RESUMO
T-2 toxin, a hazardous mycotoxin often present in cereals and products based on cereals, poses a substantial risk to humans and animals due to its high toxicity. The development of uncomplicated, quick and highly sensitive methods for detecting T-2 toxin is imperative. In this work, a portable sensing system was constructed using water column height as a readout device in combination with a controlled release system, which allows for an accurate quantitative analysis of T-2 toxin without the need for expensive instrumentation or skilled technicians. Hyaluronic acid (HA) hydrogel was constructed by double cross-linked DNA/aptamer hybrids with polyethyleneimine (PEI) and embedded with platinum nanoparticles (Pt NPs). The aptamer specifically bound to T-2 toxin in its presence, resulting in the disruption of the hydrogel and subsequent release of the Pt NPs. These Pt NPs were later mixed with a solution of H2O2 in a confined reaction flask, leading to the decomposition of H2O2 into O2. A glass capillary tube containing a column of red water had been inserted into the cap of the reaction flask, and the low solubility of O2 led to an increase in pressure within the reaction unit, causing the red water column to rise. There is a good linear correlation between the height of the capillary liquid level and the T-2 toxin concentration in the range of 20 ng/mL to 6 µg/mL. The system has been successfully used to detect T-2 toxin in samples of barley tea and corn.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Platina , Toxina T-2 , Toxina T-2/análise , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/química , Nanopartículas Metálicas/química , Platina/química , Água/química , DNA/química , DNA/análise , Hidrogéis/química , Limite de Detecção , Ácido Hialurônico/química , Polietilenoimina/químicaRESUMO
T-2 is one of the most potent cytotoxic food-borne mycotoxins. In this work, we have developed and characterized an electrochemical microfluidic immunosensor for T-2 toxin quantification in wheat germ samples. T-2 toxin detection was carried out using a competitive immunoassay method based on monoclonal anti-T-2 antibodies immobilized on the poly(methyl methacrylate) (PMMA) microfluidic central channel. The platinum wire working electrode at the end of the channel was in situ modified by a single-step electrodeposition procedure with reduced graphene oxide (rGO)-nanoporous gold (NPG). T-2 toxin in the sample was allowed to compete with T-2-horseradish peroxidase (HRP) conjugated for the specific recognizing sites of immobilized anti-T-2 monoclonal antibodies. The HRP, in the presence of hydrogen peroxide (H2O2), catalyzes the oxidation of 4-tert-butylcatechol (4-TBC), whose back electrochemical reduction was detected on the nanostructured electrode at -0.15 V. Thus, at low T-2 concentrations in the sample, more enzymatically conjugated T-2 will bind to the capture antibodies, and, therefore, a higher current is expected. The detection limits found for electrochemical immunosensor, and commercial ELISA procedure were 0.10 µg kg-1 and 10 µg kg-1, and the intra- and inter-assay coefficients of variation were below 5.35% and 6.87%, respectively. Finally, our microfluidic immunosensor to T-2 toxin will significantly contribute to faster, direct, and secure in situ analysis in agricultural samples.
Assuntos
Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Micotoxinas , Nanoporos , Toxina T-2 , Grafite/química , Imunoensaio/métodos , Microfluídica , Ouro/química , Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/química , Técnicas Eletroquímicas/métodos , Limite de Detecção , Nanopartículas Metálicas/químicaRESUMO
Aluminosilicates are adsorbents able to bind mycotoxins, and their chemical modification increases their affinity to adsorb low-polarity mycotoxins. To further investigate if the inclusion of salts in bentonite modifies its adsorptive capacity, we studied T-2 toxin adsorption in natural bentonite (NB) and when modified with quaternary ammonium salts differing in polarity and chain length: myristyl trimethyl ammonium bromide (B14), cetyl trimethyl ammonium bromide (B16) and benzyl dimethyl stearyl ammonium chloride (B18). The results showed that quaternary salts made bentonite: displace monovalent (Na+1, K+1) and divalent (Mg+2, Ca+2) ions; reduce its porosity; change its compaction and structure, becoming more crystalline and ordered; and modify the charge balance of sheets. T-2 adsorption was higher in all modified materials compared to NB (p ≤ 0.0001), and B16 (42.96%) better adsorbed T-2 compared to B18 (35.80%; p = 0.0066). B14 (38.40%) showed no differences compared to B16 and B18 (p > 0.05). We described the T-2 adsorption mechanism in B16, in which hydrogen bond interactions, Van der Waals forces and the replacement of the salt by T-2 were found. Our results showed that interaction types due to the inclusion in B16 might be more important than the hydrocarbon chain length to improve the adsorptive capacity of bentonite.
