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1.
BLV detection by the syncytial test was performed in 27 heifers experimentally and naturally infected by the enzootic bovine leukosis virus (BLV). The presence of BLV was demonstrated in 94.7% of the animals. The bovine foetal spleen cells (FBS) were found to be suitable for the syncytial test. Positive animals not reacting to infection by the production of anti-BLV antibodies were identified during the syncytial-test investigation. The importance of this finding for the programme of controlling enzootic bovine leukosis on farms is discussed. As suggested by the results, temporary occurrence of anti-BLV antibodies followed by their disappearance can be observed together with a negative result of the syncytial test in some circumstances. The discussion deals with the problems of the determination of anti-BLV antibodies in milk, and/or milk secretion, by the ELISA method.  相似文献   

2.
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3.
OBJECTIVE: To estimate the sensitivity, specificity and detection limits for two bulk-milk enzyme-linked immunosorbent assays, the Svanovir BLV-gp51-Ab and the Lactelisa BLV Ab Bi indirect tank 250, for the detection of antibody to bovine leucosis virus in milk. PROCEDURE: Milk samples from 27 cows known to have enzootic bovine leucosis (EBL) were serially diluted with milk from a herd known to be free from the disease. The dilution at which antibodies could no longer be detected by each test was determined. A total of 1959 bulk-milk samples submitted to a laboratory for the Victorian (EBL) eradication program were tested with both the Svanovir and the Lactelisa assays. A Bayesian approach was used to calculate maximum-likelihood estimates of test sensitivity and specificity. An additional 660 bulk-milk samples were tested with both the Svanovir and the Lactelisa assays. Herds that had positive results on either or both of the assays were subjected to blood or milk testing of individual cattle. RESULTS: The dilution of milk at which the Svanovir assay failed to detect enzootic bovine leucosis antibody in half of the samples was 1 in 40, whereas the comparable value for the Lactelisa was 1 in 200. Computer modeling of the operating characteristics of the Svanovir assay indicated that the sensitivity of that assay would be considerably lower than that for the Lactelisa, and the specificity was estimated to be higher. Evaluation of the assays using 660 bulk-milk samples showed that the Lactelisa assay detected four infected herds that were not detected by the Svanovir test. No false positive results were recorded for either assay. CONCLUSION: Use of the Lactelisa assay in the Victorian EBL eradication program will enhance disease detection and eradication, but may also result in an increased frequency of false positive bulk-milk test results.  相似文献   

4.
An indirect double-antibody sandwich (IDAS) enzyme-linked immunosorbent assay (ELISA) using milk samples was developed to identify cows infected with bovine leukemia virus (BLV). Two monoclonal antibodies (McAbs) were used. One, which was directed against BLV core protein p24, was used to coat ELISA plates; the other was used to prepare a horseradish peroxidase (HRP) conjugate directed against bovine immunoglobulin. The IDAS-ELISA detected antibodies directed against BLV-p24 in 97% of the milk samples collected from known seropositive cows identified by the agar gel precipitation test (AGTP). Even when milk samples were diluted 1:50, 93% of the seropositive cows were identified. Only 0.43% of the 4000 milk samples collected in The Netherlands reacted nonspecifically. Nonspecific binding disappeared, however, when these samples were diluted 50 times in BLV-negative milk. In a comparative evaluation of BLV test-kits in various European laboratories, our IDAS-ELISA using McAb directed against p24 was one of the most sensitive.  相似文献   

5.
In utero transmission of bovine leukemia virus   总被引：8，自引：0，他引：8  
In an initial study, 18 calves born to cows persistently infected with bovine leukemia virus (BLV) were tested for infective virus and antibodies at birth, and no infected or seropositive animals were found. Four of these calves were maintained in quarters where infected animals were housed, and 3 of the 4 subsequently became infected. These were probably contact infections acquired during, or at some time after, birth. The remaining 14 calves were kept in isolation pens in a building housing no infected cattle. None of this group was found to be BLV infected during 1 year of observation. In further studies, 15 pregnant cows inoculated with BLV became infected. One abortion, considered to be unrelated to the BLV inoculation, occurred 38 days later. The remaining 14 cows gave birth to 1 dead and 14 live calves. The dead calf and its live twin were seropositive for BLV at birth, indicating that they had been infected in utero. The remaining 13 calves were negative for BLV antibodies at birth and remained so during 1 year of observation.  相似文献   

