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Results A total of nine different animal-derived cell lines indigenous to North America were evaluated, including domestic ruminant (sheep and calf), wild ruminant (PDGFRB white-tailed deer, mule deer, pronghorn, elk), and non-ruminant (pig, coyote, frog) species. In both animals and humans, RVFV typically targets the liver and spleen, and can be neuroinvasive; however, it is also known to replicate in a variety of other tissues and cells (Ikegami and Makino, 2011). Cell lines derived from kidney, brain, and lung tissues were compared due to the availability of these tissues across multiple animal species. The attenuated RVFV MP-12 strain was used for initial comparisons of virus replication in various cell lines, as it does not require work at a high biosecurity level facility. Viral growth kinetics were then performed with a representative set of cell lines with the virulent SA01-1322 strain RVFV. The brain cell lines from domestic and wild ruminants supported the most efficient viral replication of the MP-12 strain. Using a MOI of 0.1, viral titers in the ruminant-derived brain cell lines peaked around 24 hpi and MP-12 titers ranged between 105 and 107 pfu/ml (Figure ?Figure1A1A). Replication kinetics of MP-12 were more rapid in the calf, white-tailed, and mule deer cell lines, with titers nearly 102 to 103 pfu/ml higher at 6 hpi compared to titers from sheep, pronghorn, and elk cell cultures (Figure ?Figure1A1A). Replication kinetics of MP-12 in the pig brain cell line was significantly delayed compared to the ruminant brain cells. Expansion of MP-12 in the pig brain cells did not occur until after 12 hpi and only a titer around 103 pfu/ml was achieved by 48 hpi which was more than 102 pfu/ml lower than the lowest titer observed at the same time point among the ruminant-derived brain cell lines (Figure ?Figure1A1A). Conversely, replication kinetics and titers of the SA01-1322 strain were more consistent among the white-tailed deer, sheep, and pig brain cell lines tested (Figure ?Figure1B1B). Titers averaged around 103 pfu/ml by 6 hpi and then peaked around 105 to 106 pfu/ml between 24 and 36 hpi, with the white-tailed deer cell line supporting slightly higher virus titers (Figure ?Figure1B1B). FIGURE 1 Rift Valley fever virus (RVFV) replication kinetics in North American livestock and wildlife cell lines. Cell cultures derived from brain (A,B), kidney (C,D), and lung (E) were infected with MP-12 (A,C,E) or SA01-1322 (B,D) strains at 0.1 multiplicity ... The replication kinetics of the MP-12 strain were slightly slower in the kidney cell lines compared to the brain cell lines with titers peaking after 36�C48 hpi (Figure ?Figure1C1C); however, high titers were still achieved in these cell lines. Titers of MP-12 in the white-tailed deer and sheep kidney cell lines reached 107 pfu/ml, while the pig and frog kidney cell lines supported MP-12 titers of 105 and 106 pfu/ml, respectively (Figure ?Figure1C1C).