Assuntos
Toxina T-2 , Poluentes Químicos da Água , Bentonita/química , Adsorção , Sais , Cátions , Poluentes Químicos da Água/químicaRESUMO
Mycotoxins represent substantial challenges to the farming industry. These include toxins produced by Fusarium fungi, particularly trichothecenes (toxin T-2) and fumonisin (FB1). In the present study, we determined the effects of addition on Saccharomyces cerevisiae lysate (SCL) added to feed contaminated with T-2 and FB1 in terms of health, productive efficiency, and egg laying quality. We used 60 Hy-line Brown laying hens, and divided them into five groups with four repetitions per group and three birds per repetition. There was one group with no contamination with toxin (NoC). The four other groups included combinations of mycotoxin (4 ppm T-2, and 20 ppm FB1): A contamination group was used as control (the C+ group), and another two contained 500 g/ton of SCL (Detoxa Plus®) (the C + D500 group) or 1000 g/ton of SCL (the C + D1000 group). Finally, one group received feed containing 500 g/ton of Detoxa Plus® and 1000 g/ton of Uniwall® MOS 25 (the C + D500 + U1000 group). The experimental period was 84 days, divided into three productive cycles of 28 days each. The NoC hens had greater egg production than the other groups. Hens that consumed feed with SCL had greater egg production than did the C+ group. The NoC hens produced eggs with greater weights than did the C hens; however, C + D1000 and C + D500 + U1000 birds produced greater egg weights than did the C+ group. The C+ group produced lower egg masses than did the NoC and C + D500 + U1000 groups. The feed intake (FI) was lower in hens that ingested mycotoxin. The use of SCL in feed minimized the negative effects of mycotoxin on feed conversion ratio (FI/dozen). Effects of treatment were detected for feed conversion ratio (kg/kg). The hens that consumed mycotoxin had lower shell resistance and thickness compared to those in the NoC group. The red color of egg yolk was greater in the control groups. There were fluctuations in levels of liver enzymes when birds consumed mycotoxin (sometimes reduced and sometimes increased); nevertheless, the cumulative effect increased the activity of alanine aminotransferase. The serum concentration of reactive oxygen species was greater in hens that ingested mycotoxin only on d 84 compared to the NoC group. Serum glutathione S-transferase activity was greater on d 56 in C + D500 and C + D1000 hens than in the others. We conclude that, in general, the consumption of mycotoxin impaired the performance and quality of the eggs of the hens; the addition of the S. cerevisiae lysate and the addition organic acids, yeast cell wall and mineral carrier minimized some of the negative effects caused by T-2 and FB1.
