title: Use of different cell lines for in vitro cultures of bovine respiratory syncytial virus.
abstract: This study compared the use of different cell lines for in vitro cultures of bovine respiratory syncytial virus (BRSV). The BRSV 375 strain and 3 nasal swabs obtained from Simmental calves were used for this study. The culture was performed on 3 cell lines: bovine kidney cells (LLC-PK1), bovine tracheal cells (TBTR) and primary chicken embryo-related cells (CER). A comparative analysis of titres was performed using a microplate agglutination test with human group O erythrocytes and bovine erythrocytes. The presence of BRSV in all cell lines was confirmed using the reverse transcriptase polymerase chain reaction (RT-PCR) method. The first small refractile changes in the LLC-PK1 cells occurred at 48h after infection. Syncytial changes were noted 4 days after incubation. Large refractile cell changes were observed on day 3 of growth in the TBTR culture. Syncytia were observed on the second day after infection in subsequent passages. The cytopathic effect in the CER cells occurred 24h after infection, and syncytia appeared after 3 passages. Changes in syncytia indicate an adaptation of the virus for the infection of cells other than tracheal cells in primary and secondary cultures. The highest viral titre was obtained using the TBTR line. The titres obtained in the LLC-PK1 and CER cultures averaged 10(1.86)/ml. The low virus titres in all culture types suggest the need for research aimed at the optimisation of culture conditions.
authors.affiliation: Sub-Department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland. Electronic address: renata.urban@up.lublin.pl.
authors.forename: Renata
authors.initials: R
authors.lastname: Urban-Chmiel
authors.affiliation: Sub-Department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Wernicki
authors.affiliation: Sub-Department of Veterinary Microbiology, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Majer-Dziedzic
authors.affiliation: Sub-Department of Veterinary Microbiology, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Gnat
authors.affiliation: Sub-Department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Puchalski
authors.affiliation: Sub-Department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033 Lublin, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Dec
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D002642:Chick Embryo; D004912:Erythrocytes; D006386:Hemagglutination Tests; D006801:Humans; D018112:Respiratory Syncytial Virus, Bovine / Q000254:growth & development*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019562:Viral Load; D014776:Virus Cultivation / Q000379:methods
keywords: Bovine respiratory syncytial virus; Calves; Cell cultures; Respiratory syndrome