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1.
Int J Biol Macromol ; 192: 369-378, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34634329

RESUMO

Trichothecene toxins cause serious hazard towards human health and economical crops. However, there are no sufficient molecular strategies to reduce the hazard of trichothecene toxins. Thus it is urgent to exploit novel approaches to control the hazard of trichothecenes. In this study, four trichothecene toxin-resistance genes including mfs1, GNAT1, TRP1 and tri12 in Paramyrothecium roridum were excavated based on genome sequencing results, and then expressed in toxin-sensitive Saccharomyces cerevisiae BJ5464, the toxin resistance genes pdr5, pdr10 and pdr15 of which were firstly knocked out simultaneously by the introduction of TAA stop codon employing CRISPR/Cas9 system. Therefore, three novel hazardous toxin-resistance genes mfs1, GNAT1, TRP1 in P. roridum were firstly excavated by the co-incubation of DON toxin and toxin resistant genes-containing BJ5464 strains. The in vitro function and properties of novel toxin-resistance genes coding proteins including GNAT1, MFS1 and TRP1 were identified by heterologous expression and cellular location analysis as well as in vitro biochemical reaction. The excavation of novel trichothecene toxin-resistance genes provide novel molecular clues for controlling the harm of trichothecenes, meanwhile, this study will also pave a new way for the yield improvement of trichothecenes by heterologous expression to facilitate the development of trichothecenes as anti-tumor lead compounds.


Assuntos
Antibiose , Proteínas Fúngicas/metabolismo , Hypocreales/metabolismo , Toxinas Biológicas/antagonistas & inibidores , Tricotecenos/antagonistas & inibidores , Antibiose/genética , Proteínas Fúngicas/genética , Expressão Gênica , Loci Gênicos , Hypocreales/genética , Proteínas Recombinantes de Fusão , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Tricotecenos/metabolismo
2.
Res Vet Sci ; 136: 66-73, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33588096

RESUMO

The intestinal tract is a target for the deoxynivalenol (DON), which has adverse effects in animals and humans' health by affecting intestinal functions. Phenethyl isothiocyanate (PEITC) is an important degradation product of glucosinolates (GSLs), belonging to an anti-nutritional factor that affects the digestion and absorption of nutrients in the animals' intestinal. However, little attention has been paid to the interaction and its mechanism between DON and PEITC. Therefore, the purpose of this study was to assess the effects of PEITC on DON-induced cytotoxicity and inflammation, and explore the potential mechanisms in IPEC-J2 cells. Our results showed that DON exposure could decrease the cell viability and pro-inflammatory cytokine expression in IPEC-J2 cells in a dose-dependent manner. PEITC treatment at the concentrations of 1.25-5 µM had no significant effect on IPEC-J2 cells viability, but above 10 µM of PEITC treatment significantly reduced the cell viability. Interestingly, 1.25-5 µM of PEITC treatment could suppress 4 µM of DON-induced decrease in cell viability and increase in pro-inflammatory cytokine expression. Meanwhile, the protein ratios of p-p65/p-65 and p-IκBα/IκBα were markedly decreased in the groups treated with 1.25-5 µM PEITC compared to DON exposure alone. However, the protective effects of PEITC treatment were significantly blocked after pre-treatment with LPS, NF-κB activator, in IPEC-J2 cells. In conclusion, these findings indicated that the nontoxic dose of PEITC could alleviate DON-induced cytotoxicity and inflammation responses via suppressing the NF-κB signaling pathway in IPEC-J2 cells. Our results provide a new theoretical basis for the rational addition of rapeseed meal in animal feedstuff.


Assuntos
Mucosa Intestinal/efeitos dos fármacos , Isotiocianatos/farmacologia , Tricotecenos/antagonistas & inibidores , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Inflamação/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Suínos , Doenças dos Suínos/prevenção & controle , Tricotecenos/toxicidade
3.
Toxins (Basel) ; 12(5)2020 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-32397551

