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A hematologic key for bovine leukemia virus screening in Japanese black cattle.

ABSTRACT: The European Community's leukosis key (EC key) is a well-known hematologic method for detecting bovine leukemia virus (BLV) infection in dairy cattle. The key identifies infected cattle with persistent lymphocytosis via a combination of lymphocyte count (LC) and age. Using the EC key to identify BLV-infected Japanese black (JB) cattle is problematic, however, given the inherently lower LCs of JB cattle compared to dairy cattle. We analyzed the LC in BLV-positive and -negative JB cattle and estimated LC cutoff values by age using receiver operating characteristic curve analysis. Among the 716 JB blood samples collected, 452 (63%) JB cattle were confirmed as BLV-positive by an antibody ELISA for ?1-y-old cattle and by real-time PCR for <1-y-old cattle. The cutoff values for the LC in each age group were calculated as 6.3 × 10⁹/L for <1?y, 5.9 × 10⁹/L for 1 to <2?y, 5.5 × 10⁹/L for 2 to <3 y, 4.5 × 10⁹/L for 3 to <6?y, 4.3 × 10⁹/L for 6 to ?10?y, and 3.7 × 10⁹/L for >10?y. The sensitivity and specificity of the estimated cutoff values were 0.49 (95% confidence interval: 0.44-0.53) and 0.81 (0.75-0.85), whereas those of the EC key were 0.20 (0.16-0.24) and 0.99 (0.97-1.00). Our LC cutoff values for screening JB cattle for BLV infection appear to be preferable to those of the EC key.

SUBMITTER: Akagami M

PROVIDER: S-EPMC6857024 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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A hematologic key for bovine leukemia virus screening in Japanese black cattle.

Akagami Masataka M Oya Shoko S Kashima Yuki Y Seki Satoko S Ouchi Yoshinao Y Hayama Yoko Y

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 20190503 4

The European Community's leukosis key (EC key) is a well-known hematologic method for detecting bovine leukemia virus (BLV) infection in dairy cattle. The key identifies infected cattle with persistent lymphocytosis via a combination of lymphocyte count (LC) and age. Using the EC key to identify BLV-infected Japanese black (JB) cattle is problematic, however, given the inherently lower LCs of JB cattle compared to dairy cattle. We analyzed the LC in BLV-positive and -negative JB cattle and estim ...[more]

PMID: 31046652

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Project description:BackgroundThe absence of virus expression during the chronic stage of bovine leukemia virus (BLV) infection and its reactivation upon ex vivo culture has become a long-lived Dogma. During the chronic stage of BLV infection the immune response limits viral replication and the mitotic division of latently infected cells, carrying BLV provirus, allows viral expansion and disease progression towards a lymphoproliferative disorder. Several stressor factors have been associated with animal production and handling. As natural mediator of stress, glucocorticoids are strong immunosuppressive agents; moreover, they can bind long-terminal repeat region of retroviruses and induce viral expression. In the present study, we present a case report describing the spontaneous reactivation of BLV infection in naturally infected cattle.Case presentationIn order to investigate if virus reactivation occurred in vivo during the course of BLV infection, we followed up for 328 days one Holstein cow (> 3 years) chronically infected with BLV which presented high-proviral loads. This animal was neither lactating nor pregnant. Furthermore, we investigated if a stressor stimulus, in this case the administration of a synthetic glucocorticoid (dexamethasone), could impact the course of BLV infection in three additional cattle. For the first time, we observed a high level of BLV transcripts in a total of four cattle chronically infected with BLV. The detection of viral transcripts corresponding to pol gene strongly suggests virus reactivation in these animals. Interestingly, this simultaneous virus reactivation was unrelated to dexamethasone treatment.ConclusionsWe reported for the first time spontaneous and high level of BLV transcriptional activation in cattle chronically infected with BLV. Although virus reactivation was unrelated to dexamethasone treatment, other stressor stimuli might have influenced this outcome. Future studies will be necessary to understand these observations, since the spontaneous virus reactivation presented here might have implications on BLV pathogenesis and transmission.

Project description:Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL), a disease that has worldwide distribution. Whilst it has been eradicated in most of Western Europe and Scandinavia, it remains a problem in other regions, particularly Eastern Europe and South America. For this study, in 2013, 24 cattle from three farms in three regions of Moldova were screened by ELISA and nested PCR. Of these cattle, 14 which were PCR positive, and these were molecularly characterized based on the nucleotide sequence of the env gene and the deduced amino acid sequence of the encoded gp51 protein. Our results demonstrated a low level of genetic variability (0-2.9%) among BLV field strains from Moldova, in contrast to that observed for other retroviruses, including human immunodeficiency virus (HIV) (20-38%) Mason IL (Trudy vologod moloch Inst 146-164, 1970) and equine infectious anemia virus (EIAV) (~40%) Willems L et al (AIDS Res Hum Retroviruses 16(16):1787-1795, 2000), where the envelope gene exhibits high levels of variation Polat M et al (Retrovirology 13(1):4, 2016). Sequence comparisons and phylogenetic analysis revealed that BLV genotype 7 (G7) is predominant in Moldova and that the BLV population in Moldovan cattle is a mixture of at least three new sub-genotypes: G7D, G7E and G4C. Neutrality tests revealed that negative selection was the major force operating upon the 51-kDa BLV envelope surface glycoprotein subunit gp51, although one positively selected site within conformational epitope G was detected in the N-terminal part of gp51. Furthermore, two functional domains, linear epitope B and the zinc-binding domain, were found to have an elevated ratio of nonsynonymous to synonymous codon differences. Together, these data suggest that the evolutionary constraints on epitopes G and B and the zinc-binding domains of gp51 differ from those on the other domains, with a tendency towards formation of homogenous genetic groups, which is a common concept of global BLV diversification during virus transmission that may be associated with genetic drift.

Dataset Information

A hematologic key for bovine leukemia virus screening in Japanese black cattle.

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A hematologic key for bovine leukemia virus screening in Japanese black cattle.

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