Effect in vitro of bovine viral diarrhea virus on bovine embryos with the zona pellucida intact, damaged and removed

1988 ◽  
Vol 12 (1) ◽  
pp. 19-24 ◽  
Author(s):  
A. Bielanski ◽  
W. C. D. Hare
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Zona Pellucida ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

154 THE QUANTITY OF BOVINE VIRAL DIARRHEA VIRUS ASSOCIATED WITH SINGLE ZONA PELLUCIDA-INTACT IN VITRO-PRODUCED BOVINE EMBRYOS FOLLOWING ARTIFICIAL EXPOSURE

2008 ◽  
Vol 20 (1) ◽  
pp. 157
Author(s):  
J. A. Gard ◽  
M. D. Givens ◽  
P. K. Galik ◽  
K. P. Riddell ◽  
M. S. D. Marley ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Bovine Viral Diarrhea Virus ◽  
Zona Pellucida ◽  
Quantitative Polymerase Chain Reaction ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

The primary objective of this study was to determine the percentage of individual, preimplantation, in vitro-produced bovine embryos which maintained association with virus despite washing following artificial exposure to a high affinity strain of bovine viral diarrhea virus (BVDV). Another objective of this study was to determine the quantity of virus associated with these embryos. A total of eighty-seven zona pellucida-intact, Day 7, in vitro-produced bovine embryos were exposed for 1 h to 2 � 106 cell culture infected doses per mL to the 50 percent endpoint (CCID50 mL–1) of a type 1 noncytopathic strain of BVDV (SD-1). Following exposure, the embryos were washed according to International Embryo Transfer Society standards for in vitro-produced bovine embryos; they then underwent sonication, RNA extraction, and freezing at –80�C until assayed for virus. A real-time quantitative polymerase chain reaction (QPCR) was run in duplicate on each of the 87 embryos. Forty-two percent (39/87) of the embryos assayed were determined to be positive for virus. The quantity of virus associated with the embryos averaged 0.55 viral copies per 5 µL (SD = 0.89 copies/5 µL, SEM = 0.14 copies/5 µL). Assessment of data using tolerance intervals (P = 0.05) indicates that 90% of contaminated embryos were associated with ≤2.40 viral copies per 5 µL while 99% of contaminated embryos were associated with ≤3.44 viral copies per 5 µL. These findings show that there is a low level of virus associated with in vitro-produced embryos but virus is associated with a significant number of exposed embryos. In conclusion, this study indicates that the potential for transmission of BVDV via embryo transfer of in vitro-produced embryos is small given the amount of virus that was found to associate with individual embryos.

Noncytopathic bovine viral diarrhea virus in a system for in vitro production of bovine embryos

1994 ◽  
Vol 41 (4) ◽  
pp. 841-853 ◽  
Author(s):  
O.V. Zurovac ◽  
D.A. Stringfellow ◽  
K.V. Brock ◽  
M.G. Riddell ◽  
J.C. Wright
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Vitro Production ◽  
Viral Diarrhea Virus

Washing and trypsin treatment of in vitro derived bovine embryos exposed to bovine viral diarrhea virus

1998 ◽  
Vol 50 (5) ◽  
pp. 717-726 ◽  
Author(s):  
E Trachte ◽  
D Stringfellow ◽  
K Riddell ◽  
P Galik ◽  
M Riddell ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
Trypsin Treatment ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Alteration in ultrastructural morphology of bovine embryos following subzonal microinjection of bovine viral diarrhea virus (BVDV)

Zygote ◽  
2008 ◽  
Vol 16 (3) ◽  
pp. 187-193 ◽  
Author(s):  
E. Kubovičová ◽  
A. V. Makarevich ◽  
J. Pivko ◽  
P. Chrenek ◽  
P. Grafenau ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Developmental Stages ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
Cell Organelles ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Stock ◽  
Viral Diarrhea Virus

