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Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer

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Abstract
The behavior of BTV-8 in cattle is different from most other serotypes not only with regards to clinical signs but certainly with respect to virus transmission (transplacental, contact). Therefore, the possibility of virus transmission by means of embryo transfer was examined by in vitro exposure of in vitro produced and in vivo derived bovine blastocysts to BTV-8 followed by different washing protocols, including longer exposure times (up to 120 s) to 0.25% trypsin at room temperature or at 37 degrees C. None of the washing protocols used was successful in removing the viral genome completely from the in vitro produced and in vivo derived embryos as was demonstrated by real-time PCR. Moreover, BTV-8 virus was transmitted to recipient cows after embryo transfer of in vivo derived BTV8-exposed embryos, which had been subjected to routine decontamination as recommended by IETS, consisting of 5 washes in PBS followed by a double treatment of 0.25% trypsin for 45s at 37 degrees C, and an additional 5 washes in PBS with 2% FCS. This study clearly demonstrates the necessity of vigorous application of the directives for screening of potential donors and the collected embryos, especially in regions with BTV-8, to prevent transmission of the disease.
Keywords
bovine embryo, IETS guidelines, Bluetongue virus, BTV-8, transmission, TIME RT-PCR, VIRAL-INFECTIONS, CLINICAL SIGNS, CATTLE, SHEEP, EPIDEMIC, BELGIUM, PERFORMANCE, CALVES, RNA

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MLA
Haegeman, Andy, et al. “Failure to Remove Bluetongue Serotype 8 Virus (BTV-8) from in Vitro Produced and in Vivo Derived Bovine Embryos and Subsequent Transmission of BTV-8 to Recipient Cows after Embryo Transfer.” FRONTIERS IN VETERINARY SCIENCE, vol. 6, 2019.
APA
Haegeman, A., Vandaele, L., De Leeuw, I., Oliveira, A., Nauwynck, H., Van Soom, A., & De Clercq, K. (2019). Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer. FRONTIERS IN VETERINARY SCIENCE, 6.
Chicago author-date
Haegeman, Andy, Leen Vandaele, Ilse De Leeuw, Andre Oliveira, Hans Nauwynck, Ann Van Soom, and Kris De Clercq. 2019. “Failure to Remove Bluetongue Serotype 8 Virus (BTV-8) from in Vitro Produced and in Vivo Derived Bovine Embryos and Subsequent Transmission of BTV-8 to Recipient Cows after Embryo Transfer.” FRONTIERS IN VETERINARY SCIENCE 6.
Chicago author-date (all authors)
Haegeman, Andy, Leen Vandaele, Ilse De Leeuw, Andre Oliveira, Hans Nauwynck, Ann Van Soom, and Kris De Clercq. 2019. “Failure to Remove Bluetongue Serotype 8 Virus (BTV-8) from in Vitro Produced and in Vivo Derived Bovine Embryos and Subsequent Transmission of BTV-8 to Recipient Cows after Embryo Transfer.” FRONTIERS IN VETERINARY SCIENCE 6.
Vancouver
1.
Haegeman A, Vandaele L, De Leeuw I, Oliveira A, Nauwynck H, Van Soom A, et al. Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer. FRONTIERS IN VETERINARY SCIENCE. 2019;6.
IEEE
[1]
A. Haegeman et al., “Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer,” FRONTIERS IN VETERINARY SCIENCE, vol. 6, 2019.
@article{8656569,
  abstract     = {The behavior of BTV-8 in cattle is different from most other serotypes not only with regards to clinical signs but certainly with respect to virus transmission (transplacental, contact). Therefore, the possibility of virus transmission by means of embryo transfer was examined by in vitro exposure of in vitro produced and in vivo derived bovine blastocysts to BTV-8 followed by different washing protocols, including longer exposure times (up to 120 s) to 0.25% trypsin at room temperature or at 37 degrees C. None of the washing protocols used was successful in removing the viral genome completely from the in vitro produced and in vivo derived embryos as was demonstrated by real-time PCR. Moreover, BTV-8 virus was transmitted to recipient cows after embryo transfer of in vivo derived BTV8-exposed embryos, which had been subjected to routine decontamination as recommended by IETS, consisting of 5 washes in PBS followed by a double treatment of 0.25% trypsin for 45s at 37 degrees C, and an additional 5 washes in PBS with 2% FCS. This study clearly demonstrates the necessity of vigorous application of the directives for screening of potential donors and the collected embryos, especially in regions with BTV-8, to prevent transmission of the disease.},
  articleno    = {432},
  author       = {Haegeman, Andy and Vandaele, Leen and De Leeuw, Ilse and Oliveira, Andre and Nauwynck, Hans and Van Soom, Ann and De Clercq, Kris},
  issn         = {2297-1769},
  journal      = {FRONTIERS IN VETERINARY SCIENCE},
  keywords     = {bovine embryo,IETS guidelines,Bluetongue virus,BTV-8,transmission,TIME RT-PCR,VIRAL-INFECTIONS,CLINICAL SIGNS,CATTLE,SHEEP,EPIDEMIC,BELGIUM,PERFORMANCE,CALVES,RNA},
  language     = {eng},
  pages        = {9},
  title        = {Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer},
  url          = {http://dx.doi.org/10.3389/fvets.2019.00432},
  volume       = {6},
  year         = {2019},
}

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