Serological evidence of an Australian bovine lentivirus.

Recombinant 26 kDa capsid (CA) proteins of bovine lentiviruses, bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV), were expressed in Escherichia coli and utilised as antigens for an enzyme-linked immunosorbent assay (ELISA) and a western immunoblot (WIB) procedure for the detection of antibody in dairy cattle in Western Australia. A total of 690 serum samples, 30 from each of 23 farms, were tested by ELISA with a JDV CA protein antigen, and antibody was detected in 3.8% (p<0.05) of the sera. Nine sera from each farm were also tested by WIB with JDV CA protein antigens and antibody was detected in 15.9% of these samples. All ELISA-positive results were also WIB-positive, and all sera antibody-positive by WIB with JDV CA protein antigens were also antibody-positive by the WIB using recombinant BIV CA antigens. This study showed that recombinant protein antigens can be used for serological tests to detect bovine lentivirus infection in Australia.     

Detection of multiple retroviral infections in cattle and cross-reactivity of bovine immunodeficiency-like virus and human immunodeficiency virus type 1 proteins using bovine and human sera in a western blot assay.

Bovine antibovine immunodeficiency-like virus (BIV) antibodies were detected by Western blot analysis (WBA) using a chemiluminescence protocol. Bovine sera with anti-BIV activity, obtained from cows in two dairy herds, had antibodies directed against a variety of BIV-specific antigens indicating chronic infections. These sera were also tested for serological reactivity against bovine leukemia virus (BLV) and bovine syncytial virus (BSV). Cows most commonly had anti-BSV antibodies (12 of 39). Evidence for infection with BSV and BIV or BSV and BLV occurred with almost equal frequency (5 of 39 and 4 of 39, respectively) while only one instance of BIV and BLV coseropositivity was detected. The high prevalence of BSV seropositivity is consistent with a relatively infectious virus, which, as is known, may be transferred congenitally. Similar rates of coseropositivity of BIV or BLV with BSV in this population suggest that BIV is no more infectious than BLV and probably requires prolonged close contact for transmission. Seven of nine cows with anti-BIV antibodies detected primarily human immunodeficiency virus type 1 (HIV-1) p51 and p63 antigens by WBA using an alkaline phosphatase detection system, suggesting that HIV-1 proteins have potential usefulness in screening cattle for BIV seropositivity. Six human sera that showed strong reactivity against multiple HIV-1 proteins and the serum from one of three patients considered to be an "indeterminate" HIV-1 reactor, cross-reacted primarily with BIV p26. This is the first report of human sera with antibody to BIV-specific proteins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seroprevalence of bovine immunodeficiency virus in dairy and beef cattle herds in Korea.

Infection of bovine immunodeficiency virus (BIV), a lentivirus, is thought to sporadically occur throughout the world, but seroepidemiological surveys concerning the incidence of BIV are limited and have not been undertaken in Korea. A total of 266 sera from different twenty dairy (Holstein) and twenty-six Korean native beef (Hanwoo) farms of the south-western part of Korea was analyzed for the presence of anti-BIV antibodies by Western blotting. Thirty five percent and 33% of dairy and beef cattle, respectively, were BIV-seropositive. By nested polymerase chain reaction, it was confirmed that these seropositive cows had provirus in the peripheral blood mononuclear cells. To demonstrate the correlation with BIV and bovine leukemia virus (BLV) infection, these sera were also analyzed for anti-BLV antibodies by immunodiffusion test, resulting in high prevalence of BLV infection but relatively a few dual infections. We report herein the first serological detection of antibodies to BIV in Korea.     

Evidence of bovine immunodeficiency virus (BIV) infection: serological survey in Argentina

This is the first report of serological evidence for bovine immunodeficiency virus (BIV) infection in Argentina. The analysis was performed in 589 dairy bovine sera samples, applying indirect enzyme-linked immunosorbent assay (I-ELISA) using a synthetic antigen (transmembrane peptide, TM) and Immunofluorescent assay (IFA). In this study, 9 dairy herds from 4 Argentinian provinces were evaluated and 12% of the animals tested positive for BIV. Seven of the 9 herds tested were BIV seropositive and the percentage of BIV seropositive animals in the herds ranged from 2% to 42%. Direct detection of BIV provirus applying nested PCR was not conclusive. Antibody detection against bovine leukemia virus (BLV) in all sera was also performed applying immunodiffusion (ID) assay and 59% resulted seropositive. Statistical analysis of the results was carried out and possible evidence of association between BIV and BLV infection was considered. Future studies should be performed including local field isolates strains of BIV.     

