Bio-Safety Technology in Production of Bovine Herpesvirus Type 5 (BoHV-5) Using an Alternative Serum-Free Medium

Abstract

The detection and growth of bovine herpes virus type 5 (BoHV-5) was evaluated in three different media without foetal bovine serum or animal protein by infecting three different cell lines. The OPTI-PRO, VP-SFM and RPMI 1640 media were supplemented by L-glutamine, antibiotics added by 5µg of insulin-like growth factor (IGF-I) and used to adapt CER, MDBK and CRIB cells in a statistic continuous culture system. The results obtained by CRIB and MDBK cells adaptation showed steady growth after 10^th passages in OPTI-PRO medium, 20^th passages in VP-SFM medium and 30^th passages in RPMI 1640 medium, respectively. The OPTI-PRO and RPMI 1640 media were not able to support CER growth, even supplemented by IGF-1 and present apoptotic cells at 72h post-infection. The CER cells seeded after adaptation by 10^th passages in VP-SFM medium added by 5µg/20ml of IGF-1 growth factor revealed the best virus titres compared to MDBK and CRIB cell lines. It was concluded that CER, MDBK and CRIB cells infected by BoHV-5 serotype could be used for laboratory diagnosis propagated on much safer culture system of high biotechnology advances.