Genetic Heterogeneity of Bovine Viral Diarrhoea Virus In Italy

The genetic characteristics, of 38 field isolates of bovine viral diarrhoea virus (BVDV) collected in 1999 from sick or healthy and persistently infected cattle of dairy farms situated in northern Italy, were investigated. A partial 5-untranslated region (5-UTR) sequence of each isolate was determined and a phylogenetic analysis was performed. All the isolates were classified as belonging to the BVDV-1 genotype and could be assigned to different BVDV-1 groups, namely BVDV-1b (n = 20), BVDV-1d (n = 6) and BVDV-1e (n = 10). Two remaining isolates could be classified as BVDV-1f and BVDV-1h, respectively. These results provided evidence for genetic heterogeneity of BVDV in Italy, and contribute to a better knowledge of the circulation of BVDV strains, and to their classification.     

Bovine viral diarrhea virus 1 is classified into different subgenotypes depending on the analyzed region within the viral genome

Phylogenetic analyses of bovine viral diarrhea virus (BVDV) were performed based on the nucleotide sequences of the 5' untranslated region (5'-UTR) and E2-coding gene. Thirty-six BVDV detected from naturally infected cattle in the northern region of Japan were divided into three genotypes, BVDV1a, BVDV1b and BVDV2, in a 5'-UTR phylogenetic tree. In a phylogenetic tree constructed from the E2-coding gene, BVDV1c was identified and the viruses classified in BVDV1c were included in BVDV1a in the 5'-UTR phylogenetic tree. Moreover, BVDV1a and BVDV1b in the E2-phylogenetic tree clustered closer together than in the 5'-UTR tree. These results suggested that phylogenetic analysis of the E2 gene was more useful for identification of subgenotypes within BVDV1.     

Clinical, pathological and antigenic aspects of bovine viral diarrhea virus (BVDV) type 2 isolates identified in Brazil

Nucleotide sequencing and phylogenetic analysis of Brazilian bovine viral diarrhea virus (BVDV) field isolates identified four viruses belonging to the genotype 2. Comparison of 5' UTR sequences from these isolates to those of North American BVDV type 2 revealed genomic variations that correlated with the geographic origins of the isolates. Two of the Brazilian type 2 viruses were isolated from clinical cases of gastroenteric/respiratory disease and two were isolated from healthy bovine fetuses. The clinical cases affected young animals (8- and 18-months-old) and were characterized by diarrhea, respiratory signs, extensive oral and digestive tract erosions, conjunctival and vulvar congestion, occasional digestive bleeding and vulvar and heart petechial hemorrhage. Antigenic analysis of these isolates with a panel of 10 monoclonal antibodies revealed marked antigenic differences in the major envelope glycoprotein, gp53/E2, compared to standard laboratory and vaccine BVDV strains. In addition, virus-specific antisera raised to Brazilian BVDV type 2 viruses displayed very low serological cross-reactivity with standard BVDV type 1 strains. Differences up to 64-fold in cross-neutralization titers were observed between BVDV type 1 and Brazilian BVDV type 2 isolates. The identification of BVDV type 2 among Brazilian cattle may have important implications for epidemiological studies, diagnostic and immunization strategies. Furthermore, the low neutralizing activity of BVDV type 1 antisera against the recently identified Brazilian BVDV type 2 isolates raises the question about the degree of protection conferred by BVDV vaccines, most of them based on a single type 1 strain.     

Genotypic characterization of Chilean llama (Lama glama) and alpaca (Vicugna pacos) pestivirus isolates

Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities worldwide where they have been introduced. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus, in particular Bovine Viral Diarrhea (BVDV), but there is little data available on Pestivirus infections in SACs. In this study we aimed to detect and identify Pestivirus genotypes and subgroups infecting SACs in both wild and confined environments. Samples were collected from 136 llamas and 30 alpacas from different areas in the Chilean Altiplano (wild animals), and from 22 llamas and 26 alpacas diagnosed as Pestivirus positive from the Metropolitana region in Chile (confined animals). Seroneutralization tests showed titers lower than 2 in all 166 samples from Chilean Altiplano. These samples were also negative to BVDV isolation, indicating that these animals have not been exposed to Pestivirus. After reactivation of positive samples from the Metropolitana region, the 5′ non-codifying region (5′NCR) and E2 glycoprotein were amplified by RT-PCR from the Pestivirus genome. Viral sequences were pairwise compared and phylogenetic trees were constructed. The 5′NCR analysis showed that all 12 sequenced isolates belonged to BVDV-1. Of particular interest, isolates from eight llama and two alpaca were BVDV-1j and two alpacas were BVDV-1b. In agreement with these results, E2 phylogenetic analysis rendered a similar grouping indicating that all 16 isolates belong to BVDV-1. However, the lower availability of E2 sequences determines the creation of a smaller number of sub-groups than the 5′NCR sequences. Based on the E2 sequences, the 5′NCR BVDV 1j group consisting of all the llamas and 3 alpacas are completely included in the E2 BVDV 1e group. Due to the universal availability of the 5′NCR segment, we propose the classification of these Chilean llamas and alpacas Pestivirus isolates as BVDV 1j and BVDV 1b respectively. Thus, this is the first time BVDV-1j is obtained in SACs. In addition, these results indicate Pestivirus infection in llamas and alpacas is associated with bovine population as genotypes and sub-groups are the same as those affecting Chilean livestock.     

