Advanced biofilm analysis in streams receiving organic deicer runoff.

Prolific heterotrophic biofilm growth is a common occurrence in airport receiving streams containing deicers and anti-icers, which are composed of low-molecular weight organic compounds. This study investigated biofilm spatiotemporal patterns and responses to concurrent and antecedent (i.e., preceding biofilm sampling) environmental conditions at stream sites upstream and downstream from Milwaukee Mitchell International Airport in Milwaukee, Wisconsin, during two deicing seasons (2009-2010; 2010-2011). Biofilm abundance and community composition were investigated along spatial and temporal gradients using field surveys and microarray analyses, respectively. Given the recognized role of Sphaerotilus in organically enriched environments, additional analyses were pursued to specifically characterize its abundance: a consensus sthA sequence was determined via comparison of whole metagenome sequences with a previously identified sthA sequence, the primers developed for this gene were used to characterize relative Sphaerotilus abundance using quantitative real-time PCR, and a Sphaerotilus strain was isolated to validate the determined sthA sequence. Results indicated that biofilm abundance was stimulated by elevated antecedent chemical oxygen demand concentrations, a surrogate for deicer concentrations, with minimal biofilm volumes observed when antecedent chemical oxygen demand concentrations remained below 48 mg/L. Biofilms were composed of diverse communities (including sheathed bacterium Thiothrix) whose composition appeared to shift in relation to antecedent temperature and chemical oxygen demand. The relative abundance of sthA correlated most strongly with heterotrophic biofilm volume (positive) and dissolved oxygen (negative), indicating that Sphaerotilus was likely a consistent biofilm member and thrived under low oxygen conditions. Additional investigations identified the isolate as a new strain of Sphaerotilus montanus (strain KMKE) able to use deicer components as carbon sources and found that stream dissolved oxygen concentrations related inversely to biofilm volume as well as to antecedent temperature and chemical oxygen demand. The airport setting provides insight into potential consequences of widescale adoption of organic deicers for roadway deicing.

Inhibitory effect of a surfactant on pure cultures of a filamentous and a floc forming micro-organism.

Activated sludge is the most widely used biological process for wastewater treatment. Inorganic and organic compounds are removed by a biotic community in the aeration basin. Problems of these systems are loss of settleability and poor sludge compaction due to excessive growth of filamentous micro-organisms. The filamentous bulking can be controlled by the addition of chemical agents. Strong oxidants, such as chlorine, are utilized to eliminate filamentous bacteria; however, these substances also tend to attack floc-forming bacteria and to cause process breakdown. Besides, chlorine may become hazardous owing to the formation of chemical products as chloramines. Surfactant addition constitutes an interesting alternative for the control of filamentous bulking. In this work the effect of a surfactant Triton X-100 (octylphenol ethoxylate), on the respiratory activity (RA) of pure cultures of a filamentous (Sphaerotilus natans) and a floc-former microorganism (Acinetobacter anitratus) was evaluated. In the concentration range tested (60-220 mg l(-1)), the surfactant was observed to exhibit high RA specific inhibition of the filamentous micro-organism with no significant effect on the floc-forming bacteria. Light microscopy observations showed that the surfactant induced cell lysis, leaving only empty sheaths in the case of filamentous micro-organisms. A kinetic equation to predict the microbial RA fraction of a S. natans pure culture as a function of surfactant concentration and contact time was proposed. The effect of Triton X-100 on the inactivation of pure cultures of both micro-organisms was compared to that of chlorine. Triton X-100 results were adequate to eliminate filamentous bacteria emerging as an alternative for filamentous bulking treatment.

Effects of fatty acids on growth and poly-3-hydroxybutyrate production in bacteria.

The effects of saturated and unsaturated fatty acids (lauric acid, palmitic acid, steric acid, oleic acid, linoleic acid, soybean oil) on Sphaerotilus natans, 0B17 (Pseudomonas sp.), and recombinant Escherichia coli DH5(/pUC19/CAB were studied. Oleic acid enhances Poly-3-hydroxybutyrate (PHB) production in these three bacterial strains, suggesting that the single double bond of the acid activates the polyhydroxylkanoate accumulation enzymatic reaction. Under the effect of lauric acid and linoleic acid, the growth of S. natans and 0B17 were totally inhibited. However, the enhanced PHB accumulation in recombinant E. coli was observed.