RESUMEN
Three hundred and sixty 14-day-old chickens were divided into seven groups. The chickens, except for blank control group, were vaccinated with Newcastle disease vaccine, repeated at 28 days old. At the same time of the first vaccination, the chickens in three astragalus polysaccharide-oxymatrine (AP-OM) groups were orally administrated respectively with the mixture of AP-OM at high, medium and low concentrations, in astragalus polysaccharide (AP) group and oxymatrine (OM) group, with corresponding medicine, in non-medicine (NM) control group, with equal volume of physiological saline, once a day for 3 successive days. On 14, 21, 28, 35 and 42 days after the first vaccination, the changes of peripheral lymphocyte proliferation and serum antibody titers of the chickens were determined by MTT method and hemagglutination inhibition test. On 14, 28 and 42 days after the first vaccination, the serum IL-2 concentration was determined by Enzyme-linked Immunosorbent Assay (ELISA). The results showed that at most time points, the lymphocyte proliferation, antibody titers and IL-2 concentrations of 5 medicine-administrating groups were significantly higher than that of corresponding NM group. At some time points, the lymphocyte proliferation, antibody titers and IL-2 concentrations in high and medium doses of AP-OM groups were significantly or numberly higher than those in AP group and OM group. It indicated that AP-OM could significantly improve the immune efficacy of Newcastle disease vaccine, astragalus polysaccharide and oxymatrine possessed synergistical immunoenhancement.
Asunto(s)
Alcaloides/farmacología , Planta del Astrágalo/química , Factores Inmunológicos/farmacología , Virus de la Enfermedad de Newcastle/inmunología , Polisacáridos/farmacología , Quinolizinas/farmacología , Vacunas Virales/inmunología , Alcaloides/inmunología , Animales , Anticuerpos/sangre , Proliferación Celular/efectos de los fármacos , Pollos , Sinergismo Farmacológico , Medicamentos Herbarios Chinos/farmacología , Factores Inmunológicos/inmunología , Interleucina-2/sangre , Linfocitos/citología , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Masculino , Polisacáridos/inmunología , Quinolizinas/inmunologíaRESUMEN
To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA-flumequine) and to cationized ovalbumin (cOVA-flumequine). For the immunization of chickens, cBSA-flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA-flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 microg/kg). No cross-reactivity was noticed with various related quinolones.
Asunto(s)
Yema de Huevo/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Fluoroquinolonas , Leche/química , Quinolizinas/análisis , Animales , Pollos/inmunología , Inmunoglobulinas , Quinolizinas/inmunología , Sensibilidad y EspecificidadRESUMEN
We have prepared monoclonal antibodies for the fluorescent molecular rotors 9-(2-carboxy-2-cyanovinyl)julolidine (CCVJ) and 9-(dicyanovinyl)julolidine (DCVJ). Mouse monoclonal antibody (IgG2b) prepared against CCVJ-conjugated bovine serum albumin strongly bound CCVJ and DCVJ. The CCVJ (or DCVJ) binding to IgG and Fab was accompanied by a drastic increase in fluorescence quantum yield, suggesting the restriction of intramolecular rotational relaxation about the donor-acceptor bond of the fluorophores. Nonspecific IgG never changed the quantum yield of the fluorophores. From the Scatchard plots, the association constants of CCVJ to IgG and Fab were 6.8 x 10(7) and 5.4 x 10(7) M-1, respectively, and the numbers of moles of CCVJ bound per mole of IgG and Fab were calculated to be 2.0 (+/- 0.1) and 1.0 (+/- 0.05), respectively. The fluorescence spectra of the IgG-bound CCVJ were quite similar to those of Fab-bound CCVJ. The fluorescence lifetimes of the IgG-bound and Fab-bound CCVJ were 388 and 383 ps at 25 degrees C, respectively. They were 6.3 times as long as the fluorescence lifetime of CCVJ free in solution (62 ps). These results indicated that the drastic increases in quantum yields were due to the decreases of the nonradiative rate constants of the antibody-bound CCVJ, as well as due to the changes of the intrinsic radiative rate constant, and that the nonradiative internal rotations about the donor-acceptor bond of CCVJ were not dependent on the size of the bound antibody molecules.(ABSTRACT TRUNCATED AT 250 WORDS)