Transmission of bovine leukaemia virus in milk

Summary Nineteen calves born to dams free of bovine leukaemia virus (BLV) did not possess maternally derived precipitating antibody to BLV in their sera after the ingestion of colostrum. Eight of these calves remained serologically negative after being fed milk from BLV-free cows while three (27.3%) of 11 similar calves that had been fed milk from BLV-infected cows developed antibody. Forty-four of 47 calves born to BLV-infected dams acquired maternal antibody to BLV after ingesting colostrum. Two (8.7%) of the 23 calves fed milk from BLV-free cows developed antibody to BLV probably as a result of transplacental or colostrum infection whereas four (16.7%) of the 24 calves fed milk from BLV-infected cows developed antibody. It is concluded that milk transmission of BLV is responsible in part for the high rates of infection encountered in our dairy herds and that calves lacking specific maternal antibody are more susceptible to BLV infection through the ingestion of milk than are calves with maternal antibody.

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Transmission Del Virus De La Leucemia Bovina Por La Ingestion De Leche

Resumen Diezinueve bezerros nacidos de vacas libres del virus de la leucemía bovina (VLB) no poseían anticuerpos precipitantes de origen materno contra el VLB en el suero después de la ingestión de calostro. Ocho de estos bezerros permanecieron serologicamente negativos después de ser alimentados con leche de vacas libres del VLB mientras que tres (27.3 por ciento) de 11 bezerros similares que habían sido alimentados con leche de vacas infectadas con el VLB desarrollaron anticuerpos. Cuarenta y cuatro de 47 bezerros nacidos de vacas infectadas con el VLB adquirieron anticuerpos maternos contra el VLB después de ingerir calostro. Dos (8.7 por ciento) de los 23 bezerros alimentados con leche de vacas libres del VLB desarrollaron anticuerpos contra el VLB, probablemente como resultado de una infección transplacentaria o através del calostro, mientras que cuatro (16.7 por ciento) de los 24 bezerros alimentados con leche de vacas infectadas con el VLB desarrollaron anticuerpos. Se concluye que la transmisión del VLB através de la leche es responsable, en parte, por los altos índices de infección encontrados en nuestros rebaños lecheros y que bezerros sin anticuerpos específicos maternos son mas susceptibles a la infección con el VLB através de la ingestión de leche que los bezerros que poseen estos anticuerpos.

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Transmission De La Leucose Bovine Parlelait

Résumé Dix neuf veaux issus de mères indemnes de virus de la leucose bovine (VLB) ne possédaient pas d'anticorps précipitants pour le VLB dans leur sérum après ingestion de colostrum. Huit d'entre eux sont restis sérologiquement négatifs après avoir été alimentés avec du lait provenant de vaches indemnes de VLB alors que trois (27,3 p.100) des 11 veaux restants qui avaient été alimentés avec du lait provenant de vaches infectées de VLB ont produit des anticorps. Quarante quatre veaux sur quarante sept issus de mères infectées de VLB ont acquis des anticorps maternels anti VLB après ingestion de colostrum. Deux (8,7 p. 100) veaux sur vingt trois alimentés avec du lait de vaches indemnes de VLB ont produit des anticorps anti VLB probablement à la suite d'infection transplacentaire ou par le colostrum alors que quatre (16,7 p. 100) veaux sur vingt quatre recevant du lait de vaches infectées de VLB ont produit des anticorps. On conclut que la transmission du virus de la leucose bovine par le lait est en partie responsable des taux élevés d'infection rencontrés dans nos troupeaux laitiers et que les veaux manquant d'anticorps maternels spécifiques sont plus sensibles à l'infection par le VLB par ingestion de lait que ceaux ayant reçu des anticorps maternels.

