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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Ruminant Diseases and Immunology Research » Research » Publications at this Location » Publication #389264
Research Project: Identification of Disease Mechanisms and Control Strategies for Viral Respiratory Pathogens of Ruminants

Location: Ruminant Diseases and Immunology Research

Title: Virome Characterization in Commercial Bovine Serum Batches—A Potentially Needed Testing Strategy for Biological Products

Author
item Falkenberg, Shollie
item PAIM, WILLIAN - Oklahoma State University
item MAGGIOLI, MAYARA - Oklahoma State University
item RAMACHANDRAN, AKHILESH - Oklahoma State University
item WEBER, MATHEUS - Brazil University
item CANAL, CLAUDIO - Brazil University
item BAUERMANN, FERNANDO - Oklahoma State University

Submitted to: Transboundary and Emerging Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/29/2021
Publication Date: 12/3/2021
Citation: Falkenberg, S.M., Paim, W.P., Maggioli, M.F., Ramachandran, A., Weber, M.N., Canal, C.W., Bauermann, F.V. 2021. Virome Characterization in Commercial Bovine Serum Batches—A Potentially Needed Testing Strategy for Biological Products. Transboundary and Emerging Diseases. 13(12). Article 2425. https://doi.org/10.3390/v13122425.
DOI: https://doi.org/10.3390/v13122425

Interpretive Summary: Bovine serum is widely used as a supplement for cell culture media. This serum can be contaminated with adventitious agents that can increase the risk of inadvertently contaminating cell cultures due to its use. Currently, the commercial bovine serum is tested for possible viral contaminants following regional guidelines and regulatory agencies are most concerned with detecting animal origin high-consequence viruses. While testing for high-consequence viruses is important, this does not account for new and emerging viruses that may contaminate cell cultures and ultimately contaminate biological products. High-throughput sequencing (HTS) is a potential option for screening of bovine serum, as no prior knowledge of the viral targets is required, and all viruses present will be detected not just specifically targeted pathogens. In the present study, multiple batches of bovine serum were tested for adventitious viral agents. A variety of sequences were assembled representing the different viruses in each bovine serum sample. Results from this study demonstrated a variety of viruses that would not have been detected using routine testing methods such as virus isolation, immunofluorescence, and hemadsorption assays. These results support that HTS should be considered as testing method for pooled biologicals to detect viral contaminants.

Technical Abstract: Bovine serum has been widely used as a universal supplement in culture media and other applications, including the manufacturer of biological products. Currently, the commercial bovine serum is tested for possible viral contaminants following regional guidelines. Regulatory agencies established tests focus on detecting selected animal origin high-consequence viruses and are based on virus isolation, immunofluorescence, and hemadsorption assays. However, these tests may fail to detect new or emerging viruses in biological products. High-throughput sequencing (HTS) is a powerful option, as no prior knowledge of the viral targets is required. In the present study, we evaluate the virome of seven commercial batches of bovine serum from Mexico (one batch), the United States (four batches), and New Zealand (two batches) using a specific preparation and enrichment method for pooled samples, and sequencing using an Illumina platform. Sequences representing viral families with single-stranded DNA (Parvoviridae), double-stranded DNA (Polyomaviridae, and Adenoviridae), circular Rep-encoding single-stranded (CRESS) DNA (Genomoviridae, Circoviridae, and Smacoviridae), single-stranded RNA (Flaviviridae, Picornaviridae, and Retroviridae), and double-stranded RNA (Reoviridae) were identified. In addition, complete coding genomes of Bovine Parvovirus 2 (BPV-2), Bovine Parvovirus 3 (BPV-3), Bosavirus (BosaV), Bovine Hokovirus 2 (BHoV-2), and Bovine Polyomavirus 1 (BPyV-1) were retrieved. Furthermore, CrAssphage, a recently discovered group of bacteriophage correlated with fecal contamination, was identified in 85% of the tested batches. These results support that HTS associated with viral enrichment is a robust tool and should be considered as an additional layer of safety when testing pooled biologicals to detect viral contaminants.