Temperate Coastal Microbial Communities Rapidly Respond to Low Concentrations of Partially Weathered Diesel.

Diesel is frequently encountered in coastal ecosystems due to land run-off from road surfaces. The current study investigates how partially weathered diesel at environmentally relevant concentrations, as may be seen during a run-off event, affect coastal microbial communities. A mesocosm experiment using seawater from the Bedford Basin, Nova Scotia, was followed for 72 h after the addition of partially weathered diesel. Sequencing data suggests partially weathered diesel acts quickly to alter the prokaryotic community, as both opportunistic (Vibrio and Lentibacter) and oil-degrading (Colwellia, Sulfitobacter, and Pseudoalteromonas) bacteria proliferated after 24 h in comparison to the control. In addition, total prokaryotes seemed to recover in abundance after 24 h, where eukaryotes only ceased to decrease slightly at 72 h, likely because of an inability to adapt to the oil-laden conditions, unlike the prokaryotes. Considering there were no highly volatile components (benzene, toluene, ethylbenzene, and xylene) present in the diesel when the communities were exposed, the results indicate that even a relatively small concentration of diesel run-off can cause a drastic change to the microbial community under low energy conditions. Higher energy conditions due to wave action may mitigate the response of the microbial communities by dilution and additional weathering of the diesel.

Submersible optical sensors exposed to chemically dispersed crude oil: wave tank simulations for improved oil spill monitoring.

In situ fluorometers were deployed during the Deepwater Horizon (DWH) Gulf of Mexico oil spill to track the subsea oil plume. Uncertainties regarding instrument specifications and capabilities necessitated performance testing of sensors exposed to simulated, dispersed oil plumes. Dynamic ranges of the Chelsea Technologies Group AQUAtracka, Turner Designs Cyclops, Satlantic SUNA and WET Labs, Inc. ECO, exposed to fresh and artificially weathered crude oil, were determined. Sensors were standardized against known oil volumes and total petroleum hydrocarbons and benzene-toluene-ethylbenzene-xylene measurements-both collected during spills, providing oil estimates during wave tank dilution experiments. All sensors estimated oil concentrations down to 300 ppb oil, refuting previous reports. Sensor performance results assist interpretation of DWH oil spill data and formulating future protocols.

Interfacial film formation: influence on oil spreading rates in lab basin tests and dispersant effectiveness testing in a wave tank.

Test facilities such as lab basins and wave tanks are essential when evaluating the use of chemical dispersants to treat oil spills at sea. However, these test facilities have boundaries (walls) that provide an ideal environment for surface (interfacial) film formation on seawater. Surface films may form from surfactants naturally present in crude oil as well as dispersant drift/overspray when applied to an oil spill. The objective of this study was to examine the impact of surface film formation on oil spreading rates in a small scale lab basin and on dispersant effectiveness conducted in a large scale wave tank. The process of crude oil spreading on the surface of the basin seawater was influenced in the presence of a surface film as shown using a 1st order kinetic model. In addition, interfacial film formation can greatly influence chemically dispersed crude oil in a large scale dynamic wave tank.

Rates of gonorrhea and Chlamydia in U.S. military personnel deployed to Iraq and Afghanistan (2004-2009).

The increased incidence of sexually transmitted infections has historically been associated with military personnel at war. The incidence of gonorrhea and Chlamydia in personnel deployed in the current wars in Iraq and Afghanistan has not been reported. An electronic records' review of testing done from January 2004 to September 2009 revealed higher rates of Chlamydia than gonorrhea, especially among females who deploy to Iraq. Additionally, increasing Chlamydia rates were noted over the study. Overall, the rates of gonorrhea and Chlamydia were the same or lower than age- and year-matched U.S. rates reported by the Center for Disease Control and Prevention. Ongoing education with emphasis on prevention and treatment are needed, as are development of specific projects to define the risk factors and timing of acquisition of sexually transmitted infections in combat zones.

Changes in the incidences of multidrug-resistant and extensively drug-resistant organisms isolated in a military medical center.

