The Fate of Methicillin-Resistant Staphylococcus aureus in a Synthetic Turf System.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for one-third of infectious outbreaks reported among competitive athletes at high school and collegiate levels of competition, while the frequency of infections among National Football League players is nearly 400-fold higher than in the general population. The increased prevalence of MRSA infections among such athletes necessitates the study of football-specific environments as MRSA reservoirs. HYPOTHESIS: The recovery and viability of MRSA will differ between turf infill and turf fibers. STUDY DESIGN: Descriptive laboratory study. LEVEL OF EVIDENCE: Level 4. METHODS: Each of 6 infill types and 3 turf fiber types were inoculated with MRSA, sacrificed over time, and enumerated after bacteria extraction and dilution plating to determine the incubation time at which 50% of inoculated MRSA are recoverable and viable (here termed A50). The role of infill toxicity on MRSA survival was assessed using a dialysis assay. RESULTS: On average, MRSA was available for up to 96 hours on infill (average A50: 13 hours) and 24 hours on turf fibers (A50: 4 hours) (P < 0.001). The A50 for each infill type was also variable among infill groups (P < 0.001), averaging 2 hours (ethylene propylene diene monomer [EPDM] rubber), 7 hours (cork-based material), 9 hours (polymer-coated materials), 12 hours (crumb rubber), 13 hours (thermoplastic elastomer [TPE] rubber), and 27 hours (sands). MRSA remained available on slit-film and nylon fiber types for 12 hours postdeposition and for 24 hours on monofilament fibers. Toxicity assays showed that over 90% of MRSA cells remained viable after 6 hours of exposure to cork infill and sands, while 79%, 71%, 68%, and 17% of MRSA remained viable after exposure to polymer-coated materials, crumb rubber, TPE rubber, and EPDM rubber, respectively. Our data also indicated that organic and sand infills exhibited minimal toxicity to MRSA, while high relative toxicity drives limited MRSA availability in EPDM rubber infill. CONCLUSION: MRSA recovery and viability varied among infill types but not among differing turf blades. CLINICAL RELEVANCE: The results of this study can inform appropriate athlete and field management practices.

A Multicenter Trial Demonstrating Presence or Absence of Bacterial Contamination at the Screw-Bone Interface Owing to Absence or Presence of Pedicle Screw Guard, Respectively, During Spinal Fusion.

STUDY DESIGN: A prospective multicenter study. OBJECTIVE: The objective of this study was to assess bacterial contamination in current practices of pedicle screw handling and comparing it to a novel method of using an intraoperative, sterile implant guard for screws. SUMMARY OF BACKGROUND DATA: Postoperative infections occur at the higher end of 2%-13%, as cited in the literature, and are underestimated due to various reasons in such publications. Despite concerns associated with vancomycin application immediately before closure, it is theoretically impossible to irrigate the screw-bone interface postimplantation. Consequently, any contamination of pedicle screw before implantation is permanent, and has the potential to cause deep-bone infection, or hardware loosening due to encapsulation of biofilm between the bone and the screw. Therefore, continued vigilance and effective preventive measures should be undertaken if available. MATERIALS AND METHODS: Two groups of presterile individually-packaged pedicle screws, one incased in a sterile, protective guard (group 1: G) and the other without such a guard (group 2: NG), 31 samples in each group were distributed over 28 spinal fusion surgeries at 5 independent hospitals groups. Each were loaded onto the insertion device by the scrub tech and left on the sterile table. Twenty minutes later, the lead surgeon who had just finished preparing the surgical site, handles the pedicle screw, to check the fit with the insertion device. Then, instead of implantation, it was transferred to a sterile container using fresh sterile gloves for bacterial analysis. RESULTS: The standard unguarded pedicle screws presented bioburden in the range of 10 to 10 colonies forming units per screw, whereas the guarded pedicle screws showed no bioburden. CONCLUSION: Standard, current, handling of pedicle screws leads to bacterial contamination, which can be avoided if the screws are sterilely prepackaged with an intraoperative guard (preinstalled).

Genetic evidence for the offsite transport of E. coli associated with land application of Class B biosolids on agricultural fields.

