Abstract
Small ruminant lentiviruses (SRLVs) exist as populations of closely related genetic variants, known as quasispecies, within an individual host. The privileged way of SRLVs transmission in goats is through the ingestion of colostrum and milk of infected does. Thus, characterization of SRLV variants transmitted through the milk, including milk epithelial cells (MEC), may provide useful information about the transmission and evolution of SRLVs. Therefore, the aim of this study was to detect SRLVs in peripheral blood leukocytes (PBLs) and milk epithelial cells of goats naturally infected with SRLVs and perform single nucleotide variations analysis to characterize the extent of genetic heterogeneity of detected SRLVs through comparison of their gag gene sequences. Blood and milk samples from 24 seropositive goats were tested in this study. The double immunolabeling against p28 and cytokeratin demonstrated that milk epithelial cells originated from naturally infected goats were infected by SRLVs. Moreover, PCR confirmed the presence of the integrated SRLVs proviral genome indicating that MECs may have a role as a reservoir of SRLVs and can transmit the virus through milk. The blood and MEC derived sequences from 7 goats were successfully sequenced using NGS and revealed that these sequences were genetically similar. The MEC and blood-derived sequences contained from 3 to 30 (mean, 10.8) and from 1 to 10 (mean, 5.4) unique SNVs, respectively. In five out of seven goats, SNVs occurred more frequent in MEC derived sequences. Non-synonymous SNVs were found in both, PBLs and MEC-derived sequences of analyzed goats and their total number differed between animals. The results of this study add to our understanding of SRLVs genomic variability. Our data provides evidence for the existence of SRLVs quasispecies and to our knowledge, this is the first study that showed quasispecies composition and minority variants of SRLVs present milk epithelial cells.
Highlights
Small ruminant lentiviruses (SRLVs), comprising maedi-visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are RNA viruses belonging to the Lentivirus genus of the family Retroviridae which infect sheep and goats
We investigated the existence of differences in the population of SRLVs proviral DNA present in the peripheral blood leukocytes (PBLs) and colostrum cells of goats naturally infected with SRLV subtype A17
This study extends our previous investigative approach, including next-generation sequencing (NGS), applied to perform single nucleotide variations analysis of gag gene sequences derived from PBLs and milk epithelial cells (MEC) of goats naturally infected with SRLVs representing different subtypes
Summary
Small ruminant lentiviruses (SRLVs), comprising maedi-visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are RNA viruses belonging to the Lentivirus genus of the family Retroviridae which infect sheep and goats. SRLVs induce a persistent infection-causing progressive inflammatory lesions exerting arthritis, mastitis, chronic interstitial pneumonia, encephalitis, and progressive weakness [1]. Lactogenic transmission via ingestion of colostrum and milk is considered to be the most important way of infection for SRLVs in kids while horizontal transmission via respiratory secretions play important role in older animals [1,2,3]. The provirus of SRLVs contains three structural genes (gag, pol, and env) and accessory genes (vif, rev, and vpr) flanked by non-coding long terminal repeat (LTR) sequences. The pol and gag genes are relatively conserved while the env gene shows a high level of variability
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