RESUMO

The efficacy of yeast-based mycotoxin detoxifiers on health and growth performance of newly-weaned pigs (27-d-old) fed diets naturally contaminated with deoxynivalenol was investigated. Sixty pigs were individually assigned to five treatments for 34 d: NC (negative control, 1.2 mg/kg of deoxynivalenol); PC (positive control, 3.2 mg/kg of deoxynivalenol); CYC (PC + clay/yeast culture-based product, 0.2%); CYE (PC + clay/yeast cell wall/plant extracts/antioxidants-based product, 0.2%); and CYB (PC + clay/inactivated yeast/botanicals/antioxidants-based product, 0.2%). Blood and jejunal mucosa were sampled, and data were analyzed using Proc Mixed of SAS with pre-planned contrasts. Deoxynivalenol reduced the average daily gain (ADG) in phase 3. Pigs fed CYC had greater overall ADG, average daily feed intake during phase 3, and gain to feed ratio during phase 2 than PC. At d 14, deoxynivalenol reduced blood urea nitrogen/creatinine and tended to reduce blood urea nitrogen. Pigs fed CYB tended to have greater aspartate aminotransferase than PC. At d 34, pigs fed CYC and CYB tended to have lower serum creatine phosphokinase than PC. Pigs fed CYE had lower blood urea nitrogen/creatinine than PC. In jejunal mucosa, deoxynivalenol tended to increase malondialdehydes and decrease glutathione. Pigs fed CYE and CYB had lower malondialdehydes, pigs fed CYB had greater glutathione and tended to have lower immunoglobulin A than PC. Pigs fed CYC and CYE tended to have lower interleukin 8 than PC. In summary, deoxynivalenol challenge (1.2 vs. 3.2 mg/kg) mildly compromised growth performance and increased the oxidative stress of pigs. Mycotoxin detoxifiers could partially overcome deoxynivalenol toxicity enhancing liver health, whereas CYE and CYB reduced oxidative stress, and CYC and CYB reduced immune activation. In conclusion, yeast-based detoxifiers with functional components as clay/inactivated yeast/botanicals/antioxidants had increased detoxifying properties in newly-weaned pigs challenged with deoxynivalenol, potentially by enhancing adsorbability, immune function, gut health, and reducing oxidative stress.


Assuntos
Ração Animal/microbiologia , Antitoxinas/administração & dosagem , Suplementos Nutricionais , Microbiologia de Alimentos , Fungos/metabolismo , Micotoxinas/antagonistas & inibidores , Tricotecenos/antagonistas & inibidores , Animais , Antioxidantes/administração & dosagem , Argila , Feminino , Jejuno/efeitos dos fármacos , Jejuno/metabolismo , Masculino , Micotoxinas/administração & dosagem , Micotoxinas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Sus scrofa , Tricotecenos/administração & dosagem , Tricotecenos/toxicidade , Desmame , Aumento de Peso/efeitos dos fármacos , Fermento Seco/administração & dosagem
4.
Mycotoxin Res ; 35(2): 187-196, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30710317

RESUMO

Regardless of the efforts put into preventing or reducing fungal growth, extensive mycotoxin contamination has been reported in animal feeds. In the case of pigs, one of the mycotoxins of major concern is deoxynivalenol (DON). The use of adsorbents as feed additives represents one of the strategies to control mycotoxins' contamination in feedstuff. Therefore, the aim of the study was to verify the ability of chlorophyllin (CHL) to reduce the absorption rate of DON in swine mucosa explants. Intestine was obtained from routinely slaughtered adult pigs. The mucosa explants were studied by means of Ussing chamber technique. The effect of DON (10 and 30 µg/ml) on mucosa viability and permeability and CHL (100 µg/ml) impact on DON (30 µg/ml) absorption was verified. The results revealed that mucosa explants isolated from adult animals remained unaffected for 90 min in the presence of DON in the lower concentration (10 µg/ml). Mycotoxin in the higher dose (30 µg/ml) increased mucosa permeability (decreased transepithelial electrical resistance value) and enhanced paracellular transport of lucifer yellow and mannitol but did not affect lactate dehydrogenase leakage. The introduction of CHL neither diminished the absorption rate of DON across swine mucosa explants nor prevented the toxic effects of DON on intestine. In conclusion, the results confirm the negative effect of DON on pig jejunum mucosa. However, the toxic effect of DON was observed only when it was used in relatively high doses. A promising adsorbent agent, CHL, failed to reduce the intensity of DON transport across intestine under in vitro conditions.


Assuntos
Clorofilídeos/metabolismo , Mucosa Intestinal/metabolismo , Jejuno/metabolismo , Micotoxinas/antagonistas & inibidores , Micotoxinas/toxicidade , Tricotecenos/antagonistas & inibidores , Tricotecenos/toxicidade , Animais , Absorção Intestinal/efeitos dos fármacos , Modelos Biológicos , Técnicas de Cultura de Órgãos , Permeabilidade/efeitos dos fármacos , Suínos
5.
Poult Sci ; 96(12): 4236-4244, 2017 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-29053812

RESUMO

Corn with naturally occurring aflatoxin (AF), wheat with naturally occurring doxynivalenol (DON), and barley with naturally occurring zearalenone (ZEA) were used to make rations for feeding turkey hen poults to 6 weeks of age. Control rations with equal amounts of corn, wheat, and barley were also fed. The control rations did contain some DON while both sets of rations contained ZEA. Within each grain source, there were 4 treatments: the control ration plus 3 rations each with a different feed additive which were evaluated for the potential to lessen potential mycotoxin effects on bird performance and physiology. The additives were Biomin BioFix (2 lb/ton), Kemin Kallsil (4 lb/ton), and Nutriad UNIKE (3 lb/ton). The mycotoxin rations reduced poult body weight (2.31 vs. 2.08 ± 0.02 kg) and increased (worsened) poult feed conversion (1.47 vs. 1.51 ± 0.01) at 6 wk. Feeding the poults the mycotoxin feed also resulted in organ and physiological changes typical of feeding dietary aflatoxin although a combined effect of AF, DON, and ZEA which cannot be dismissed. The feed additives resulted in improved feed conversion to 6 wk in both grain treatment groups. The observed physiological effect of feeding the additives was to reduce relative gizzard weight for both groups and to lessen the increase in relative kidney weight for the birds fed the mycotoxin feed. In conclusion, the feed additives used in this study did alleviate the effect of dietary mycotoxins to some degree, especially with respect to feed conversion. Further studies of longer duration are warranted.