     

Production and related variables in bovine leukaemia virus-infected cows

A newly developed milk dot blot test was used to detect anti-bovine leukaemia virus (BLV) antibody in milk samples from 2079 lactating adult cows from among 61 herds. The milk dot blot test was highly repeatable; the concordance rate, compared with the agar gel immunodiffusion test performed on serum, was 83.5%. All herds contained BLV-positive cows; the prevalence rate was 36%. BLV-positive cows tended to come from larger herds and were older and more often later in lactation. Fourteen production and related variables (herd size, age, days open, days in milk, milk somatic cell count, milk, fat, and protein produced in the current lactation, projected production of milk, fat, and protein, and breed class average deviations for milk, fat, and protein) were compared between BLV-positive and BLV-negative cows. Although somatic cell count, milk produced, and projected production of milk and protein were related significantly to BLV status using simple tests of association, once the variables herd size, age and days in milk were controlled, these differences were removed. Further analyses using logistic (outcome: individual cow BLV status) and least-squares regression (outcome:herd proportion of BLV-positive cows) failed to show an association between any of the measured production or related variables and BLV-positivity. We concluded that the effect of BLV on production and related variables in dairy cows was below the sensitivity of our analytical techniques or was non-existent.Abbreviations ABCA herd average breed class average for milk, fat, and protein production - AVGAGE average age of the herd - ADIM herd average for days in milk - AGID agar gel immunodiffusion - AVGSCC herd average milk somatic cell count - BCA breed class average, a milk, fat and protein production index calculated by comparing a cow's actual 305-day lactation production to the corresponding BCA standard for the same breed, age, and month of calving - BLV bovine leukaemia virus - CALVINT calving interval - COWAGE cow age - DBCA breed class average deviation for milk, fat, and protein production, the difference between an individual cow's BCA and the herd average - DIM days in milk - HS herd size corresponding to the number of lactating cows in a herd - LACT actual amount of milk, fat, and protein produced in a cow's lactation - ODHIC Ontario Dairy Herd Improvement Corporation - PCTPOS percentage of herd that is BLV-positive - PROJ projected 305-day production for milk, fat, and protein by fitting to a standard lactation curve adjusted for days in milk and age at calving - RHBCA rolling herd average for breed class average for milk, fat, and protein production, the average for all cows that completed a lactation (cows must have completed a 305-day lactation) during the previous 12 months - SCC milk somatic cell count     

Measurement of telomerase activity in bovine leukaemia virus infected cows

Telomerase adds new telomeric sequences to the end of chromosomal DNA in order to overcome the end-replication problem. The upregulation of telomerase activity in tumours has been reported in humans and some mammals and is considered to be a tumour marker; however, such activity has not been investigated in cows. Therefore, we investigated telomerase activity in bovine leukaemia, the most common tumour in cows and its relationship with the bovine leukaemia virus (BLV) infection, which is the major cause of leukaemia. Telomerase activity was detected in 25 of 29 bovine leukaemia tissue samples. In peripheral blood lymphocytes (PBL) from BLV-infected cases that did not develop the tumour, telomerase activity was detected in 11 of 71 cases (15.5%). When these cases were classified based on serological tests and the peripheral blood lymphocyte count, the telomerase activity was observed to be the highest in the seropositive, non-lymphoproliferative (PBL<8000 microl(-1)) cases (three of seven cases, 42.9%), and not observed in the lymphoproliferative cases (PBL<16,000 microl(-1)) except in one case. Although the precise pathogenesis of BLV-related diseases remains obscure, persistent lymphocytosis is considered as a pre-neoplastic state. In contrast, our results suggested that given the fact that telomerase activity indicates tumour development, the aleukaemic stage could be defined as the 'pre-neoplastic state'. In conclusion, similar to many tumours in humans, telomerase activity was detected in bovine leukaemia; further, this activity can be a potentially useful prediction marker for tumour development and/or a good therapeutic target.     

Lymphosarcoma development in sheep experimentally infected with bovine leukaemia virus

Twelve sheep were experimentally infected with a phytohemagglutinin (PHA) treated short term culture of lymphocytes from a cow naturally infected with BLV at the PL stage. Five of 12 (42%) BLV infected sheep had histologically confirmed lymphosarcoma 10-16 months after infection. The PBL's were increased to leukemic levels 3-21 weeks before death due to lymphoblastic leukemia. Lymphocyte proliferation and appearance of immature lymphocytes and lymphoblastic cells in the blood were a characteristic feature of tumour development following inoculation with an Australian strain of BLV. In contrast to a number of previous studies the peripheral lymph nodes of all infected sheep were clinically normal throughout the experimental period but at death gross tumours were evident in the mesentric lymph nodes and the heart in all cases. All the other lymph nodes, liver, spleen, kidney and lung were histologically infiltrated with lymphoid tumour cells. Gross tumours were present in the abomasum (1 out of 5) in the urinary tract (2 out of 5) and in the uterus (1 out of 2). The majority of the tumour cells isolated from the various tissues were centroblastic demonstrating that the malignant leukemia in experimentally infected sheep was of a multicentric centroblastic type. The central nervous system was not involved in any case.     

