Bovine leukaemia virus infection in New Zealand cattle

Bovine Leukaemia Virus (BLV) infection in New Zealand cattle was investigated. In a national survey of 5000 sera from 500 herds, BLV antibody was not detected. An additional 1062 sera from 140 herds were tested and 3 sera were positive. In the herd of origin of one of these 3 sera, 22.6% of cattle were serologically positive for BLV. Where cases of bovine lymphosarcoma had been diagnosed, 38 of 39 herds tested were negative for BLV antibody. Within the remaining herd, 36% of cows tested were serologically-positive. BLV was isolated from 2 serologically positive cows in this herd.     

Survey for antibodies to leukemia (C-type) virus in cattle.

Serums from 4,394 dairy cattle in 100 herds and from 2,794 beef cattle in 50 herds were tested for antibody to the bovine (C-type) leukemia virus (BLV), using the agar gel immunodiffusion test. Reactors were found in 66% of the dairy herds (10.2% of the cattle) and in 14% of the beef herds (1.2% of the cattle). The prevalence of reactors was examined with respect to age, herd size, and sex. Few of the reactors were less than 2 years old. There was a high percentage of reactors in small dairy herds (less than 50 cattle). In 22 dairy herds (1,354 cows and 96 bulls), the rate of infection in cows was compared with that in bulls. In those herds, 13.5% of the cows and 10.4% of the bulls were reactors.     

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.     

Seroprevalence of infection with Mycobacterium avium subspecies paratuberculosis, bovine leukemia virus, and bovine viral diarrhea virus in maritime Canada dairy cattle

The purpose of this study was to survey the seroprevalence of infection with the agents of production-limiting diseases in dairy cattle in New Brunswick, Nova Scotia, and Prince Edward Island. In 30 randomly selected herds per province, 30 cattle per herd were randomly selected and tested for antibodies to bovine leukemia virus (BLV) and Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis), while 5 unvaccinated cattle over 6 months of age were tested for antibodies to bovine viral diarrhea virus (BVDV). For BLV, 20.8% (15.8% to 27.0%) of cows were positive, and 70.0% (60.3% to 79.7%) of herds had at least one positive cow. In BLV-positive herds, the average BLV prevalence was 30.9% (24.8% to 37.2%). For M. paratuberculosis, 2.6% (1.8% to 3.9%) of cows were positive, and 16.7% (8.8% to 24.5%) of herds had at least 2 M. paratuberculosis-positive cows. In M. paratuberculosis-positive herds, the average M. paratuberculosis prevalence was 8.5% (6.9% to 10.1%). For BVDV, 46.1% (35.5% to 56.7%) of herds had at least 1 BVDV-positive animal with a titer greater than or equal to 1:64.     

Evaluation of a bulk-milk ELISA test for the classification of herd-level bovine leukemia virus status

The results of a commercial bulk-milk enzyme-linked immunosorbent assay (ELISA) test for herd-level bovine leukemia virus (BLV) status were compared to results obtained from individual agar-gel immunodiffussion (AGID) testing on sampled cattle. A positive herd was defined as a herd having one or more AGID-positive animals. The estimated true herd status was based on the sensitivity and specificity of the AGID test and the number of cattle sampled per herd. Ninety-seven herds were used, with a mean of 13 cows sampled per herd. The AGID test indicated an apparent herd prevalence of 70.1%. After accounting for the number of cows sampled and the sensitivity and specificity of the AGID test, the estimated true herd prevalence of BLV was 52.3%. The ELISA test identified 79.4% of herds as positive for BLV, and had an apparent sensitivity and specificity of 0.97 and 0.62, respectively. However, after accounting for the sensitivity and specificity of the AGID test in individual animals, the specificity of the ELISA test was 0.44. The ELISA test was useful for identifying BLV-negative herds (i.e., ruling out the presence of BLV infection in test negative herds). With the moderately low specificity, herds identified as positive by the ELISA test would require further testing at the individual or herd level to definitively establish their BLV status.     

