ABSTRACT
In this study, a Specific Pathogen-Free (SPF) Bama miniature pig herd was established.The standards for breeding experimental pigs, genetic test and microbiological quality control had been drafted.The local standards of Heilongjiang province SPF technical specifications for microbiological monitoring of pigs had been formulated.The genetic and microbiological quality criterion had been used to control of the SPF pig herd.Classical swine fever virus (CSFV) and highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) infection models of Bama miniature pigs were established, especially the infection miniature pig model of PRRSV was used to evaluated commercially vaccine.The comparison of the cytokine homology between Bama miniature pigs and domestic pigs showed that the Bama miniature pigs can be used to instead of domestic pigs as an ideal experiental animal.At present the SPF Bama miniature pig colonies have been widely used to study the mechanism of prevention and pathogenic in the classical swine fever virus, porcine reproductive and respiratory syndrome virus and porcine transmissible gastroenteritis virus and porcine circovirus and so on.The completion of the project solved the bottleneck problem of the experimental pig usage which provided a new ideal experimental animal for animal disease and life science research.
ABSTRACT
The VP60 protein of rabbit hemorrhagic disease virus (RHDV) is a structural protein with important roles in viral replication and assembly. In this study, we immunized BALB/c mice with the RHDV-TP strain. Six monoclonal antibodies (mAbs) were selected and characterized by enzyme-linked immunosorbent assay, Western blotting, and indirectly immunofluorescence analysis (IFA). All six mAbs (AD4, AG10, BC9, BE8, BH3, and DE2) had positive reactions with recombinant VP60 as analyzed by IFA, but only two (AG10 and DE2) reacted with denatured RHDV by Western blotting. Fifty-four partially overlapping fragments of the VP60 gene were expressed with His or Glutathione S-transferase (GST) tags to identify the epitopes recognized by AG10 and DE2. These two epitopes were located at the C-terminal of VP60 and were longer (64 and 53 amino acids, respectively) than normal B cell epitopes. However, both AG10 and DE2 also interacted with RHDV2 VP60 expressed in insect cells. Amino acid alignments of the AG10 and DE2 epitope regions between RHDV and RHDV2 VP60 indicated several mutations, suggesting that the epitopes recognized by the mAbs AG10 and DE2 were discontinuous. Epitope immunogenicity was evaluated by inoculating specific pathogen-free rabbits with saline, purified DE2 epitope, or RHDV inactive vaccine. Rabbits immunized with the DE2 epitope developed high levels of RHDV-specific antibodies but no cellular immune response and died after challenge with RHDV-HYD isolate. Despite their lack of neutralizing activity, these mAb reagents and epitopes may have useful clinical applications and will be valuable tools in further studies of the structure and function of the RHDV VP60 protein.
Subject(s)
Antibodies, Viral/immunology , Caliciviridae Infections/veterinary , Epitopes, B-Lymphocyte/immunology , Hemorrhagic Disease Virus, Rabbit/immunology , Viral Structural Proteins/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/isolation & purification , Antibodies, Viral/isolation & purification , Blotting, Western , Caliciviridae Infections/immunology , Caliciviridae Infections/virology , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Epitope Mapping , Fluorescent Antibody Technique, Indirect , Mice, Inbred BALB C , RabbitsABSTRACT
Objective To understand the genetic background of the specific pathogen-free Yorkshire and Landrace pigs, imported from Canada by Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences.Methods The population genetics were investigated by using 19 pairs of microsatellite primers.Results In the Yorkshire pigs and Landrace pigs, 84 and 89 alleles, respectively, were detected at 19 microsatellites loci.The average polymorphic informa-tion content and mean heterozygosity in the Yorkshire pigs were 0.5271 and 0.5877, and in the Landrace population were 0.5652 and 0.6066, respectively.Because of the significant ( P<0.01) differences of alleles in different loci such as S0155,S0143,S0178,Sw857 and Sw936, it made them possible to be used to identify Yorkshire and Landrace pigs′breed. F-statistics showed that the differentiation within the population was small and genetic structure was stable.Conclusions Compare with the domestic pedigree large White and Landrace pigs, the SPF pedigree pigs imported from Canada are more stable in genetic structure, and can be used as laboratory animal models in animal science research.