Slow virus infections with particular reference to maedi-visna and enzootic bovine leukaemia

The slow virus infections include the four neurological diseases: scrapie, transmissible mink encephalopathy, kuru and Creutzfeldt-Jakob's disease in which a non-classified agent is involved. The group also includes diseases caused by infectious-exogenous — RNA tumour viruses, such as maedi-visna virus and bovine leukaemia virus.There have been no indications so far that maedi-visna or bovine leukaemia are transmitted by true vertical transmission with an integrated DNA-provirus, but there is an increasing tendency to recognise that vertical transmission, across the placenta or through the milk, does occur with viral information transmitted as infectious particles.Colostral antibodies give rise to positive serological reaction in the offspring and these antibodies may also interfere with the propagation of viruses in the newborn animal and thus enable them to escape detection in tests for both virus and selfsynthesized antibodies. The tests used at present are suitable only as herd tests and until more sensitive tests have been developed it will not be possible to remove infected animals completely from infected herds and flocks.

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Kurzfassung Zu den Slow Virus Infektionen gehören vier neurologische Erkrankungen: die Traberkrankheit, die übertragbare Nerz-Enzephalopathie, die KuruKrankheit und die Jacob-Creutzfeldt Pseudosklerose, bei denen ein unbekannter Faktor beteiligt ist. Zur gleichen Gruppe gehören weiterhin Krankheiten, die durch infektiöse, exogene RNA-Tumorviren, wie Maedi-Visna Virus und den Rinderleukosevirus hervorgerufen werden.Die beiden letztgenannten werden horizontal übertragen; es besteht jedoch zunehmend die Neigung, auch die vertikale Ubertragung als wichtigen Aspekt in der Epidemiologie in Betracht zu ziehen. Bei diagnostischen Tests nach hämatologischen oder serologischen Verfahren werden einzelne Tiere unter Umständen nicht herausgefunden, wenn sie als Jungtiere vertikal infiziert wurden. Ausserdem zeigt sich, dass auch horizontal infizierte Einzeltiere unter Umständen nicht herausgefunden, wenn sie als Jungtiere vertikal infiziert wurden. Ausserdem zeigt sich, dass auch horizontal infizierte Einzeltiere unter Umständen serologisch oder hämatologisch oder hämatologisch lange Zeit nicht reagieren und daher nicht identifiziert werden können. Die geganwärtig benutzten Tests eignen sich nur als Herdentests, und erst wenn empfindlichere Tests entwickelt worden sind, wird es möglich sein, sämtliche infizierte Tiere aus Herden auszusondern.

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Resume Les infections à virus lents comprennent les quatre maladies neurologiques suivantes: la tremblante, l'encéphalopathie transmissible du vison, le kuru et la maladie de Creutzfeldt-Jacob, dans laquelle intervient un agent non classé. Ce groupe englobe en outre les maladies provoquées par des virus ARN tumoraux, agents infectieux éxogènes tels que le virus de la maladie de Maedi-Visna et le virus de la Leucose bovine.Ces deux dernières maladies sont transmises horizontalement mais on a de plus en plus tendance à considérer la transmission verticale comme un aspect important de leur épidémiologie. Il arrive que les tests de dépistage basés sur les méthodes hématologiques ou sérologiques soient incapables de détecter certains animaux s'ils sont contaminés en bas âge par la voie verticale. On s'est également rendu compte que, parfois, des animaux contaminée par la voie horizontale ne réagissent pas pendant une période prolongée aux tests sérologiques et hématologiques et qu'il risquent ainsi d'échapper au dépistage. Les tests utilisés actuellement conviennent uniquement pour les examens de troupeaux, et ce n'est que lorsqu'on aura mis au point des tests plus sensibles qu'on pourra épurer complètement les troupeaux contaminés.

