Bovine Viral Diarrhea- Parvovirus Disease Bivalent Vaccine Development

Bovine viral diarrhea(Bovine viral diarrhea, BVD), also known as bovine viral diarrhea- a mucosal disease, caused by bovine viral diarrhea virus(Bovine viral diarrhea virus, BVDV) of cattle often made between the more common infections disease. The main clinical features of the disease can occur in herds of persistent infection, manifested as calf diarrhea, a cow multiple mucosal disease, while producing severe immune suppression, dams abortion and other symptoms.Bovine Parvovirus disease is caused by Bovine Parvovirus(Bovine parvovirus,BPV), can be an important infectious disease spread when cattle contact with each other. The disease is characterized by major clinical pregnancy, miscarriage and newborn calf heifers to respiratory and digestive tract disorders.In the US the top ten all kinds of veterinary vaccine products have nine products for multivalent or combined vaccine, combined vaccine in more foreign cattle vaccine concern, but also the direction of development. But there is no current domestic and against bovine viral diarrhea, bovine parvovirus of bivalent vaccine development reported by this study, in vitro cell culture method successfully isolated two bovine parvovirus, the establishment of bovine parvovirus strains of bovine viral diarrhea strain virus seed vaccine development with libraries and library exists on the virus seed exogenous factor pollution, the two strains and strains of replicator dynamics immunogenicity in different cells were studied. Preparation of parvovirus bivalent vaccine research process, were analyzed by the virus culture process parameters, the virus was inactivated harvest conditions of the experimental data,optimization of vaccine- bovine viral diarrhea. Finally, the vaccine’s safety and immunogenicity evaluation confirmed the vaccine prepared by the optimized process is safe and effective.First, this study from the domestic bovine serum successfully isolated two bovine parvovirus, named NB0801, NB1202. This two Bovine Parvovirus has aggregated guinea pig, horse, human red blood cell characteristics. Virus virulence determined using Reed-Muench method, virulence NB0801, NB1202 two strains of bovine parvovirus, respectively, 1.5 × 106 TCID50 / ml, 1.2 × 109 TCID50 / ml, this experiment saved NM0901 strains of bovine viral diarrhea virus Virulence was 1.6 ×107TCID50 / ml.Established NB1202 strain, NM0901 with virus seed strains for vaccine development library for virus seed library research exogenous factor detection and immunogenicity of the toxic species. Exogenous factor test results show NB1202 strain of bovine parvovirus, bovine viral diarrhea virus strain NM0901 without internal, exogenous factor pollution. Virus seed immunogenicity results show strains in vitro bovine parvovirus NB1202 antibody titer of 1: 160, bovine viral diarrhea virus strains in vitro NM0901 antibody neutralizing titers of 1: 40, confirmed that two strains possess relatively good immunogenicity. By bovine parvovirus, bovine viral diarrhea virus research replicator dynamics in different cells, and ultimately determine bovine turbinate cells(Bovine turbinate cells, BT) cell lines as bovine parvovirus vaccine production cells, Ma- Darby bovine kidney cells(Madin-Darby Bovine Kidney Cells, MDBK) as bovine viral diarrhea virus vaccine production cells. The results show NB1202 strain, NM0901 virus seed strains libraries comply with bovine viral diarrhea- Parvovirus bivalent vaccine R & D use.Bovine viral diarrhea- Parvovirus bivalent vaccine research process, NB1202 strain of bovine parvovirus, bovine viral diarrhea virus strain NM0901 3L spinner flasks process parameters at different temperatures, different access agents related to the amount of virus propagation culture sexual conduct studies to determine the NB1202 strain of bovine parvovirus, bovine viral diarrhea virus strain NM0901 3L roller bottle culture process parameters and optimal cultivation temperature virus inoculum. Viral inactivation by formalin verification test, to determine the optimum concentration of formaldehyde inactivated 0.7mg / ml. By screening test dose of aluminum adjuvant, aluminum hydroxide adjuvant determine the optimum dosage of1 mg / ml. Through the above experimental study to optimize the bovine viral diarrhea- Preparation of parvovirus bivalent vaccine.In this study, evaluation of vaccine safety, while using the SPF method in mice and guinea pigs law bovine viral diarrhea- Bovine Parvovirus bivalent vaccine bovine viral diarrhea vaccine security check, the results of this study demonstrate the preparation of vaccine safety sex meet the requirements. In the evaluation of the effectiveness of the vaccine, this study established a method of SPF mice detect bovine viral diarrhea- Parvovirus bivalent vaccine efficacy methods. The method is based on diphtheria vaccine efficacy trials(Vero cell method) based on theestablishment for bovine viral diarrhea- Bovine Parvovirus bivalent vaccine immunogenicity. The advantage of this method is to detect antibodies to attack instead of animal toxicity tests, with high accuracy and stability. Bovine viral diarrhea- Bovine Parvovirus bivalent vaccine immunogenicity evaluation showed that the bovine viral diarrhea vaccine potency liquid can reach 1:80, bovine parvovirus vaccine stock solution titer of 1: 160, indicating that the vaccine It has a good protective.The research results of our bovine viral diarrhea, bovine parvovirus disease prevention and control provides a scientific experiment data, and lay a solid technical foundation.