Halophilic and halotolerant bacteria from river waters and shallow groundwater along the Rouge River of southeastern Michigan

The use of sodium chloride to melt highway and road snow is believed to have a significant effect on the groundwater ecosystem of the rivers where the salt from the roads drain. As the river composition changes, the bacterial population also changes to favour those bacteria that are more suited to the higher salt concentrations. In this experiment, we surveyed the cultivable salt-loving organisms (halophiles) on three sites that encompass the Rouge River (Lotz; site 1, Lilly, site; 8, and Ford Field, site 9). A total of 125 isolates were surveyed. Representative isolates of distinct morphologies were subjected to physiological test, using API strips and identified by 16 rDNA sequence analysis. The 16S rDNA sequences were analyzed and compared with sequences from Genbank. Results indicated that the SSU rRNA sequences of the bacterial isolates were similar to six major genera, Bacillus, Staphylococcus, Halobacillus, Paenabacillus, Halomonas, and Clostridium. Half of the isolates sequenced were similar to Bacillus spp. The API assay showed that the majority of the isolates were positive for the enzymes tryptophane deaminase, gelatinase and P-galactosidase. Indole production, acetoin production and citrate utilization were not observed for any isolates. Fermentation of carbohydrates was observed for very few isolates. The primary enzyme found in all isolates was arginine dihydrolase, which might be an indicator of the presence of such enzyme in halophilic and halotolerant bacteria present in the Rouge River.