strains. Enterococcus faeciumGJ40 removes 24-27% and 17-24%, and Ent. faeciumMF4 removes 36-42% and 27-32% of AFB1 (50 and 100 ppb, respectively) throughout a 48 h incubation period. In general, the removal of AFB1 was highest at pH 7.00 for both strains. The stability of the bacteria-AFB1 complex formed was found to be high (up to 50% of AFB1 remained bounded in bacterial cell after three washes with phosphate buffered saline). CONCLUSION: The Ent. faecium strains assayed are capable of removing AFB1 under different conditions in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first AFB1 binding assay performed with Ent. faecium strains isolated from dog faeces, being an interesting strategy for AFB1 decontamination of pet food.
Assuntos
Aflatoxina B1/metabolismo , Enterococcus faecium/metabolismo , Animais , Cães , Enterococcus faecium/isolamento & purificação , Fezes/microbiologiaRESUMO
The main objective of this study was to determine if the competitive adsorption of tryptophan (Trp) and aflatoxin B1 (AFB1) could potentially affect the ability of a sodium bentonite (NaB) to prevent aflatoxicosis in monogastric animals. The adsorption of Trp and AFB1 on this adsorbent is fast and could be operating on the same time-scale making competition feasible. In vitro competitive adsorption experiments under simulated gastrointestinal conditions were performed. A high affinity of the clay for Trp and NaB was observed. The effect of an excess of KCl to mimic the ionic strength of the physiological conditions were also investigated. A six-times decrease in the Trp surface excess at saturation was observed. A similar behaviour was previously found for AFB1 adsorption. Taking into account the amount of Trp adsorbed by the clay and the usual adsorbent supplementation level in diets, a decrease in Trp bioavailability is not expected to occur. Tryptophan adsorption isotherms on NaB were 'S'-shaped and were adjusted by the Frumkin-Fowler-Guggenheim model. The reversibility of the adsorption processes was investigated in order to check a potential decrease in the ability of NaB to protect birds against chronic aflatoxicoses. Adsorption processes were completely reversible for Trp, while almost irreversible for AFB1. In spite of the high affinity of the NaB for Trp, probably due to the reversible character of Trp adsorption, no changes in the AFB1 adsorption isotherm were observed when an excess of the amino acid was added to the adsorption medium. As a consequence of the preferential and irreversible AFB1 adsorption and the reversible weak binding of Trp to the NaB, no changes in the aflatoxin sorption ability of the clay are expected to occur in the gastrointestinal tract of birds.
Assuntos
Aflatoxina B1/química , Bentonita/química , Carcinógenos Ambientais/química , Quelantes/química , Modelos Químicos , Triptofano/química , Adsorção , Aflatoxina B1/antagonistas & inibidores , Aflatoxina B1/metabolismo , Ração Animal , Animais , Argentina , Bentonita/metabolismo , Ligação Competitiva , Carcinógenos Ambientais/metabolismo , Quelantes/metabolismo , Estudos de Viabilidade , Aditivos Alimentares/química , Aditivos Alimentares/metabolismo , Contaminação de Alimentos/prevenção & controle , Conteúdo Gastrointestinal , Cinética , Concentração Osmolar , Aves Domésticas , Triptofano/metabolismoRESUMO
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.
Assuntos
Probióticos/toxicidade , Saccharomyces cerevisiae , Administração Oral , Aflatoxinas/toxicidade , Ração Animal , Animais , Dano ao DNA , Masculino , Ratos , Ratos Wistar , Testes de Toxicidade SubcrônicaRESUMO
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.
Assuntos
Aspergilose/microbiologia , Aspergilose/veterinária , Aspergillus fumigatus/classificação , Grão Comestível/microbiologia , Gliotoxina/metabolismo , Mastite Bovina/microbiologia , Silagem/microbiologia , Animais , Aspergillus fumigatus/genética , Aspergillus fumigatus/isolamento & purificação , Aspergillus fumigatus/metabolismo , Técnicas de Tipagem Bacteriana , Sequência de Bases , Brasil , Bovinos , Feminino , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
Aflatoxins (AF) are the most important mycotoxins produced by toxigenic strains of various Aspergillus spp. Biological decontamination of mycotoxins using microorganisms is a well-known strategy for the management of mycotoxins in feeds. Saccharomyces cerevisiae strains have been reported to bind aflatoxin B1 (AFB1). The aim of this study was to evaluate the ability of S. cerevisiae CECT 1891 in counteracting the deleterious effects of AFB1 in broiler chicks. Experimental aflatoxicosis was induced in 6-d-old broilers by feeding them 1.2 mg of AFB1/kg of feed for 3 wk, and the yeast strain was administrated in feed (10(10) cells/kg), in the drinking water (5 × 10(9) cells/L), or a combination of both treatments. A total of 160 chicks were randomly divided into 8 treatments (4 repetitions per treatment). Growth performance was measured weekly from d 7 to 28, and serum biochemical parameters, weights, and histopathological examination of livers were determined at d 28. The AFB1 significantly decreased the BW gain, feed intake, and impaired feed conversion rate. Moreover, AFB1 treatment decreased serum protein concentration and increased liver damage. The addition of S. cerevisiae strain to drinking water, to diets contaminated with AFB1, showed a positive protection effect on the relative weight of the liver, histopathology, and biochemical parameters. Furthermore, dietary addition of the yeast strain to drinking water alleviated the negative effects of AFB1 on growth performance parameters. In conclusion, this study suggests that in feed contaminated with AFB1, the use of S. cerevisiae is an alternative method to reduce the adverse effects of aflatoxicosis. Thus, apart from its excellent nutritional value, yeast can also be used as a mycotoxin adsorbent.
Assuntos
Aflatoxinas/química , Ração Animal/análise , Galinhas , Contaminação de Alimentos , Micotoxicose/veterinária , Saccharomyces cerevisiae/fisiologia , Animais , Masculino , Micotoxicose/prevenção & controle , Doenças das Aves Domésticas/induzido quimicamente , Doenças das Aves Domésticas/prevenção & controle , ProbióticosRESUMO
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.
Assuntos
Aflatoxina B1/farmacologia , Bentonita/química , Trato Gastrointestinal/metabolismo , Adsorção , Aflatoxina B1/química , Aflatoxina B1/metabolismo , Animais , Soluções Tampão , Cromatografia Líquida de Alta Pressão , Concentração de Íons de Hidrogênio , Técnicas In Vitro , RuminantesRESUMO
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.
Assuntos
Fungos/isolamento & purificação , Micotoxinas/isolamento & purificação , Silagem/microbiologia , Animais , Bovinos , Fungos/metabolismo , Micotoxicose/veterinária , Micotoxinas/metabolismoRESUMO
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.