A phylogenetic analysis of Bovine Viral Diarrhoea Virus (BVDV) isolates from six different regions of the UK and links to animal movement data

Bovine Viral Diarrhoea Virus (BVDV) is a pestivirus which infects cattle populations worldwide and is recognised as a significant source of economic loss through its impact on health and productivity. Studies investigating the molecular epidemiology of BVDV can give invaluable information about the diversity of viral strains present in a population and this, in turn, can inform control programs, drive vaccine development and determine likely infection sources. The current study investigated 104 viral isolates from forty farms across the UK. Through phylogenetic and nucleotide sequence analysis of the 5′UTR and N^pro regions of the isolates investigated, it was determined that BVDV 1a was the predominant sub-genotype. However, BVDV 1b, 1e and 1i were also identified and, for the first time in the UK, BVDV 1d. Through analysis of animal movement data alongside the phylogenetic analysis of these BVD isolates, it was possible to link animal movements to the viral isolates present on several premises and, for the first time, begin to elucidate the routes of viral transmission. With further work, this type of analysis would enable accurate determination and quantification of the true biosecurity risk factors associated with BVDV transmission.     

Genetic typing of bovine viral diarrhea virus isolates from Argentina

Genetic typing of 29 Bovine Viral Diarrhea Virus (BVDV) isolates from Argentina was carried out by sequencing 245 nucleotides of the RT-PCR products of the 5'-UTR region. Sequence analysis shows that these Argentinean BVDV include types 1 and 2. The majority (26/29) of the isolates are type 1, which comprises subtypes 1a and 1b, together with an additional subgroup within subtype 1a. This subgroup is close to the South African subgroup Ic of 1a viruses, and to the deer pestivirus strain "Deer". The three type 2 BVDV were isolated from fetal tissues or serum during the 7-8 years before a clinical outbreak in Argentina had been reported. Only inactivated vaccines are used in bovines of the country, thus the analysed viruses are authentic field strains. The long term circulation of type 2 BVDV (situation similar to that of North America before the epidemic of 1993), and the existence of viral populations which differ from the reference strains commonly used in vaccine elaboration should be considered by manufacturers of diagnostic reagents and vaccines.     

Clinical and epidemiologic observations of bovine viral diarrhea virus in the northwestern United States

Retrospective analyses of cases from which bovine viral diarrhea virus (BVDV) was isolated from 1980 to 2000 were conducted. These cases originated from the northwestern US and included both beef and dairy cattle. The results indicated that there was a shift in diseases associated with BVDV infection and in the animal age at onset of disease. Comparative results from the 1980 data indicated a low fetal infection rate (<5%), followed by steady increases of clinical cases and peaking at 6 months (30%). By 2000, the shift of BVDV cases was noticeable and indicated a biphasic occurrence of disease. The first phase was fetal infections, which increased to >25%, followed by a second phase at 6 months (>35%). Phylogenetic analysis was conducted on selected isolates from the time period 1998-2000 (n = 54). There were representative viral isolates from the two genotypes (BVDV1 and BVDV2), as well as subgenotypes, BVDV1a and BVDV1b. The types were further correlated with the clinical manifestation, which were reported as mucosal disease, persistently infected (PI)-poor doer, and abortion-open cows. The results indicated that BVDV were distributed throughout the clinical spectrum of disease, with BVDV2 representing the greatest frequency of isolation, and the greatest association with abortion-open cows. When the BVDV genotypes and subgenotypes were categorized into early (<100 days gestation) versus late (>100 days gestation) fetal infections, there was an inverse relationship noted. It was observed that BVDV1a was associated least with early infection (14%) and most with late infections (86%). BVDV1b was intermediate, followed by BVDV2, which was associated more with early infections (45%) and less with late infections (55%) when compared with BVDV1a and BVDV1b.     

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.     

Phylogenetic, antigenic and clinical characterization of type 2 BVDV from North America

Bovine viral diarrhea virus (BVDV) infection continues to have a significant impact upon US cattle producers despite the availability of more than 140 federally licensed vaccines. Detection and control is hampered by viral heterogeneity that results in differences in neutralizing epitopes, cytopathology and virulence. Recently it was found that there are two different genotypes, BVDV1 and BVDV2, among BVDV. BVDV2 isolates make up a significant proportion of the BVDV isolated in North America. Serologically BVDV2 viruses can be distinguished from BVDV1 and border disease viruses. Mab binding also distinguishes between BVDV1, BVDV2 and BDV. Like the BVDV1 viruses, BVDV2 viruses may exist as one of two biotypes, cytopathic or noncytopathic, based on their activity in cultured cells. Cytopathogenic effects on cultured cells does not correlate with virulence in vivo, as BVDV2 associated with hemorrhagic syndrome (HS) are noncytopathic. Variation among BVDV1 and BVDV2 in the 5' UTR is similar. Phylogenetic analysis and differences in virulence suggest that BVDV2 are heterogeneous. Symptoms resulting from BVDV2 infections may range from clinically inapparent to clinically severe. Recently, disease outbreaks associated with acute uncomplicated BVDV infection have been reported in the US and Canada. These outbreaks of clinically severe disease, termed HS, were all associated with viruses from the BVDV2 genotype. Not all BVDV2 isolates cause clinically severe disease. Avirulent BVDV2 isolates do exist and may predominate over virulent BVDV2 in nature. When virulent BVDV2 viruses are inoculated into calves they induce a disease characterized by fever, diarrhea, leukopenia, lymphopenia, neutropenia, thrombocytopenia, and death. Infection with avirulent BVDV2 results in a reduction of luekocytes that may be accompanied by a low-grade fever. These viruses do not cause clinical disease or a clinical leukopenia.