This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID50,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×108.0 TCID50/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×108.0TCID50/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China. 相似文献
Lambs sucking non-immunised ewes or ewes immunised 4-5 weeks before lambing with live attenuated, aromatic-dependent (aroA) Salmonella typhimurium (strain CS 332) were challenged orally at either 2, 4 or 7 days of age with virulent S. typhimurium (strain CS 94) at doses ranging from 109 to 1013 colony forming units. No lambs displayed signs of clinical salmonellosis and all survived challenge but those sucking immunised ewes had organisms of the challenge strain in their faeces for much shorter periods of time than lambs of the control ewes. High titres of specific antibodies were measured in colostrum and milk of immunised ewes in comparison with very low titres measured in samples from control ewes; these differences were reflected by the titres of antibodies in the sera of corresponding lambs. At 2 days after lambing, the major antibody isotype in the colostrum of immunised ewes and sera of their lambs was IgM whereas at 7 days IgG1 was the predominant isotype. While it was clear that vaccination of pregnant ewes with the live attenuated vaccination conferred protection against experimentally-induced salmonellosis in their lambs, considerable protection was observed in control lambs in spite of there being very low titres of antibodies in the mammary secretion of their dams. The latter observation could be related to the presence of contain non-antibody potent bactericidal factors previously described in colostrum and milk.
Résumé
Des agneaux qui tètent des brebis non immunisées, ou bien immunisées, 4 à 5 semaines avant la mise bas avec un vaccin atténué constitué d'une souche de Salmonelle typhimurium dépendente pour sa croissance de la presence de composés aromatiques (souche aro A CS 332) ont reçu, à l'âge de 2, 4 ou 7 jours, par voie orale, des doses allant de 109 à 1013 Salmonella (souche virulente CS 94). Aucun agneau n'a présenté de signes cliniques et tous ont survécu à cette administration, mais les agneaux des brebis immunisées ont excrété la souche d'épreuve dans leur fécès pendent moins longtemps que les agneaux des brebis non vaccinées. Des titres plus élevés d'anticorps sont détectés dans le colostrum et le lait des brebis vaccinées et comparés aux brebis témoins. Cette différence est également notée pour les titres d'anticorps présents dans le sérum des agneaux issus de brebis vaccinées ou non. Deux jours après la mise bas, la classe des anticorps majoritaires, tant au niveau du colostrum des brebis vaccinées que du sérum de leurs agneaux, est la classe des IgM, alors qu'après 7 jours, les IgG1 prédominent. Bien qu'il soit clair que des brebis pleines vaccinées à l'aide d'un vaccin vivant atténué, apportent à leurs agneaux une protection vis à vis d'une Salmonella expérimentale, une réelle protection est également observée chez les agneaux témoins, malgré les titres faibles d'anticorps présente dans les sécrétions mammaires de leur mères. Cette dernière observation peut être mise en rapport avec la présence d'agents bactériodes ne contenant pas d'anticorps, déjà décrits dans le colostrum et le lait. 相似文献
To estimate the potency of a porcine parvovirus (PPV) vaccine, three vaccinated and three non-vaccinated pregnant gilts were infected with PPV and the distribution of the virus was studied in the tissues of their 51 fetuses. Virus detection was attempted using haemagglutination (HA) and immunofluorescence (IF) assays, as well as by standard (single) and nested polymerase chain reactions (PCR). None of the detection methods yielded positive results when used to test for the presence of virus in suspensions of organs from the fetuses from the vaccinated gilts. However, the virus was detected in the fetuses from non-vaccinated gilts as follows: HA was positive in 14 cases out of 23 (60.8%), IF in 16/23 (69.5%), standard PCR in 12/20 (60%), and the nested PCR in 19/23 (82.6%). Although the correlation among the results of various methods of virus detection was rather close (r<0.83), the sensitivity of the nested PCR was the highest, both when testing dilutions of PPV and when analysing the fetal organs. The nested PCR therefore provides a reliable approach for studies of virus distribution in fetal organs, with special reference to potency tests on vaccines. 相似文献