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Microbial catabolic diversity in soils contaminated with hydrocarbons and heavy metals.

Understanding indigenous microbial function in contaminated soil is crucial to the successful development and use of bioremediation technologies. We measured the catabolic diversity of indigenous microbial communities in soils with a 30-yr history of Pb, Cr, and hydrocarbon (HC) contamination using a modified substrate-induced respiration method. There were characteristic differences of microbial respirations in the response of highly versus less contaminated soils to the range of organic substrates used. The catabolic response to glucose as compared to succinic acid was approximately 1:5 in less contaminated soils, but 1:25 in highly contaminated soils. In contrast, the response ratio to glucose versus aromatics was about 1:0.4 in less contaminated soils and 1:1 in highly contaminated soils. Principal components analysis (PCA) of the responses confirmed that catabolic diversity differed between highly and less contaminated soils. Univariate analysis also indicated that catabolic diversity was reduced in highly contaminated soils. This catabolic difference was strongly associated with the alteration of microbial community composition. Statistical analyses suggested that the variation in microbial community catabolic diversity was attributed to HCs more than to Pb and Cr.

Bacteria↗

The relevance of speciation in the remediation of soils and sediments contaminated by metallic elements--an overview and examples from Central Scotland, UK.

The environmental impact of metallic contaminants in soils and sediments is dependent both on the chemical speciation of the metal and the response of the matrix to biological and physicochemical conditions. These factors are responsible for the mobilisation of the metal from the solid into the aquatic phase and hence transport within the immediate vicinity, impacting on the rate of dispersal, dilution, uptake and transfer into living systems. The impact of changing environmental conditions on the contaminant inventory can be to enhance or moderate these phenomena, with subsequent consequences for the broader risk assessment of the contaminants. Remediation of metallic contaminants can only be brought about by their removal from the site or by establishing conditions which favour their retention in the solid phase. A wide range of in situ and ex situ approaches are available and a summary overview is presented. The examples show assessment at both the field and laboratory scale and demonstrate an equally wide range of success in achieving remediation targets. This can be attributed to limitations in ensuring that the desired conditions for the initial removal or immobilisation process are met and maintained over a suitable period of time. Three areas are reviewed which include: the transport and release of metallic contaminants in estuarine sediments and the assessment of their potential to impact on biota; terrestrial contamination systems involving the release of chromium from waste ore contaminating urban environments; the response of metal-contaminated wastes to changing environmental conditions and the impact of natural bioremediation. The focus of the discussion is to highlight the generation of reliable speciation information and the problems associated with impact and risk assessment. Particular issues of concern are the laboratory to field scale evaluation of contaminant behaviour and the approach used to assess the reliability of remediation options. In conclusion, part of a recent initiative in risk assessment and the development of pilot scale experimental systems to study long-term behaviour are addressed as future goals to fill gaps in current research.

Animals↗

The effect of saliva contamination on shear bond strength of orthodontic brackets when using a self-etch primer.