Assuntos
Fumonisinas , Fusarium , Micotoxinas , Toxina T-2 , Ração Animal/análise , Animais , Galinhas , Dieta/veterinária , Feminino , Aves Domésticas , Saccharomyces cerevisiaeRESUMO
Fusarium Head Blight (FHB) is a destructive disease of small grain cereals and a major food safety concern. Epidemics result in substantial yield losses, reduction in crop quality, and contamination of grains with trichothecenes and other mycotoxins. A number of different fusaria can cause FHB, and there are significant regional differences in the occurrence and prevalence of FHB pathogen species and their associated mycotoxins. Information on FHB pathogen and mycotoxin diversity in Mexico has been extremely limited, but is needed to improve disease and mycotoxin control efforts. To address this, we used a combination of DNA sequence-based methods and in-vitro toxin analyses to characterize FHB isolates collected from symptomatic wheat in Mexico during the 2013 and 2014 growing seasons. Among 116 Fusarium isolates, we identified five species complexes including nine named Fusarium species and 30 isolates representing unnamed or potentially novel species. Significant regional differences (Pâ¯<â¯0.001) in pathogen composition were observed, with F. boothii accounting for >90% of isolates from the Mixteca region in southern Mexico, whereas F. avenaceum and related members of the F. tricinctum species complex (FTSC) accounted for nearly 75% of isolates from the Highlands region in Central Mexico. F. graminearum, which is the dominant FHB pathogen in other parts of North America, was not present among the isolates from Mexico. F. boothii isolates had the 15-acetyldeoxynivalenol toxin type, and some of the minor FHB species produced trichothecenes, such as nivalenol, T-2 toxin and diacetoxyscirpenol. None of the FTSC isolates tested was able to produce trichothecenes, but many produced chlamydosporol and enniatin B.
Assuntos
Depsipeptídeos/análise , Grão Comestível/microbiologia , Fusarium/metabolismo , Micotoxinas/análise , Toxina T-2/análise , Tricotecenos/análise , Triticum/microbiologia , Grão Comestível/química , Contaminação de Alimentos/análise , Fusarium/classificação , Fusarium/genética , Fusarium/isolamento & purificação , México , Doenças das Plantas/microbiologiaRESUMO
Diversas micotoxinas amplamente distribuídas na natureza possuem propriedades tóxicas acentuadas e podem contaminar culturas de milho destinadas à alimentação humana e animal. A aflatoxina e as fusariotoxinas (principalmente fumonisina e zearalenona) são as que mais comumente provocam prejuízos na suinocultura. A produção das fusariotoxinas ocorre principalmente antes da colheita, mas assim como a aflatoxina, também pode ocorrer pós-colheita caso não seja tratada e seca corretamente. Os efeitos relacionados à ingestão destes compostos são dependentes de fatores relacionados à toxina (estrutura química, dose) e ao animal (espécies, raça, sexo, idade, condições nutricionais). Ainda, a possibilidade de ocorrência simultânea de duas ou mais micotoxinas, pode acarretar na potencialização de seus efeitos tóxicos sobre o organismo susceptível e, dependendo do grau de intoxicação, os sintomas desencadeados são representados principalmente pela baixa produtividade animal, maior susceptibilidade a doenças, o que gera grandes prejuízos econômicos. Esta revisão apresenta aspectos gerais relacionados à toxicologia, como mecanismos de ação e efeitos em suínos, além de dados de ocorrência de aflatoxinas, fumonisinas e zearalenona no milho brasileiro.(AU)
Several mycotoxins widely distributed in nature have accentuated toxic properties and can contaminate corn crops for human and animal consumption. Aflatoxin and fusariotoxins (mainly zearalenone and fumonisin) are the most common causes of losses in pig farming. The production of fusariotoxins mainly occur before harvest, but just as aflatoxin, can also occur post-harvest if not treated and dried properly. The effects related to the ingestion of these compounds are dependent on factors related to the toxin (chemical structure, dose) and animal (species, breed, sex, age, nutritional conditions). Moreover, the possibility of simultaneous occurrence of two or more mycotoxins, may result in potentiation of their toxic effects on the organism susceptible and depending on the degree of intoxication, the triggered symptoms are primarily represented by the low animal productivity, increased susceptibility to diseases, which generates great economic losses. This review presents general aspects related to toxicology, as mechanisms of action and effects in pigs, as well as the occurrence of aflatoxin, fumonisin and zearalenone in Brazilian corn.(AU)
