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During environmental monitoring of our institution's rodent watering systems, one vivarium was found to have high bacterial loads in the reverse-osmosis (RO) automatic water system. These findings prompted evaluation of the entire RO water production and distribution system. Investigation revealed insufficient rack and RO system sanitization, leading to heavy biofilm accumulation within the system. Approximately 2 wk after discovery in the water system, one of the bacterial organisms isolated in the water supply, Sphingomonas paucimobilis, was isolated from a peritoneal abscess of a severely immunodeficient B6. Cg-Slc11a1(r) Rag1(tm1Mom)/Cwi mouse housed in the same vivarium, suggesting that rodents drinking from this system were being exposed randomly to fragments of biofilm. Plans were developed to sanitize the entire system. Hypercholorination was used first, followed by treatment with a combination of peracetic acid and hydrogen peroxide. Between system sanitizations, a low-level chlorine infusion was added to the system as a biocide. Heterotrophic plate counts and bacterial isolation were performed on water samples obtained before and after sanitization procedures. We here discuss the process of identifying and correcting this important water-quality issue.

This study was performed in order to characterize the relationship between adhesion and biofilm formation abilities of drinking water-isolated bacteria (Acinetobacter calcoaceticus, Burkholderia cepacia, Methylobacterium sp., Mycobacterium mucogenicum, Sphingomonas capsulata and Staphylococcus sp.). Adhesion was assessed by two distinct methods: thermodynamic prediction of adhesion potential by quantifying hydrophobicity and the free energy of adhesion; and by microtiter plate assays. Biofilms were developed in microtiter plates for 24, 48 and 72 h. Polystyrene (PS) was used as adhesion substratum. The tested bacteria had negative surface charge and were hydrophilic. PS had negative surface charge and was hydrophobic. The free energy of adhesion between the bacteria and PS was > 0 mJ/m(2) (thermodynamic unfavorable adhesion). The thermodynamic approach was inappropriate for modelling adhesion of the tested drinking water bacteria, underestimating adhesion to PS. Only three (B. cepacia, Sph. capsulata and Staphylococcus sp.) of the six bacteria were non-adherent to PS. A. calcoaceticus, Methylobacterium sp. and M. mucogenicum were weakly adherent. This adhesion ability was correlated with the biofilm formation ability when comparing with the results of 24 h aged biofilms. Methylobacterium sp. and M. mucogenicum formed large biofilm amounts, regardless the biofilm age. Given time, all the bacteria formed biofilms; even those non-adherents produced large amounts of matured (72 h aged) biofilms. The overall results indicate that initial adhesion did not predict the ability of the tested drinking water-isolated bacteria to form a mature biofilm, suggesting that other events such as phenotypic and genetic switching during biofilm development and the production of extracellular polymeric substances (EPS), may play a significant role on biofilm formation and differentiation. This understanding of the relationship between adhesion and biofilm formation is important for the development of control strategies efficient in the early stages of biofilm development.

The effects of solution ionic strength on the collision efficiency (alpha) distribution of a Sphingomonas sp. were investigated using multiple sand columns of varying lengths and analyzing the bacteria clean-bed breakthrough concentrations using a distributed colloid filtration theory (D-CFT). Five different probability density functions (PDFs) were investigated and all accurately replicated the lab-scale experimental data, whereas a single alpha value could not. The alpha distribution shifted toward smaller values with decreasing ionic strength and the PDF parameters were strongly correlated to the Debye length, indicating that electrostatic interactions had a direct impact on the alpha distribution. The results indicate that while ionic strength has a large impact on bacterial transport distances for a concentration reduction of a few orders of magnitude, as occurs at the laboratory scale, due to the distributed nature of the collision efficiency, it has a minor effect on predicted transport distances required to achieve concentration reductions on the order of 10(6), which occurs at the field scale. Because of this, bacterial inactivation (e.g., death), rather than physically removing the bacteria from solution via filtration, is likely the key process impacting the transport of viable bacteria at the field scale. Overall, for systems with a distributed alpha, the results indicate that ionic strength has a strong influence on the transport of bacteria at the lab-scale (centimeters to one meter), both ionic strength and bacterial inactivation are important at the meso-scale (tens of meters), and inactivation becomes the dominant mechanism for reducing the transport of viable bacteria at the field scale (hundreds of meters).

In the present study, water was circulated in a simulated dental unit water line with electrifying a small current. The morphology of the biofilm developed on inner surface of the water line and the number of heterotrophic bacteria were investigated to elucidate the effect of a low level electric current on the biofilms formation associated with bacteria reproduction. Destruction and malconformation of biofilms by electrification was observed using SEM, in addition to deformation and hypertrophy of the bacteria. By naked eye observation, small pieces, which were possibly exfoliated biofilms, were detected in electrified water. While an adherent, yellow gel was demonstrated on the inner surface of the water line without electrification. With electrification, the number of bacteria decreased during the first week, however the bacteria increased gradually after that. The number of bacteria without electrification was consistently greater than that with electrification and the difference was statistically significant (P<0.05). The predominant bacteria were identified as Sphingomonas paucimobilis. The excess chlorine levels decreased to a minimum value within one week. The small current appeared to have effects on biofilm formation of heterotrophic bacteria that resulted in enhanced chlorine sterilization of dental unit water. Thus, electrification has considerable potential for the extermination of bacterial biofilms in dental unit water lines.

The oceans of the world are nutrient-limited environments that support a dynamic diversity of microbial life. Heterotrophic prokaryotes proliferate in oligotrophic regions and affect nutrient transformation and remineralization thereby impacting directly on the all marine biota. An important challenge in studying the microbial ecology of oligotrophic environments has been the isolation of ecologically important species. This goal has been recognized not only for its relevance in defining the dynamics of community composition, but for enabling physiological studies of competitive species and inferring their impact on the microbial food web. This review describes the successful isolation attempts of the ultramicrobacterium, Sphingopyxis alaskensis (formerly described as Sphingomonas alaskensis) using extinction dilution culturing methods. It then provides a comprehensive perspective of the unique physiological and genetic properties that have been identified that distinguish it from typical copiotrophic species. These properties are described through studies of the growth phase and growth rate control of macromolecular synthesis, stress resistance and global gene expression (proteomics). We also discuss the importance of integrating ecological and physiological approaches for studying microorganisms in marine environments.