Evidence for bovine immunodeficiency virus infection in cattle in Zambia

We report herein on the first evidence for the presence of bovine immunodeficiency virus (BIV) in Zambia. Serological surveillance of BIV and bovine leukemia virus (BLV) was conducted in traditional cattle herds in Zambia. Out of a total of 262 sera analyzed, 11.4% were found positive for anti-BIV p26 antibodies as determined by Western blot analysis, while 5.0% were positive for anti-BLV gp51 antibodies as detected by immunodiffusion test. Peripheral blood mononuclear cells from BIV seropositive cattle were found to have BIV-provirus DNA, as detected by nested polymerase chain reaction. A nucleotide sequence corresponding to a 298 bp fragment of the BIV pol gene was also analyzed. Amino acid sequences of these Zambian pol gene products showed 98.0 to 100% homology to the American strain BIV R29, 97.0 to 99.0% to Japanese BIV isolates, and divergence ranged from 0.0 to 2.0% among Zambian BIV isolates.     

Seroprevalence and molecular evidence for the presence of bovine immunodeficiency virus in Brazilian cattle

Data on the worldwide distribution of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) is limited. A prevalence study of antibodies to BIV and BLV was conducted in six different cattle herds in Brazil. Out of a total of 238 sera analyzed, 11.7% were found positive for anti-BIV p26 antibodies as determined by Western blot analysis, 2.1% were positive for anti-BLV gp51 antibodies as detected by immunodiffusion test. Peripheral blood mononuclear cells from BIV seropositive cattle were found to have BIV-provirus DNA, as detected by nested polymerase chain reaction. A nucleotide sequence corresponding to a 298 bp fragment of the BIV pol gene was also analyzed. Amino acid sequences of these Brazilian pol gene products showed 98.0 to 100% homology to the American strain BIV R29, 97.0 to 99.0% to Japanese BIV isolates, and divergence ranged from 0 to 4.0% among Brazilian BIV isolates. This evidence of the presence of BIV and BLV infections in Brazil should be considered a health risk to Brazilian cattle populations and a potential causative agent of chronic disease in cattle.     

Seroprevalence to bovine immunodeficiency virus and lack of association with leukocyte counts in Italian dairy cattle

We report herein on the first serological detection of antibodies to bovine immunodeficiency virus (BIV) in Italy. According to criteria of a stratified-random sampling of dairy cattle reared in the Parma area (a province in the Po Valley, Northern Italy), sera from 3166 cows belonging to 272 herds were collected. In addition, sera of 138 bulls from eight artificial-insemination (AI) centres were sampled. Seventy-eight cows (2.5%) from 16 herds (5.8%) and seven bulls (5.1%) from two AI centres were positive for BIV-R29 antibodies in the IFA-test. IFA-positive sera assayed by Western blot had reaction to different viral proteins: 81 out of 85 sera showed antibody to p26 (considered the BIV major internal core protein); four sera reacted to other viral proteins but not to p26. Peripheral blood leukocytes of 60 seropositive and 60 seronegative animals, belonging to eight BIV-infected herds, were enumerated to assess any effect of BIV infection on white-blood cells. No significant differences were detected between the two groups. These data indicate that BIV infection is present in Italian dairy cattle – but the role of BIV in inducing disease remains unclear.     

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.     