Assuntos
Ração Animal/análise , Dieta/veterinária , Micotoxinas/antagonistas & inibidores , Substâncias Protetoras/farmacologia , Perus/metabolismo , Aflatoxinas/antagonistas & inibidores , Animais , Suplementos Nutricionais/análise , Feminino , Distribuição Aleatória , Tricotecenos/antagonistas & inibidores , Perus/crescimento & desenvolvimento , Zearalenona/antagonistas & inibidores
6.
Pestic Biochem Physiol ; 138: 1-7, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28456298

RESUMO

Trichothecene mycotoxins often accumulate in apparently normal grains of cereal crops. In an effort to develop an agricultural chemical to reduce trichothecene contamination, we screened trichothecene production inhibitors from the compounds on the chemical arrays. By using the trichodiene (TDN) synthase tagged with hexahistidine (rTRI5) as a target protein, 32 hit compounds were obtained from chemical library of the RIKEN Natural Product Depository (NPDepo) by chemical array screening. At 10µgmL-1, none of the 32 chemicals inhibited trichothecene production by Fusarium graminearum in liquid culture. Against the purified rTRI5 enzyme, however, NPD10133 [progesterone 3-(O-carboxymethyl)oxime amide-bonded to phenylalanine] showed weak inhibitory activity at 10µgmL-1 (18.7µM). For the screening of chemicals inhibiting trichothecene accumulation in liquid culture, 20 analogs of NPD10133 selected from the NPDepo chemical library were assayed. At 10µM, only NPD352 [testosterone 3-(O-carboxymethyl)oxime amide-bonded to phenylalanine methyl ester] inhibited rTRI5 activity and trichothecene production. Kinetic analysis suggested that the enzyme inhibition was of a mixed-type. The identification of NPD352 as a TDN synthase inhibitor lays the foundation for the development of a more potent inhibitor via systematic introduction of wide structural diversity on the gonane skeleton and amino acid residues.


Assuntos
Carbono-Carbono Liases/antagonistas & inibidores , Fungicidas Industriais/química , Fungicidas Industriais/farmacologia , Fusarium/metabolismo , Tricotecenos/antagonistas & inibidores , Inibidores Enzimáticos , Bibliotecas de Moléculas Pequenas
7.
Arch Toxicol ; 91(1): 495-507, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26979077

RESUMO

Food contamination by the trichothecene mycotoxin deoxynivalenol (DON, vomitoxin) has the potential to adversely affect animal and human health by suppressing food intake and impairing growth. In mice, the DON-induced anorectic response results from aberrant satiety hormone secretion by enteroendocrine cells (EECs) of the gastrointestinal tract. Recent in vitro studies in the murine STC-1 EEC model have linked DON-induced satiety hormone secretion to activation of calcium-sensing receptor (CaSR), a G-coupled protein receptor, and transient receptor potential ankyrin-1 (TRPA1), a TRP channel. However, it is unknown whether similar mechanisms mediate DON's anorectic effects in vivo. Here, we tested the hypothesis that DON-induced food refusal and satiety hormone release in the mouse are linked to activation of CaSR and TRPA1. Oral treatment with selective agonists for CaSR (R-568) or TRPA1 (allyl isothiocyanate (AITC)) suppressed food intake in mice, and the agonist's effects were suppressed by pretreatment with corresponding antagonists NPS-2143 or ruthenium red (RR), respectively. Importantly, NPS-2143 or RR inhibited both DON-induced food refusal and plasma elevations of the satiety hormones cholecystokinin (CCK) and peptide YY3-36 (PYY3-36); cotreatment with both antagonists additively suppressed both anorectic and hormone responses to DON. Taken together, these in vivo data along with prior in vitro findings support the contention that activation of CaSR and TRPA1 contributes to DON-induced food refusal by mediating satiety hormone exocytosis from EEC.