The purpose of this study was to assess the effect of saliva contamination on the shear bond strength of orthodontic brackets, at various stages of the bonding procedure using a new self-etch primer. Brackets were bonded to 52 extracted human molars according to one of the following four protocols. Group I (uncontaminated control): A self-etch acidic primer, Angel I (3M/ESPE Minneapolis, Minn) was placed on the enamel for 15 seconds, gently dried with air, and light cured for 10 seconds. Precoated brackets APC II (3M Unitek, Monrovia, Calif) were then placed on the teeth and light cured for 20 seconds. Group II: The enamel surface was first contaminated with human saliva for 10 seconds, blown off with an air syringe for five seconds. The bonding procedure was then repeated as in group I. Group III: The self-etch primer was applied for 15 seconds, gently dried with air, and light cured for 10 seconds. The surface was then contaminated with human saliva for 10 seconds, blown off with an air syringe for five seconds. The precoated brackets were then bonded as in groups I and II. Group IV: The enamel surface was contaminated with human saliva for 10 seconds, blown off with an air syringe for five seconds. The self-etch primer was applied. The surface was then re-contaminated with human saliva for 10 seconds, blown off with an air-syringe for five seconds. The precoated brackets were then bonded as in groups I, II, and III. The results of the analysis of variance (F = 4.79) indicated that the shear bond strengths of the four groups were significantly different (P = .005). Tukey HSD tests indicated that contamination both before and after the application of the acid-etch primer resulted in a significantly lower (=1.7+/-1.4 MPa) shear bond strength than either the control group (=6.0+/-3.5 MPa) or the groups where contamination occurred either before (=4.8+/-3.3 MPa) or after (=4.8+/-3.3 MPa) the application of the primer. The new acid-etch primer can maintain adequate shear bond strength if contamination occurs either before or after the application of the primer. On the other hand, contamination both before and after the application of the primer significantly reduced the shear bond strength of orthodontic brackets.

Acid Etching, Dental↗

Efficacy of using self-etching primer with a 4-META/MMA-TBB resin cement in bonding orthodontic brackets to human enamel and effect of saliva contamination on shear bond strength.

The objective of this study was (1) to evaluate the effectiveness of Megabond when used with Superbond C&B, a 4-methacryloloxyethyl trimellitate anhydride (4-META)/methyl methacrylate (MMA)-tri-n-butyl borane (TBB) resin, to bond orthodontic metal brackets to human enamel and (2) to examine the influence of saliva contamination on shear bond strength. Metal brackets were bonded to phosphoric acid-etched or Megabond-treated human premolars using Superbond C&B resin cement. The effects of saliva contamination after acid etching or self-etch priming, and the effect of re-etching or self-etch priming after saliva contamination on shear bond strength were also assessed. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours. Data were analyzed using two-way analysis of variance and Fisher's protected least significant difference test for multiple comparisons. There were no significant differences in shear bond strength between phosphoric acid etching and self-etch priming for no contamination, saliva contamination, and repeat treatment (etching or priming) after saliva contamination. With phosphoric acid etching, saliva contamination significantly decreased the shear bond strength. Repeat phosphoric acid etching after saliva contamination did not significantly improve the bond strengths. With self-etching primer treatment, however, saliva contamination did not cause any decrease of bond strength. Phosphoric acid etching produced more enamel fracture than self-etching primer treatment. Field-emission scanning microscopy revealed less dissolution of enamel surface resulted from self-etching primer compared with phosphoric acid. These results suggest that Megabond when used with Superbond C&B resin cement may be a good candidate for bonding orthodontic brackets to human enamel.

Acid Etching, Dental↗

A prospective microbiologic surveillance program to detect and prevent the transfusion of bacterially contaminated platelets.

After two patients received bacterially contaminated platelet transfusions, a prospective surveillance program was instituted to perform Gram staining and microbiologic culturing of platelets at the time of transfusion. In 12 months, 3141 random-donor platelet pools (prepared from 14,481 units) and 2476 single-donor apheresis units were cultured. All single-donor apheresis units were sterile, but 6 (0.19%) of the random-donor pools were found to be bacterially contaminated, with 1 unit of 5 in the pool being the source in each case. Contaminants were Staphylococcus epidermidis (4 cases), Bacillus cereus (1), and Staphylococcus aureus (1) at counts of 0.5 x 10(2) to 10(11) colony-forming units per mL in platelet pools and 10(3) to 10(13) colony-forming units per mL in source units. The contamination rate for units transfused at < or = 4 days (1.8/10,000) was significantly lower than that at 5 days (11.9/10,000; p < 0.05), as was the magnitude of contamination (p < 0.05). Use of the pretransfusion Gram stain on 4- and 5-day-old platelet pools was 100 percent sensitive (4/4 true positives) and 99.93 percent specific (1 false positive) in detecting contaminated pools. These data define the extent and magnitude of platelet bacterial contamination and demonstrate the efficacy of the pretransfusion Gram stain on platelet units stored for 4 and 5 days in preventing the transfusion of heavily contaminated units. It is concluded that the risk of platelet contamination is related to the duration of component storage.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Surface contamination to UV-curable acrylates in the furniture and parquet industry.

