Chemical and antimicrobial treatments change the viscoelastic properties of bacterial biofilms.

Changes in the viscoelastic material properties of bacterial biofilms resulting from chemical and antimicrobial treatments were measured by rheometry. Colony biofilms of Staphylococcus epidermidis or a mucoid Pseudomonas aeruginosa were subjected to a classical creep test performed using a parallel plate rheometer. Data were fit to the 4-parameter Burger model to quantify the material properties. Biofilms were exposed to the chloride salts of several common mono-, di-, and tri- valent cations, and to urea, industrial biocides, and antibiotics. Many of these treatments resulted in statistically significant alterations in the material properties of the biofilm. Multivalent cations stiffened the P. aeruginosa biofilm, while ciprofloxacin and glutaraldehyde weakened it. Urea, rifampin, and a quaternary ammonium biocide weakened the S. epidermidis biofilm. In general, there was no correspondence between the responses of the two different types of biofilms to a particular treatment. These results underscore the distinction between the killing power of an antimicrobial agent and its ability to alter biofilm mechanical properties and thereby influence biofilm removal. Understanding biofilm rheology and how it is affected by chemical treatment could lead to improvements in biofilm control.

Seasonal influence on sulfate reduction and zinc sequestration in subsurface treatment wetlands.

To characterize the effects of season, temperature, plant species, and chemical oxygen demand (COD) loading on sulfate reduction and metals removal in treatment wetlands we measured pore water redox potentials and concentrations of sulfate, sulfide, zinc and COD in subsurface wetland microcosms. Two batch incubations of 20 day duration were conducted in each of four seasons defined by temperature and daylight duration. Four treatments were compared: unplanted controls, Typha latifolia (broadleaf cattail), and Schoenoplectus acutus (hardstem bulrush), all at low COD loading (267 mg/L), plus bulrush at high COD loading (534 mg/L). Initial SO4-S and zinc concentrations were 67 and 24 mg/L, respectively. For all treatments, sulfate removal was least in winter (4 degrees C, plant dormancy) greatest in summer (24 degrees C, active plant growth) and intermediate in spring and fall (14 degrees C), but seasonal variation was greater in cattail, and especially, bulrush treatments. Redox measurements indicated that, in winter, plant-mediated oxygen transfer inhibited activity of sulfate reducing bacteria, exacerbating the reduction in sulfate removal due to temperature. Doubling the COD load in bulrush treatments increased sulfate removal by only 20-30% when averaged over all seasons and did not alter the basic pattern of seasonal variation, despite tempering the wintertime increase in redox potential. Seasonal and treatment effects on zinc removal were broadly consistent with sulfate removal and presumably reflected zinc-sulfide precipitation. Results strongly suggest that interactive effects of COD loading rate, temperature, season, and plant species control not only sulfate reduction and zinc sequestration, but also the balance of competition between various microbial consortia responsible for water treatment in constructed wetlands.

Chlorination of model drinking water biofilm: implications for growth and organic carbon removal.

The influence of chlorine on biofilm in low organic carbon environments typical of drinking water or industrial process water was examined by comparing biomass and kinetic parameters for biofilm growth in a chlorinated reactor to those in a non-chlorinated control. Mixed-population heterotrophic biofilms were developed in rotating annular reactors under low concentration, carbon-limited conditions (< 2 mg/L as carbon) using three substrate groups (amino acids, carbohydrates and humic substances). Reactors were operated in parallel under identical conditions with the exception that chlorine was added to one reactor at a dose sufficient to maintain a free chlorine residual of 0.09-0.15 mg/L in the effluent. The presence of free chlorine resulted in development of less biofilm biomass compared to the control for all substrates investigated. However, specific growth and organic carbon removal rates were on the average five times greater for chlorinated biofilm compared to the control. Observed yield values were less for chlorinated biofilm. Although chlorinated biofilm's specific organic carbon removal rate was high, the low observed yield indicated organic carbon was being utilized for purposes other than creating new cell biomass. The impacts of free chlorine on mixed-population biofilms in low-nutrient environments were different depending upon the available substrate. Biofilms grown using amino acids exhibited the least difference between control and chlorinated kinetic parameters; biofilm grown using carbohydrates had the greatest differences. These findings are particularly relevant to the fundamental kinetic parameters used in models of biofilm growth in piping systems that distribute chlorinated, low-carbon-concentration water.