BACKGROUND: Multidrug-resistant (MDR) Acinetobacter baumannii and Pseudomonas aeruginosa have emerged as the causes of nosocomial infections in critically ill patients. OBJECTIVE: To characterize the incidence of these MDR bacteria over time in the military healthcare system, comparing isolates recovered from overseas combat casualties with isolates recovered from local military and civilian patients. METHODS: Retrospective electronic records review of culture and/or susceptibility testing results of patients admitted to a military level I trauma center in San Antonio, Texas, during the period from January 2001 through December 2008. Multidrug resistance was defined as the first isolated organism resistant to 3 or more classes of antimicrobial agents. RESULTS: Over time, the percentage of MDR A. baumannii isolates increased from 4% to 55%, whereas the percentage of MDR P. aeruginosa isolates increased from 2% to 8%. Respiratory tract specimens had a higher percentage of MDR A. baumannii isolates (49%), compared with specimens obtained from blood (30%), wound sites (24%), or urine (19%). No difference in the percentages of MDR P. aeruginosa isolates was observed with regard to source of specimen. The percentage of MDR A. baumannii isolates recovered was higher among patients who had been deployed overseas (52%) than among local patients (20%). When isolates recovered from patients in the burn intensive care unit (53% of MDR A. baumannii isolates) were removed from analysis, the percentage of MDR A. baumannii isolates decreased from 38% to 30% while the percentage of MDR P. aeruginosa isolates remained unaffected. CONCLUSION: The percentage of MDR A. baumannii isolates increased in this facility among combat casualties and among local patients, which indicates nosocomial transmission; however, there was no significant increase in the percentage of MDR P. aeruginosa isolates. Isolated changes in the MDR pathogens within a facility can occur. Possible interventions to limit the spread of these organisms could include implementing aggressive infection control practices, controlling antibiotic use, and using active culture surveillance.

Incidence and bacteriology of burn infections at a military burn center.

Considerable advancements in shock resuscitation and wound management have extended the survival of burned patients, increasing the risk of serious infection. We performed a 6-year review of bacteria identification and antibiotic susceptibility records at the US Army Institute of Surgical Research Burn Center between January 2003 and December 2008. The primary goal was to identify the bacteria recovered from patients with severe burns and determine how the bacteriology changes during extended hospitalization as influenced by population and burn severity. A total of 460 patients were admitted to the burn ICU with 3507 bacteria recovered from 13,727 bacteriology cultures performed. The most prevalent organisms recovered were Acinetobacter baumannii (780), Pseudomonas aeruginosa (703), Klebsiella pneumoniae (695) and Staphylococcus aureus (469). A. baumannii was most often recovered from combat-injured (58%) and S. aureus the most frequent isolate from local (46%) burn patients. Culture recovery rate of A. baumannii and S. aureus was highest during the first 15 hospital days (73% and 71%); while a majority of P. aeruginosa and K. pneumoniae were recovered after day 15 (63% and 53%). All 4 pathogens were recovered throughout the course of hospitalization. A. baumannii was the most prevalent pathogen recovered from patients with total body surface area (TBSA) burns less than 30% (203) and 30-60% (338) while P. aeruginosa was most prevalent in patients with burns greater than 60% TBSA (292). Shifting epidemiology of bacteria recovered during extended hospitalization, bacteriology differences between combat-injured and local burn patients, and impact of % TBSA may affect patient management decisions during the course of therapy.

Aminoglycoside resistance and susceptibility testing errors in Acinetobacter baumannii-calcoaceticus complex.

Antimicrobial resistance is depleting the pharmacopeia of agents clinically useful against Gram-negative bacilli. As the number of active agents diminishes, accurate susceptibility testing becomes critical. We studied the susceptibilities of 107 isolates of the Acinetobacter baumannii-calcoaceticus complex to amikacin, gentamicin, and tobramycin using disk diffusion, Etest, as well as the Phoenix, Vitek 2, and MicroScan automated systems, and compared the results to those obtained by broth microdilution. Genes encoding aminoglycoside-modifying enzymes (AMEs) were detected by multiplex PCR, and clonal relationships were determined by pulsed-field gel electrophoresis. Tobramycin was the most active aminoglycoside (27.1% of isolates were susceptible). Disk diffusion and Etest tended to be more accurate than the Vitek 2, Phoenix, and MicroScan automated systems; but errors were noted with all methods. The Vitek 2 instrument incorrectly reported that more than one-third of the isolates were susceptible to amikacin (a very major error). Isolates were polyclonal, with 26 distinct strains, and carried multiple AME genes unrelated to the strain type. The presence of the ant(2")-Ia gene was statistically associated with resistance to each aminoglycoside. The AME genotype accounted for the resistance profile observed in a minority of isolates, suggesting the involvement of multiple resistance mechanisms. Hospital pharmacy records indicated the preferential use of amikacin over other aminoglycosides in the burn intensive care unit, where aminoglycoside resistance is prevalent. The resistance in that unit did not correlate with a predominant strain, AME genotype, or total annual aminoglycoside consumption. Susceptibility to tobramycin increased, even though susceptible isolates carried AME genotypes predicting the inactivation of tobramycin. Determination of the relative contribution of multiple concurrent resistance mechanisms may improve our understanding of aminoglycoside resistance in the Acinetobacter baumannii-calcoaceticus complex.