Varias micotoxinas ampliamente distribuidas en la naturaleza tienen acentuadas propiedades tóxicas y pueden contaminar los cultivos de maíz para el consumo humano y animal. La aflatoxina y las fusariotoxinas (principalmente zearalenona y fumonisinas) son las que más comúnmente causan pérdidas en la cría de cerdos. La producción de fusariotoxinas ocurre principalmente antes de la cosecha, pero igual de aflatoxina, también puede ocurrir después de la cosecha si no se trata y se seca correctamente. Los efectos relacionados con la ingestión de estos compuestos dependen de factores relacionados con la toxina (estructura química, la dosis) y animal (especie, raza, sexo, edad, estado nutricional). Sin embargo, la posibilidad de ocurrencia simultánea de dos o más micotoxinas, puede dar lugar a la potenciación de sus efectos tóxicos sobre el organismo susceptible y, dependiendo del grado de intoxicación, los síntomas provocados están representados principalmente por la baja productividad de los animales, aumento de la susceptibilidad a las enfermedades, lo que genera grandes pérdidas económicas. Esta revisión presenta los aspectos generales relacionados con la toxicología, como mecanismos de acción y efectos en los cerdos, así como dados de ocurrencia de aflatoxina, zearalenona y fumonisinas en el maíz brasileño.(AU)
Assuntos
Animais , Micotoxicose/veterinária , Suínos , Aflatoxinas/toxicidade , Toxina T-2/toxicidade , Zearalenona/toxicidadeRESUMO
A trial was conducted to evaluate a feed additive containing epoxidase activity from a bacterium (Mycofix-S) as a potential protection against the adverse effects of 2.5 ppm dietary T-2 toxin in male growing broiler chickens. A total of 144 one-day-old Ross 308 male chicks were individually wing-banded and allotted into each of the four experimental groups. Group 1: negative control, no T-2 toxin or additive; group 2: Mycofix-S, 2.5 g/kg; group 3: positive control, 2.5 ppm T-2 toxin; group 4: 2.5 ppm T-2 toxin + 2.5 g/kg Mycofix-S. Feed and water were provided ad libitum for 28 days (days 1 to 28 of age). Each experimental treatment was replicated 6 times, with 6 birds per replicate pen. Response variables included performance parameters, serum activity of alkaline phosphatase (ALP) and amylase, relative weight of selected organs and histology of the upper digestive system. T-2 toxin at 2.5 ppm significantly (P = 0.016) decreased the 28-day body weight gain and cumulative feed intake without affecting feed conversion. The feed additive counteracted these adverse effects. Serum enzyme activities were not significantly (P>0.05) affected for the four experimental groups but when data from the groups receiving T-2 toxin was pooled and compared against the pooled data from groups without the toxin a significant decrease in amylase activity was observed in chickens receiving T-2 toxin. The histological examination of the upper digestive system revealed lesions in mouth, esophagus, proventriculus, gizzard and duodenum in the chickens fed T-2 toxin without the additive. Chickens fed T-2 toxin plus the additive showed lesions in the same tissues except in the duodenum. The results of the present study show that the addition of 2.5 g/kg of the feed additive tested protects against adverse effects on performance and also the integrity of the duodenal mucosa.(AU)