Association between bovine-leukosis virus seroprevalence and herd-level productivity on US dairy farms

Bovine-leukosis virus (BLV; also termed ‘bovine-leukemia virus’) is a retrovirus that primarily affects lymphoid tissue of dairy and beef cattle. Our objective was to investigate the association between BLV infection and annual value of production (AVP) on dairy herds within the United States, as part of the USDA National Animal Health Monitoring System’s 1996 dairy study. 1006 herds (in 20 states) with at least 30 dairy cows were interviewed during 1996. The agar-gel immunodiffusion test was used to detect serum antibodies to BLV. 10–40 cows from each herd were tested and each tested cow was classified as negative or positive based on results of a single test.

A multivariable regression model was used with the 976 herds with complete data for analysis. When compared to herds with no test-positive cows, herds with test-positive cows produced 218 kg per cow (i.e. 3%) less milk. The average reduction in AVP was $ 59 per cow for test-positive herds relative to test-negative herds. For the dairy industry as a whole, BLV seropositivity was associated with loss to producers of $ 285 million and $ 240 million for consumers. Most of this $ 525 million industry loss was due to reduced milk production in test-positive herds.     

Serological detection of multiple retroviral infections in cattle: bovine leukemia virus, bovine syncytial virus and bovine visna virus

Individual experimental animals used in our studies on bovine leukemia virus (BLV) are routinely screened for the presence of antibodies to the three bovine lymphotropic retroviruses. We utilized these screening methods to examine frozen sera from eight herds for antibodies to BLV, bovine visna virus (BVV) and bovine syncytial virus (BSV). Serum samples from 235 animals in four dairy and four beef herds were analyzed. Detection methods used included indirect fluorescent antibody tests of virus-infected cell cultures (BLV, BSV, BVV) and agar gel immunodiffusion (BLV). Sera from the BLV-infected animals in the dairy herds showed the highest single (50%, 49/97) and multiple (30%, 29/97) infections compared with 5% (7/138) and less than 1% (1/138), respectively in the beef herds. Single BVV infections were not detected in the dairy herds, but 11% (11/97) of the sera contained antibodies to BVV plus BLV or BSV. Five sera from beef cattle had antibodies only to BVV and four were obtained from one herd. Only one beef serum of the 138 tested demonstrated multiple antibodies (BLV, BVV).     

Prevalence and regional distribution of paratuberculosis in dairy herds in The Netherlands

In the Netherlands a survey was conducted to estimate the prevalence of paratuberculosis in dairy herds. In total 15822 cows of at least 3 years of age, belonging to 378 herds were tested using an absorbed ELISA. Of these herds, 55% (n=207) had one or more serologically positive cows. Of the positive non-vaccinated herds, most had one (n=98) or two (n=49) positive cows. The percentage positive cows per herd was 2.5+/-3.2%.The true prevalences on cow and herd levels, based on a test sensitivity that ranged from 0.3 to 0.4 and a specificity that ranged from 0.985 and 0.995, were estimated at 2. 7-6.9% and 31-71%.Seven herds had been vaccinated against paratuberculosis and these herds had a significantly higher percentage of serologically positive cows (23%) than the non-vaccinated herds (2.5%).In conclusion, a small percentage of the dairy cows and a high percentage of the dairy herds in the Netherlands is serologically positive. The percentages true infected cows and herds are difficult to estimate precisely due to uncertainties in test sensitivity and specificity.     

Risk factor analysis of bovine leukemia virus infection in dairy cattle in Egypt

Identification of the risk factors associated with Enzootic bovine leukosis (EBL) is essential for the adoption of potentially prevention strategies. Accordingly, our objectives were to determine the geographic distribution of Bovine Leukemia Virus (BLV) infection and identify the risk factors associated with cow-level BLV infection in the Egyptian dairy cattle. A cross-sectional study was conducted on 1299 mixed breed cows distributed over four provinces in the Nile Delta of Egypt in 2018. The randomly selected cows on each farm were serologically tested for BLV, and the cow’s information was obtained from the farm records. Four variables (geographic location, herd size, number of parities, and age) were used for risk analysis. A total of 230 serum samples (17.7 %) were serologically positive for BLV. The highest prevalence of BLV infection was associated with parity (OR = 3.4, 95 %CI 2.4–4.9) with 80 % probability of being BLV-positive at parity ≥5, followed by herd size (OR = 1.8, 95 %CI 1.4–2.2). However, geographic location seems to have no impact on the prevalence of BLV infection in Egypt. Our findings strongly indicate that the intensive surveillance and effective prevention strategies against BLV infection in Egypt should be provided to multiparous cows with ≥5 parities and live in large farm with more than 200 cows.     