SummaryThe aim of the present study was to evaluate the development and ultrastructure of preimplantation bovine embryos that were exposed to bovine viral diarrhea virus (BVDV) in vitro.The embryos were recovered from superovulated and fertilized Holstein-Friesian donor cows on day 6 of the estrous cycle. Compact morulae were microinjected with 20 pl of BVDV suspension (105.16 TCID50/ml viral stock diluted 1:4) under the zona pellucida (ZP), then washed in SOF medium and cultured for 24–48 h. Embryos were evaluated for developmental stages and then processed immunocytochemically for the presence of viral particles, using fluorescent anti-BVDV–FITC conjugate. Ultrastructure of cellular organelles was analysed by transmission electron microscopy (TEM).After microinjection of BVDV under the ZP, significantly more (p < 0.001) embryos (83.33%) were arrested at the morula stage compared with the intact control (30.33%). Immunocytochemical analysis localized the BVDV–FITC signal inside the microinjected embryos. TEM revealed: (i) the presence of virus-like particles in the dilated endoplasmic reticulum and in cytoplasmic vacuoles of the trophoblast and embryoblast cells; (ii) the loss of microarchitecture: and (iii) abnormal disintegrated nuclei, which lacked reticular structure and the heterochromatin area. In all, the embryo nuclear structure was altered and the microarchitecture of the nucleolus had disappeared when compared with the nuclei from control embryos. Dilatation of the intercellular space and the loss of the intercellular gap junctions were often observed in bovine BVDV-exposed embryos.These findings provide evidence for the adverse effect of BVDV virus on the development of bovine embryos, which is related to irreversible changes in the ultrastructure of cell organelles.

234 QUANTITATION OF BOVINE VIRAL DIARRHEA VIRUS ASSOCIATED WITH SINGLE ZONA PELLUCIDA-INTACT IN VIVO-DERIVED BOVINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 233 ◽  
Author(s):  
J. A. Gard ◽  
M. D. Givens ◽  
P. K. Galik ◽  
D. A. Stringfellow ◽  
K. P. Riddell ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Bovine Viral Diarrhea Virus ◽  
Zona Pellucida ◽  
Bovine Viral Diarrhea ◽  
Bovine Embryos ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
High Affinity ◽  
Viral Diarrhea Virus

Quantitation of bovine viral diarrhea virus (BVDV) associated with individual transferable embryos is prerequisite to a thorough assessment of the risk for transmission of BVDV via embryo transfer. One objective of this study was to determine the proportion of in vivo-derived bovine embryos that remained virus-positive after artificial exposure to a high-affinity strain of BVDV and thorough washing. A second objective was to determine the quantity of virus associated with these individual embryos. A total of 87 zona pellucida-intact, Day 7, in vivo-derived bovine embryos were exposed to a type 1 noncytopathic strain of BVDV (SD-1) and washed according to International Embryo Transfer Society recommendations. Subsequently, individual embryos were sonicated, and the RNA was extracted from the sonicate fluids and stored at -80�C until assayed using a real-time quantitative polymerase chain reaction (QPCR). Twenty-six percent (23/87) of the embryos contained virus. The average quantity of virus associated with individual embryos after viral exposure and washing was 1.12 viral copies per 5 �L (SD = 1.57 copies 5 per �L-1; SEM = 0.33 copies 5 per �L-1). Assessment of data using tolerance intervals (P = 0.05) indicates that 90% of contaminated embryos will be associated with ≤4.64 viral copies per 5 �L, whereas 99% of contaminated embryos will be associated with ≤6.62 copies per 5 �L. Obviously, only extremely small quantities of virus were associated with less than one-third of the embryos tested. Based on previous research, it is presumed that this virus is associated with the outer layers of the zona pellucida. A logical next step in the risk assessment would be to determine if these quantities of zona-associated virus are sufficient to infect na�ve recipients and/or embryonic cells after embryos are transferred. Further, similar efforts should be made to estimate the quantity of virus associated with in vitro-derived, zona pellucida-intact, bovine embryos after exposure to the same high-affinity strain of virus and washing.

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