Surface contamination to ultraviolet radiation curable coatings (UV coatings), used increasingly in the parquet and furniture industry, is a matter of concern as a source for skin contamination. UV coatings contain chemically and biologically reactive acrylates, well known as skin contact irritants and sensitizers. Surface contamination may spread secondarily to equipment and other unexpected areas even outside the workplace. Yet, studies concerning this type of contamination are lacking due to lack of suitable sampling methods. Surface contamination of the work environment with risk for skin exposure to UV coating was measured employing a quantitative adhesive tape sampling method developed for this purpose. A pilot study was first performed at three workplaces to evaluate the contamination. In the main study, we wanted to locate and identify in detail the surface contamination of areas where problems exist, and to determine the extent of the problem. Measurements were performed at seven workplaces on two separate workdays (round 1 and 2) within a six-month period. Samples were collected from the workplaces based on the video monitoring of skin contact frequency with the surfaces and categorized into three groups to analyze risk. The pilot study indicated that surface contamination to TPGDA containing UV coatings was common, found in 76 percent of the surfaces, and varied with a maximum of 909 microg TPGDA 10 cm(-2) sampling area. In the main study TPGDA was found in 153 out of 196 collected samples (78.1%); for round one 78.1 percent (82 out of 105 samples) and for round two 78.0 percent (71 out of 91 samples). The average TPGDA mass on positive surface samples was on the first round 2,247 +/- 7,462 microg, and on the second round 2,960 +/- 4,590 microg. We conclude that surface contamination to uncured UV coatings at UV-curing lines is common and this involves a risk for harmful, unintentional skin exposure to acrylates.

Acrylates↗

Regulation of translation initiation in mussels (Mytilus galloprovincialis, lam.) following contamination stress.

The use of mussels (Mytilus sp.) as bioindicators of the aquatic environment is a valuable approach to monitor environmental contamination. Applying batteries of biomarkers is an essential prerequisite, since the complexity of environmental contaminants can induce in mussels a variety of structural and functional responses, which are not necessarily correlated. In an attempt to correlate translation responses to contamination stress, the sedimentation profiles of runoff ribosomes isolated from digestive gland cells of control or contaminated Mediterranean mussels (Mytilus galloprovincialis, Lam.) were examined and the efficiency of these ribosomes to accomplish protein synthesis was determined. While the major species of ribosomal material was 80S monomers, native 60S and 40S ribosomal subunits were also detected independently of the contamination level in the surrounding waters. However, concomitant with the increase of contamination stress, the level of 80S ribosomes was reduced in favor of free ribosomal subunits. In addition, ribosomes isolated from contaminated mussels and programmed with poly(U) were less efficient to enzymatically bind AcPhe-tRNA, compared with ribosomes from control samples. These results suggest that the contamination stress causes stoichiometric aberrations in the ribosomal particle pool and reduction of translation machinery capability to initate protein synthesis. Data support the notion that downregulation of translation is an important component of the cellular stress response and may be exploited as a biomarker of environmental contamination.

Animals↗

Evaluation of three methods for curing hybridomas from mycoplasma contamination.