Seroprevalence of bovine immunodeficiency virus and bovine leukemia virus in draught animals in Cambodia

Since bovine immunodeficiency virus (BIV), known as bovine lentivirus, has been detected in dairy and beef cattle in various countries around the world, a prevalence study of antibodies to BIV and bovine leukemia virus (BLV) was conducted in draught animals in five provinces in Cambodia, where protozoan parasite infections were suspected in some animals. To clarify the status of draught animals including Haryana, Brahman, mixed-breed, local breed cattle and muscle water buffaloes, a total of 544 cattle and 42 buffaloes were tested, and 26.3 and 16.7%, respectively, were found positive for anti-BIV p26 antibodies determined by Western blotting. There were 5.3% positive for anti-BLV antibodies detected by immunodiffusion test among the cattle, but no reactors among buffaloes and no dual infection for both BIV and BLV was determined in this study. Peripheral blood mononuclear cells from BIV-seropositive cattle were found to have BIV-provirus DNA, as detected by polymerase chain reaction and subsequent Southern blot hybridization. This is the first evidence for the presence of BIV and BLV infections in draught animals in tropical countries such as Cambodia. This wide distribution of BIV suggests its association with problems in animal health as reported worldwide, and that a primary BIV infection can predispose death of affected animals by other aggressive pathogens or stresses.     

Serological survey for bovine immunodeficiency virus in dairy cattle from Poland

A seroprevalence study of bovine immunodeficiency virus (BIV) was undertaken on 1,541 serum samples from Holstein cattle from 23 herds, located in different geographical regions of Poland. The analysis was performed using ELISA, with recombinant Gag protein of BIV as antigen. The average BIV prevalence was 4.9% in individual cattle, while the percentage of herds harboring at least one seropositive animal, was 82.6%. To demonstrate the correlation of BIV and bovine leukemia virus infection, all sera were analysed for BLV antibodies and there was only a slight association between both infections. Overall, these results show that BIV infection is present in dairy cattle in Poland at a prevalence rate found in other European countries.     

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.     

Evidence of bovine immunodeficiency virus in cattle in Turkey

A seroepidemiological study of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) infections was conducted in four different cattle herds in Turkey. A total of 300 blood samples were analyzed and 12.3% were found to be positive for anti-BIV p26 antibodies by Western blot analysis and 1.6% positive for anti-BLV gp51 antibodies by an immunodiffusion test. BIV infection was confirmed with the detection of BIV-provirus DNA using the nested polymerase chain reaction. This is the first evidence for the presence of BIV in cattle in Turkey.     

Performance characteristics and optimisation of cut-off values of two enzyme-linked immunosorbent assays for the detection of antibodies to Neospora caninum in the serum of cattle

AIM: To determine the performance characteristics of two enzyme-linked immunosorbent assays (ELISAs) manufactured by Institut Pourquier (IP) for the detection of antibodies against Neospora caninum in bovine sera. METHODS: Sera from 526 cattle were assayed in two ELISAs (IP) for the detection of anti-N. caninum antibodies. Results from a further ELISA (IDEXX) were used to provide the "gold standard"N. caninum infection status of the cattle and the ELISA results assessed by two-graph receiver operating characteristic (TG-ROC) analysis. RESULTS: TG-ROC analysis suggested changes to one of the IP ELISA protocols, arriving at a cut-off threshold that was different to the one recommended by the manufacturer. With that change, both of the ELISAs performed with high sensitivity and specificity (in excess of 98%) for bovine sera. CONCLUSIONS: The analysis of the two IP ELISAs when used on individual bovine sera demonstrated high sensitivity and specificity. TG-ROC analyses optimised the cut-off point suggested by the manufacturer for one of these commercial diagnostic assays and found agreement with the manufacturer's cut-off regarding the other assay. This will help with the accurate identification of infected animals and thereby contributing to the control of neosporosis.     

Serological evidence of bovine immunodeficiency-like virus infection in a sheep.

A six month-old sheep was entered into a control group in an experiment designed to study the effects of exposure to the bovine immunodeficiency-like virus (BIV). Anti-BIV antibodies were detected in the serum of this sheep prior to the start of the study; these antibodies persisted for 12 months at which time the animal was destroyed. The sheep was normal clinically and was grossly normal at postmortem examination. Blood from this sheep was inoculated into a recipient sheep which subsequently showed a transient anti-BIV antibody response beginning two months postinoculation. Sheep have been previously shown to produce anti-BIV antibodies after experimental inoculation with infected cell culture material or infected bovine blood and BIV infection was found in a sheep pastured with BIV-infected cattle. In the present case there was no contact with cattle; the source of the infection was not identified.     