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Riassunto Le infezioni striscianti da virus comprendono quattro affezioni neurologiche: scrapie, encefalopatia contagiosa dei visoni, kuru e malattia di Creutzfeldt-Jacob, dovuta ad un agente sconosciuto. Il gruppo comprende inoltre affezioni causate da virus tumorali esogeni RNA, come il virus maedi-visna e quello della leucemia bovina.La trasmissione degli ultimi due virus avviene in senso orizzontale, ma vi è una tendenza sempre crescente ad ammettere la trasmissione in senso verticale quale importante caratteristica della loro epidemiologia. I test diagnostici sul sangue o sul siero non sempre possono identificare i singoli animali contagiati in precedenza in senso verticale. E'quindi evidente che di tanto in tanto gli animali contagiati per via orizzontale non presentino reazioni all'esame serologico o a quello ematologico per un lungo periodo di tempo e che pertanto non vengano neppure scoperti. I test attualmente in uso sono adatti soltanto ad usi di massa. Fino a quando non saranno stati scoperti metodi più precisi non sarà possibile disinfestare completamente le mandrie e le greggi contagiate.

     

Evaluation of alternative methods for the detection of bovine leukaemia virus in cattle

AIM: To evaluate commercially available enzyme-linked immunosorbent assays (ELISAs) and the polymerase chain reaction (PCR) for their ability to detect antibodies against or nucleic acid of the bovine leukaemia virus (BLV), the causal agent of enzootic bovine leukosis (EBL), and to assess their usefulness in a national eradication programme. METHODS: Eighty-two well-defined sera (including 18 from an OIE reference laboratory) and 399 field sera from New Zealand cattle were tested in five ELISAs and the results compared with the agar gel immunodiffusion (AGID) test and electrophoretic immunoblotting (EIB) results. A polymerase chain reaction-based technique, which could detect BLV-RNA and proviral-DNA, was also evaluated on a subsample of the field cases. RESULTS: Two commercial ELISAs classified 99% of the defined sera correctly, with the other three ranging in their correct classification between 88% and 95%. The ELISAs agreed in their general classification on the majority of the 399 blood samples (91.7%), and with the AGID for more than 95 % of the sera. In a dilution series of the international reference serum E4, the highest dilution with a positive (or suspicious) result ranged from 1:80 to 1:5120. A dilution series of 202 field positive samples tested in the preferred ELISA detected 98% of positive sera at a 15 and 1: 10 dilution, reducing to 78% at a 1:80 dilution of the sera. Agreement between serological tests and PCR was poor, mainly due to failure of the PCR to detect a number of serologically positive animals. CONCLUSION: ELISA tests detected about 10% more reactors than the AGID and the EIB combined. Some ELISA-positive animals were not detected by PCR, raising doubts about the usefulness of PCR-based technology in EBL eradication programmes.     

Changes in B cell and T cell subsets in bovine leukaemia virus-infected cattle

Direct immunofluorescence and fluorescence-activated cell sorter techniques were used for the detection of surface immunoglobulin positive (SIg+) cells in peripheral blood lymphocytes (PBL's) of bovine leukaemia virus (BLV) infected cattle with or without persistent lymphocytosis (PL+, PL-) and in BLV-free cattle. The percentage of SIg+ cells was more than twice as high in BLV+PL+ cattle than in BLV-free and BLV+PL- cattle. Bovine T cells, and T cell subsets were identified indirectly by the same techniques using three monoclonal antibodies (MAb's) specific for all T cells (IL-A43), T helper (BoT4) cells (IL-A12) and T cytotoxic (BoT8) cells (IL-A17). The major histocompatibility complex (MHC) determinants of both class II (BoT4) and class I (BoT8) as well as all T cells were significantly reduced in BLV+PL+ compared to BLV-free cattle. The actual decrease in the BoT8 cell subset or the dilution effect that would change effector:target cell ratio suggests that a resultant decrease in cytotoxic activity in BLV+PL+ cattle may play an important role in the progress of BLV infection in cattle.     