Hybridomas are as susceptible to mycoplasma contamination as animal cell cultures. Two in vitro methods and one in vivo passage into mice were compared for efficiency of curing hybridomas from mycoplasma contaminations. Four contaminated clones were treated with the combined action of 5-bromouracil (5-BrUra) and the Hoechst 33258 followed by photosensitization. The other in vitro method involved the use of BM-cycline. The success of overcoming the mycoplasma contamination was dependent on the level of the initial contamination of the individual hybridoma. BM-cycline was more efficient than the photosensitization method. For the most contaminated hybridomas, 10 successive treatments with 5-BrUra were necessary as compared to six treatments with BM-cycline. Moreover, the use of BM-cycline reduced the contamination by as much as 50% after the first treatment. After twenty passages following the curing of hybridomas with BM-cycline, cells were stable and retained their specificity and secretion of their respective immunoglobulins. Whereas, treatment with 5-bromouracil, recurrence of contamination was observed in one of the four hybridomas after 10 passages following treatment. Decontamination after one passage in peritoneal cavity in mice was not always sufficient since one of four hybridomas remained contaminated. BM-cycline appears to be the method of choice since it is more efficient, less time consuming, simpler and less expensive. Mycoplasma strains that could be identified were of bovine origin: Mycoplasma arginini and Acholeplasma laidlawii.

Animals↗

Hair analysis for cocaine: the requirement for effective wash procedures and effects of drug concentration and hair porosity in contamination and decontamination.

It has been proposed that issues of external contamination of hair can be resolved by the presence of metabolites. This is true for some drugs in some circumstances. However, even if some metabolites such as cocaethylene are sufficiently definitive to indicate certainty of use, others such as benzoylecgonine can occur as contaminants. In addition, for phencyclidine, only the parent compound is determined. Therefore, effective washing procedures are needed to rule out contamination as the source of the parent or metabolite. The protection provided by cutoffs is also dependent on washing to remove surface contamination, which can range from very little to greater than 20 times the amount in the hair. In cocaine-positive head hair samples from 67 subjects with cocaine-positive urine tests, contamination of positive hair samples ranged from 0.7 to 8009 ng/10 mg hair, demonstrating that analyses of such samples to determine use is meaningless without removal of contamination. Wash procedures have been a focus of this laboratory for many years. In this paper, the effectiveness of our wash procedure was further challenged by application to negative hair (blonde, auburn, brown, and black) contaminated by soaking for 1 h in 1000, 10,000, and 50,000 ng cocaine/mL water. The uptake of the cocaine was approximately linear with increasing concentrations of cocaine in the soaking solutions. By application of the wash criterion, all samples were clearly identified as contaminated (i.e., negative). The effects of hair porosity on uptake of cocaine were also studied with 10 hair samples of all colors. Permed and unpermed samples were soaked in 10,000 ng cocaine/mL for 1 hour. All hair responded to the perm with increased uptake, and all were decontaminated or identified as contaminated (i.e., negative for cocaine use). Porosity, not hair color, determined the rate of penetration of hair by cocaine in solution, and, likewise, washing characteristics were unrelated to hair color.

Cocaine↗

The effect of common clinical contaminants on amniotic fluid fluorescence polarization results.

OBJECTIVE: To determine the effect of blood, meconium, and vaginal secretions on amniotic fluid (AF) fluorescence polarization results. METHODS: Amniotic fluid was collected by transabdominal amniocentesis from women at 20-41 weeks of gestation and contaminated with blood, meconium, and vaginal secretions to concentrations of 0.5, 1, 2, 5, and 10%. An additional 20% concentration was performed with meconium and vaginal secretions. Fluorescence polarization was determined by a TDx Analyzer with the NBD-PC fluorescent probe. Results were compared for each contaminant by concentration level using repeated-measures analysis of variance. RESULTS: Forty-eight samples from women at a mean gestational age of 35 weeks (range 20-41.5 weeks) were evaluated. Before contamination, 16 (33%) samples had fluorescence polarization values greater than 290 mPol (immature), 10 (21%) were 260- 289 mPol (transitional), and 22 (46%) were less than 260 mPol (mature). Contamination with blood significantly altered fluorescence polarization values in AF samples with baseline values in the immature and mature categories such that values trended toward the transitional range. Contamination of baseline immature samples with vaginal secretions at 20% contamination level resulted in more mature fluorescence polarization values. Contamination with meconium more than 2% in the baseline immature category or more than 20% in the baseline transitional category also resulted in significantly more mature fluorescence polarization values. CONCLUSION: Amniotic fluid contamination with blood can result in more transitional range fluorescence polarization values, whereas contamination with meconium and vaginal secretions can result in more mature fluorescence polarization values.