Evaluation of two commercial enzyme-linked immunosorbent assays for detection of bovine viral diarrhoea virus in serum and skin biopsies of cattle

AIM: To assess the ability of two commercial bovine viral diarrhoea (BVD) virus (BVDV) antigen-capture enzyme-linked immunosorbent assays (ELISAs) to detect virus in serum and skin biopsies. METHODS: Thirty cattle persistently infected (PI) with BVDV were identified using routine diagnostic laboratory testing. Additional ear-notch skin biopsies and blood samples were collected from these animals to confirm the diagnosis, and from 246 cohorts, to determine their BVDV status. Skin biopsies were soaked overnight in buffer and the eluate collected. All sera and eluate were tested using two commercially available ELISAs for detecting BVDV antigen, and a subsample of positive and negative sera was tested using a polymerase chain reaction (PCR) test. A study was also performed to ascertain the risk of cross contamination occurring during the collection and processing of skin biopsies. RESULTS: Both serum and skin samples tested using either ELISA resulted in the detection of all cattle identified as PI and no non-infected cattle were incorrectly classified as infected using either method. Agreement between all assays (ELISAs, whether performed on serum or skin, and PCR) was 100%. No cross-contamination of skin samples between animals was evident using routine biopsy methods. CONCLUSIONS: Viraemic cattle infected with BVDV were accurately identified using either of the two commercial ELISAs evaluated on either serum or skin samples. CLINICAL RELEVANCE: Either skin biopsies or serum samples can be collected from cattle to determine their BVDV status. This should overcome problems in accurately identifying the infection status of young calves in which colostral antibodies might interfere with the antigen-capture ELISA.     

Decline of maternal antibodies to small ruminant lentivirus in goat kids

We carried out this study to determine for how long small ruminant lentivirus (SRLV)‐specific antibodies can be detected by three commercial ELISA kits in goat kids after suckling infected does in field conditions. Forty‐one kids born to SRLV‐seropositive asymptomatic does were blood sampled prior to colostrum consumption, and then weekly for 6 months in total. The sera were screened with three commercial ELISA kits: whole‐virus ELISA (wELISA), recombinant transmembrane and capsid antigen ELISA (TM/CA‐ELISA), and surface antigen ELISA (SU‐ELISA). All but one kid were seronegative in all three ELISAs right after birth. At the age of 1 week all kids turned seropositive in wELISA, 39 kids (95%) in TM/CA‐ELISA, and 35 kids (85%) in SU‐ELISA. All seropositive kids turned seronegative in wELISA by the 15th week, and in SU‐ELISA by the 19th week (median of 8 weeks in both ELISA), whereas in TM/CA‐ELISA five kids (13% of 39 initially seropositive) were still seropositive at the age of 6 months (median of 11 weeks). Antibody levels at the age of 1 week proved significantly linked to the duration of maternal antibodies in all three ELISAs and could be employed to predict for how long maternal antibodies would remain detectable.     

Seroprevalence to bovine virus diarrhoea virus and other viruses of the bovine respiratory complex in Venezuela (Apure State)

Six hundred and fifteen serum samples obtained from cows in five districts of Apure State, Venezuela, were tested by ELISA for antibodies to bovine virus diarrhoea virus (BVDV). The same samples were also ELISA-tested for antibodies to bovine herpesvirus type 1 (BHV-1) and bovine respiratory syncytial virus (BRSV). Additionally, the haemagglutination-inhibition (HI) test was used for detecting antibodies to parainfluenza virus type 3 (PIV-3). Overall, seroprevalence to BVDV was 36±7% (SE); seroprevalence varied by district (19–42%). BHV-1 seroprevalence was 67±4%; variation by district was similar to that of BVDV. However, the first 80 serum samples tested by BHV-1 ELISA all had a strong background reaction with the control antigen. Therefore, these sera were adsorbed to a homogenate of non-infected bovine kidney cell line (MDBK) and re-tested by ELISA. The non-specific reactivity was significantly reduced (p < 0.001 by Wilcoxon's signed-rank test). Compared to the virus-neutralisation (VN) test, the adsorbed BHV-1 ELISA showed 94% agreement and gave a κ value of 0.84, indicating that the adsorption did not interfere with test accuracy. Seroprevalence against BRSV was 85±3%, and showed differences across districts. Most of the cows (94±2%) were seropositive to PIV-3, and there were no significant differences among districts.     