Assuntos
Anorexia/induzido quimicamente , Depressores do Apetite/toxicidade , Poluentes Ambientais/toxicidade , Modelos Biológicos , Receptores Acoplados a Proteínas G/agonistas , Canais de Potencial de Receptor Transitório/agonistas , Tricotecenos/toxicidade , Animais , Anorexia/metabolismo , Anorexia/prevenção & controle , Depressores do Apetite/química , Estimulantes do Apetite/uso terapêutico , Comportamento Animal/efeitos dos fármacos , Colecistocinina/agonistas , Colecistocinina/antagonistas & inibidores , Colecistocinina/sangue , Quimioterapia Combinada , Ingestão de Energia/efeitos dos fármacos , Poluentes Ambientais/antagonistas & inibidores , Feminino , Fragmentos de Peptídeos/agonistas , Fragmentos de Peptídeos/antagonistas & inibidores , Fragmentos de Peptídeos/sangue , Peptídeo YY/agonistas , Peptídeo YY/antagonistas & inibidores , Peptídeo YY/sangue , Distribuição Aleatória , Receptores de Detecção de Cálcio , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/metabolismo , Resposta de Saciedade/efeitos dos fármacos , Canal de Cátion TRPA1 , Canais de Potencial de Receptor Transitório/antagonistas & inibidores , Canais de Potencial de Receptor Transitório/metabolismo , Tricotecenos/antagonistas & inibidores
8.
Appl Biochem Biotechnol ; 182(1): 411-427, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27878744

RESUMO

The aims of our study were to evaluate the effects of Saccharomyces boulardii (S. boulardii) on deoxynivalenol (DON)-induced injury in porcine alveolar macrophage cells (PAMCs) and to explore the underlying mechanisms. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometric analysis, ELISA, qRT-PCR, and western blot were performed to assess whether S. boulardii could prevent DON-induced injury by p38 mitogen-activated protein kinase (p38 MAPK) signal pathway. The results showed that pretreatment with 8 µM DON could decrease the viability of PAMC and significantly increase the apoptosis rate of PAMC, whereas S. boulardii could rescue apoptotic PAMC cells induced by DON. Further experiments revealed that S. boulardii effectively reversed DON-induced cytotoxicity via downregulating the expression of TNF-α, IL-6, and IL-lß. In addition, S. boulardii significantly alleviated DON-induced phosphorylation and mRNA expression of p38 and further increased the expression of apoptosis regulation genes Bcl-xl and Bcl-2 and inhibited the activation of Bax. Our results suggest that S. boulardii could suppress DON-induced p38 MAPK pathway activation and reduce the expression of downstream inflammatory cytokines, as well as promote the expression of anti-apoptotic genes to inhibit apoptosis induced by DON in PAMC.


Assuntos
Macrófagos Alveolares/efeitos dos fármacos , Fatores de Proteção , Saccharomyces boulardii/metabolismo , Transdução de Sinais , Tricotecenos/toxicidade , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Regulação da Expressão Gênica , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Macrófagos Alveolares/citologia , Macrófagos Alveolares/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Saccharomyces boulardii/crescimento & desenvolvimento , Suínos , Tricotecenos/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
Toxicology ; 341-343: 17-27, 2016 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-26783879

RESUMO

The worldwide contamination of grains designated to human and animal feeding with Fusarium mycotoxins is a significant problem. Among Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZEA) are the most prevalent mycotoxins found in cereals. Co-occurrence of DON and ZEA is also very frequent and indicates that these mycotoxins might be involved in a wide range of synergistic or additive interactions. Both mycotoxins have been linked to various male reproduction problems including downregulation of steroidogenesis. In this study, the impact of DON and ZEA alone or in combination on the viability and steroid production of Leydig cell line MA-10 was determined. The ability of vitamin E, sesamin and their combination to prevent oxidative stress and restore progesterone secretion in DON- and ZEA-exposed cells was also determined. Results showed that MA-10 cells were more sensitive to the effect of DON compared to ZEA. DON and ZEA also significantly reduced MA-10 progesterone secretion after forskolin activation but no significant interactions between DON and ZEA were detected. Preventive treatment with the combination of vitamin E and sesamin significantly reduced ROS production and increased cell survival after exposition to DON and ZEA. However this treatment failed to restore normal progesterone secretion. In conclusion, both DON and ZEA are deleterious to steroidogenesis in Leydig cells. Prevention of oxidative stress caused by DON and ZEA was effective to restore cell viability but failed to restore other functions of Leydig cells suggesting that ROS production is not the main cause of steroidogenic failure in DON and ZEA treated MA-10 cells.


Assuntos
Células Intersticiais do Testículo/efeitos dos fármacos , Micotoxinas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Tricotecenos/toxicidade , Zearalenona/toxicidade , Animais , Antioxidantes/farmacologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Colforsina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos , Micotoxinas/antagonistas & inibidores , Progesterona/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Esteroides/biossíntese , Tricotecenos/antagonistas & inibidores , Zearalenona/antagonistas & inibidores
11.
Biosci Biotechnol Biochem ; 80(1): 43-54, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26376763

RESUMO

Mycotoxin contamination of crops is a serious problem throughout the world because of its impact on human and animal health as well as economy. Inhibitors of mycotoxin production are useful not only for developing effective methods to prevent mycotoxin contamination, but also for investigating the molecular mechanisms of secondary metabolite production by fungi. We have been searching for mycotoxin production inhibitors among natural products and investigating their modes of action. In this article, we review aflatoxin and trichothecene production inhibitors, including our works on blasticidin S, methyl syringate, cyclo(L-Ala-L-Pro), respiration inhibitors, and precocene II.


