Maedi-Visna virus was detected in association with virally exposed IVF-produced early ewes embryos.
The objective of this study was to determine whether MVV can be transmitted by ovine embryos produced in vitro and whether the zona pellucida (ZP) provides any protection against MVV infection. Zona pellucida (ZP)-intact and ZP-free embryos, produced in vitro, at the 8-16 cell stage, were cocultured for 72h in an insert over an ovine oviduct epithelial cell (OOEC)-goat synovial membrane (GSM) cell monolayer that had been previously infected with MVV (K1514 strain). The embryos were then washed and transferred to either direct contact or an insert over a fresh GSM cell monolayer for 6 h. The presence of MVV was detected using RT-PCR on the ten washing fluids and by the observation of typical cytopathic effects (CPE) in the GSM cell monolayer, which was cultured for 6 weeks. This experiment was repeated 4 times with the same results: MVV viral RNA was detected using RT-PCR in the first three washing media, while subsequent baths were always negative. Specific cytopathic effects of MVV infection and MVV-proviral DNA were detected in GSM cells that were used as a viral indicator and cocultured in direct contact or as an insert with MVV-exposed ZP-free embryos. However, no signs of MVV infection were detected in cells that were cocultured with exposed ZP-intact or non-exposed embryos. This study clearly demonstrates that (i) in vitro, ZP-free, early ovine embryos, which had been exposed to 10(3) TCID(50)/m MVV in vitro, are capable of transmitting the virus to susceptible GSM target cells, and that (ii) the IETS recommendations for handling in vivo produced bovine embryos (use of ZP-intact embryos without adherent material and performing ten washes) are effective for the elimination of in vitro MVV infection from in vitro produced ovine embryos. The absence of interaction between ZP-intact embryos and MVV suggests that the in vitro produced embryo zona pellucida provides an effective protective barrier.
Romero
C Cortez
CC
ONIRIS, (Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering), Department of Research into the Health Risks and Biotechnology of Reproduction, UPSP 5301 DGER, France.
Pellerin
J L
JL
Poulin
N
N
Cognié
Y
Y
Chebloune
Y
Y
Pépin
M
M
Fieni
F
F
D000818:Animals; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000821:virology*; D005307:Fertilization in Vitro; D006041:Goats / Q000821:virology; D011021:Pneumonia, Progressive Interstitial, of Sheep / Q000635:transmission* / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D012756:Sheep / Q000196:embryology; D013583:Synovial Membrane / Q000821:virology; D016182:Visna / Q000635:transmission* / Q000821:virology; D014790:Visna-maedi virus / Q000235:genetics / Q000302:isolation & purification*; D015044:Zona Pellucida / Q000502:physiology
D012367:RNA, Viral