Evaluation of Gallium Citrate Formulations against a Multidrug-Resistant Strain of Klebsiella pneumoniae in a Murine Wound Model of Infection.

Skin and soft tissue infections (SSTIs) are a common occurrence in health care facilities with a heightened risk for immunocompromised patients. Klebsiella pneumoniae has been increasingly implicated as the bacterial agent responsible for SSTIs, and treatment can be challenging as more strains become multidrug resistant (MDR). Therefore, new treatments are needed to counter this bacterial pathogen. Gallium complexes exhibit antimicrobial activity and are currently being evaluated as potential treatment for bacterial infections. In this study, we tested a topical formulation containing gallium citrate (GaCi) for the treatment of wounds infected with K. pneumoniae. First, the MIC against K. pneumoniae ranged from 0.125 to 2.0 µg/ml GaCi. After this in vitro efficacy was established, two topical formulations with GaCi (0.1% [wt/vol] and 0.3% [wt/vol]) were tested in a murine wound model of MDR K. pneumoniae infection. Gross pathology and histopathology revealed K. pneumoniae-infected wounds appeared to close faster with GaCi treatment and were accompanied by reduced inflammation compared to those of untreated controls. Similarly, quantitative indications of infection remediation, such as reduced weight loss and wound area, suggested that treatment improved outcomes compared to those of untreated controls. Bacterial burdens were measured 1 and 3 days following inoculation, and a 0.5 to 1.5 log reduction of CFU was observed. Lastly, upon scanning electron microscopy analysis, GaCi treatment appeared to prevent biofilm formation on dressings compared to those of untreated controls. These results suggest that with more preclinical testing, a topical application of GaCi may be a promising alternative treatment strategy for K. pneumoniae SSTI.

Rare taxa have potential to make metabolic contributions in enhanced biological phosphorus removal ecosystems.

Enhanced biological phosphorus removal (EBPR) relies on diverse but specialized microbial communities to mediate the cycling and ultimate removal of phosphorus from municipal wastewaters. However, little is known about microbial activity and dynamics in relation to process fluctuations in EBPR ecosystems. Here, we monitored temporal changes in microbial community structure and potential activity across each bioreactor zone in a pilot-scale EBPR treatment plant by examining the ratio of small subunit ribosomal RNA (SSU rRNA) to SSU rRNA gene (rDNA) over a 120 day study period. Although the majority of operational taxonomic units (OTUs) in the EBPR ecosystem were rare, many maintained high potential activities based on SSU rRNA : rDNA ratios, suggesting that rare OTUs contribute substantially to protein synthesis potential in EBPR ecosystems. Few significant differences in OTU abundance and activity were observed between bioreactor redox zones, although differences in temporal activity were observed among phylogenetically cohesive OTUs. Moreover, observed temporal activity patterns could not be explained by measured process parameters, suggesting that other ecological drivers, such as grazing or viral lysis, modulated community interactions. Taken together, these results point towards complex interactions selected for within the EBPR ecosystem and highlight a previously unrecognized functional potential among low abundance microorganisms in engineered ecosystems.

Production and characterization of foam in the anoxic zone of a membrane-enhanced biological phosphorus removal process.

A pilot-scale membrane-enhanced biological phosphorus removal process accumulated substantial quantities of stable foam on the surface of the anoxic zone. The foam contained 4 to 6% dry matter, with specific nitrogen and phosphorus contents that were similar to those of the underlying anoxic zone mixed liquor. Kinetic studies demonstrated that the specific rate of phosphorus release from the foam was only 25 to 30% of that observed with mixed liquor from the aerobic zone. Molecular techniques demonstrated that the calculated similarity of the microbial communities in the foam and the underlying mixed liquor was approximately 80%, with two phylotypes (Gordonia amarae and Microthrix parvicella) being uniquely enriched in the foam and one phylotype (Epistylis sp.) more abundant in the underlying mixed liquor. The production of foam was demonstrated to be a consistent phenomenon that depended on the concentration of the suspended solids in the bioreactor.