Prevalence of multidrug-resistant organisms recovered at a military burn center.

Infections caused by multidrug-resistant (MDR) pathogens are associated with significant morbidity and mortality in patients with burn injuries. We performed a 6-year antibiotic susceptibility records review from January 2003 to December 2008 to assess the prevalence of MDR isolates by pathogen at the US Army Institute of Surgical Research Burn Center. During the study period Acinetobacter baumannii (780 isolates [22%]) was the most prevalent organism recovered, followed by Pseudomonas aeruginosa (703 isolates [20%]), Klebsiella pneumoniae (695 isolates [20%]), and Staphylococcus aureus (469 isolates [13%]). MDR prevalence rates among these isolates were A. baumannii 53%, methicillin-resistant S. aureus (MRSA) 34%, K. pneumoniae 17% and P. aeruginosa 15%. Two isolates, 1 A. baumannii and 1 P. aeruginosa, were identified as resistant to all 4 classes of antibiotics tested plus colistin. A. baumannii isolates recovered from patients with burns greater than 30% of total body surface area (TBSA) were more likely to be MDR (61%) with no significant difference for P. aeruginosa and K. pneumoniae. A higher proportion of MDR P. aeruginosa isolates were recovered from respiratory specimens compared to blood specimens (24% vs. 9%) while the opposite was true for MRSA (35% vs. 54%). A comparison of A. baumannii recovered during hospitalization days 1-5 and 15-30 revealed higher MDR levels as length of stay increased (48% vs. 75%) while no significant trends were observed for P. aeruginosa and K. pneumoniae. A similar pattern was observed for MDR A. baumannii levels for the facility between 2003 and 2005 and 2006-2008 (39% vs. 70%), with no significant increase in MDR P. aeruginosa and MDR K. pneumoniae. Increasing antibiotic resistance patterns of the most prevalent isolates recovered during extended hospitalization, impact of % TBSA and other clinical parameters may affect empirical antimicrobial therapy and patient management decisions during treatment.

Biodegradation of endocrine disrupting compounds in harbour seawater and sediments.

The biodegradation of three endocrine disrupting compounds was examined using samples of seawater and sediment collected from Halifax Harbour, Nova Scotia, Canada, an urbanized harbour impacted by over two centuries of anthropogenic contamination. Flask experiments, where the samples were mixed to form a slurry were used to monitor the aerobic biodegradation of the synthetic plasticizer bisphenol A (BPA), the natural hormone 17beta-estradiol (E2), and the pharmaceutical and contraceptive ethinylestradiol (EE2). Degradation rates followed the order E2>EE2>BPA with half-lives of up to 1, 5 and 14 days in seawater, respectively. A rapid initial degradation rate for all three compounds with no apparent lag phase indicated the ability of the microbial community to readily catabolise the chemicals. The formation of unidentified non-persistent intermediate metabolites was observed during the E2 degradation experiments. These degradation rates are more rapid and complete than reported in previous studies, indicating the adaptation of native microbial communities to these contaminants.

Effects of changing land use on the microbial water quality of tidal creeks.

Population growth along the southeastern United States coast has precipitated the conversion of forested watersheds to suburban and urban ones. This study sampled creeks representing forested, suburban, and urban watersheds along a longitudinal gradient for indicators of water quality, including traditional indicator bacteria (fecal coliforms and enterococci) and alternative viral indicators (male-specific and somatic coliphages). Tested microorganisms were generally distributed with highest concentrations in creek headwaters and in more developed watersheds. The headwaters also showed the strongest predictive relationship between indicator concentrations and urbanization as measured by impervious cover. A seasonal pattern was observed for indicator bacteria but not for indicator viruses. Coliphage typing indicated the likely source of contamination was nonhuman. Results suggest that headwater creeks can serve as sentinel habitat, signaling early warning of public health concerns from land-based anthropogenic activities. This study also implies the potential to eventually forecast indicator concentrations under land use change scenarios.

