Genetic typing of bovine viral diarrhoea virus isolates from India

Thirteen BVDV isolates collected in four geographic regions of India between 2000 and 2002 were typed in 5'-UTR. To confirm results of genetic typing, selected viruses were also analysed in the N(pro) region. Phylogenetic analysis revealed that all Indian BVDV isolates belong to BVDV-1b (Osloss-like group). Despite a long distance between the farms from which the viruses were isolated there was no correlation between the origin of viral isolates and their position in a phylogenetic tree. Higher genetic similarity of Indian BVDV isolates was observed most probably due to the uncontrolled movement of cattle as well as the uncontrolled use of semen from bulls for breeding of local and farm cattle in different states of India.     

Genetic variety of bovine viral diarrhea virus 2 strains isolated from sheep

Bovine viral diarrhea virus 2 (BVDV-2) strains, isolated from sheep showing clinical symptoms of border disease, have been evaluated by the palindromic nucleotide substitution (PNS) method at the three variable loci (V1, V2 and V3) in the 5'-untranslated region (UTR) of genomic RNA. The characteristic two base-pairings common to the BVDV-2 species, a C-G pairing which was common to the V1 locus, and a G*U pairing common to the V2 locus, were observed in all tested strains. Strains BD-78 and C413 were identified by a unique C-G pairing at position 4 from the bottom of the V2 stem region, which is characteristic to BVDV-2b. BVDV-2d characteristic U-A pairing at position 18 of the V1 stem region was observed in five strains, Lees, 167 237, 168 149, 173 157 and 175 375. No strains have been assigned to the genotypes BVDV-2a or BVDV-2c. Furthermore, the investigation at the level of the 5'-UTR excluded the application in sheep of the proposed BVDV-2 genetic virulence markers described in cattle. The two specific positions of uracil and cytosine nucleotides related to low or high virulence where indifferently present in the ovine BVDV-2 strains responsible of border disease.     

Comparative pathogenicity of selected bovine viral diarrhea virus isolates in gnotobiotic lambs

The infectivity and pathogenicity of selected bovine viral diarrhea virus (BVDV) isolates were determined in gnotobiotic, colostrum-deprived neonatal lambs. Five-day-old cesarean-derived gnotobiotic lambs were exposed to 1 of 10 BVDV isolates via aerosol suspension. These isolates were from tissues or secretions of calves or lambs affected with respiratory tract disease, weak neonatal calves, aborted bovine fetuses, or reference Singer or Draper BVDV. The pathogenicity of each isolate, relative to the others, was evaluated in lambs by measurement of the neutralizing antibody response, virus isolation from nasal secretions or tissues, and postmortem lesions. The BVDV isolates varied in their infectivity and pathogenicity. Singer, the cytopathic reference strain, was the most lymphotrophic isolate and stimulated the greatest neutralizing antibody response. Encephalitis was the most consistent lesion observed and was used as the final determinant of relative pathogenicity of the viruses. The most neuropathogenic isolates were the 2 viruses originating from lambs affected with respiratory tract disease, the 2 weak neonatal calf isolates, and 1 isolate from an aborted bovine fetus. The least pathogenic isolates were the 2 reference isolates, Draper and Singer; the 2 mucosal disease isolates; and 1 isolate originating from an aborted bovine fetus.     

Differentiation of cytopathic and noncytopathic isolates of bovine viral diarrhea virus by virus neutralization

Soluble antigens of cytopathic and noncytopathic isolates of bovine viral diarrhea virus were resolved by high-performance liquid gel-permeation chromatography into 4 major and 3 minor peaks. The 2 peaks with the larger molecular weights (240,000 and 140,000 daltons) were immunogenic when inoculated into rabbits. Virus neutralizing antibodies were specific for the homologous virus. The soluble antigens were determined to be greater than 100,000 daltons by filtration.     

Genetic analysis of bovine viral diarrhea virus in pre-weaned native Korean calves

Tropical Animal Health and Production - Bovine viral diarrhea virus (BVDV), a prominent viral pathogen worldwide, causes substantial economic losses in the cattle industry. BVDV comprises two...     