6.
A microbiological cancer test, previously verified in men and dogs using a clostridium strain (Clostridium butyricum CNRZ 528), was applied to cattle infected with bovine leucosis virus (BLV). An extended period of time was allowed to pass after infection with BLV, which had been checked up through specific serological and virological examinations. The cattle belonged to different age groups and stages of infection (with and without haematological alterations [preleukosis], with incipient tumour development [swelling of externally visible and palpable lymph nodes]). Controls included BLV-infected cows as well as test animals to which isotonic saline had been applied or healthy BLV-free cattle in which the clostridium strain had been used. The serological investigation was carried out in a blind test. 3 of 6 BLV-infected spore-treated heads of cattle responded positively to the cancer test, while the other 3 were negative. The 3 cows with positive cancer test were haematologically and serologically leucosis-positive animals with clinically detectable enlargement of lymph nodes. The 3 negative ones of this group, also serologically and haematologically leucosis-positive, were younger animals without signs of tumorous process. 3 spore-treated BLV-free cows and 2 BLV-infected animals, treated with isotonic saline, were cancer test-negative, as well. Finally, 4 BLV-infected and 2 BLV-free cattle, all of them without spore injection, were completely cancer test-negative. 1 cow of the BLV-infected group did not produce spore antibodies after spore treatment, while 1 cow of the BLV-free untreated control group developed spore antibodies.  相似文献   

7.
OBJECTIVE: To develop a blocking ELISA for detection of bovine leukemia virus (BLV) antibodies that is comparable to a radioimmunoprecipitation (RIP) assay, to evaluate use of this ELISA for identification of BLV-infected herds, and to develop a polymerase chain reaction (PCR) assay for direct diagnosis of infection with BLV. SAMPLE POPULATION: Serum samples and pooled bulk-tank milk samples from cattle. PROCEDURE: The blocking ELISA was developed, using BLV gp51 as antigen, captured by a selected bovine polyclonal serum. A nested PCR was conducted with primers specific for a segment of the pol region of the BLV genome. RESULTS: Sensitivity and specificity of the ELISA were comparable to those of the RIP assay. Use of the ELISA on pooled milk samples allowed identification of herds in which prevalence of BLV infection among lactating cows was as low as 2.5%. Pooled milk samples from BLV-free herds did not react in the ELISA. All cattle that had positive results for the nested PCR had BLV antibodies, but cattle with consistantly low antibody titers required examination of sequential DNA samples to detect viral sequences. None of the 63 antibody-negative cattle had positive results for the PCR. CONCLUSIONS AND CLINICAL RELEVANCE: This ELISA is a highly specific and sensitive assay for the detection of BLV antibodies in serum and milk samples of cattle. Examination of pooled milk samples with the ELISA provides a reliable, practical, and economic procedure for identification of BLV-infected herds. The nested PCR also constitutes a specific procedure for direct diagnosis of infection with BLV.  相似文献   

