ABSTRACT
The first isolation and partial characterization of bovine foamy virus (BFV), also known as bovine syncytial virus, in Poland is described. This virus was isolated by co-cultivation of peripheral blood leukocytes from infected cattle with permissive Cf2Th cells. The new isolate, called BFV100 was identified using several techniques: electron microscopy, western blotting, PCR and sequencing of a part of the gag and pol/env genes. Based on syncytia induction, antigenic determinants, primer binding sites and sequence analysis, it can be concluded that isolate BFV100 is bovine foamy virus and is related to the known American and German BFV isolates by sequence homology and antigenic relatedness.
Subject(s)
Cattle Diseases/virology , Retroviridae Infections/veterinary , Spumavirus/isolation & purification , Animals , Cattle , Cattle Diseases/epidemiology , Cell Line , Cytopathogenic Effect, Viral , Dogs , Leukocytes/cytology , Leukocytes/virology , Poland/epidemiology , Retroviridae Infections/virology , Spumavirus/geneticsABSTRACT
A set of primers was developed to detect by polymerase chain reaction (PCR) the proviral DNA of bovine immunodeficiency virus (BIV). A short fragment of 101 bp BIV gene was selected as a target for primers; sequences of proviral DNA isolated from both a cell culture with BIV and from lymphocytes of an experimentally infected animal were known for the fragment. An amplicon of an expected size was detected by standard PCR in a transformed cell series of bovine testicles with Florida 112 BIV DNA, and in a plasmid DNA with a cloned proviral DNA of R29 BIV. Described in the paper are the results of a theoretical comparison of primers used in the detection of BIV by PCR. The presence of non-complementary nucleotides in the set of "primer-single stranded amplicon" was shown to bring about false positive results in the detection of BIV by PCR. No 1500 bp PCR product was detected after PCR with a synthesized pair of primers and with 100% homology for all known BIV isolates complementary to env gene. Finally, the issue of how to detectVIR in clinical samples obtained from experimentally and naturally infected is discussed.
Subject(s)
Immunodeficiency Virus, Bovine/isolation & purification , Polymerase Chain Reaction/veterinary , Proviruses/isolation & purification , Animals , Base Sequence , Cattle , Cattle Diseases/blood , Cattle Diseases/virology , DNA Primers , DNA, Viral/analysis , Gene Products, env/genetics , Immunodeficiency Virus, Bovine/genetics , Lentivirus Infections/veterinary , Lentivirus Infections/virology , Lymphocytes/virology , Molecular Sequence Data , Polymerase Chain Reaction/methods , Proviruses/genetics , Sequence AlignmentABSTRACT
The gag gene encoded protein, p24 of bovine leukemia virus (BLV), was cloned and expressed as thioredoxin-6xHis-p24 protein in Escherichia coli. The bacterial cells carrying plasmid pT7THis-p24 expressed the protein of 38 kDa that was detected by immunoblotting analysis using anti-p24 monoclonal antibodies and sera from BLV infected cattle and sheep. The purified p24 fusion protein was shown to be sensitive and specific for detection of BLV antibodies in the infected cattle.
Subject(s)
Escherichia coli/metabolism , Viral Core Proteins/biosynthesis , Viral Core Proteins/chemistry , Animals , Blotting, Western , Cattle , Gene Products, gag/metabolism , Immunoblotting/methods , Leukemia Virus, Bovine/immunology , Plasmids/metabolism , Recombinant Fusion Proteins/metabolism , Sheep , Thioredoxins/metabolismABSTRACT
Sera from chickens affected by Marek's disease or developing Rous sarcoma were investigated. There were changes in the protein fractions, and the amount of alpha and beta fractions was consistently increased. At the same time, immunosuppressive factors were found to inhibit the number of plaque-forming cells in the spleen of mice immunized with sheep red blood cells.