Factors affecting the natural transmission of bovine leukaemia virus infection in Queensland dairy herds

Natural transmission of bovine leukaemia virus (BLV) infection in south-eastern Queensland dairy herds was slow in 2 herds with a low to moderate (13 to 22%) prevalence of infection. Infection spread much more rapidly in a herd that had a higher prevalence (42%) when first tested. In a 13 month study of this herd, the cumulative incidence of infection was 24%. In one herd new infections were confined almost entirely to calves of uninfected dams. Following the end of feeding bulk milk to calves, a common practice in dairy herds, no more calves in this herd became infected. In laboratory experiments, neither prolonged housing of susceptible calves with infected cattle, consumption of drinking water contaminated with infected blood, nor inoculation of sheep with saliva from infected cattle resulted in transmission of BLV infection. Sheep were infected by subcutaneous inoculation of a suspension of purified lymphocytes from an infected heifer. The minimum infective dose was 10(3) lymphocytes, equivalent to the number of lymphocytes in approximately 0.1 microliter blood. Thus, procedures involving the transfer of a very small volume of blood from animal-to-animal have the potential to transmit infection.     

Decreased expression of L-selectin (CD62L) on lymphocytes in enzootic bovine leukaemia

Expression of L-selectin was determined by single- and two-colour immunofluorescence on granulocytes, peripheral blood mononuclear cells (PBMC) and blasts of bovine origin by means of a monoclonal antibody IVA94 which recognizes bovine L-selectin (CD62L). Cells were separated from peripheral blood of healthy cattle and colleagues infected with bovine leukaemia virus (BLV). BLV-infected animals comprised lymphocytotic and non-lymphocytotic cows. L-selectin was expressed on 90-98% of granulocytes in all tested animals. The percentage of PBMC expressing L-selectin was lower in cattle with persistent lymphocytosis than in non-lymphocytotic or BLV-free cattle, and inversely correlated with lymphocyte counts. The ratio of B lymphocytes stained for L-selectin was significantly decreased from 60.2 +/- 1.9% in BLV-free cattle to 43.8 +/- 3.6 and 22.5 +/- 5.7% in non-lymphocytotic and lymphocytotic cattle, respectively. B-lymphocytes stained for L-selectin exhibited about 50% reduction in L-selectin expression in BLV-infected cattle compared with BLV-free cattle, as judged by the mean fluorescence intensity (MFI). The percentage of L-selectin-positive PBMC not bearing surface immunoglobulin M (predominantly T lymphocytes) was comparable in BLV-free and BLV-infected cattle. However, L-selectin expression on T lymphocytes was reduced (about 50%) in BLV-infected cattle, as judged by the MFI. We suppose that BLV infection results in a decreased L-selectin expression on lymphocytes, and accordingly, it may contribute to deregulation of the host immune system.     

Partial sequencing of env gene of bovine leukaemia virus from Brazilian samples and phylogenetic analysis

Analysis of the partial bovine leukaemia virus (BLV) env gp51 gene sequences obtained from three BLV strains isolated in three different regions of Brazil was carried out. The Brazilian BLV env gp51 sequences were compared with seven other corresponding sequences of BLV strains isolated in different countries and with consensus sequence as well. The obtained data point on qualitative and quantitative differences among the analysed strains as far as the occurrence of single point mutations is concerned. Two Brazilian strains show significantly higher mutation rate than other analysed strains. Amino acid analysis did not show, however, any substantial changes of the primary protein structure coded by well conserved region of BLV env gp51 gene. Based on the obtained data, the putative dendogram image of possible phylogenetic relations among the studied BLV strains is presented as well.     

Prevalence of the plasma bovine leukaemia virus blocking factor in cattle from a commercial dairy herd

The prevalence of the plasma bovine leukaemia virus blocking factor was examined in a commercial herd. The detection of the blocking factor activity depended on the nature of the infected target cells used in the assay. Using the sensitive bovine leukaemia virus-infected target lymphocytes, the factor was found to be significantly more prevalent in infected cattle than in virus-free cattle. Furthermore, in infected cattle, no correlation was observed between the level of bovine leukaemia virus antibody and the presence of the blocking factor.