Global warming and anthropogenic disturbances significantly influence the biosphere, tremendously increasing species extinction rates. In central Alabama we analyzed Drosophilidae species composition change 94 years after the previous survey. We found ten Drosophilid species that were not reported during the last major biodiversity survey, two of which are invasive pests. In addition, we analyzed the influence of environmental variables characteristic of the subtropical climate zone on Drosophila abundance and biodiversity. We found a significant correlation between temperature and abundance of total Drosophila as well as for six out of the seven most represented species individually, with a maximum abundance at intermediate temperatures (18-26°C). In addition, temperature was positively correlated with biodiversity of Drosophila. Precipitation produced a significant effect on the abundance of five species of Drosophila, with different optima for each species, but did not affect overall biodiversity. We concluded that in the subtropical climate zone of central Alabama, temperature and precipitation produce a significant effect on Drosophila abundance and biodiversity, while local land use also impacts fly abundance, contributing to a substantial shift in species composition over the last century. We expect global climate change and other anthropogenic factors to further impact Drosophila species composition in the subtropical climate zone into the future.