Genetic typing of bovine viral diarrhoea virus: most Slovenian isolates are of genotypes 1d and 1f

A selection of 43 bovine viral diarrhoea viruses isolated from mainly persistently infected cattle on 23 Slovenian farms between 1997 and 2001 were characterised genetically. Viral RNA was extracted from infected cell cultures, reverse transcribed and amplified by PCR with primers targeting the 5'-UTR and the N(pro) gene, followed by direct sequencing of purified PCR products obtained for both genomic regions. The N(pro) sequences provided the best genetic resolution, and gave also higher statistical support for phylogenetic classification of the viruses. Thirty-eight of the Slovenian isolates were of genetic subtypes 1d and 1f, four were 1b, and one subtype 1g. No BVDV type 2 viruses were found. This genetic prevalence matched those previously reported for neighbouring countries, as opposed to findings reported for more distant European countries, e.g. France, Spain and the UK. From eight cattle herds several virus isolates were analysed; with one exception all isolates from each herd were of the same genetic group. Extended sequencing of the N(pro) and part of the C gene of virus isolates with identical 5'-UTR sequences allowed differentiation between isolates obtained at different times from one herd.     

Response of cattle persistently infected with noncytopathic bovine viral diarrhea virus to vaccination for bovine viral diarrhea and to subsequent challenge exposure with cytopathic bovine viral diarrhea virus

Nine steers persistently infected with noncytopathic bovine viral diarrhea (BVD) virus were allotted into 3 groups (3 cattle/group). Cattle in group A were vaccinated with a modified-live BVD virus vaccine of porcine cell origin, cattle in group B with a modified-live BVD virus vaccine of bovine cell origin, and cattle in group C with a killed BVD virus vaccine of bovine cell origin. Detrimental effects due to vaccination were not seen. Six weeks after vaccination, the steers were challenge exposed with a cytopathic BVD virus. All steers developed mucosal disease after challenge exposure, produced antibodies that neutralized various isolates of BVD virus, and remained persistently infected until death. Steers given killed virus vaccine had a minimal neutralizing-antibody response and developed mucosal disease as quickly as reported for challenge-exposed, nonvaccinated, persistently infected cattle. Steers given modified-live virus vaccines had higher neutralizing-antibody response and longer intervals from challenge exposure to development of mucosal disease. The specificity of the neutralizing-antibody response differed between groups of vaccinated cattle.     

Evolution of bovine viral diarrhea virus vaccines.

Control of bovine viral diarrhea virus (BVDV) infection is economically important to the cattle industry because the virus causes a variety of clinical diseases that adversely affect essentially all stages of the production cycle. Production losses primarily stem from reproductive failure and from immunosuppression during acute BVDV infection, which predisposes calves to respiratory or enteric diseases. Control is achieved by implementing herd health pro-grams focused on limiting exposure by avoiding persistently infected (PI) carrier cattle and by optimizing protective immunity through immunization. Vaccination cannot be relied upon solely to protect against fetal infection and losses due to BVD. This is because no single BVDV vaccine has been shown to give complete fetal protection. In addition to strategic use of vaccines, herd management practices should also be implemented to identify and eliminate PI carrier cattle and to avoid exposure to BVDV infection.     

Structural proteins of bovine viral diarrhea virus.

A procedure for the purification of radioactively labeled bovine viral diarrhea virus was critically evaluated. Purification of virus from artificial mixtures of unlabeled infected and labeled noninfected cells indicated that the extent of purification was approximately 100-fold with respect to host proteins. Residual host proteins were found to contaminate the viral preparation even after extensive purification by differential and isopycnic zonal centrifugation. Co-electrophoresis of 3H-labeled virus with 14C-labeled host cell material in neutral sodium dodecyl sulfate-7.5% polyacrylamide gels provided a means to distinguish viral specific proteins from host cell protein contaminants. Four major electrophoretic components were identified as being of viral origin; molecular weights of the components were estimated from their migration rates relative to protein markers of known molecular weight. Two viral components (VC), VC 1 and VC 3, migrated heterogeneously and had molecular weights of 93,000 to 110,000 and 50,000 to 59,000 daltons, respectively. Molecular weights of VC 2 and VC 4 were 70,000 and 25,000 daltons, respectively.     

Comparison of cytopathic and noncytopathic isolates of bovine viral diarrhea virus by oligonucleotide fingerprinting.

The molecular technique of RNA fingerprinting was used to characterize the genomes of 5 isolates of bovine viral diarrhea virus (BVDV): 2 viral pairs from the same animal, BVD-ILN/BVD-ILC and BVD-TGAN/BVD-TGAC, and the cytopathic viral prototype, BVD-NADL. Oligonucleotide patterns from the viruses were compared, and unique and overlapping oligonucleotides were identified. A comparison of the fingerprints indicated that the genome of each virus was distinguishable by the T1 RNase oligonucleotide fingerprinting technique. The greatest similarity observed was between oligonucleotides from BVD-ILC and BVD-ILN. Eighteen large oligonucleotides were conserved in all 5 BVDV isolates studied. We found that within a pair of BVDV, the cytopathic fingerprint was different from the noncytopathic fingerprint, indicating that cytopathic and noncytopathic BVDV may be distinct viruses.     