Comparison of four serologic tests for the detection of antibodies to bovine leukemia virus

Four tests for detection of antibodies to bovine leukemia virus (BLV) were compared. The sera that were tested came from cattle in naturally infected commercial dairy herds, cattle that were infected under experimental conditions, and cattle in an isolated BLV-free herd. The tests that were compared included a radioimmunoprecipitation assay (RIA) with p24 antigen, a RIA with glycoprotein (gp) antigen, an agar-gel immunodiffusion (AGID) test with gp antigen, and a virus-neutralization (VN) test that was based on inhibition of BLV-induced syncytia in cell culture. Results of the 4 serologic tests agreed for 96.8% of the sera from cattle in commercial herds. The gp RIA detected the greatest number of positive sera (188); it was followed in turn by the p24 RIA (187), the VN test (183), and the AGID test (176). The gpd RIA titers of the 12 sera that gave negative AGID results were 175 or less. In RIA, the percentage of precipitation of labeled antigen by positive sera was almost always higher with gp antigen than with p24 antigen. Satisfactory sensitivity in the p24 RIA required the acceptance of a low level of antigen precipitation, 15%, as a positive test. In the gp RIA, however, almost all positive sera precipitated at least 50% of the labeled antigen. Nonspecific precipitation of antigen in the RIA by sera from BLV-free cattle ranged from 4% to 10%. Examination of sequential serum samples from 17 experimentally infected cattle showed that BLV antibody was first detected 2 to 8 weeks after inoculation. In 9 cattle, seroconversion was detected simultaneously by all of the tests. Results from the other 8 cattle indicated that seroconversion could be detected first by p24 RIA, followed by the gp RIA and the VN test. The longest interval between RIA seroconversion and AGID seroconversion was 10 days. Monthly tests of sera from 10 laboratory cattle that were infected by contact exposure showed that 7 animals seroconverted in all tests at the same time. Two cattle were positive first in RIA, but the next month they were also positive in the VN and AGID tests. One animal was positive in the RIA and the VN test for 2 months before antibody was detected by AGID.     

Eradication of Prototheca zopfii infection in a dairy cattle herd

Protothecosis is a severe form of mastitis in dairy cows caused by colorless algae of the genus Prototheca. Since P. zopfii is highly resistant to all known chemotherapeutics, infected cows must be removed from the herd. Eradication measures are difficult since many chronically infected cows may become intermittent shedders. Therefore, cultural methods are insufficient for control measures. In order to eradicate Prototheca zopfii-mastitis in dairy cattle herds, two isotype specific indirect ELISA for detection of IgA and IgG1 in whey were used in a dairy herd highly affected with protothecal mastitis. All cows (n = 313) were tested four times in intervals of six months. Milk specimens were examined in parallel by cultivation and serologically using two indirect ELISA systems for specific IgA and IgG1 in whey. Cows tested Prototheca positive were consequently separated from the herd and slaughtered. At the first examination, 15.6% of the animals were found positive by culture, and 23.3% were positive in at least one of the ELISA systems. Within two years, protothecal prevalence and incidence decreased to zero indicating that the eradication strategy used was successful. In summary, serological identification of P. zopfii-infected lactating cows is an useful tool to eradicate protothecal bovine mastitis in infected herds.     

Study on prevalence of bovine viral diarrhoea virus (BVDV) antibodies in 29 Italian dairy herds with reproductive problems

An epidemiological survey on prevalence distribution of antibodies to BVDV was carried out in dairy cattle herds during 1995-1996 in northern Italy. A total of 704 serum samples from 29 non-vaccinated herds reported to have reproductive problems were tested for serum neutralising antibodies. In each herd, sampling was based on the stratification by age into five classes (< 6 months old calves, 6-12 months old calves, pregnant heifers, uniparous, pluriparous). Overall, 53.3% of samples were serologically positive, with the lowest ratio in 6-12 months old calves (37.9%) and the highest in pluriparous cows (71.2%).     

