ovalbumin and deoxynivalenol

Deoxynivalenol (DON), a highly prevalent contaminant of grain-based products, is known to induce reproductive- and immunotoxicities. Considering the importance of immune development in early life, the present study investigated the effects of perinatal DON exposure on allergy development and vaccine responsiveness in the offspring. Pregnant mice received control or DON-contaminated diets (12.5 mg/kg diet) during pregnancy and lactation. After weaning, female offspring were sensitized to ovalbumin (OVA) by oral administration of OVA with cholera toxin (CT). Male offspring were injected with Influvac vaccine. OVA-specific acute allergic skin response (ASR) in females and vaccine-specific delayed-type hypersensitivity (DTH) in males were measured upon intradermal antigen challenge. Immune cell populations in spleen and antigen-specific plasma immunoglobulins were analyzed. In female CT+OVA-sensitized offspring of DON-exposed mothers ASR and OVA-specific plasma immunoglobulins were significantly higher, compared to the female offspring of control mothers. In vaccinated male offspring of DON-exposed mothers DTH and vaccine-specific antibody levels were significantly lower, compared to the male offspring of control mothers. In both models a significant reduction in regulatory T cells, Tbet

Topics: Animals; Antibodies, Viral; Cells, Cultured; Cholera Toxin; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunogenicity, Vaccine; Influenza Vaccines; Lactation; Male; Maternal Exposure; Mice, Inbred C3H; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Spleen; T-Lymphocytes, Regulatory; Th1 Cells; Trichothecenes; Vaccination; Vaccine Efficacy

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 mg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 mg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Topics: Animal Feed; Animals; Antibodies, Fungal; Antibodies, Monoclonal; Chemistry Techniques, Analytical; Enzyme-Linked Immunosorbent Assay; Female; Food Contamination; Fusarium; Imidazoles; Magnetics; Mice; Mice, Inbred BALB C; Mycotoxins; Nanoparticles; Ovalbumin; Trichothecenes

Deoxynivalenol (DON), one of the trichothecene mycotoxins, is a worldwide contaminant of wheat and barley, especially when infected by Fusarium graminearum, the causative agent of an epidemic wheat disease called Fusarium Head Blight. Because of the high risk of DON ingestion and the possibility of frequent exposure, it is important to develop a rapid and highly sensitive method for easy identification and quantification of DON in grain samples. In this study, we have developed an indirect competitive enzyme-linked immunosorbent assay (ELISA) to detect DON in wheat. We conjugated 3-O-Hemisuccinyl-DON (3HS-DON) to Bovine serum albumin (BSA) and Ovalbumin (OVA), and obtained DON-specific mice antisera. The indirect competitive ELISA revealed that the optimal concentration of mice serum and the coated antigen was 1/1600 and 1/1500, respectively. The antiserum cross-reacted with the trichothecenes 3-acetyl-DON and T-2 toxin, reaching about 55.2% and 6.3%, respectively, as compared with DON. Results showed that the assay could be performed satisfactorily using an extraction buffer containing less than 15% methanol. Recovery from DON was 82-93% in grains. The linear detection range of DON in grains was between 0.01 and 100 μg/mL.

Topics: Animals; Cattle; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Female; Fusarium; Hordeum; Immune Sera; Mice; Mice, Inbred BALB C; Ovalbumin; Serum Albumin, Bovine; T-2 Toxin; Trichothecenes; Triticum

Single Comb White Leghorn hens at 58 weeks of age were given control (C) and vomitoxin (V)-contaminated feed for 4 weeks; then the V treatment was changed to C for 2 subsequent weeks. Fusarium graminearum-infected corn was substituted for sound corn to attain a practical extreme of 38 ppm V. Hen-day production, feed consumption, body weight, and gross pathology were the same between treatments. Egg weight, internal quality, and shell strength were not adversely affected; however, dietary V led to a small reduction in the percentage of yolk while albumen increased. Solids content of both egg components remained unchanged, and no V as such could be detected (less than .2 ppm). Presence of toxic V metabolites in the egg were indicated by increased (although still low) embryonic mortality upon incubation. Improvement in yolk yield and relief from germ losses occurred 1 week after the change from V to C feed. Overall responses to present extreme circumstances were no greater than variation occurring between weeks, and problems in practice seem remote.

Topics: Animals; Body Weight; Chick Embryo; Chickens; Eating; Egg Shell; Egg Yolk; Eggs; Female; Ovalbumin; Oviposition; Sesquiterpenes; Trichothecenes