Assuntos
Aflatoxinas/antagonistas & inibidores , Aspergillus/efeitos dos fármacos , Contaminação de Alimentos/prevenção & controle , Fungicidas Industriais/farmacologia , Fusarium/efeitos dos fármacos , Tricotecenos/antagonistas & inibidores , Aflatoxinas/biossíntese , Aspergillus/patogenicidade , Aspergillus/fisiologia , Benzopiranos/farmacologia , Produtos Agrícolas/efeitos dos fármacos , Produtos Agrícolas/microbiologia , Fusarium/patogenicidade , Fusarium/fisiologia , Ácido Gálico/análogos & derivados , Ácido Gálico/farmacologia , Humanos , Nucleosídeos/farmacologia , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Peptídeos Cíclicos/farmacologia , Doenças das Plantas/microbiologia , Relação Estrutura-Atividade , Tricotecenos/biossíntese
12.
PLoS One ; 10(8): e0135031, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26248339

RESUMO

Precocene II, a constituent of essential oils, shows antijuvenile hormone activity in insects and inhibits trichothecene production in fungi. We investigated the molecular mechanism by which precocene II inhibits trichothecene production in Fusarium graminearum, the main causal agent of Fusarium head blight and trichothecene contamination in grains. Voltage-dependent anion channel (VDAC), a mitochondrial outer membrane protein, was identified as the precocene II-binding protein by an affinity magnetic bead method. Precocene II increased the superoxide level in mitochondria as well as the amount of oxidized mitochondrial proteins. Ascorbic acid, glutathione, and α-tocopherol promoted trichothecene production by the fungus. These antioxidants compensated for the inhibitory activity of precocene II on trichothecene production. These results suggest that the binding of precocene II to VDAC may cause high superoxide levels in mitochondria, which leads to stopping of trichothecene production.


Assuntos
Antifúngicos/farmacologia , Benzopiranos/farmacologia , Proteínas Fúngicas/metabolismo , Fusarium/efeitos dos fármacos , Proteínas Mitocondriais/metabolismo , Tricotecenos/antagonistas & inibidores , Canais de Ânion Dependentes de Voltagem/metabolismo , Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Benzopiranos/antagonistas & inibidores , Proteínas Fúngicas/genética , Fusarium/genética , Fusarium/metabolismo , Regulação Fúngica da Expressão Gênica , Glutationa/farmacologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Ligação Proteica , Superóxidos/agonistas , Superóxidos/metabolismo , Tricotecenos/biossíntese , Canais de Ânion Dependentes de Voltagem/genética , alfa-Tocoferol/farmacologia
13.
Food Chem Toxicol ; 83: 174-82, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26115597

RESUMO

This study was conducted to prepare and characterize activated carbon (AC) and to evaluate its protective effect against deoxynivalenol (DON) toxicity in rats compared to Egyptian montmorillonite (EM). AC was prepared using a single-step chemical activation with phosphoric acid (H3PO4). The resulted AC has a high surface area and a high total pore volume. Male Sprague-Dawley rats were divided into 6 groups (n = 10) and treated for 3 weeks as follow: the control group, the groups fed AC or EM-supplemented diet (0.5% w/w), the group treated orally with DON (5 mg/kg b.w.) and the groups fed AC or EM-supplemented diet and treated with DON. Blood and liver samples were collected for different analyses. Treatment with DON increased liver function enzymes, lipid peroxidation, tumor necrosis factor α, DNA fragmentation, decreased hepatic glutathione content, up regulating mRNA Fas and TNF-α genes expression and increased micronucleated polychromatic erythrocytes and normochromatic erythrocytes in bone marrow. Co-treatment of DON plus AC or EM succeeded to normalize the levels of the biochemical parameters, reduced the cytotoxicity of bone marrow and ameliorated the hepatic genotoxicity. Moreover, AC was more effective than EM and has a high affinity to adsorb DON and to reduce its cytotoxicity and genotoxicity.