A comparison of bacterial populations in enhanced biological phosphorus removal processes using membrane filtration or gravity sedimentation for solids-liquid separation.

In an earlier phase of this study, we compared the performances of pilot scale treatment systems operated in either a conventional enhanced biological phosphorus removal (CEBPR) mode, or a membrane enhanced biological phosphorus removal (MEBPR) mode. In the present investigation, we characterized the bacterial community populations in these processes during parallel operation with the same municipal wastewater feed. The objectives of the study were (1) to assess the similarity of the bacterial communities supported in the two systems over time, (2) to determine if distinct bacterial populations are associated with the MEBPR and CEBPR processes, and (3) to relate the dynamics of the community composition to changes in treatment process configuration and to treatment process performance. The characteristics of the bacterial populations were first investigated with ribosomal intergenic spacer analysis, or RISA. To further understand the bacterial population dynamics, important RISA phylotypes were isolated and identified through 16S RNA gene sequencing. The parallel MEBPR and CEBPR systems developed bacterial communities that were distinct. The CEBPR community appeared to exhibit greater diversity, and this may have been the primary reason why the CEBPR treatment train demonstrated superior functional stability relative to the MEBPR counterpart. Moreover, the more diverse bacterial population apparent in the CEBPR system was observed to be more dynamic than that of the MEBPR process. Several RISA bands were found to be characteristic of either the membrane or conventional biological system. In particular, the MEBPR configuration appeared to be selective for the slow-growing organism Magnospira bakii and for the foam-associated Microthrix parvicella and Gordonia sp., while gravity separation led to the washout of M. parvicella. In both pilot trains, sequence analysis confirmed the presence of EBPR-related organisms such as Accumulibacter phosphatis. The survey of the CEBPR system also revealed many uncultured organisms that have not been well characterized. The study demonstrated that a simple replacement of a secondary clarifier with membrane solids-liquid separation is sufficient to shift the composition of an activated sludge microbial community significantly.

A comparative study of fouling-related properties of sludge from conventional and membrane enhanced biological phosphorus removal processes.

The physical and biochemical properties of activated sludge mixed liquor, including floc size distribution, zeta potential, relative hydrophobicity, and bound and unbound (soluble) extracellular polymeric substances (EPS), were examined in this study to evaluate their relationship to membrane fouling. Mixed liquors from a membrane enhanced biological phosphorus removal (MEBPR) process and a conventional enhanced biological phosphorus removal (CEBPR) process were compared. It was found that the floc size distribution and the amount of soluble EPS in the mixed liquor were the most important properties that significantly influenced the fouling propensity of sludge. Contrary to the literature, the content of EPS bound in activated sludge flocs was not found to be directly associated with membrane fouling, and sludge surface properties such as zeta potential and relative hydrophobicity were not closely related to the observed differences in the fouling tendencies of the two types of sludge.

Toward a high-rate enhanced biological phosphorus removal process in a membrane-assisted bioreactor.

A membrane enhanced biological phosphorus removal (MEBPR) process was studied to determine the impact of hydraulic retention time (HRT) and solids retention time (SRT) on the removal of chemical oxygen demand (COD), nitrogen, and phosphorus from municipal wastewater. The MEBPR process was capable of delivering complete nitrification independent of the prevailing operating conditions, whereas a significant improvement in COD removal efficiency was observed at longer SRTs. In the absence of carbon-limiting conditions, the MEBPR process was able to achieve low phosphorus concentrations in the effluent at increasingly higher hydraulic loads, with the lowest HRT being 5 hours. The MEBPR process was also able to maintain optimal phosphorus removal when the SRT was increased from 12 to 20 days. However, at higher suspended solids concentrations, a substantial increase was observed in carbon utilization per unit mass of phosphorus removed from the influent. These results offer critical insights to the application of membrane technology for biological nutrient removal systems.