Amniocentesis↗

The prevalence and risk factors for donor corneal button contamination and its association with ocular infection after transplantation.

BACKGROUND: One of the most serious complications of corneal transplantation is postoperative ocular infection, which may result in loss of the eye. Contamination of the donor corneal button before transplantation may result in such complication. PURPOSE: To evaluate the prevalence of donor corneal button contamination, the spectrum of the contaminating microorganisms, and their sensitivity to antimicrobial agents. To investigate the risk factors for contamination of the donor corneal buttons and the effect of corneal button contamination on the prevalence of ocular infection in corneal transplanted patients. SETTING: Tertiary referral medical center. MATERIALS AND METHODS: Four hundred sixty-nine corneal transplantations were included in the study. Microbial cultures from the corneoscleral rims of the donor corneal buttons were obtained for isolation of bacteria and fungi and for their sensitivity to antimicrobial agents. Ocular microbial cultures were also obtained from corneal transplanted patients with clinical signs of ocular infection (ie, corneal scrapes from corneal ulcers and vitreous tap from eyes with endophthalmitis). RESULTS: Seventy-nine donor corneal buttons (16.8%) had positive bacterial cultures, and none had positive fungal culture. Staphylococci (63.7%) and streptococci (11.3%) were the most common isolated bacteria. Sensitivity to vancomycin, amikacin, and gentamicin was found in 71.3%, 28.5%, and 22.3% of all isolated bacteria, respectively. Malignancy and cardiac diseases as causes of donor death were associated with donor button contamination (P = 0.043 and P = 0.011, respectively), and septicemia was a marginally significant risk factor (P = 0.059). Age and gender of the donor, duration from death to corneal button harvesting, and time from harvesting to transplantation were not found significant risk factors for contamination. Six of the corneal transplanted patients (1.27%) had infected corneal graft ulcer, and 1 (0.22%) had endophthalmitis. The infected corneal ulcer appeared between 3 and 14 days (average 5 days), and endophthalmitis was disclosed 8 months after transplantation. Two (33%) of the 6 patients with corneal ulcer had the same species as the donor corneal rim. Postoperative ocular infection occurred in 2 (2.5%) patients out of 79 who received contaminated corneal buttons compared with 5 (1.3%) out of 390 patients who received sterile corneal buttons (P = 0.335). CONCLUSIONS: Postkeratoplasty infection of the recipient eye is infrequent despite relatively high prevalence of microbial contamination of the corneal buttons, suggesting that other risk factors for postoperative ocular infection are involved.

Adolescent↗

Risk perception of heavy metal soil contamination and attitudes toward decontamination strategies.

Contaminated soils are a common environmental risk all over the world. One major source of risk is heavy metal soil contamination caused by industrial emissions. This quasiexperimental study investigated the perception of these risks by exposed and nonexposed people, their attitudes toward bioremediation methods using hyperaccumulating plants, and the influence of long-term aspects of sustainability on the acceptance of bioremediation methods. Major findings were that people living in a contaminated area perceived the risk of the heavy metal soil contamination as higher than the general risk of contamination. Second, a factor analysis showed that the factors dread, control, and catastrophic potential were relevant for the perception and valuation of low-dose environmental risks such as the contamination of the investigated area. In addition, a cluster analysis showed that the risk of heavy metal soil contamination was perceived as similar to that of oil contamination, ozone layer, preservatives and genetic technology. It was perceived indifferently with regard to dread. The uncontrollability of heavy metal soil contamination was estimated as medium, and its catastrophic potential as low. Third, exposed and nonexposed participants preferred bioremediation methods to classical methods (e.g., excavation and chemical treatment of the soil), because they perceived the environmental and esthetical performance of the bioremediation as important criteria. Sustainability or precautionary issues, such as the prevention of harm for future generations, were highly correlated with the acceptance of the use of bioremediation methods in people's residential areas.

