First introduced into the United States in 1999,West Nile virus (WNV) has become endemic and has established itself as the predominant mosquito-borne arbovirus in North America. Transmitted by mosquitoes of the genus Culex, regional landscapes influence local vector species abundance, creating different mosquito ecologies that drive local transmission dynamics. In central Iowa, two mosquito species, Culex restuansTheobald and Culex pipiens Linnaeus, serve as the predominant mosquito vectors. Importantly, these mosquito populations are influenced by seasonal patterns in their abundance, with Cx. restuans preferring cool, early spring temperatures, while Cx. pipiens prefer the warmer, mid- to late-summer months. The point of the season at which Cx. pipiens becomes the dominant species is generally referred to as a ‘crossover’ period. To better understand the influence of crossover dynamics on WNV transmission, we examined environmental and mosquito abundance data, as well as mosquito infection rates and human disease cases from 2016 to 2018. We demonstrate that temperature influences the timing and duration of the crossover period, influencing mosquito abundance and human disease transmission. Together, these results suggest that Culex species crossover is an important variable of WNV transmission dynamics, which may provide an early warning indicators of increased WNV transmission.