Assuntos
Carbono/uso terapêutico , Carcinógenos Ambientais/toxicidade , Quelantes/uso terapêutico , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Fígado/efeitos dos fármacos , Mutagênicos/toxicidade , Tricotecenos/antagonistas & inibidores , Animais , Bentonita/uso terapêutico , Carbono/química , Carbono/economia , Carcinógenos Ambientais/química , Quelantes/química , Quelantes/economia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Egito , Indústria de Processamento de Alimentos/economia , Indicadores e Reagentes/química , Resíduos Industriais/análise , Resíduos Industriais/economia , Fígado/metabolismo , Masculino , Mutagênicos/química , Phoeniceae/química , Ácidos Fosfóricos/química , Porosidade/efeitos dos fármacos , Ratos Sprague-Dawley , Sementes/química , Propriedades de Superfície/efeitos dos fármacos , Tricotecenos/toxicidade
14.
Artigo em Inglês | MEDLINE | ID: mdl-25230728

RESUMO

An immunogen synthesis strategy was designed to develop anti-deoxynivalenol (DON) monoclonal antibodies with low cross-reactivity against structurally similar trichothecenes. A total of eight different DON immunogens were synthesised, differing in the type and position of the linker on the DON molecule. After immunisation, antisera from mice immunised with different DON immunogens were checked for the presence of relevant antibodies. Then, both homologous and heterologous enzyme-linked immunosorbent assays (ELISAs) were performed for hybridoma screening. Finally, three monoclonal antibodies against DON and its analogues were generated. In addition, monoclonal antibody 13H1 could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyldeoxynivalenol (15-ADON) > DON, with IC50 ranging from 1.14 to 2.13 µg ml⁻¹. Another monoclonal antibody 10H10 manifested relatively close sensitivities to DON, 3-acetyldeoxynivalenol (3-ADON) and 15-ADON, with IC50 values of 22, 15 and 34 ng ml⁻¹, respectively. Using an indirect ELISA format decreases the 10H10 sensitivity to 15-ADON with 92%. A third monoclonal antibody 2A9 showed to be very specific and sensitive to 3-ADON, with IC50 of 0.38 ng ml⁻¹. Using both 2A9 and 10H10 monoclonal antibodies allows determining sole DON contamination.


Assuntos
Anticorpos Monoclonais/metabolismo , Modelos Imunológicos , Modelos Moleculares , Venenos/análise , Tricotecenos/análise , Vacinas Sintéticas/química , Animais , Anticorpos Monoclonais/biossíntese , Afinidade de Anticorpos , Especificidade de Anticorpos , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Feminino , Contaminação de Alimentos , Inspeção de Alimentos/métodos , Hibridomas/metabolismo , Cinética , Camundongos Endogâmicos BALB C , Estrutura Molecular , Venenos/química , Venenos/metabolismo , Tricotecenos/antagonistas & inibidores , Tricotecenos/química , Vacinas Sintéticas/metabolismo
15.
Foodborne Pathog Dis ; 11(6): 433-8, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24660841

RESUMO

The walls of yeast cells, which contain ß-D-glucan biopolymers, have an active role in reducing mycotoxins in animal feed. This study aimed to evaluate the ß-D-glucan biopolymers as a mycotoxin binder for fumonisin (FUM) and deoxynivalenol (DON) toxins as well as their effect on the nutritional value of soybean, which is considered one of the important feed row materials. The evaluation was carried out using some toxigenic Fusarium isolates (Fusarium solani, F. oxysporum, and F. verticillioides) in vitro and in vivo. The FUM and DON levels were determined by immune affinity column. The F. verticillioides was the most toxigenic, followed by F. oxysporum and lastly F. solani, while secretion of DON toxin was determined to be greater than FUM with all the tested fungi. The effectiveness of ß-D-glucan biopolymers on FUM and DON absorption was greater than clay and calcium propionate. In vivo, treating soybean seeds with ß-D-glucan biopolymers led to reduction in the level of FUM and DON toxins in seeds artificially inoculated by F. verticillioides. ß-D-glucan treatment also has a low effect on nutritional components of the seeds compared to untreated ones. In conclusion, this study found a new approach to reduce Fusarium mycotoxins in feed to an allowable safe limit and at the same time maintaining the nutritional value of these materials.


Assuntos
Ração Animal/análise , Antídotos/química , Contaminação de Alimentos/prevenção & controle , Conservantes de Alimentos/química , Glycine max/química , Micotoxinas/antagonistas & inibidores , beta-Glucanas/química , Absorção Fisico-Química , Ração Animal/efeitos adversos , Ração Animal/microbiologia , Animais , Antídotos/efeitos adversos , Parede Celular/química , Carboidratos da Dieta/análise , Gorduras na Dieta/análise , Egito , Conservantes de Alimentos/efeitos adversos , Doenças Transmitidas por Alimentos/prevenção & controle , Doenças Transmitidas por Alimentos/veterinária , Fumonisinas/análise , Fumonisinas/antagonistas & inibidores , Fumonisinas/metabolismo , Fumonisinas/toxicidade , Fusarium/crescimento & desenvolvimento , Fusarium/isolamento & purificação , Fusarium/metabolismo , Micotoxinas/análise , Micotoxinas/metabolismo , Micotoxinas/toxicidade , Venenos/análise , Venenos/metabolismo , Venenos/toxicidade , Saccharomyces cerevisiae/química , Sementes/efeitos adversos , Sementes/química , Sementes/microbiologia , Proteínas de Soja/análise , Proteínas de Soja/química , Glycine max/efeitos adversos , Glycine max/microbiologia , Especificidade da Espécie , Tricotecenos/análise , Tricotecenos/antagonistas & inibidores , Tricotecenos/metabolismo , Tricotecenos/toxicidade , beta-Glucanas/efeitos adversos
16.
Int J Environ Res Public Health ; 11(1): 1094-105, 2014 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-24441510