Adult↗

Microbial contamination of cosmetics and personal care items in Egypt--shaving creams and shampoos.

We examined a total of 192 samples, including eight different brands of shaving cream and eight brands of shampoo, for their total aerobic bacterial, coliforms and fungal counts. Shaving creams were more heavily contaminated with bacteria than shampoos. Viable bacterial were not recovered from 57% and 10% of shampoos and shaving creams, respectively. Only 3% of shaving creams were heavily contaminated with more than 10(4) c.f.u./g, while none of the shampoos contained such a high number of bacteria. With regard to the medium range contamination levels, 52% of shaving creams showed bacterial counts ranging from 10(2) to 10(3) c.f.u./g or ml, compared to 15% of shampoos which were contaminated to the same level. Fourteen per cent of shaving creams were contaminated with greater than 10(3)-10(4) c.f.u./g or ml, compared to 1% of the shampoos. No coliforms were recovered from either the shaving creams or the shampoos; however, Staphylococcus spp. were detected in six samples of both shampoos and shaving creams. Some of these Staphylococci, were aureus type. One isolate of Pseudomonas aeruginosa was also detected in a sample of shampoo. The incidence of fungal contamination was much less than the bacterial contamination. No viable fungi were recovered from 88% and 76% of the shaving creams and shampoos, respectively. The majority of the remaining samples, for both products, were contaminated with less than 100 fungal cell/g or ml. The pH of all the tested samples was alkaline (pH 7.2-9), which is well known to inhibit fungal contamination.

Bacteria↗

Sequential electron acceptor model for evaluation of in situ bioremediation of petroleum hydrocarbon contaminants in groundwater.

Mathematical development and model application is provided for a multiple substrate, sequential electron acceptor model, accounting for hydrodynamic transport, adsorption, and sequential oxygen/iron(III)-based biodegradation. Equations for iron(III)-based biodegradation of petroleum hydrocarbons are developed based on oxygen-inhibited Monod kinetics. The iron(III)-based biodegradation expressions were combined with earlier work by Widdowson and Aelion, to develop the two-dimensional, multiple substrate, oxygen/iron(III) sequential electron acceptor biodegradation model presented here. In addition to mathematical model development, simulations demonstrating the advantages of sequential electron acceptor and multiple substrate biodegradation models are provided. These simulations show that commonly-used single electron acceptor models may underpredict natural, in situ biodegradation potential at sites where indigenous microorganisms are capable of using multiple electron acceptors. Additional simulations show that, for contaminant plumes composed of constituents which biodegrade at different rates and under varying electron acceptor conditions, a multiple substrate model may allow more accurate prediction of both individual contaminant concentrations and the total amount of biodegraded contaminant. Considering that typical contaminant plumes are composed of multiple constituents with varying biodegradation properties and health risks, multiple substrate sequential electron acceptor models have the potential to provide more accurate tracking of individual constituent migration. The model was applied to a leaking UST site in Laurel Bay, South Carolina. Laboratory and monitoring well data presented in Landmeyer et al. have established that the petroleum hydrocarbon contaminants are present in the groundwater and are undergoing sequential oxygen-iron(III)-based biodegradation. Model simulations proved capable of reproducing the trends observed at the Laurel Bay site in that BTX contaminants were removed by sequential biodegradation, occurring first aerobically and subsequently anaerobically, and that iron(III)-reducing organisms biodegrade contaminants only in the absence of oxygen. The BTX compounds were individually but simultaneously modeled, allowing explicit modeling of specific contaminant biodegradation properties (e.g., toluene and xylene are assumed to degrade sequentially and benzene is assumed to degrade aerobically only). Although simulations presented here can reproduce trends observed at the Laurel Bay site, inclusion of additional electron acceptors and additional model calibration to data from this and other sites is necessary to improve and verify the model's capability to predict the efficacy of intrinsic biodegradation of petroleum hydrocarbon contaminants in groundwater.

