Detection of ovine Lentivirus in the cumulus cells, but not in the oocytes or follicular fluid, of naturally infected sheep
ABSTRACT The aim of this study was to examine the Maedi-Visna virus (MVV) infection status of oocytes, cumulus cells, and follicular fluid taken from 140 ewes from breeding flocks. MVV proviral-DNA and MVV RNA were detected using nested-PCR and RT-PCR MVV gene amplification, respectively in the gag gene. Nested-PCR analysis for MVV proviral-DNA was positive in peripheral blood mononuclear cells in 37.1% (52/140) of ewes and in 44.6% (125/280) of ovarian cortex samples. The examination of samples taken from ovarian follicles demonstrated that 8/280 batches of cumulus cells contained MVV proviral-DNA, whereas none of the 280 batches of oocytes taken from the same ovaries and whose cumulus cells has been removed, was found to be PCR positive. This was confirmed by RT-PCR analysis showing no MVV-viral RNA detection in all batches of oocytes without cumulus cells (0/280) and follicular fluid samples taken from the last 88 ovaries (0/88). The purity of the oocyte fraction and the efficacy of cumulus cell removal from oocytes was proved by absence of granulosa cell-specific mRNA in all batches of oocytes lacking the cumulus cells, using RT-PCR. This is the first demonstration that ewe cumulus cells harbor MVV genome and despite being in contact with these infected-cumulus cells, the oocytes and follicular fluid remain free from infection. In addition, the enzymatic and mechanical procedures we used to remove infected-cumulus cells surrounding the oocytes, are effective to generate MVV free-oocytes from MVV-infected ewes.
SourceAvailable from: Yahia Chebloune[Show abstract] [Hide abstract]
ABSTRACT: Reproductive biotechnologies are essential to improve the gene pool in small ruminants. Although embryo transfer (ET) and artificial insemination (AI) greatly reduce the risk of pathogen transmission, few studies have been performed to quantify this risk. The aim of this review is to contribute to the elements needed to evaluate the risk of lentivirus transmission in small ruminants (SRLV) during ET, from embryos produced in vitro or in vivo, and with the use of the semen destined for AI. The purpose is to consider the genetic possibilities of producing uninfected embryos from infected females and males or bearers of the SRLV genome. We have reviewed various studies that evaluate the risk of SRLV transmission through genital tissues, fluids, cells, and flushing media from female and male animals. We have only included studies that apply the recommendations of the International Embryo Transfer Society, to obtain SRLV-free offspring from infected female animals using ET, and the justification for using healthy male animals, free from lentivirus, as semen donors for AI. As such, ET and AI will be used as routine reproductive techniques, with the application of the recommendations of the International Embryo Transfer Society and World Organization for Animal Health.Theriogenology 11/2012; 79(1). DOI:10.1016/j.theriogenology.2012.09.021 · 1.85 Impact Factor
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ABSTRACT: Small ruminant lentiviruses include members that infect sheep (ovine lentivirus [OvLV]; also known as ovine progressive pneumonia virus/maedi-visna virus) and goats (caprine arthritis encephalitis virus [CAEV]). Breed differences in seroprevalence and proviral concentration of OvLV had suggested a strong genetic component in susceptibility to infection by OvLV in sheep. A genetic marker test for susceptibility to OvLV has been developed recently based on the TMEM154 gene with validation data from over 2,800 sheep representing nine cohorts. While no single genotype has been shown to have complete resistance to OvLV, consistent association in thousands of sheep from multiple breeds and management conditions highlight a new strategy for intervention by selective breeding. This genetic marker-assisted selection (MAS) has the potential to be a useful addition to existing viral control measures. Further, the discovery of multiple additional genomic regions associated with susceptibility to or control of OvLV suggests that additional genetic marker tests may be developed to extend the reach of MAS in the future. This review will cover the strengths and limitations of existing data from host genetics as an intervention and outline additional questions for future genetic research in sheep, goats, small ruminant lentiviruses, and their host-pathogen interactions.Viruses 06/2013; 5(6):1466-99. DOI:10.3390/v5061466 · 3.28 Impact Factor
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ABSTRACT: Caprine arthritis encephalitis is a worldwide, multisystemic disease caused by a small ruminant lentivirus. Although the main route of transmission is oral, detection of proviral DNA of the caprine arthritis encephalitis virus (CAEV) in caprine semen has been previously described. However, the presence of viral antigens in the male reproductive tract has apparently never been reported. The objective was to study lesions in the buck reproductive system and to detect, in these tissues, the presence of proviral DNA, viral RNA and CAEV antigens. Tissues from eight CAEV-infected bucks (one naturally and seven experimentally infected) were analyzed by histopathology, nested polymerase chain reaction, reverse transcriptase-polymerase chain reaction, and immunohistochemistry. Interstitial pneumonia, synovitis, and lesions in the male reproductive tract were detected in some of the bucks. Proviral DNA was detected in the lungs and joints as well as in the reproductive systems of all animals, whereas viral RNA was detected only in the genital tract of the naturally infected buck. Viral antigens were immunostained in most of the organs of the male reproductive tract. This report was apparently the first to clearly demonstrate CAEV antigen expression in the male reproductive tract, which indicates the possibility of venereal transmission of CAEV.Theriogenology 08/2013; DOI:10.1016/j.theriogenology.2013.07.021 · 1.85 Impact Factor