Impact of temperature on the extrinsic incubation period of Zika virus in Aedes aegypti

Since Zika virus (ZIKV) emerged as a global human health threat, numerous studies have pointed to Aedes aegypti as the primary vector due to its high competence and propensity to feed on humans. The majority of vector competence studies have been conducted between 26-28 degrees C, but arboviral extrinsic incubation periods (EIPs), and therefore transmission efficiency, are known to be affected strongly by temperature. To better understand the relationship between ZIKV EIPs and temperature, we evaluated the effect of adult mosquito exposure temperature on ZIKV infection, dissemination, and transmission in Ae. aegypti at four temperatures: 18 degrees C, 21 degrees C, 26 degrees C, and 30 degrees C. Mosquitoes were exposed to viremic mice infected with a 2015 Puerto Rican ZIKV strain, and engorged mosquitoes were sorted into the four temperatures with 80% RH and constant access to 10% sucrose. ZIKV infection, dissemination, and transmission rates were assessed via RT-qPCR from individual mosquito bodies, legs and wings, and saliva, respectively, at three to five time points per temperature from three to 31 days, based on expectations from other flavivirus EIPs. The median time from ZIKV ingestion to transmission (median EIP, EIP50) at each temperature was estimated by fitting a generalized linear mixed model for each temperature. EIP50 ranged from 5.1 days at 30 degrees C to 24.2 days at 21 degrees C. At 26 degrees C, EIP50 was 9.6 days. At 18 degrees C, only 15% transmitted by day 31 so EIP50 could not be estimated. This is among the first studies to characterize the effects of temperature on ZIKV EIP in Ae. aegypti, and the first to do so based on feeding of mosquitoes on a live, viremic host. This information is critical for modeling ZIKV transmission dynamics to understand geographic and seasonal limits of ZIKV risk; it is especially relevant for determining risk in subtropical regions with established Ae. aegypti populations and relatively high rates of return travel from the tropics (e.g. California or Florida), as these regions typically experience cooler temperature ranges than tropical regions. Author summary Understanding how the environment affects Zika virus transmission is critical for protecting public health. Warmer temperatures accelerate mosquitoes' transmission of disease-causing pathogens, but this is not well-characterized for Zika virus in its primary vector, Aedes aegypti. In this study we measure the effect of a range of temperatures on Zika virus infection, dissemination, and transmission in Ae. aegypti and provide a mathematical equation that can be used in risk models to predict Zika virus transmission dynamics. We also explore how the effect of temperature on Zika virus transmission compares to the effects of temperature on other mosquito-borne viruses.