Proviral detection and serology in bovine leukemia virus-exposed normal cattle and cattle with lymphoma.

Twenty-seven cattle with lymphoma and 46 cows from a known bovine leukemia virus (BLV)-infected herd were tested for anti-BLV antibody by the agar gel immunodiffusion (AGID) test and an enzyme-linked immunosorbent assay (ELISA). The polymerase chain reaction (PCR) and Southern hybridization were used to detect BLV provirus in the tumor DNA of the 27 cattle with lymphoma. The PCR was used to detect BLV provirus in the peripheral blood mononuclear cell DNA of the 46 normal known-exposed cattle. Two presumed false negative AGID test results compared to ELISA were found. Of ten cattle three years of age or less with "sporadic" forms of lymphoma, four had BLV provirus in tumor DNA, detectable by PCR. In two of these four, BLV provirus was clonally integrated based on digestion of tumor DNA with restriction enzymes followed by Southern hybridization. The BLV provirus was not detected by PCR in 5 of 17 cattle with "enzootic" lymphoma and two of these five were seronegative. Among normal BLV-exposed cows, 6.5% (3 of 46) were serologically positive and PCR negative; serologically negative and PCR positive cows occurred with the same frequency. Serological and PCR test results, when considered in all cattle (n = 73), had a concordance rate of 83.6%. Discordant test results occurred with approximately equal frequency between serologically positive and PCR negative (7 of 73, 9.6%) and serologically negative and PCR positive (5 of 73, 6.8%) groups. These data suggest that the role of BLV in some "sporadic" bovine lymphomas, previously unassociated with BLV, should be reexamined. The BLV provirus was not demonstrable in the tumor DNA from five adult cattle with lymphoma, suggesting that BLV may not be the etiological agent in all adult bovine lymphomas. The findings of persistently seronegative PCR positive and seropositive PCR negative cattle indicate that further work is needed to more fully understand the host-virus interaction. Present serological screening methods may not have sufficient sensitivity for determining BLV status in some circumstances.     

Vertical transmission of bovine leukemia virus and bovine immunodeficiency virus in dairy cattle herds.

Vertical transmission of bovine leukemia virus (BLV) and bovine immunodeficiency virus (BIV) was investigated in five dairy cattle herds in Hokkaido, where 36.1 and 17.0% of cattle were BLV and BIV seropositive, respectively, and 9.9% of dams were co-infected with both BIV and BLV. Twenty six cases of offspring born from dams infected with only BLV (17 cases) or with both BIV and BLV (9 cases) were examined for the presence of BLV and BIV before and after colostrum feeding by polymerase chain reaction (PCR) and syncytium assay. After birth, all calves were separated immediately from their dams. The offspring born from BLV-positive dams were BLV-negative before colostrum feeding, suggesting that no transplacental transmission had occurred. Thereafter, these offspring were fed colostrum or milk from their dams, but still remained BLV-negative. The other offspring born from BLV-positive dams were fed with BLV-negative colostrum, or with pasteurized BLV-positive colostrum. All these calves remained negative for BLV infection, suggesting that in utero transmission of BLV is negligible. In the case of offspring born from dams co-infected with BLV and BIV, calves were BIV-positive before colostrum feeding at 1 day after the birth, indicating in utero transmission of BIV. After colostrum feeding from their dams, newborn calves became BLV-positive. In addition, one calf was BLV-positive even before colostrum feeding. These results suggest that BIV can be transmitted to offspring in utero, and that BLV can be transmitted through colostrum or milk if dams are infected with both BIV and BLV.     

Seroprevalence of bovine immunodeficiency-like virus and bovine leukemia virus in a dairy cattle herd.