RESUMO

Fusarium graminearum is the main causal pathogen affecting small-grain cereals, and it produces deoxynivalenol, a kind of mycotoxin, which displays a wide range of toxic effects in human and animals. Bacterial strains isolated from peanut shells were investigated for their activities against F. graminearum by dual-culture plate and tip-culture assays. Among them, twenty strains exhibited potent inhibition to the growth of F. graminearum, and the inhibition rates ranged from 41.41% to 54.55% in dual-culture plate assay and 92.70% to 100% in tip-culture assay. Furthermore, eighteen strains reduced the production of deoxynivalenol by 16.69% to 90.30% in the wheat kernels assay. Finally, the strains with the strongest inhibitory activity were identified by morphological, physiological, biochemical methods and also 16S rDNA and gyrA gene analysis as Bacillus amyloliquefaciens. The current study highlights the potential application of antagonistic microorganisms and their metabolites in the prevention of fungal growth and mycotoxin production in wheat kernels. As a biological strategy, it might avoid safety problems and nutrition loss which always caused by physical and chemical strategies.


Assuntos
Arachis/microbiologia , Bacillus/fisiologia , Fusarium , Controle Biológico de Vetores , Triticum/microbiologia , Bacillus/isolamento & purificação , Fusarium/crescimento & desenvolvimento , Fusarium/metabolismo , Tricotecenos/antagonistas & inibidores , Tricotecenos/metabolismo
17.
Mycotoxin Res ; 29(3): 203-8, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23813191

RESUMO

As contradictory results have been reported on the immunotoxic properties of deoxynivalenol (DON) in animal studies, we introduced a lymphoblast cell culture model in order to examine the effects of DON on lymphoblastic cell growth and metabolism as well as the preventive properties of free radical scavenger molecules against the DON-induced cell damage. Murine YAC-1 lymphoma cells were used because lymphoblasts have been shown to be sensitive to DON-induced immunotoxicity. Cells were quantified and their proliferative activity was measured by a proliferation test. Lipid peroxidation and protein oxidation were determined using assays quantifying thiobarbituric acid reactive substances (TBARS) and carbonylated proteins. Severely reduced cell counts were detected in DON-treated samples, confirmed by a 5-10 times lower proliferative activity. Significant increases in lipid peroxidation and protein oxidation were found in parallel incubated samples. The pre-incubation with free radical scavengers significantly reduced DON-induced changes to proteins and lipids as well as the tarnished cell viability and cell proliferation. These results suggest that YAC-1 lymphoma cells are a suitable model to investigate and elucidate the basic molecular and cellular mechanisms for possible immunotoxic effects of DON. With regard to the impact of free radical scavengers, the applied in-vitro model might enable the investigation of potential prophylactic and therapeutic effects before or even without harmful animal experiments and cost- and time-intensive expenses.


Assuntos
Antioxidantes/metabolismo , Linfócitos/efeitos dos fármacos , Tricotecenos/antagonistas & inibidores , Tricotecenos/toxicidade , Animais , Contagem de Células , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Peroxidação de Lipídeos , Camundongos , Oxirredução , Proteínas/metabolismo
18.
J Toxicol Sci ; 38(3): 495-502, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23719927

RESUMO

The in vitro effects of 2 representative mycotoxins, T-2 toxin and deoxynivalenol (DON), of trichothecene group on the electron transport system (ETS) of mitochondria in rat cardiomyocytes were investigated by measuring oxygen consumption rates (OCR). The ATP-linked OCR and the reserve capacity (RC) of the mitochondria ETS were quantified by a "mitochondria stress test" which was estimated by the OCR responses to oligomycin and carbonyl cyanide-p-trifluoromethoxyphenylhydrazone, with an extracellular flux analyzer. The basal OCR was significantly inhibited by the application of T-2 toxin at concentrations of 6 × 10⁻¹ to 6 × 10⁻5 µM and DON at concentrations of 0.78 to 100 µM for 24 hr. The threshold of cardiomyocyte toxicity was estimated to be between 6.0 × 10⁻6 and 6.0 × 10⁻5 µM for T-2 toxicity on both ATP-linked OCR and RC and between 0.39 and 0.78 µM on ATP-linked OCR or between 1.56 and 3.13 µM on RC for DON. The decrease in OCR of cardiomyocytes exposed to T-2 toxin with a concentration of 6.0 × 10⁻³ and 6.0 × 10⁻4 µM was significantly inhibited by antioxidants, catalase and vitamin C. In conclusion, the present study demonstrated, through the direct and real-time measurement of respiratory function in mitochondria, that a marked inhibition of mitochondrial ETS function in cardiomyocytes was induced by T-2 toxin and DON and that the mitochondrial dysfunction by T-2 toxin was largely associated with oxidative stress.


