Cultured early goat embryos and cells are susceptible to infection with caprine encephalitis virus.

Department of Research into the Health Risk and Biotechnology of Reproduction ENVN/DGER, National Veterinary School, BP 40706, 44307 Nantes Cedex 03, France.
Virology (Impact Factor: 3.37). 10/2006; 353(2):307-15. DOI: 10.1016/j.virol.2006.06.007
Source: PubMed

ABSTRACT Zona-pellucida-free embryos at 8-16 cell stage were co-cultured for 6 days in an insert over a mixed cell monolayer infected with CAEV-pBSCA. Embryos were washed and transferred to an insert on CAEV indicator goat synovial membrane cells for 6 h, then they were washed and cultivated in B2 Ménézo for 24 h, finally, embryo cells were dissociated and cultivated on a feeder monolayer for 8 days. After 5 weeks, multinucleated giant cells typical of CAEV infection were observed in indicator GSM cell monolayers. In the acellular medium, the early embryonic cells produced at least 10(3.25) TCID50/ml over 24 h. The monolayer of cultivated embryonic cells developed cytopathic lesions within 8 days, and CAEV RNA, CAEV proviral DNA and protein p28 of the capsid were detected. All of these results clearly demonstrate that caprine early embryonic cells are susceptible to infection with CAEV and that infection with this virus is productive.

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    ABSTRACT: The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.
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    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; · 2.08 Impact Factor
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    ABSTRACT: Caprine arthritis-encephalitis (CAE) is an infectious disease caused by the caprine arthritis-encephalitis virus (CAEV), belonging to the lentivirus genus. The presence of the virus has been observed in the nervous system, respiratory tract and mammary gland, and also in the male and female genital tract. The objective of this study is to identify the virus in oocyte and uterine fluid of infected goats by molecular diagnostic techniques, in order to assess the possibility of CAEV transmission with reproduction. Thirteen infected goats were selected and submitted to euthanasia for the collection of the reproductive system, aspiration of the uterine fluid and dissection of ovaries for oocyte collection. In order to identify the CAEV in the collected material, in the protovirus and free forms, it was submitted to the nRT-PCR and nPCR techniques, respectively. As a result, it was observed that 53.8% of oocytes were positive to nRT-PCR, while only 9.1% were positive to nPCR. The nRT-PCR also identified the virus in the uterine fluid of 46.1% of the tested females. Even though the 13 goats had CAEV, 30.8% presented negative results in nPCR and nRT-PCR in all of the analyzed samples (oocyte and uterine fluid). This work concludes that nRT-PCR and nPCR can be used in the diagnosis of CAE for the analysis with oocytes and uterine fluid, and that the presence of CAEV in these materials points out to the risk of CAEV transmission through reproductive technologies used in females.
    Arquivos do Instituto Biológico. 12/2012; 80(4):381-386.

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