A serological survey for bovine immunodeficiency-like virus in Ontario dairy cattle and associations between test results, production records and management practices.

A chemiluminescence Western blot analysis (WBA) for detecting antibovine immunodeficiency-like virus (BIV) antibodies, had good repeatability. The test was subsequently applied to a bank of serum samples from 928 adult cows from 265 herds in Ontario; the number of cows sampled within each herd ranged from 1 to 13. The overall prevalence of anti-BIV antibodies among cows was 5.5% with a 95% confidence interval of 4.2% to 7.2%. In contrast, 18.1% of herds had at least one reactor among cows tested, resulting in a herd-prevalence confidence interval of 13.8% to 23.4%. These estimates of prevalence were in the same range as previous reports from the US and Europe. Bovine immunodeficiency-like virus may have a worldwide distribution. Unfortunately, BIV test sensitivity and specificity are difficult to estimate because virus isolation is inefficient. Therefore, the apparent prevalences could not be adjusted for test sensitivity and specificity, to estimate the true prevalence of infection. The serum samples had previously been tested for antibodies to bovine leukemia virus (BLV). There were no significant associations between BIV and BLV test results. Least squares regression was used to investigate potential associations between BIV test results and selected production indices.(ABSTRACT TRUNCATED AT 250 WORDS)     

A single dilution enzyme-linked immunosorbent assay for the quantitative detection of antibodies to bovine herpesvirus type 1

Three serological assays were compared for detection of antibodies to bovine herpes-virus type 1. These were virus neutralization (VN), enhanced complement fixation (CF) and enzyme-linked immunosorbent assay (ELISA). The ELISA was developed using an infected cell lysate antigen and purified virus and was optimized in relation to antigen and antisera dilutions. The CF assay was enhanced by the addition of bovine complement. These 3 assays were compared for detection of: specific virus antibody titers; sero-conversions; early antibody response in experimentally-infected cattle. Both ELISA end-point titers and single dilution values were found to be more sensitive than the CF or VN assays for specific antibody level quantitation. With a single dilution ELISA test procedure a correlation was obtained between ELISA values and VN titers. Using the single dilution ELISA test the assay also detected antibodies in experimentally-infected cattle before either the VN or CF assays, and agreed with the VN test in 35/38 seroconversions found by 4-fold or more VN changes between acute and convalescent paired sera from naturally-infected animals. The single dilution ELISA was a rapid and sensitive test for routine antibody detection in bovine sera.     

Natural transplacental infection of dairy calves with bovine immunodeficiency virus and estimation of effect on neonatal health

Many experimental infection studies with bovine immunodeficiency virus (BIV) have been conducted, but neither virus transmission under natural conditions nor longitudinal clinical effects of naturally occurring infections in non-experimental populations are well explored. We tested the hypotheses that BIV is transmitted across the placenta during gestation and that intragestionally infected calves are at increased risk of neonatal disease. A cohort of 59 dairy cows on one farm were enrolled at parturition and the BIV serostatus of the cows and their pre-colostral calves determined with an indirect fluorescent-antibody assay. Moreover, the enrolled calves were monitored thrice weekly for specific clinical signs through the duration of the 30 day neonatal period and the occurrence of clinical signs analyzed for association with calf pre-colostral BIV serostatus and dam BIV serostatus. Confounding due to calf passive immunity and season of birth were also explored. Forty percent of seropositive cows (14/35) gave birth to seropositive calves but no seropositive calves (0/19) were born to seronegative dams (estimated relative risk 16, 95% exact confidence interval 2.6–5.8×10²⁹). Calf pre-colostral BIV serostatus was not associated with the occurrence or frequency of clinical signs — but dam BIV serostatus was associated with the odds of occurrence of calf hyperthermia and with the frequency of occurrence of calf hyperthermia and hyperventilatory events. This study is inconclusive about the effects of prenatal BIV infection on neonatal health — but it does provide evidence for the natural occurrence of transplacental BIV infection.