The land-application of Class B biosolids is tightly regulated to allow for natural attenuation of co-applied pathogens. Since many agricultural fields that receive biosolids are artificially drained through subsurface tiles, it is possible that under scenarios of excessive drainage associated with heavy rainfall events, co-applied pathogens might be carried offsite to contaminate nearby surface waters. To address this concern, we used genetic as well as traditional methods to investigate the impact of rainfall on the offsite drainage of Escherichia coli from agricultural fields during biosolids application. Water samples from field drain tiles and a reference field (no biosolids applied) were collected pre-, during and post-biosolids application, while samples of applied biosolids were collected on site during application. The samples were analyzed for E. coli-density and community- and isolate-fingerprinting to assess the genetic link between E. coli in drainage water and those co-applied with biosolids. In contrast to E. coli densities present in the reference field drainage, our results revealed that post-application drainage water collected from biosolids treated fields contained significantly higher E. coli densities following heavy rainfall events, as compared to light rainfall events. Also, in contrast to the reference field, heavy rainfall correlated significantly with increased similarity of E. coli community fingerprints occurring in biosolids to those draining from treated field. Fingerprinting of individual E. coli revealed a high similarity (>94%) between some isolates collected from biosolids and post-application drainage water. Using a combination of enumeration and genetic typing methods, we show that heavy rainfall following biosolids application to agricultural fields induced the offsite transport of biosolids-associated E. coli, potentially compromising the quality of water draining through the watershed.

Short-term effect of capping on microbial communities in freshwater sediments.

Because biogas bubbles can influence cap integrity, the effect of capping and cap material on the ebullition potential in sediments must be studied. The goal of this comprehensive study was to determine the short-term effect of capping regime on the activity, metabolic potential, and community structure of sediment microorganisms. To evaluate the effect of capping (sand, synthetic aggregate, and no cap) on microbial communities (i.e., nitrifiers and methanogens), sediments were collected from the Anacostia River (Washington, D.C.). Microbial communities in sand-capped sediments exhibited the highest activity (tetrazolium redox dye, fluorescein diacetate hydrolysis assay, and biogas production), while communities in uncapped sediments exhibited the highest metabolic diversity. Substantial changes in microbial community structure (denaturing gradient gel electrophoresis) did not occur as a result of capping. Our data showed that the nature and magnitude of the effect that capping can have on microbial activity (biogas production) will likely be dependent on the capping materials chosen.

Effect of elevated CO2 and drought on soil microbial communities associated with Andropogon gerardii.

Our understanding of the effects of elevated atmospheric CO2, singly and in combination with other environmental changes,on plant-soil interactions is incomplete. Elevated CO2 effects on C4 plants, though smaller than on C3 species, are mediated mostly via decreased stomatal conductance and thus water loss. Therefore, we characterized the interactive effect of elevated CO2 and drought on soil microbial communities associated with a dominant C4 prairie grass, Andropogon gerardii Vitman. Elevated CO2 and drought both affected resources available to the soil microbial community. For example, elevated CO2 increased the soil C:N ratio and water content during drought, whereas drought alone decreased both. Drought significantly decreased soil microbial biomass. In contrast, elevated CO2 increased biomass while ameliorating biomass decreases that were induced under drought. Total and active direct bacterial counts and carbon substrate use (overall use and number of used sources) increased significantly under elevated CO2. Denaturing gradient gel electrophoresis analysis revealed that drought and elevated CO2, singly and combined, did not affect the soil bacteria community structure.We conclude that elevated CO2 alone increased bacterial abundance and microbial activity and carbon use, probably in response to increased root exudation. Elevated CO2 also limited drought-related impacts on microbial activity and biomass,which likely resulted from decreased plant water use under elevated CO2. These are among the first results showing that elevated CO2 and drought work in opposition to modulate plant-associated soil-bacteria responses, which should then influence soil resources and plant and ecosystem function.

Optimization of DGGE community fingerprinting for characterizing Escherichia coli communities associated with fecal pollution.

We evaluated the use of DGGE fingerprinting to differentiate communities of Escherichia coli from animal and geographic sources. An initial screening of 15 gene candidates revealed the ability of three target genes (mdh, phoE and uidA-4) to effectively differentiate E. coli communities originating in horses, pigs, geese and goats. Cluster and jackknife analyses performed on the communities from a more extensive number of hosts (n=150) including humans (via raw sewage), horses, pigs, geese and cows revealed that the internal accuracy of classification of E. coli community fingerprints to their origin was similar for each of the three genes (85-86%). Each of the three genes were tested for their ability to associate E. coli source- and sink communities in two settings featuring contaminated water; (i) a stream receiving municipal wastewater effluent and (ii) a pond inhabited by geese. For each gene, DGGE fingerprints effectively matched effluent- and downstream E. coli communities (98-100% similarity) and excluded upstream communities, while communities from goose fecal material were 77-79% similar to communities in pond water, indicating fecal inputs from geese. Furthermore, each gene discriminated against E. coli communities from hosts non-indigenous to either setting. DGGE analysis of E. coli communities appears to be a promising tool to augment existing efforts aiming to address the dynamics of bacteria pollution in complex, natural environments.

A multiphasic characterization of the impact of the herbicide acetochlor on freshwater bacterial communities.