8.
BACKGROUND: The relation between calf bovine leukosis virus (BLV) infection status and colostrum ingestion is unclear. Two conclusions have been drawn from previous studies. One suggests that colostrum ingestion transmits BLV to neonatal calves. The second suggests that colostral antibodies are protective. HYPOTHESIS: Colostrum from BLV-positive cattle is protective in naturally exposed calves. ANIMALS: Twelve colostrum-deprived Holstein calves and 20 colostrum-fed Holstein calves born to BLV-infected cows. METHODS: Prospective study. Colostrum-deprived calves were tested weekly by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) tests for BLV antibody and provirus for 12 weeks or until the animal became positive for BLV infection. Colostrum-fed calves were fed colostrum derived from BLV-positive cows. Thereafter, ELISA and PCR tests for BLV antibody and provirus were performed every other week until 2 consecutive negative ELISA tests or 1 positive PCR test was achieved. The proportion of calves that converted to BLV-positive status was calculated for each group and compared between groups by using the Fisher exact test. RESULTS: Four of 12 colostrum-deprived calves (33%) became BLV positive, whereas 0 of 20 colostrum-fed calves (0%) became BLV positive. The proportion of calves that became infected was significantly higher in the colostrum-deprived group (P = .014). CONCLUSIONS AND CLINICAL RELEVANCE: Calves born to BLV-positive cows are exposed during parturition, and a proportion of these calves will become infected with BLV. Administration of colostrum from BLV-positive cows greatly decreases the risk of infection.  相似文献   

9.
Haematological parameters and reactivity of lymphocyte antigens to monoclonal antibodies were studied over a 10-month period in sheep experimentally infected with bovine leukaemia virus (BLV). BLV-inoculated animals seroconverted within 1 month and showed a significant lymphocytosis 2-6 weeks after infection. Control animals inoculated with BLV-free lymphocytes showed a stronger and more immediate neutrophil response than those inoculated with BLV-positive lymphocytes. One month after infection, BLV-inoculated sheep showed a relative increase of cells bearing antigens T4, T6, T8 and T19, and 10 months into the trial, MHC II lymphocytes increased, T6 remained elevated, but T4 helper cells were significantly decreased in number. Lymphoma tissue showed the presence of T8 cells, and lymph nodes from seroconverted sheep had areas of concentrated T4 staining cells. These results demonstrate responses in cellular immune mechanisms to infection with BLV.  相似文献   

10.
We have established experimental models of bovine leukemia virus (BLV) infection followed by progression to persistent lymphocytosis (PL) positive (BLV+PL+) or PL negative (BLV+PL-) stages of infection. Two out of six BLV infected animals developed PL+ 4 weeks after BLV infection. One other animal became PL+ late in the course of infection and three infected animals stayed PL-. These animals (PL-) exhibited transient lymphocytosis 3-4 weeks after infection and sustained PL- lymphocyte counts up to 24 weeks after infection. Competitive RT-PCR analysis of IFN-gamma mRNA expression revealed that peripheral blood mononuclear cells (PBMC) of animals with PL+ status developed by 4 weeks after infection had augmented IFN-gamma mRNA expression 3-4 weeks after BLV infection. However PBMC of animals that sustained a long-termed PL- lymphocyte count had elevated IFN-gamma mRNA expression 1-24 weeks after infection. Competitive RT-PCR analysis of IL-2 mRNA expression showed an increase in the levels of IL-2 mRNA in PL animals. Interleukin-10 (IL-10) mRNAs expression were elevated both in PL+ and PL- animals from 3 and 12 weeks after infection respectively. We suggest that early and extended expression of cellular response cytokines may delay the progression to PL+ in enzootic bovine leukemia.  相似文献   

11.
Radioimmunoassay (RIA), using the virion glycoprotein antigen, was applied in an attempt to eradicate bovine leukemia virus (BLV) infection from a herd in which virtually all the adult cattle are infected. Considering that most calves born to BLV-infected cows are negative for BLV at birth and remain negative for the first several months of life, the eradication program was based on the identification and isolation of the BLV-free calves born to infected cows. Twenty-five calves raised on colostrum and milk from their infected dams were classified as BLV-free on the basis of negative results in the RIA at 6 to 8 and 9 to 11 months of age. These animals were maintained in either complete (10 calves) or partial (15 calves) isolation from infected cattle and were examined at regular intervals for BLV and BLV antibodies. With the exception of 1 calf in the group raised in partial isolation, the animals have remained free of BLV up to the time of the last evaluation, when they were 32 to 35 months old. At these ages, more than 90% of the nonisolated cattle in the herd are BLV-positive. The data also show that this eradication trial would have failed if, in the initial procedure used to classify the calves as BLV-free, the agar gel immunodiffusion test instead of the RIA had been used. Inasmuch as the 25 calves in this study were fed colostrum and milk from their dams, the fact that only 1 of the calves became infected during the 26 to 29 months of observation provides further evidence that milk-borne transmission of BLV is infrequent and perhaps inconsequential.  相似文献   