Estrogenic compounds in seawater and sediment from Halifax Harbour, Nova Scotia, Canada.

Abstract-Samples of seawater and surface sediment were collected from seven locations around Halifax Harbour, Nova Scotia, Canada, and analyzed for the presence of the organic estrogenic contaminants, bisphenol A (BPA), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2). Samples were extracted using solid phase extraction (seawater) or sonication (sediments), followed by fractionation on a two-layer alumina/silica gel column prior to analysis by liquid chromatography-tandem mass spectrometry (LCMS/MS) with negative-ion electrospray ionization. Levels of the three compounds consistently ranked as BPA > E2 > EE2. The least potent compound and plasticizer BPA reached levels of up to 2.6 ng/L in seawater and 9.5 ng/g in sediments; the natural product E2 was detected at concentrations up to 0.57 ng/L and 0.86 ng/g; while the synthetic estrogen EE2 was in most cases below the method detection limit (0.14 ng/L and 0.28 ng/g). The highest levels were observed in the influent of a secondary treatment plant that discharges into the harbor, with concentrations of 32.4 ng/L for BPA and 5.3 ng/L for E2. Overall, the results indicate that these compounds readily associate with suspended particles rather than remaining in the soluble phase. Measurement of the octanol-water partition coefficient (log K(OW)) confirmed these results, with values of 3.41, 3.89, and 4.16 for BPA, E2, and EE2, respectively. Partitioning experiments using spiked field samples further confirmed these findings, with sorption directly related to sediment total organic content and following the order EE2 > E2 > BPA.

A statistical appraisal of disproportional versus proportional microbial source tracking libraries.

Library-based microbial source tracking (MST) can assist in reducing or eliminating fecal pollution in waters by predicting sources of fecal-associated bacteria. Library-based MST relies on an assembly of genetic or phenotypic "fingerprints" from pollution-indicative bacteria cultivated from known sources to compare with and identify fingerprints of unknown origin. The success of the library-based approach depends on how well each source candidate is represented in the library and which statistical algorithm or matching criterion is used to match unknowns. Because known source libraries are often built based on convenience or cost, some library sources may contain more representation than others. Depending on the statistical algorithm or matching criteria, predictions may become severely biased toward classifying unknowns into the library's dominant source category. We examined prediction bias for four of the most commonly used statistical matching algorithms in library-based MST when applied to disproportionately-represented known source libraries; maximum similarity (MS), average similarity (AS), discriminant analyses (DA), and k-means nearest neighbor (k-NN). MS was particularly sensitive to disproportionate source representation. AS and DA were more robust. k-NN provided a compromise between correct prediction and sensitivity to disproportional libraries including increased matching success and stability that should be considered when matching to disproportionally-represented libraries.

Acquisition and Maintenance of Endoscopic Skills: Developing an endoscopic dexterity training system for anesthesiologists.

The acquisition and maintenance of essential psychomotor skills that are only required sporadically is a significant problem in medical training and practice. It is of particular relevance to anesthesiologists with regard to fibreoptic intubation, a technique that may be under-utilized despite its central role in the management of the difficult airway. Dexterity deficit due to current training models, dexterity decay due to lack of practice, and situational stress related to the clinical environment may combine to impede effective training and confident use of endoscopes in airway management. An educational resource (DexterÔ) has been developed to overcome these problems. Dexter is a non-anatomical, endoscopic dexterity training system designed to encourage practice and help establish and maintain a state of procedural readiness, even if clinical exposure to difficult airway situations is sporadic.

Phenotypic library-based microbial source tracking methods: efficacy in the California collaborative study.