Acetochlor is the third most frequently detected herbicide in natural waters; however, it is unknown if exposure to environmentally relevant concentrations of acetochlor will impact bacterial community structure and function. This study examined the impact of acetochlor on freshwater heterotrophic bacteria number, and community structure and function using direct counting, community level physiological profiling (CLPP) and denaturing gradient gel electrophoresis (DGGE) analysis. Acetochlor concentration did not appear to correlate with the number of total (P=0.69) and viable (P=0.80) bacteria, even at concentrations up to 500 microg l(-1). However, CLPP indicated that acetochlor increased functional diversity as shown by (i) an increase in the number of carbon sources utilized by the microbial community, relative to nonexposed controls and (ii) increased functional evenness within the heterotrophic bacterial community. Conversely, DGGE fingerprints suggested that exposure to acetochlor generally decreased the community complexity, as the average number of DGGE bands in most treatments was significantly less than in the control treatment. Cluster analysis of DGGE fingerprints revealed three distinct, dose-dependent clusters (i) communities exposed to 0, 1 and 5 microg l(-1); (ii) 50 and 100 microg l(-1) and (iii) 500 microg l(-1), indicating a relationship between acetochlor concentration bacterial community changes. This study indicated that while exposure to environmentally relevant concentrations of acetochlor resulted in no significant impact to the number of freshwater bacteria, impacts to the function and structure of the community were revealed by adopting a multiphasic approach.

Public computer surfaces are reservoirs for methicillin-resistant staphylococci.

The role of computer keyboards used by students of a metropolitan university as reservoirs of antibiotic-resistant staphylococci was determined. Putative methicillin (oxacillin)-resistant staphylococci isolates were identified from keyboard swabs following a combination of biochemical and genetic analyses. Of 24 keyboards surveyed, 17 were contaminated with staphylococci that grew in the presence of oxacillin (2 mg l(-1)). Methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA), -S. epidermidis (MRSE) and -S. hominis (MRSH) were present on two, five and two keyboards, respectively, while all three staphylococci co-contaminated one keyboard. Furthermore, these were found to be part of a greater community of oxacillin-resistant bacteria. Combined with the broad user base common to public computers, the presence of antibiotic-resistant staphylococci on keyboard surfaces might impact the transmission and prevalence of pathogens throughout the community.

A comparison of the benthic bacterial communities within and surrounding Dreissena clusters in lakes.

The impact of Dreissena (Dreissena polymorpha and D. bugensis) on the benthic bacterial community in lakes is largely unknown. Therefore, we quantified differences in the structure and activity of bacterial communities living in sediments (1) associated with Dreissena clusters, and (2) unassociated with established clusters (lake bottom sediments). Dreissena clusters and sediments were collected from locations in Lake Erie, Lake Ontario, and several inland lakes. Denaturing gradient gel electrophoresis (DGGE) analysis of the benthic bacterial community showed that the bacterial populations selected for by Dreissena represent a subset of the bottom communities and are geographically distinct. Community-level physiological profiling (CLPP) showed that overall bacterial activity and metabolic diversity were enhanced by the presence of clusters in all samples, with the exception of those harvested from the two Lake Erie sites. Therefore, Dreissena appears to affect both structure and metabolic function of the benthic bacterial community and may have yet unexplored ecosystem and food web consequences.

Evaluation of denaturing gradient gel electrophoresis to differentiate Escherichia coli populations in secondary environments.

The development of methodology to differentiate mixed populations of Escherichia coli in the secondary habitat might improve monitoring of fecal pollution indicators and facilitate the development of strategies to mitigate bacterial pollution. The objective of this study was to determine the ability of denaturing gradient gel electrophoresis (DGGE) to differentiate mixed assemblages of E. coli in the natural environment. After confirming the identity of 184 environmental bacterial isolates as E. coli, each was subjected to polymerase chain reaction (PCR) of the beta-glucuronidase gene (uidA) followed by DGGE fingerprinting. The ability of DGGE to discriminate individual isolates at the strain level was determined by comparing fingerprints to those resulting from a standard, library-dependent fingerprinting method, BOX-PCR. Computerized analysis of fingerprints indicated that DGGE and BOX-PCR identified 15 and 21 unique phylotypes respectively. Rank-abundance plots comparing the numerical distribution of unique E. coli phylotypes detected by both methods revealed no difference in resolution at the population level. In water and sediment samples from two beaches, DGGE effectively distinguished indigenous E. coli populations with an average rate of correct classification (site-based) of 83%. Denaturing gradient gel electrophoresis of uidA genes isolated and PCR-amplified from environmental samples appears to be an effective tool to differentiate unique E. coli populations and should be useful to characterize E. coli dynamics in the secondary environment.