Risk factors for Brucella spp. infection In dairy cattle farms in Asmara, State of Eritrea

A cross-sectional study was conducted to identify risk factors for herd infection by Brucella spp. in dairy cattle in the suburbs of Asmara, Eritrea. Data were collected from 64 herds, randomly selected from a total of 99 herds with a minimum herd size of 9 cows. A questionnaire was used to gather data on management, hygiene and herd structure. Serum samples collected from all pregnant heifers, cows and bulls, were screened for Brucella infection by the Rose Bengal test (RBT), and all RBT-positive sera re-tested with the complement-fixation test (CFT) for confirmation. A seropositive herd was defined as one in which at least one animal tested positive in the CFT. There were 23 (36%) positive herds among the 64 studied. Both multiple logistic and multiple betabinomial regression modeling were used to analyze the data. Mixed-breed herds, compared to single (exotic)-breed herds, were found to be independently associated with increased herd seroprevalence (OR=5.2, 95% confidence interval 1.4–18.7) in the multiple logistic model with the herd infection status as the dependent variable. The importance of this variable was supported by the multiple betabinomial regression model (OR=3.3, 1.4–7.6) with animal-level prevalence within herd as the outcome variable. Both models also revealed the presence of a negative association between seropositivity and cattle stocking density.     

Diagnosis of naturally occurring Fasciola hepatica infections in cattle with an enzyme-linked immunosorbent assay

Sera from 100 herds of cattle located in the state of Washington were examined with an enzyme-linked immunosorbent assay for antibody to Fasciola hepatica in a screening procedure that included 5 to 10 samples/herd. Twenty-eight herds contained infected cattle and F hepatica was most prevalent in 3 distinct geographic areas. Subsequent retesting of all sera available from 14 herds (mean of 109 samples/herd) revealed that the screening procedure correctly detected 7 of 7 operations in which greater than 40% of samples were positive or suspect and 3 of 3 operations in which 12% to 13% of the samples were positive or suspect. One of 3 herds considered negative after screening was found to contain a few (7%) positive samples and 1 herd considered possibly infected was negative on retest. These results were compared with those obtained by fecal examination for F hepatica eggs in 9 of the 14 herds. A good correlation (5 of 5) was found in which a high percentage (48% to 85%) of sera were positive or suspect. Fasciola eggs were not found in samples from 2 herds with few (7% to 12%) positive or suspect sera or in 2 herds that were negative by an enzyme-linked immunosorbent assay.     

The vaginal mucus agglutination test in the diagnosis of bovine brucellosis

Of 1140 vaginal mucus agglutination tests (VMAT) on specimens obtained in 1971-72 from 663 dairy cows in seven herds infected with brucellosis, 97 were positive. When the VMAT was positive one or more serological tests were also positive. Of the 97 corresponding serum agglutination tests 80 sera had titres of more than 533 international units. Only 69.8 per cent of VMAT from serologically positive cows were positive. No evidence was found of non-specific agglutinins in vaginal mucus and positive VMAT reactions appeared to be specific for field infection. Three cows showed evidence of local agglutinins in the vagina. Hence herd testing by VMAT has no advantage over tests of blood serum but the test could be an aid in establishing whether individual cattle are infected.     

Risk factors associated with Mycobacterium avium subspecies paratuberculosis seropositivity in Canadian dairy cows and herds

Our objective was to determine the risk factors associated with the seroprevalence of Mycobacterium avium subspecies paratuberculosis (MAP) in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Neospora caninum (NC). Serum samples from 30 randomly selected cows, where available, in 315 herds from seven provinces were tested for antibodies against BLV, MAP and NC using commercially available enzyme-linked immunosorbant assay (ELISA) test kits, while five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV. We used a zero-inflated negative-binomial (ZINB) multivariable model to determine simultaneously the risk factors associated with the count of MAP-seropositive cows in a herd, and the odds of herds having no MAP-seropositive cows as compared to having one or more MAP seropositive cows in a herd. The following factors were significantly positively associated with the count of MAP-seropositive cows: "more than one cow in the maternity pen", "group-housing for pre-weaned calves in winter", "open heifers purchased during the last 12 months", "beef cattle direct (nose-to-nose) contact", "BVDV-seropositive herds (> or = 1 animal with > or = 1:64 titer)" and "BVD vaccination not done properly in calves" (i.e. after 6 months old, animals were not boostered 2-4 weeks after their first killed vaccine, or not given modified live vaccine), with count ratios of 1.7, 2.0, 2.3, 1.9, 1.4 and 1.8, respectively. The variable "BVDV vaccination (modified live) done properly in calves" (i.e. received another modified live vaccination after 6 months as well) was associated with 0.4 times fewer MAP-seropositive cows.     