Bacteria↗

Controlling microbial contamination on beef and lamb meat during processing.

The microbiological quality of carcases, meat and environmental surfaces was evaluated in commercial boning rooms processing beef and lamb. There was considerable variation in the level of microbial contamination on both carcases and meat, with counts ranging from less than 20 to 10(8)/cm2 on carcases and to 2 x 10(7)/cm2 on meat. The level of microbial contamination on meat was influenced by the level of carcase contamination at boning and by the boning process itself. Carcase contamination was the major determinant of microbiological quality, as more than 70% of carcase had microbial counts greater than 10(3)/cm2. Cutting boards were a major source for microbial dissemination during boning, particularly when carcase counts were less than 10(3)/cm2. If carcases were heavily contaminated, the contamination of processing surfaces was irrelevant in determining microbial loads on meat. Where carcase contamination was at low to moderate levels, the contribution of the boning process to the contamination on meat assumed increased significance. Under these conditions, improved sanitation of cutting surfaces in the boning room resulted in a significant reduction in microbial contamination on the surface of meat. These results can form the basis for ensuring that improvements made in carcase management before boning, to improve microbiological quality, will be preserved through attention to cutting board hygiene during boning.

Animals↗

Enumeration and characterization of iron(III)-reducing microbial communities from acidic subsurface sediments contaminated with uranium(VI).

Iron(III)-reducing bacteria have been demonstrated to rapidly catalyze the reduction and immobilization of uranium(VI) from contaminated subsurface sediments. Thus, these organisms may aid in the development of bioremediation strategies for uranium contamination, which is prevalent in acidic subsurface sediments at U.S. government facilities. Iron(III)-reducing enrichment cultures were initiated from pristine and contaminated (high in uranium, nitrate; low pH) subsurface sediments at pH 7 and pH 4 to 5. Enumeration of Fe(III)-reducing bacteria yielded cell counts of up to 240 cells ml(-1) for the contaminated and background sediments at both pHs with a range of different carbon sources (glycerol, acetate, lactate, and glucose). In enrichments where nitrate contamination was removed from the sediment by washing, MPN counts of Fe(III)-reducing bacteria increased substantially. Sediments of lower pH typically yielded lower counts of Fe(III)-reducing bacteria in lactate- and acetate-amended enrichments, but higher counts were observed when glucose was used as an electron donor in acidic enrichments. Phylogenetic analysis of 16S rRNA gene sequences extracted from the highest positive MPN dilutions revealed that the predominant members of Fe(III)-reducing consortia from background sediments were closely related to members of the Geobacteraceae family, whereas a recently characterized Fe(III) reducer (Anaeromyxobacter sp.) and organisms not previously shown to reduce Fe(III) (Paenibacillus and Brevibacillus spp.) predominated in the Fe(III)-reducing consortia of contaminated sediments. Analysis of enrichment cultures by terminal restriction fragment length polymorphism (T-RFLP) strongly supported the cloning and sequencing results. Dominant members of the Fe(III)-reducing consortia were observed to be stable over several enrichment culture transfers by T-RFLP in conjunction with measurements of Fe(III) reduction activity and carbon substrate utilization. Enrichment cultures from contaminated sites were also shown to rapidly reduce millimolar amounts of U(VI) in comparison to killed controls. With DNA extracted directly from subsurface sediments, quantitative analysis of 16S rRNA gene sequences with MPN-PCR indicated that Geobacteraceae sequences were more abundant in pristine compared to contaminated environments,whereas Anaeromyxobacter sequences were more abundant in contaminated sediments. Thus, results from a combination of cultivation-based and cultivation-independent approaches indicate that the abundance/community composition of Fe(III)-reducing consortia in subsurface sediments is dependent upon geochemical parameters (pH, nitrate concentration) and that microorganisms capable of producing spores (gram positive) or spore-like bodies (Anaeromyxobacter) were representative of acidic subsurface environments.

