Molecular methods to detect and monitor dissimilatory arsenate-respiring bacteria (DARB) in sediments

Dissimilatory arsenate-respiring bacteria (DARB) reduce arsenate to arsenite and may play a significant role in arsenic mobilization in aquifers and anoxic sediments. Many studies have been conducted with pure cultures of DARB to understand their involvement in arsenic contamination. However, few studies have examined uncultured DARB in the environment. In order to investigate uncultured DARB in anoxic sediments, genes encoding arsenate respiratory reductases (arr) were targeted as a genetic marker. Degenerate primers for the alpha-subunit of arr genes were designed and used with PCR amplification to detect uncultured DARB in the sediments collected from three stations (upper, mid and lower bay) in the Chesapeake Bay. Phylogenetic analysis of putative arrA genes revealed the diversity of DARB with distinct community structures at each of the three stations. Arsenate reduction in sediment communities was confirmed using enrichment cultures established with sediment samples from the upper bay. In addition, terminal restriction fragment length polymorphism analysis of the putative arrA genes showed changes in the community structure of DARB in the enrichment cultures while reducing arsenate. This was also confirmed by cloning and sequence analysis of the arrA genes obtained from the enrichment cultures. Thus, we were able to detect diverse uncultured DARB in sediments, as well as to describe changes in DARB community structure during arsenic reduction in anoxic environments.