As part of a larger microbial source tracking (MST) study, several laboratories used library-based, phenotypic subtyping techniques to analyse fecal samples from known sources (human, sewage, cattle, dogs and gulls) and blinded water samples that were contaminated with the fecal sources. The methods used included antibiotic resistance analysis (ARA) of fecal streptococci, enterococci, fecal coliforms and E. coli; multiple antibiotic resistance (MAR) and Kirby-Bauer antibiotic susceptibility testing of E. coli; and carbon source utilization for fecal streptococci and E. coli. Libraries comprising phenotypic patterns of indicator bacteria isolated from known fecal sources were used to predict the sources of isolates from water samples that had been seeded with fecal material from the same sources as those used to create the libraries. The accuracy of fecal source identification in the water samples was assessed both with and without a cut-off termed the minimum detectable percentage (MDP). The libraries (approximately 300 isolates) were not large enough to avoid the artefact of source-independent grouping, but some important conclusions could still be drawn. Use of a MDP decreased the percentage of false-positive source identifications, and had little effect on the high percentage of true-positives in the most accurate libraries. In general, the methods were more prone to false-positive than to false-negative errors. The most accurate method, with a true-positive rate of 100% and a false-positive rate of 39% when analysed with a MDP, was ARA of fecal streptococci. The internal accuracy of the libraries did not correlate with the accuracy of source prediction in water samples, showing that one should not rely solely on parameters such as the average rate of correct classification of a library to indicate its predictive capabilities.

The role of strain gauge plethysmography in the assessment of patients with suspected deep vein thrombosis.

Suspicion of deep vein thrombosis (DVT) is a common reason for medical referral to hospital. Clinical signs and symptoms are notoriously unreliable, hence there is the need for objective testing. Strain gauge plethysmography (SGP) has been marketed as a technique for excluding lower limb DVT. We therefore set out to evaluate this screening tool. Over a 2 year period, 437 consecutive patients referred with suspected DVT were assessed using both plethysmography and Doppler ultrasound. When the two techniques were compared, plethysmography was found to have a negative predictive value of 90%. We conclude that strain gauge plethysmography has a role in the screening of patients with suspected DVT but should not be used as the sole method in patient assessment.

Vascular responsiveness to brachial artery infusions of phenylephrine during isoflurane and desflurane anesthesia.

UNLABELLED: Compared with equi-minimum alveolar anesthetic concentration (MAC) isoflurane, desflurane is associated with greater levels of sympathetic nerve activity in humans but similar reductions in blood pressure. To explore these divergent effects, we evaluated vascular alpha(1)-adrenoceptor responses in the human forearm during isoflurane and desflurane anesthesia to determine if alpha(1)-adrenoceptor responses were more substantially attenuated during desflurane administration. Bilateral forearm venous occlusion plethysmography was used to examine arterial blood flow and to determine changes in forearm vascular resistance during brachial artery infusions of saline and phenylephrine (0.2, 0.4, 0.8, and 1.6 microg/min) in 22 conscious subjects and during anesthesia with 0.65 and 1.3 MAC isoflurane or desflurane. Infusion of phenylephrine into the brachial artery increased the forearm vascular resistance in a dose-dependent manner. The arterial response to phenylephrine was significantly attenuated by 0.65 and 1.3 MAC desflurane and similarly attenuated during 1.3 MAC isoflurane (P < 0.05). Impaired arterial alpha(1)-adrenoceptor responsiveness occurred during desflurane. However, this effect was statistically similar (P > 0.05) to the impaired responses during isoflurane. Blood pressure decreases during volatile anesthesia may be, in part, caused by decreased alpha(1)-adrenoceptor responsiveness. IMPLICATIONS: alpha-receptors on blood vessels regulate constriction and dilation and therefore modulate blood pressure. This research indicates that vasoconstriction via the alpha(1)-receptor vascular response is impaired during isoflurane and desflurane anesthesia.

Acquisition and maintenance of endoscopic skills: developing an endoscopic dexterity training system for anesthesiologists.

The acquisition and maintenance of essential psychomotor skills that are only required sporadically is a significant problem in medical training and practice. It is of particular relevance to anesthesiologists with regard to fiberoptic intubation, a technique that may be under-utilized despite its central role in the management of the difficult airway. Dexterity deficit due to current training models, dexterity decay due to lack of practice, and situational stress related to the clinical environment may combine to impede effective training and confident use of endoscopes in airway management. We describe an educational resource (Dexter), which has been developed to overcome these problems. Dexter is a nonanatomical, endoscopic dexterity training system designed to encourage practice and help establish and maintain a state of procedural readiness, even if clinical exposure to difficult airway situations is sporadic.