RESUMEN
The International Veterinary Industry Test Replacement Organisation (In-VITRO) was established in 1995 with the aim of developing, validating and harmonising in vitro alternatives to replace in vivo methods for in-process and potency testing of veterinary clostridial vaccines. The emphasis has been on the reduction of animal usage in the Clostridium chauvoei potency assay and its eventual replacement by an in vitro assay. Replacement of the toxin neutralisation assay for Cl. tetani by an internationally validated indirect ELISA has already started. A validation programme involving a collaboration organised through EDQM which could ultimately lead to the standardisation of in vitro tests for all clostridial vaccines is in progress. In addition In-VITRO is now considering the setting up of a programme for Erysipelas vaccines. The collaboration between manufacturers of veterinary vaccines in the development and validation of in vitro tests is a major step towards the reduction and replacement of animal tests.
Asunto(s)
Alternativas a las Pruebas en Animales , Vacunas Bacterianas/normas , Clostridium/inmunología , Medicina Veterinaria , Animales , Anticuerpos Antibacterianos/análisis , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/toxicidad , Infecciones por Clostridium/inmunología , Infecciones por Clostridium/prevención & control , Infecciones por Clostridium/veterinaria , Ensayo de Inmunoadsorción Enzimática/métodos , Agencias Internacionales , Reproducibilidad de los ResultadosRESUMEN
A type-specific monoclonal antibody was produced by immunizing mice with purified equid herpesvirus-1 (EHV-1). The EHV-1 specific mAb reacted with all the EHV-1 strains tested so far by indirect ELISA, immunofluorescence, and immunoperoxidase tests. No reactions were detected with the EHV-4, EHV-2, or EHV-3 strains tested. The indirect immunofluorescence and immunoperoxidase tests showed that the nuclei of infected cells were predominantly stained by this mAb. Triton treatment of the virus and immunogold labeling experiments indicated that the nucleocapsid of EHV-1 was the target antigen of the mAb. Preliminary results indicated that this mAb might be a useful tool in detecting specific antibody in horses that have been exposed to EHV-1. In a blocking ELISA, antibodies in sera from hamsters, mice and a foal which had been exposed to EHV-1 were differentiated from those in sera of animals exposed to EHV-4.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Cápside/inmunología , Epítopos , Infecciones por Herpesviridae/veterinaria , Herpesvirus Équido 1/inmunología , Enfermedades de los Caballos/microbiología , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Antivirales/biosíntesis , Anticuerpos Antivirales/inmunología , Células Cultivadas , Cricetinae , Reacciones Cruzadas , Femenino , Infecciones por Herpesviridae/diagnóstico , Enfermedades de los Caballos/diagnóstico , Caballos , Técnicas Inmunológicas , Ratones , Ratones Endogámicos BALB C , Conejos , Pruebas SerológicasRESUMEN
Six monoclonal antibodies (mAb) were produced by immunizing mice with a Cryptococcus neoformans serotype A spheroplast lysate (CNSL). Two mAb were of the IgG1 isotype; the others were IgM. The results obtained with one IgM mAb (CN6) are reported herein. This mAb recognized the four serotypes of C. neoformans and no cross-reactions were observed with extracts from Cryptococcus melibiosum, Candida albicans, Saccharomyces cerevisiae, Torulopsis glabrata or Trichosporon beigelii. Fractionation of the CNSL by gel filtration revealed that mAb CN6 recognized high molecular weight substances as well as a range of smaller molecules. Indirect ELISA inhibition studies showed that this mAb recognized substances in a cryptococcal culture filtrate. Inhibition studies and agglutination tests using latex beads sensitized with purified CN6 showed that CN6 strongly reacted with the C. neoformans serotype A cell envelope galactoxylomannan-mannoprotein complex (GAlXM-MP) and only weakly with the glucuronoxylomannan (GXM) component. These tests also showed that purified galactoxylomannan (GalXM) from C. neoformans serotype A was more reactive than purified mannoprotein (MP). An anti-GalXM mAb might be a useful tool for monitoring the clinical course of cryptococcal infections.
Asunto(s)
Anticuerpos Antifúngicos/biosíntesis , Cryptococcus neoformans/inmunología , Cryptococcus/inmunología , Polisacáridos Bacterianos/inmunología , Polisacáridos , Animales , Anticuerpos Monoclonales/biosíntesis , Especificidad de Anticuerpos , Fraccionamiento Celular , Reacciones Cruzadas , Medios de Cultivo , Ensayo de Inmunoadsorción Enzimática , Filtración , Inmunoglobulina M/biosíntesis , Pruebas de Fijación de Látex , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos BALB CRESUMEN
Macrophages of 1+ rainbow trout are parasitised by an unknown protozoan. An ultrastructural study permits to associate it with the genus Dermocystidium described in many fishes. This protozoan is characterized by a large vacuole of mitochondrial origin and by concentric structured bodies.