12.
Serum, milk and urine specimens were taken from 15 bovine leukaemia virus (BLV)-positive and 20 BLV-negative cattle which had been determined previously to be infected or not by the use of a monoclonal enzyme-linked immunosorbent assay (ELISA). An ELISA was performed on the samples for the detection of IgG1 antibodies to the BLv surface glycoprotein, gp 51. The three types of samples had parallel optical density (OD) values apart from three urine samples which, although accepted as- negative for anti-BLv antibodies, had numerically higher ODS than those of control BLV-negative animals. Therefore, detection of IgG1 antibodies against BLv in the urine of naturally infected animals could be an indication for the use of urine for diagnosis of BLV infection.  相似文献   

13.
The early diagnosis of bovine leukosis virus (BLV) infection, the aetiological agent in enzootic bovine leukosis, is important for the implementation of control measures. BLV infection is currently assessed by the detection of circulating antibodies against the viral envelope protein, gp51. However, this approach has shortcomings in the time taken to detect anti-BLV antibodies (three to four weeks after infection), and in the failure to detect antibodies in some animals. Clearly a technique such as the polymerase chain reaction (PCR), which directly detects the presence of viral DNA, has advantages over methods designed to measure host antibodies. The use of PCR for the detection of proviral DNA in an affected DNA sample with as little as 10(-5) micrograms of host DNA using agarose gel electrophoresis followed by ethidium bromide staining is described here. It was possible to improve the sensitivity of this assay by using hybridisation analysis with a BLV gene probe. PCR used in combination with hybridisation analysis will provide a sensitive diagnostic assay to detect BLV when antibody tests give weakly positive or equivocal results.  相似文献   

14.
Serum and milk samples from 1229 cows on 22 Ontario dairy farms were individually tested for antibodies specific for bovine leukosis virus (BLV) and Neospora caninum by enzyme-linked immunosorbent assay (ELISA). Antibodies against BLV were present in 361 serum samples (29.4%) and 369 milk samples (30.0%). Comparing the 2 tests, agreement was almost perfect (k = 0.86; 95% CI = 0.83 to 0.90) and the proportions of samples positive were not significantly different (P = 0.56). Both tests identified the same 3 herds free of bovine leukosis virus. Antibodies against N. caninum were detected in 138 serum samples (11.2%), and 111 milk samples (9.0%). Agreement between the 2 tests was moderate (k = 0.52; 95% CI = 0.43 to 0.59). Four herds were free of neosporosis by the serum test, while 10 herds were negative by the milk test. The ELISA on milk samples facilitates sample collection to classify herds free of BLV; the milk N. caninum ELISA was less reliable in predicting herd-level infection.  相似文献   