Foi realizado um experimento com o objetivo de avaliar um aditivo alimentar contendo atividade de epoxidase de uma bactéria (Mycofix-S) como proteção potencial contra os efeitos adversos de uma dieta com 2,5ppm de toxina T-2 em frangos de corte machos. Um total de 144 pintos machos Ross 308 de um dia de idade foram marcados na asa individualmente e alocados em um de quatro grupos experimentais: grupo 1: controle negativo, sem toxina T-2 ou aditivo; grupo 2: 2,5g/kg de Mycofix-S; grupo 3: controle positivo, 2,5ppm de toxina T-2; grupo 4: 2,5ppm de toxina T-2 + 2,5g/kg de Mycofix-S. Alimento e água foram fornecidos ad libitum por 28 dias (dias um a 28 de idade). Cada tratamento experimental foi replicado seis vezes, com seis pintos por gaiola de replicação. As variáveis de resposta incluíram parâmetros de desempenho, atividade sérica de fosfatase alcalina (ALP) e amilase, peso relativo de órgãos selecionados e histologia do sistema digestivo superior. A toxina T-2 a 2,5ppm diminuiu significativamente (P = 0.016) o ganho de peso corporal aos 28 dias e o consumo de alimento acumulado, sem afetar a conversão alimentar. O aditivo diminuiu os efeitos adversos. As atividades séricas das enzimas não foram afetadas significativamente (P>0.05) nos quatro grupos experimentais, porém, quando os dados dos grupos que receberam a toxina T-2 foram combinados e comparados com o pool de dados dos grupos sem toxina, foi observado um decréscimo significativo da atividade de amilase nos frangos que receberam a toxina T-2. O exame histológico do sistema digestivo superior revelou lesões em boca, esôfago, pró-ventrículo, moela e duodeno nos frangos alimentados com toxina T-2 sem aditivo. Frangos alimentados com toxina T-2 mais aditivo mostraram lesões nos mesmos tecidos, exceto no duodeno. Os resultados do presente estudo mostram que a adição de 2,5g/kg do aditivo alimentar testado protege contra os efeitos adversos sobre o desempenho e a integridade da mucosa duodenal.(AU)
Assuntos
Animais , Amilases , Galinhas , Sistema Digestório , Aditivos Alimentares , Tricotecenos , Dieta/veterinária , Duodeno , Micotoxinas , Toxina T-2RESUMO
A trial was conducted to evaluate a feed additive containing epoxidase activity from a bacterium (Mycofix-S) as a potential protection against the adverse effects of 2.5 ppm dietary T-2 toxin in male growing broiler chickens. A total of 144 one-day-old Ross 308 male chicks were individually wing-banded and allotted into each of the four experimental groups. Group 1: negative control, no T-2 toxin or additive; group 2: Mycofix-S, 2.5 g/kg; group 3: positive control, 2.5 ppm T-2 toxin; group 4: 2.5 ppm T-2 toxin + 2.5 g/kg Mycofix-S. Feed and water were provided ad libitum for 28 days (days 1 to 28 of age). Each experimental treatment was replicated 6 times, with 6 birds per replicate pen. Response variables included performance parameters, serum activity of alkaline phosphatase (ALP) and amylase, relative weight of selected organs and histology of the upper digestive system. T-2 toxin at 2.5 ppm significantly (P = 0.016) decreased the 28-day body weight gain and cumulative feed intake without affecting feed conversion. The feed additive counteracted these adverse effects. Serum enzyme activities were not significantly (P>0.05) affected for the four experimental groups but when data from the groups receiving T-2 toxin was pooled and compared against the pooled data from groups without the toxin a significant decrease in amylase activity was observed in chickens receiving T-2 toxin. The histological examination of the upper digestive system revealed lesions in mouth, esophagus, proventriculus, gizzard and duodenum in the chickens fed T-2 toxin without the additive. Chickens fed T-2 toxin plus the additive showed lesions in the same tissues except in the duodenum. The results of the present study show that the addition of 2.5 g/kg of the feed additive tested protects against adverse effects on performance and also the integrity of the duodenal mucosa.(AU)