To determine the prevalence of single vs. dual infection with bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV), sera (n = 95) from a dairy cattle herd were analyzed for anti-BIV and anti-BLV antibodies by an enzyme linked immunosorbent assay. Twenty-one percent (20/95) of samples were BIV-seropositive, while 52% (49/95) of the same samples were BLV-seropositive. A significantly greater percentage of BIV-seronegative samples were BLV-seropositive, 57% (43/75), than were BIV-seropositive samples, 30% (6/20). There was no significant correlation between data ranked from least to greatest amount of anti-viral antibody. Five cattle had persistent lymphocytosis (PL); all five were BLV-seropositive and two were BIV-positive. The mean anti-BLV titer was significantly greater in PL cattle, as compared at non-PL cattle, whereas there was no significant difference between the mean anti-BIV titer in PL cattle, as compared with non-PL cattle. These results provide additional information on the seroprevalence of naturally occurring BIV infection, and indicate that BIV can exist independent of other common infectious agents, such as BLV. Further, the results suggest that infection with BIV is not associated with an increased rate of infection with other infectious agents such as BLV.     

Rectal palpation and transmission of bovine leukemia virus in dairy cattle.

To determine whether rectal palpation, using common obstetrical sleeves, serves as a mode of transmission of bovine leukemia virus in dairy cattle, field studies were conducted at 2 dairies. At a commercial dairy, significant difference was not observed in rate of seroconversion in heifers and cows in which the same sleeve or new sleeves were used for palpations. At a university dairy, where cattle were used to teach dairy husbandry and veterinary procedures, significantly (P less than 0.02) greater rate of seroconversion was observed in heifers and cows palpated with unwashed common sleeves than that observed in heifers and cows palpated with sleeves washed between use. Although rectal transmission of bovine leukemia virus under field conditions was documented, it was related to frequency of palpation and age of cattle.     

Rectal transmission of bovine leukemia virus in cattle and sheep

Bovine leukemia virus (BLV) was transmitted by rectal inoculation of BLV-infective whole blood into cattle and sheep. Two cows and 2 sheep each were given 500 ml and 50 ml of blood, respectively, by rectal infusion. Two sheep which served as positive controls each were given 1 ml of the same blood, IV. All animals became seropositive to BLV by postinoculation week 5. Although relatively large volumes of blood were used for rectal inoculation, a base line for infectivity was established for the rectal route.     

Effects of bovine leukemia virus infection on production and reproduction in dairy cattle.

The purpose of this study was to determine the effects of bovine leukemia virus (BLV) infection on production, reproduction and longevity in dairy cattle. The study population was a commercial Holstein dairy herd of approximately 400 milking cows. Cattle were tested for antibodies to BLV at least annually for three years and when culled. Four groups of culled cows were compared: seronegative cows (n = 79), seropositive cows without lymphocytosis (n = 176), seropositive cows with lymphocytosis (> or = 9,000 lymphocytes/microliter) (n = 74), and seropositive cows with lymphosarcoma (n = 29). Seropositive groups of cows were bred more times and had longer calving intervals than seronegative cows. The seropositive groups had greater 305-day ME (mature equivalent) FCM (3.5% fat-corrected milk) per lactation and were older when culled than seronegative cows. However, the percent fat per lactation was greater in seronegative cows. In the last complete lactation, differences in 305-day ME FCM, days open and cull age between groups were reduced and none were significant (p > 0.05). In the cull lactation, only cows with lymphocytosis had reduced milk production relative to seronegative cows, although this difference was not significant. After adjustment for initial production and reproductive values, only seropositive nonlymphocytotic cows were culled at a significantly older age than seronegative cattle. Lymphocytotic cows were culled four months younger on average than nonlymphocytotic seropositive cows. Hence, BLV infected cows had greater milk production on average than uninfected cows. Adverse effects of BLV infection were primarily limited to lymphocytotic cows which were culled earlier and had reduced milk production in the cull lactation.     

Circulating immune complexes in bovine leukemia virus (BLV)-infected cattle.

Circulating immune complexes (ICs) were detected in the sera of bovine leukemia virus (BLV)-seropositive cattle. Immune complexes were precipitated in 2.5% polyethylene glycol (PEG) and further dissociated. Bovine leukemia virus antigens, IgG and IgM molecules were detected after solubilization in the presence of sodium dodecyl sulphate, and quantitated by enzyme-linked immunosorbent assay (ELISA) assays. Mean values of IgG and IgM in BLV-containing ICs did not significantly differ from those obtained from ICs originating from BLV-seronegative animals. However, differences were found in the composition of ICs from older BLV-positive animals as compared to those obtained from young animals. The ratio of IgG/IgM was 5.02 in animals aged 5-10 years, while this ratio was 11.66 in animals of less than 5 years of age and 10.19 in controls. This might indicate a possible increase in the contribution of IgM molecules to the structural composition of ICs in BLV-infected cattle as related to age or stage of infection.     