15.
OBJECTIVE: To determine the interval to provirus and serum antibody detection (via PCR assay and ELISA, respectively) in calves after experimental inoculation with bovine leukemia virus (BLV). ANIMALS: 8 colostrum-deprived, BLV-negative Holstein bull calves (> or = 6 weeks old). PROCEDURES: Via IM injection, each calf received a fresh whole-blood inoculum (day 0) calculated to contain 2 x 10(6) lymphocytes. Blood samples for the ELISA and PCR assay were collected from calves immediately prior to inoculation and weekly thereafter for 7 weeks. Mean and median number of weeks to PCR-detected conversion of BLV status and seroconversion were calculated. Point sensitivity and cumulative sensitivity of the 2 assays were calculated at each sample collection. At each sampling time, the proportion of calves identified as infected by the cumulative weekly ELISA and PCR assay results was compared by use of a Fisher exact test. RESULTS: In 5 calves, conversion of BLV status was detected via PCR assay before seroconversion was identified. However, seroconversion preceded PCR-detected conversion in 2 calves. In 1 calf, both assays yielded positive results at the same test date. These differences were not significant. CONCLUSIONS AND CLINICAL RELEVANCE: In experimentally inoculated BLV-negative calves, conversion of BLV status was detected via PCR assay more quickly than via ELISA; this difference was not significant and probably not clinically important. The PCR assay may be useful as a confirmatory test in animals of exceptional value; tests based on viral identification may become critically important if vaccines against BLV infection are developed and marketed.  相似文献   

16.
Lymphoblastic leukaemia, preceded by a significantly increasing percentage of prolymphocytes in peripheral blood smears for from 12 to 68 weeks before death was a feature of sheep which developed lymphosarcoma following inoculation with the Australian strain of bovine leucosis virus (BLV). Lymphocytosis and/or the appearance of immature cells were a reliable predictor of tumour formation in sheep, but not in cattle. There was a terminal lymphoblastic leukaemia in only 43 of 84 cattle with lymphosarcoma. Differences in the morphological appearance and glycogen content of the leukaemic lymphoblasts of sheep and cattle were observed. In spite of these differences the high frequency of lymphocytosis and lymphosarcoma in experimentally infected sheep suggests that they could be a useful model for studying the pathological and immunological responses to BLV infection.  相似文献   

17.
The objective of this study was to investigate the associations between herd-level bovine leukemia virus (BLV) status and herd-level management and production variables. The study population consisted of 1330 cows sampled from 102 Ontario dairy herds. The individual-cow prevalence of BLV infection in the population (based on AGID testing) was 23%, with 69.6% of herds having one or more positive animals. The herd-level explanatory variables were divided into two datasets containing winter housing variables and all non-seasonal variables, and summer housing variables and all non-seasonal variables. In both datasets, multivariable analyses found a negative association between herd-level milk production and BLV status, and positive associations between weaning age and purchasing animals from outside sources, and BLV status. Housing pre-weaned calves in hutches or separate calf buildings in either season was associated with an increased risk of BLV. The model containing winter housing variables also included positive associations between contact with older animals and BLV status, and between BLV status and the facilities used to house dry cows in the winter.  相似文献   

18.
The bovine lymphoblastoid BL 20 cell line derived from a case of sporadic bovine leukosis when inoculated into sheep did not induce an antibody response directed against bovine leukosis virus (BLV) structural proteins. Sheep were inoculated twice with the BL 20 cell line and then challenged with BLV infected lymphocytes. Three out of four sheep challenged four weeks after BL 20 inoculation did not develop BLV antibodies. Of the 12 sheep challenged later, three sheep did not develop BLV antibodies. BLV was isolated from all the seropositive animals and from none of the seronegative animals.  相似文献   

19.
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.  相似文献   

20.
Bovine leukaemia virus (BLV) is the causative agent in enzootic bovine leukosis a disease occurring worldwide. This virus is normally detected by the agar gel immunodiffusion or ELISA assays which rely on the appearance of antibodies to a major surface protein of the virus, gp51, present in the serum of infected cattle. We have used the polymerase chain reaction, which depends on the amplification of specific DNA sequences as a sensitive assay for the detection of BLV. It was possible to detect proviral DNA in 100 pg of tumour DNA from an infected host using agarose gel electrophoresis followed by ethidium bromide staining. The sensitivity of the assay was increased by two log orders when hybridization analysis, using a BLV proviral DNA probe, was used in combination with amplification of the DNA. Proviral DNA was detected in both lymphocytic and tumour DNA and at all stages of infection in cattle.  相似文献   

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