Foi realizado um experimento com o objetivo de avaliar um aditivo alimentar contendo atividade de epoxidase de uma bactéria (Mycofix-S) como proteção potencial contra os efeitos adversos de uma dieta com 2,5ppm de toxina T-2 em frangos de corte machos. Um total de 144 pintos machos Ross 308 de um dia de idade foram marcados na asa individualmente e alocados em um de quatro grupos experimentais: grupo 1: controle negativo, sem toxina T-2 ou aditivo; grupo 2: 2,5g/kg de Mycofix-S; grupo 3: controle positivo, 2,5ppm de toxina T-2; grupo 4: 2,5ppm de toxina T-2 + 2,5g/kg de Mycofix-S. Alimento e água foram fornecidos ad libitum por 28 dias (dias um a 28 de idade). Cada tratamento experimental foi replicado seis vezes, com seis pintos por gaiola de replicação. As variáveis de resposta incluíram parâmetros de desempenho, atividade sérica de fosfatase alcalina (ALP) e amilase, peso relativo de órgãos selecionados e histologia do sistema digestivo superior. A toxina T-2 a 2,5ppm diminuiu significativamente (P = 0.016) o ganho de peso corporal aos 28 dias e o consumo de alimento acumulado, sem afetar a conversão alimentar. O aditivo diminuiu os efeitos adversos. As atividades séricas das enzimas não foram afetadas significativamente (P>0.05) nos quatro grupos experimentais, porém, quando os dados dos grupos que receberam a toxina T-2 foram combinados e comparados com o pool de dados dos grupos sem toxina, foi observado um decréscimo significativo da atividade de amilase nos frangos que receberam a toxina T-2. O exame histológico do sistema digestivo superior revelou lesões em boca, esôfago, pró-ventrículo, moela e duodeno nos frangos alimentados com toxina T-2 sem aditivo. Frangos alimentados com toxina T-2 mais aditivo mostraram lesões nos mesmos tecidos, exceto no duodeno. Os resultados do presente estudo mostram que a adição de 2,5g/kg do aditivo alimentar testado protege contra os efeitos adversos sobre o desempenho e a integridade da mucosa duodenal.(AU)
Assuntos
Animais , Galinhas , Aditivos Alimentares , Amilases , Tricotecenos , Histologia , Sistema Digestório , Dieta/veterinária , Micotoxinas , Toxina T-2 , DuodenoRESUMO
Moulds are capable of reducing the nutritional value of feedstuff as well as elaborating several mycotoxins. Mycotoxin-contaminated feed has adverse effects on animal health and productivity. Also, mycotoxins may be carried over into meat and eggs when poultry are fed with contaminated feed. In a point prevalence study feedstuff used for poultry nutrition in Argentina was analyzed for fungal flora, natural incidence of selected mycotoxins, and nutritional quality. Ten mould genera were recovered, six of them known to be mycotoxigenic. More than 28 species were determined. Fumonisins were detected in all the samples (median 1,750 ppb). Forty-four out of 49 samples (90%) were contaminated with DON (median 222 ppb) and OTA (median 5 ppb). Also, 44 out of 49 samples were contaminated with aflatoxins (median 2.685 ppb), 42 samples (86%) with ZEA (median 50 ppb), and 38 samples (78%) with T2-toxin (median 50 ppb). Ninety percent of the samples had at least one type of nutritional deficiency. This study indicates the need for continuous assessment of the mycological status of animal feed production, in order to feed animals for optimal performance ensuring food safety.
Assuntos
Ração Animal/microbiologia , Ração Animal/normas , Contaminação de Alimentos/análise , Fungos/química , Micotoxinas/análise , Valor Nutritivo , Aflatoxinas/análise , Ração Animal/análise , Animais , Argentina , Fumonisinas/análise , Aves Domésticas , Especificidade da Espécie , Estatísticas não Paramétricas , Toxina T-2/análiseRESUMO
A total of 120 pelleted poultry feed samples from Entre Ríos Province, Argentina, were evaluated. The aims were to investigate (1) the presence of relevant toxigenic fungi, as well as to determine the ability to produce aflatoxins (AFs) by Aspergillus section Flavi isolated strains; and (2) the natural co-occurrence of AFs, fumonisins (FBs), gliotoxin, diacetoxyscirpenol (DAS), HT-2 and T-2 toxin by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Total fungal counts were below the established value (1 × 104 CFU g⻹). Aspergillus flavus and A. parasiticus were the only aflatoxigenic species isolated. Co-occurrence of fumonisin B1 (FB1), HT-2 and T-2 toxin was detected in 100% of the feeds, with mean levels from 4502 to 5813; 6.7 to 21.6 and 19.6 to 30.3 µg kg⻹, respectively. A large number of starter samples were co-contaminated with aflatoxin B1 (AFB1), FB1, HT-2 and T-2 toxins. Gliotoxin and DAS were not found in this survey.