Assuntos
Transporte de Elétrons/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Miócitos Cardíacos/ultraestrutura , Consumo de Oxigênio/efeitos dos fármacos , Toxina T-2/toxicidade , Tricotecenos/toxicidade , Trifosfato de Adenosina/fisiologia , Animais , Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Catalase/farmacologia , Células Cultivadas , Relação Dose-Resposta a Droga , Ratos Sprague-Dawley , Toxina T-2/antagonistas & inibidores , Tricotecenos/antagonistas & inibidores
19.
Mycotoxin Res ; 29(3): 169-75, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23536360

RESUMO

Fusarium toxin-contaminated ground maize was hydrothermally treated in the presence of different combinations of chemicals in order to simultaneously reduce zearalenone (ZEA) and deoxynivalenol (DON) concentrations. Treatments were carried out in a laboratory conditioner at 80 °C and 17 % moisture. Six different treatments were performed, consisting of 3 doses of methylamine (MMA; 2.5, 5 and 10 g/kg maize) at a constant dose of 5 g sodium metabisulfite (SBS)/kg, either with or without the addition of 20 g calcium hydroxide (Ca(OH)2)/kg. The used maize was contaminated with approximately 45.99 mg DON/kg and 3.46 mg ZEA/kg. Without the addition of Ca(OH)2, DON reductions reached approximately 82% after 1-min treatment and the toxin disappeared nearly completely after 10 min when 2.5 or 5 g MMA were applied. ZEA concentrations were only marginally affected. In the presence of Ca(OH)2, reductions in DON concentrations were lower, but were enhanced by increasing doses of MMA. ZEA concentrations were reduced by 72, 85 and 95% within the first 5 min of the treatment at MMA dosages of 2.5, 5 and 10 g/kg maize, respectively. The application of SBS in combination with a strong alkaline during hydrothermal treatment seems to be a promising approach to simultaneously decontaminate even high amounts of DON and ZEA in ground maize and may contribute to reduce the toxin load of diets.


Assuntos
Descontaminação/métodos , Contaminação de Alimentos/análise , Tricotecenos/antagonistas & inibidores , Zea mays/química , Zea mays/efeitos da radiação , Zearalenona/antagonistas & inibidores , Hidróxido de Cálcio/metabolismo , Temperatura Alta , Metilaminas/metabolismo , Sulfitos/metabolismo , Tricotecenos/análise , Zearalenona/análise
20.
Poult Sci ; 91(9): 2089-95, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22912441

RESUMO

The current experiment was conducted to determine the effect of mycotoxin-contaminated diets with aflatoxin (AFLA) and deoxynivalenol (DON) and dietary inclusion of deactivation compound on layer hen performance during a 10-wk trial. The experimental design consisted of a 4 × 2 factorial with 4 toxin levels: control, low (0.5 mg/kg AFLA + 1.0 mg/kg DON), medium (1.5 mg/kg AFLA + 1.5 mg/kg DON), and high (2.0 mg/kg AFLA + 2.0 mg/kg DON) with or without the inclusion of deactivation compound. Three hundred eighty-four 25-wk-old laying hens were randomly assigned to 1 of the 8 treatment groups. Birds were fed contaminated diets for a 6-wk phase of toxin administration followed by a 4-wk recovery phase, when all birds were fed mycotoxin-free diets. Twelve hens from each treatment were subjected to necropsy following each phase. Relative liver and kidney weights were increased (P < 0.05) at the medium and high toxin levels following the toxin phase, but the deactivation compound reduced (P < 0.05) relative liver and kidney weights following the recovery period. The high toxin level decreased (P < 0.05) feed consumption and egg production during the toxin period, whereas the deactivation compound increased (P < 0.05) egg production during the first 2 wk of the toxin phase. Egg weights were reduced (P < 0.05) in hens fed medium and high levels of toxin. An interaction existed between toxin level and deactivation compound inclusion with regard to feed conversion (g of feed/g of egg). High inclusion level of toxins increased feed conversion compared with the control diet, whereas deactivation compound inclusion reduced feed conversion to a level comparable with the control. These data indicate that deactivation compound can reduce or eliminate adverse effects of mycotoxicoses in peak-performing laying hens.


Assuntos
Aflatoxinas/toxicidade , Ração Animal/análise , Galinhas/fisiologia , Dieta/veterinária , Contaminação de Alimentos , Tricotecenos/toxicidade , Aflatoxinas/antagonistas & inibidores , Aflatoxinas/química , Animais , Suplementos Nutricionais , Esquema de Medicação , Feminino , Rim/anatomia & histologia , Rim/efeitos dos fármacos , Fígado/anatomia & histologia , Fígado/efeitos dos fármacos , Baço/anatomia & histologia , Baço/efeitos dos fármacos , Tricotecenos/antagonistas & inibidores , Tricotecenos/química
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