Seroprevalence of bovine immunodeficiency virus and bovine leukemia virus in dairy and beef cattle in hokkaido

Serological survey of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) infection was conducted in dairy cattle from 10 different regions of Hokkaido, Japan. Among 390 cattle, 11.0% of cattle were BIV-seropositive and 3.3% were BLV-seropositive. Moreover, in two dairy farms, where bovine leukosis has been reported, prevalence of BIV infections were 6.4 and 9.1%, respectively. In contrast, among 150 beef cattle, 16.6% were BIV-seropositive while none was BLV-seropositive. Dual infections with BLV and BIV in dairy cattle were tested by using 107 BLV-seropositive sera, and 20 sera were found BIV-positive (18.7%). These results indicate that BIV infection was widespread in Hokkaido.     

An immunoblotting procedure for detection of antibodies against bovine leukemia virus in cattle.

A sensitive protein immunoblotting (Western blot) procedure has been developed for detecting anti-BLV antibodies in cattle sera. The antibodies against most of the major viral proteins could be detected. This procedure does not give any non-specific background staining and there is absence of any erroneous results due to utilisation of purified viral preparations. The procedure has been applied for detection of antibodies to BLV in a set of 74 sera samples and it has been compared with other commonly used serological tests like ELISA and agar gel immunodiffusion test.     

Infection of bovine immunodeficiency virus and bovine leukemia virus in water buffalo and cattle populations in Pakistan

A survey of antibodies to bovine immunodeficiency virus (BIV) known as bovine lentivirus and bovine leukemia virus (BLV) was conducted with samples from water buffalo and cattle populations in Pakistan. A total of 370 water buffaloes and 76 cattle were tested, and 10.3% and 15.8%, respectively, were found positive for anti-BIV p26 antibodies determined by Western blotting, while 0.8% of water buffaloes and no cattle were positive for anti-BLV antibodies determined by immunodiffusion test. BIV-seropositive water buffaloes and cattle were found to have BIV proviral DNA in the peripheral blood mononuclear cells determined by nested polymerase chain reaction. This is the first report of BIV infections in water buffaloes.     

Antibody titers in cattle clinically and subclinically infected with bovine leukemia virus

Antibody titers to protein (p) and glycoprotein (gp) antigens of bovine leukemia virus were studied by the immunodiffusion test in two groups of cattle. One group showed evidence of enzootic bovine leukosis (BL+ cattle), whilst the second group, possessing antibodies to gp antigen, were adjudged as subclinical cases (BL?). All 165 BL? and 97% (108/111) BL+ cattle possessed antibodies to gp antigen but those to p antigen were evident in only 67% BL? cattle in contrast to 89% in BL+ cattle. Mean antibody titers to both antigens were statistically higher in BL+ cattle.     

Serum immunoglobulin concentrations in cattle naturally infected with bovine leukemia virus

In order to elucidate whether natural infection of BLV in cattle might induce humoral immunological responses, changes in IgG1, IgG2, and IgM concentrations in the sera of infected cattle were determined. Twelve BLV-infected cattle were used. Cattle of different breeds were classified serologically and haematologically into BLV + PL+, BLV + PL- and BLV-free groups. Ig concentrations in the sera of the three groups were quantitated using a commercial single radial immunodiffusion assay. The findings were compared to those of BLV-free cattle. The serum IgM concentrations were significantly lower in cattle with PL (P less than 0.001) than in BVL + PL- and BLV-free cattle. The IgM concentrations tended to be lower in BLV+ PL- than those of BVL-free cattle. There were no significant differences in IgG1 and IgG2 serum concentrations between the three experimental cattle groups. IgG1 was the predominant subtype in the sera of all cattle groups.