Application of single-strand conformation polymorphism to the study of bovine viral diarrhea virus isolates.

Single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products is a genetic screening technique for rapid detection of nucleotide substitutions in PCR-amplified genomic DNA or cDNA. It is based on the observation that partially formamide-denatured double-stranded DNA migrates as 2 single-stranded DNA molecules when electrophoresed in nondenaturing polyacrylamide gels. The mobility depends on the 3-dimensional conformation of the strand under the conditions used. It is possible to discriminate between DNA strands differing in only 1 nucleotide. The method was applied to the analysis of Bovine Viral Diarrhea Virus (BVDV) isolates. Reference and Argentinian strains were assessed for variations in their 5' untranslated region (5'-UTR). The PCR products of the 5'-UTR ends were formamide denatured and compared by SSCP analysis in nondenaturing 15% polyacrylamide and 15% polyacrilamide-5% glycerol gels. The reference strains SD-1, Singer, and Oregon C24V had differences in electrophoretic patterns. Despite the high conservation among the 5'-UTR of pestiviruses, the method allowed discrimination among all 9 Argentinian isolates. The 5'-UTR of a fetal kidney-derived isolate (1R93) was PCR amplified and cloned in a plasmid vector; the SSCP analysis of 30 PCR products obtained by direct amplification over randomly selected clones produced 5 different banding patterns, indicating the existence of viral quasispecies. The results show that SSCP may be used to identify and differentiate among BVDV isolates.     

Genetic typing of recent classical swine fever virus isolates from Croatia

During a period of 5 years (1997-2001) several outbreaks of classical swine fever (CSF) were recorded in Croatia. For genetic typing, fragments of 150 nucleotides within the 5'-non-translated region (5'-NTR) and 190 nucleotides within the E2 glycoprotein coding gene of nine field isolates that were derived from domestic pigs and wild boars were used. For better epizootiological understanding, isolates from other European countries were included in the study. The results show that the isolates belong to subgroups 2.1 and 2.3 of CSF virus. Isolates from subgroup 2.1 were collected from domestic pigs during sporadic outbreaks in June 1997 and are genetically closely related. A genomic similarity between these isolates and CSF virus isolates from pigs in other European countries from the same year could also be confirmed. In contrast, the isolate from October 1997 was found to be a member of subgroup 2.3, and is closely related to European CSF virus isolates from outbreaks in the last decade in Western and Central European countries. These results show that two different sources of CSF virus caused outbreaks in Croatia during the same year. Furthermore, a close relationship was found between an isolate from a domestic pig in 1999 and isolates of subgroup 2.3 that originated from Croatian wild boars.     

牛病毒性腹泻疫苗的研究进展

牛病毒性腹泻(bovine viral diarrhea,BVD)作为一种病毒性腹泻黏膜病,给养殖业造成了巨大的经济损失,应用疫苗来防控牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)依然是重要的方法。直至目前,我国还没有成熟的BVDV疫苗,本研究主要综述了近几年来国外BVDV疫苗的研究进展,旨在为国内BVDV疫苗的研制及应用提供参考。     

Isolation of cytopathic and noncytopathic bovine viral diarrhea virus from the spleen of cattle acutely and chronically affected with bovine viral diarrhea

Both cytopathic and noncytopathic bovine viral diarrhea virus (BVDV) were isolated from 16 of 17 bovine spleens representing 11 herds that had experienced acute BVD and from 12 of 21 bovine spleens from 1 herd affected with chronic BVD. It was concluded that isolation of cytopathic and noncytopathic BVDV from the same spleen probably indicates that an animal with a persistent, noncytopathic BVDV infection was superinfected with a cytopathic BVDV. The prevalence (greater than 70%) of 2 viruses in the spleen of cattle with acute or chronic BVD suggested that persistent infection with noncytopathic BVDV may be an important factor in the pathogenesis of BVD.     

Efficacy of bovine viral diarrhea vaccine used in Japan against bovine viral diarrhea virus type 2 strain 890

Bovine viral diarrhea virus (BVDV) has been segregated into two genotypes, type 1 and type 2. To determine the efficacy of the commercially available bovine viral diarrhea type 1 vaccine used in Japan against BVDV type 2, calves were infected with BVDV type 2 strain 890 4 weeks after administration of the vaccine. The vaccinated calves did not develop any clinical signs and hematological changes such as observed in unvaccinated calves after the challenge. Furthermore, the challenge virus was not recovered from the vaccinated calves throughout the duration of the experiment, whereas it was recovered from all unvaccinated calves. The bovine viral diarrhea vaccine used in Japan is efficacious against infection with BVDV type 2 strain 890.