Document Type
Thesis - Open Access
Award Date
2026
Degree Name
Master of Science (MS)
Department / School
Biology and Microbiology
Second Advisor
Radhey Kaushik
Abstract
The emergence of avian metapneumovirus (aMPV) subgroups A and B in the United States poultry industry has created an urgent need for standardized, host-specific diagnostic platforms. Historically, reliance on continuous, multi-species cell lines has limited the primary isolation of field strains and introduced potential adaptation artifacts. To address these challenges, this research first established and characterized a reproducible primary cell culture system derived from the tracheal tissues of 1-day-old turkey poults. This system was maintained in an optimized DMEM/F-12 formulation supplemented with 5% FBS, EGF, and ITS, with early-passage cultures achieving 90% confluence within 24 to 48 hours. Morphological and immunocytochemical validation confirmed a transition from a heterogeneous population to a uniform, homogenous mesenchymal/myofibroblastic monolayer characterized by the expression of alpha-smooth muscle actin (ɑ-SMA) and vimentin. Leveraging this standardized methodology, the study further developed and validated an embryonic chicken tracheal (ECT) cell model to directly isolate, adapt, and characterize the comparative dynamics of U.S. aMPV-A and aMPV-B strains. Primary ECT monolayers were inoculated with circulating strains to assess host susceptibility, replication kinetics, and protein expression. Using an optimized four-day harvest protocol, infection was confirmed and visualized via indirect immunofluorescence assay (IFA). Results demonstrate that this system effectively supports viral entry, replication, and protein synthesis for both aMPV-A and aMPV-B subgroups, with distinct fluorescence patterns: aMPV-A exhibited intense, punctate branching networks, whereas aMPV-B displayed heavily granular focal clusters. These findings validate these primary cell models as biologically relevant, host-homologous platforms for future virological applications, pathogen isolation, and diagnostic modeling of emerging respiratory threats.
Publisher
South Dakota State University
Recommended Citation
Serrao, Tricia, "Primary Avian Tracheal Cell Models for the Isolation of Emerging Avian Metapneumovirus Strains" (2026). Electronic Theses and Dissertations. 2143.
https://openprairie.sdstate.edu/etd2/2143
Rights Statement
