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Transmission of Mouse Minute Virus (MMV) but Not Mouse Hepatitis Virus (MHV) Following Embryo Transfer with Experimentally Exposed In Vivo-Derived Embryos¹

Department of Comparative Medicine,³ GSF–National Research Center for Environment and Health, D-85764 Neuherberg, Germany The Microbiology Laboratories,⁴ North Harrow, Middlesex HA2 7RE, United Kingdom Departments of Obstetrics, Reproduction, and Herd Health;⁵ and Parasitology, Virology, and Immunology,⁶ Faculty of Veterinary Medicine, University of Ghent, 9820 Merelbeke, Belgium

ABSTRACT

The present study investigated the presence and location of fluorescent microspheres having the size of mouse hepatitis virus (MHV) and of mouse minute virus (MMV) in the zona pellucida (ZP) of in vivo-produced murine embryos, the transmission of these viruses by embryos during embryo transfer, and the time of seroconversion of recipients and pups. To this end, fertilized oocytes and morulae were exposed to different concentrations of MMVp for 16 h, while 2-cell embryos and blastocysts were coincubated for 1 h. In addition, morulae were exposed to MHV-A59 for 16 h. One group of embryos was washed, and the remaining embryos remained unwashed before embryo transfer. Serological analyses were performed by means of ELISA to detect antibodies to MHV or MMV in recipients and in progeny on Days 14, 21, 28, 42, and 63 and on Days 42, 63, 84, 112, 133, and 154, respectively, after embryo transfer. Coincubation with a minimum of 10⁵/ml of fluorescent microspheres showed that particles with a diameter of 20 nm but not 100 nm crossed the ZP of murine blastocysts. Washing generally led to a 10-fold to 100-fold reduction of MMVp. Washed MMV-exposed but not MHV-exposed embryos led to the production of antibodies independent of embryonic stage and time of virus exposure. Recipients receiving embryos exposed to a minimum of 10⁷ mean tissue culture infective dose (TCID₅₀)/ml of MHV-A59 and 10² TCID₅₀/ml of MMVp seroconverted by Day 42 after embryo transfer. The results indicate that MMV but not MHV can be transmitted to recipients even after washing embryos 10 times before embryo transfer.

assisted reproductive technology, embryo transfer, health monitoring, mouse, mouse hepatitis virus, mouse minute virus

¹Supported in part by the Gesellschaft für Versuchstierkunde–Society of Laboratory Animal Science and the National Genome Research Network, Germany.

Correspondence: ²Esther Mahabir, Department of Comparative Medicine, GSF–National Research Center for Environment and Health, Ingolstädter Landstrasse 1, D-85764 Neuherberg, Germany. FAX: 49 89 3187 3321; e-mail: mahabir{at}gsf.de

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				E Mahabir, D Bulian, R Schmoller, J Needham, and J Schmidt Production of Virus-Free Seronegative Pups from Murine Embryos Arising from In Vitro Fertilization with Mouse Minute Virus-Exposed Spermatozoa Biol Reprod, January 1, 2008; 78(1): 53 - 58. [Abstract] [Full Text] [PDF]