Molecular characterization of bovine foamy virus and its association with bovine leukemia virus infection in Vietnamese cattle

The detection of bovine foamy virus (BFV) in Vietnamese cattle was performed using conventional PCR targeting pol and gag genes. Out of 243 tested samples, ten (4.1%) and eight (3.3%) samples were positive for BFV gag and pol DNA, respectively. The prevalence of bovine leukemia virus (BLV) estimated by detection of proviral DNA using nested PCR targeting env gene was 26.7% (65/243). The results of nucleotide sequence alignment and the phylogenetic analysis suggested that Vietnamese BFV strains showed high homology to isolates belonging to either European or non-European clades. There was no significant correlation between BLV and BFV. This study provides information regarding BFV infection and confirms the existence of two BFV clades among Vietnamese cattle for the first time.     

Isolation and genetic characterization of bovine parainfluenza virus type 3 from cattle in China

Bovine parainfluenza virus type 3 (BPIV3) is one of the most important of the known viral respiratory pathogens of both young and adult cattle. However BPIV3 has not been detected or isolated in China prior to this study. In 2008, four BPIV3 strains were isolated with MDBK cells from cattle in China and characterized by RT-PCR, nucleotide sequence analysis, transmission electron microscope observation, hemadsorption and hemagglutination tests. Nucleotide phylogenetic analysis of partial hemagglutinin-neuraminidase (HN) gene for four isolates and the complete genome for the SD0835 isolate implicated that the four Chinese BPIV3 strains were distinct from the previously reported genotype A (BPIV3a) and genotype B (BPIV3b) and might be a potentially new genotype, which was tentatively classified as genotype C (BPIV3c). This is the first study to report the isolation and genetic characterization of BPIV3 from cattle in China.     

Genomic characterization and distribution of bovine foamy virus in Japan

Bovine foamy virus (BFV) is distributed through worldwide cattle herds. Although the biological features of BFV are not well understood, appearance of clinical manifestation by superinfection with other microorganisms is inferred. In Japan, reports of genomic characterizations and epidemiology of this virus are limited. In this study, we performed whole genomic sequencing of BFV strains Ibaraki and No.43, which were isolated in this country. Additionally, we investigated BFV in geographically distant four daily farms in Japan, to estimate the distribution of BFV and its correlation to bovine leukemia virus (BLV). BFV was distributed throughout Japan; the average positive rate was 12.7%. The nucleotide sequence identities of the isolates were 99.6% when compared with BFV strain isolated in the USA. The phylogenetic tree using env gene sequence showed strains Ibaraki, No.43 and Kagoshima were sorted in the same cluster including the USA and Chinese strains, while Hokkaido strain was in the other cluster including European strains. Although no clear correlation between BFV and BLV could be found, BFV and BLV infections were likely to increase with ages. Our data on epidemiology and characteristics of BFV will provide important information to reveal biological features of BFV.     

Isolation of bovine syncytial virus from mesenteric lymph nodes of cattle

Bovine syncytial virus was isolated from mesenteric lymph nodes of young dairy calves, beef cattle, and mature dairy cattle. Isolations were made in cocultures of lymph node specimens and bovine embryonic lung cells or by subpassage of cocultures in which cytopathic effects were not initially detected. The bovine syncytial viral isolates after storage at -65 C for 3 months in either infected cell cultures of supernate fluids from suspensions or sonically treated infected cell monolayers were usually recovered. Bovine syncytial virus also was recovered from cell cultures after storage for 6 months at -25 C.     

Isolation and characterization of epizootic hemorrhagic disease virus from sheep and cattle in Colorado

Epizootic hemorrhagic disease virus was isolated from cattle and sheep in northeastern Colorado during July and August 1984. The isolates were identified as serotype 2 by plaque-inhibition serotyping, genome electropherotyping, and protein analysis.     

Genetic and antigenic characterization of bovine viral diarrhoea virus type 2 isolated from cattle in India

Previous studies have shown that bovine viral diarrhoea virus type 1 (BVDV-1) subtype b is predominantly circulating in Indian cattle. During testing for exotic pestiviruses between 2007 and 2010, BVDV-2 was identified by real time RT-PCR in two of 1446 cattle blood samples originating from thirteen states of India. The genetic analysis of the isolated virus in 5′ UTR, N^pro, entire structural genes (C, E^rns, E1 and E2), nonstructural genes NS2-3 besides 3′ UTR demonstrated that the nucleotide and amino acid sequences showed highest similarity with BVDV-2. The entire 5′ and 3′ UTR consisted of 387 and 204 nucleotides, respectively, and an eight nucleotide repeat motif was found twice within the variable part of 3′ UTR that may be considered as a characteristic of BVDV-2. The phylogenetic analysis revealed that the cattle isolate and earlier reported goat BVDV-2 isolate fall into separate clades within BVDV-2a subtype. Antigenic typing with monoclonal antibodies verified the cattle isolate also as BVDV-2. In addition, cross-neutralization tests using antisera raised against Indian BVDV strains circulating in ruminants (cattle, sheep, goat and yak) displayed significant antigenic differences only between BVDV-1 and BVDV-2 strains. This is the first identification of BVDV-2 in Indian cattle that may have important implications for immunization strategies and molecular epidemiology of BVD.     

Isolation and partial characterization of a calicivirus from calves

A calicivirus was isolated from 3 dairy calves in a herd with persistent calf respiratory tract problems. This virus, named Tillamook calicivirus, was not neutralized by 18 different calicivirus-typing serums available. The agent caused only minimal lesions in 2 experimentally exposed calves, but did establish a persistent infection with virus shedding for 45 days, after which time the experiment was terminated. Experimentally exposed swine developed clinical vesicular lesions. The possible origins, disease potential, and relationships to the exotic animal disease agent, vesicular exanthema of swine are discussed for this first calicivirus isolate of bovine origin.     

