The impact of highways on local snakes was examined on the new and old sections of Xiaomo Highway in Xishuangbanna between Yunlu Rubber Factory and Manfen and a rural road nearby. We cruised along the three roads in search of signs of live and dead snakes from December 2007 to December 2008. A total of 19 sites were founded with snakes alive or dead on the roads. At each location where snake was found we recorded information of the surrounding vegetation type, forest coverage and the distance to the nearest village. To compare to the background environment, we set 68 transect points at a 100m interval along the roads and recorded the same parameters. We also collected meteorological data including temperature, precipitation, and humidity of the day snake signs were found and of the three days prior to the survey date. Data collected from both the snake-present and background control sites were analyzed in a forward logistic regression model to identify key environment variables affecting the presence of snakes on roads. We then treated each survey day as a single sample unit, and summed up the number of snakes recorded on all three roads in a day as the dependent variable. A multivariate stepwise linear regression was applied to evaluate the correlation between the meteorological variables and the timing of snakes found on roads. A total of 20 individuals of 10 snake species were recorded on roads during this 52-day survey. The detection likelihood of snakes on roads with natural forest nearby was significantly lower than roads where roadside was the farmland or the rubber tree plantation. The primary meteorological factors leading to the presence of snakes on roads may be the lower maximal temperature and the higher maximal humidity two days prior to the survey date. It is evident that humidity and temperature plays important roles on the activeness of sub-tropical snakes, but not precipitation. The likelihood of snakes being crushed by cars varies among species, which may due to species′ differences in body size and behavioral patterns. At last, among the 14 species of snakes found there were some of the rarest species in China such as Boiga cyanea and Chrysopelea ornate. Our findings urged conservation measures including building culverts or pathways for wildlife along the Xiaomo Highway to reduce the road mortality of snakes and other reptiles such as Varanus salvator.