Pestivirus infections in Norway. Serological investigations in cattle, sheep and pigs.

Serum samples from 1,133 dairy cows (187 herds), 3,712 ewes (103 flocks) and 1,317 adult pigs (877 herds), were tested for neutralizing antibodies against the NADL strain of bovine virus diarrhoea virus. The prevalence rate of seropositive animals was 18.5% in cattle, 4.5% in sheep and 2.2% in pigs, such seroreactors being found in 28% of the cattle herds and 18% of the sheep flocks. In all three species the rate showed considerable herd and geographical variation. In cattle the seroreactor rate was similar in herds with normal reproduction and in 62 herds with problems of repeat breeding. Of 31 pig sera containing antibodies against the NADL strain, 27 were also positive in a neutralization test for antibodies against swine fever virus (Baker strain). However, all sera showed a higher titre of antibodies against the bovine strain than against the swine fever virus. It was concluded that the immune response of the pigs had been induced by ruminant pestivirus, and not by swine fever virus.     

Bovine mastitis in Ontario due to Mycoplasma agalactiae subsp. bovis.

Bovine mastitis caused by Mycoplasma agalactiae subsp. bovis was first diagnosed in 16 of 55 cows in an Ontario herd in Feburary 1972. A total of 182 of 598 (30.4%) cows from 33 of 64 (51.5%) farms in widely separated areas of the province were culturally positive. Herd incidence varied from 15 to 40% with one closed herd having an incidence of 61%. Four herds were investigated culturally and serologically by the growth inhibition test for 15 months. In the acute phase the organism was present in the milk in extremely high numbers and could still be isolated from a few cows after eight to 12 months. The sera from 89.5% of the animals with clinical mycoplasma mastitis produced a zone of surface "film" and/or colony inhibition and some cows remained positive for six to 12 months. The disease was experimentally reproduced with a pure culture of the organism isolated from the milk of a cow from one of the herds.     

Seroprevalence survey for Anaplasma marginale-infection of Austrian cattle.

A serologic survey study of 5,076 Austrian cattle farming herds was carried out in the period of December 1988 till March 1990. One animal was randomly selected from each herd and the antibody titer against Anaplasma marginale in blood serum samples was evaluated by means of the complement fixation test. The number of these tested blood samples was 3.6% of 140,081 cattle herd farms of Austria. 109 (2.1%) of the tested animals showed positive titers (1:10) against Anaplasma marginale, in relation to the 140,081 cattle herds 0.08%, 4,786 (94.3%) blood serum samples were sero-negative, 188 (3.8%) reacted anticomplementary. The highest number of antibody-positive animals of 8 tested Austrian districts could be found in Carinthia (46 = 5.7%). In Burgenland all tested sera turned out to be negative. Concerning the distribution of sero-positive animals in Austria it can be stated that a decrease of positive reactors from southern to northern region is evident. A connection between the occurrence of anaplasmosis in Italy, Yugoslavia, Switzerland and Hungary, is postulated as a result of the different systems of keeping cattle in the provinces and the regional increase of tick invasion. Possibly an intensive animal transportation is of importance due to the introduction of the disease mentioned before. The results obtained show that anaplasmosis does occur in different areas of Austria. For control of this disease in Austria it is proposed that all imported cattle should be tested serologically for antibodies against Anaplasma marginale. Other diseases in connection with anemia should be excluded by clinical, serological, blood-, as well as pathological examinations.     

Seroprevalence of enzootic bovine leukosis in Trakya district (Marmara region) in Turkey

In this study, 481 cattle belonging to 77 farms from nine localities in Trakya district in the Marmara region of Turkey were blood sampled and serologically tested for enzootic bovine leukosis (EBL). Antibodies to bovine leukosis virus (BLV) were detected in 51 cattle sera (11%) belonging to nine farms in five localities. Cattle tested were mostly female Holstein or Brown Swiss of ages ranging between 18 months and 10 years. Analysis of the relationships between age, breed or sex and seropositivity to EBL in seropositive herds indicated no significant associations (p>0.05). The relationship between seropositivity and haematological changes was also studied, and seropositive cattle had higher lymphocyte percentage and lower neutrophil percentage than seronegative cattle (p<0.001).