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.     

Isolation of bovine syncytial virus from lymphocytes recovered from fluids used to flush uterus and oviducts of superovulated cattle.

A Ficoll-gradient method was applied to isolate lymphocytes from fluids used to flush the uterus and oviducts of superovulated cows. Bovine syncytial virus antigens were demonstrated in 15 of 19 cows by cocultivation of lymphocytes with fetal lamb spleen cells and examining them with direct immunofluorescence. Viral serum antibodies were found in the same 15 of 19 cows as above by the modified direct complement-fixation test. The virus was also detected in one of four uterotubal cell cultures.     

Molecular and phylogenetic characterization of bovine coronavirus virus isolated from dairy cattle in Central Region,Thailand

Tropical Animal Health and Production - Bovine coronavirus (BCoV) is involved mainly in enteric infections in cattle. This study reports the first molecular detection of BCoV in a diarrhea outbreak...     

Isolation of bovine adenovirus type 4 from cattle in Oregon.

Four isolates of bovine adenovirus type 4 were recovered from Oregon cattle. One isolate was recovered from a 1-week-old calf with pneumoenteritis, and 1 isolate from an 8-month-old bull with fever and respiratory disease. Two isolates were recovered as latent viruses in testicular cell cultures. All 4 isolates of the virus were shown to have certain physical, chemical, biologic, and antigenic characteristics similar to previously described strains of bovine adenovirus type 4. Of 246 adult beef and dairy cattle in Oregon, 51% had serum-viral neutralizing antibodies to the type 4 virus at the 1:8 dilution level.     

Cultivation and partial characterization of bovine astrovirus

Bovine astrovirus serotype 2 (US2) was adapted to primary neonatal kidney cell (NBK) cultures by the addition of 50 micrograms ml-1 of trypsin in the medium. Infectious virus was released from the cells within 7 days post-infection in early passages and within 3 days in later passages. In the absence of trypsin, neither passage of infected cells nor release of infectious virus occurred. The virus was shown to be similar to the fecal astrovirus by a neutralization test and by ultrastructural studies of infected cells. Primary embryo bovine kidney (EBK) and NBK cell cultures supported infection with both fecal and tissue culture adapted (TCA) astrovirus. The time-related development of infection, as studied by immunofluorescence, was similar for both fecal and TCA astrovirus and for both cell culture types. The first indication of viral infection and expression of viral antigens occurred at 7 h post-infection and was characterized by the appearance of a diffuse faint immunofluorescence (IF) of the cytoplasm. Soon after, two or three brilliant IF granules were observed in the nucleus, which appeared to involve the nucleoli. Subsequently, dense granular IF was seen in the perinuclear region of the cytoplasm, which later extended to involve all the cytoplasmic area. In both EBK and NBK cultures infected with either fecal or tissue culture adapted astrovirus, only a minority of cells became infected, even when the multiplicity of infection exceeded one. Occasionally 10-20% of cells were infected, but in most cultures the proportion did not exceed 2% and in NBK cultures, from 3/9 calves, no infected cells were observed. The virus did not infect bovine cell lines. Infectivity of the virus was not removed by treatment with chloroform, and iododeoxyuridine and actinomycin D when added to the medium, did not block replication. Masses of virions were observed by electron microscopy in discrete areas in the cytoplasm, with similar distributions as the viral antigen foci as seen by IF. The mean diameter of the virions was 34 nm. In conclusion, bovine astrovirus lacks both essential lipids and an envelope, probably has an RNA genome, may have a nuclear phase of replication involving the nucleoli which is not blocked by DNA inhibitors, and has a selective cell tropism.     

Isolation and characterization of sulfur oxidizing bacteria from cattle manure compost

Hydrogen sulfide causes offensive odors. We attempted to isolate sulfur oxidizing bacteria (SOB) from cattle manure compost. A most probable number assay could not detect SOB by using a Cha medium which had been applied in order to isolate the SOB from active sewage. Cultivation using a Cha plate medium revealed 5.75 × 10⁷ colony forming unit/g of bacteria. A single strain of SOB was isolated from a colony formed on the plates and identified as Halothiobacillus neapolitanus. This is the first report that H. neapolitanus has been found in cattle manure compost.     

Studies on bovine herpesviruses. Part 1. Isolation and characterization of viruses isolated from the genital tract of cattle

Herpesviruses, previously isolated from cattle (Theodoridis, 1978), were further studied and provisionally placed in the bovid herpesvirus 4 (BHV-4) group. Major differences were found between IBR-IPV (BHV-1) and BHV-4 virus strains. In MDBK cells, all BHV-4 strains started growing at the edges of the culture, the process progressing slowly until destruction of the cells was complete by the 10th day. BHV-4 strains failed to induce neutralizing antibodies in cattle, goats and rabbits. Only the addition of mineral oil adjuvant induced neutralizing and complement fixing antibodies in goats. BHV-1 strains, in contrast, produced very potent antisera in all these systems. Cross-neutralization tests indicated the existence of 2 distinct serological groups representing BHV-1 and BHV-4. The BHV-4 strains appear to be interrelated and they could not be grouped. A BHV-1 strain showed fixation of complement with the antisera of 6 BHV-4 strains. Electron micrographs showed an accumulation of nucleocapsids in the cytoplasm and an early release of virus particles due to cell destruction. Variation in incubation temperature had a significant effect on the particle formation. At lower temperatures, the number of enveloped particles in the cytoplasm increased. On the basis of the characteristics uncovered in this study, it is possible that all the BHV-4 strains represent one and the same virus which has undergone certain biological changes, thus illustrating a phenomenon which appears to be a characteristic of the herpesviruses.