Bacillaceae↗

Blood culture contamination in Tanzania, Malawi, and the United States: a microbiological tale of three cities.

We conducted retrospective, comparative analyses of contamination rates for cultures of blood obtained in the emergency rooms of Muhimbili National Hospital (MNH) in Dar es Salaam, Tanzania; Lilongwe Central Hospital (LCH) in central Malawi; and the Duke University Medical Center (DUMC) in the United States. None of the emergency room patients had indwelling intravascular devices at the time that the blood samples for cultures were obtained. In addition, we reviewed the contamination rates for a cohort of patients already hospitalized in the DUMC inpatient medical service, most of whom had indwelling intravascular devices. The bloodstream infection rates among the patients at MNH (n=513) and LCH (n=486) were similar (approximately 28%); the contamination rates at the two hospitals were 1.3% (7/513) and 0.8% (4/486), respectively. Of 54 microorganisms isolated from cultures of blood collected in the DUMC emergency room, 26 (48%) were identified as skin contaminants. Cultures of blood collected in the DUMC emergency room were significantly more likely to yield growth of contaminants than the cultures of blood collected in the emergency rooms at MNH and LCH combined (26/332 versus 11/1,003; P<0.0001) or collected in the DUMC inpatient medical service (26/332 versus 7/283; P<0.01). For the MNH and LCH blood cultures, lower contamination rates were observed when skin was disinfected with isopropyl alcohol plus tincture of iodine rather than isopropyl alcohol plus povidone-iodine. In conclusion, blood culture contamination was minimized in sub-Saharan African hospitals with substantially limited resources through scrupulous attention to aseptic skin cleansing and improved venipuncture techniques. Application of these principles when blood samples for culture are obtained in U.S. hospital emergency rooms should help mitigate blood culture contamination rates and the unnecessary microbiology workup of skin contaminants.

2-Propanol↗

Polymerase chain reaction based human leucocyte antigen genotyping for the investigation of suspected gastrointestinal biopsy contamination.

BACKGROUND: Mislabelling or contamination of surgical specimens may lead to diagnostic inaccuracy, particularly within gastrointestinal pathology when multiple small mucosal biopsy specimens are commonly taken, and where a tiny fragment of foreign tissue may be indistinguishable from true biopsy material using histological assessment alone. AIMS: To assess the utility of polymerase chain reaction (PCR) based human leucocyte antigen (HLA) genotyping techniques for the investigation of potentially mislabelled or contaminated gastrointestinal biopsy specimens. PATIENTS: Ten cases (28 samples) in which mislabelling or contamination was suspected, comprising four upper gastrointestinal tract biopsies and six colonoscopic biopsy series. METHODS: Direct and nested PCR-sequence specific primer (SSP) based HLA class II genotyping was performed on DNA extracted from formalin fixed and paraffin wax embedded tissue (23 samples) or peripheral blood leucocytes (five samples). RESULTS: A full HLA-DRB1 genotype was determined in all 28 samples. In seven cases the HLA-DRB1 genotype of the putative contaminant was different to that of the corresponding reference tissue, confirming different individual origins for the contaminant and reference material. In one case the contaminant tissue was shown to possess the same HLA-DRB1 alleles as a second patient (probable source). In the remaining three cases the same HLA-DRB1 alleles were detected within the potential contaminant and reference tissues. CONCLUSIONS: PCR based HLA class II genotyping is a valuable tool for investigating potential contamination or mislabelling within gastrointestinal biopsy specimens and this report has confirmed contamination in seven of ten cases studied.

Adult↗