Microbial penetration and utilization of organic aircraft fuel-tank coatings

Microorganisms have been found as contaminants in various types of aircraft fuel tanks. Their presence introduces problems in the operation of the aircraft, including destruction of components such as the organic coatings used as protective linings in the fuel tanks. Microbial penetration and utilization of the currently used organic coatings, EC 776, DV 1180, PR 1560, and DeSoto 1080, were determined by changes in electrical resistances of the coatings; mycelial weight changes; growth counts of the bacteria; and manometric determinations on Pseudomonas aeruginosa (GD-FW B-25) and Cladosporium resinae (QMC-7998). The results indicate EC 776 and DV 1180 to be less resistant to microbial degradation than the other coatings. Organic coatings, serving as a source of nutrition, would be conducive to population buildups in aircraft fuel tanks.     

Corrosion of aluminum alloy 2024 by microorganisms isolated from aircraft fuel tanks

Microorganisms frequently contaminate jet fuel and cause corrosion of fuel tank metals. In the past, jet fuel contaminants included a diverse group of bacteria and fungi. The most common contaminant was the fungus Hormoconis resinae. However, the jet fuel community has been altered by changes in the composition of the fuel and is now dominated by bacterial contaminants. The purpose of this research was to determine the composition of the microbial community found in fuel tanks containing jet propellant-8 (JP-8) and to determine the potential of this community to cause corrosion of aluminum alloy 2024 (AA2024). Isolates cultured from fuel tanks containing JP-8 were closely related to the genus Bacillus and the fungi Aureobasidium and Penicillium. Biocidal activity of the fuel system icing inhibitor diethylene glycol monomethyl ether is the most likely cause of the prevalence of endospore forming bacteria. Electrochemical impedance spectroscopy and metallographic analysis of AA2024 exposed to the fuel tank environment indicated that the isolates caused corrosion of AA2024. Despite the limited taxonomic diversity of microorganisms recovered from jet fuel, the community has the potential to corrode fuel tanks.     

Identification of microorganisms isolated from jet fuel systems

Seventy-two samples from jet aircraft fuel systems were examined for microbial contamination. Ten contaminated samples yielded 43 microorganisms which were classified into nine genera of bacteria and three genera of fungi. The predominant types, comprising about 37% of the isolated cultures, were identified as Bacillus spp. The remaining cultures were distributed among 11 genera, each of which represented 2 to 9% of the total isolates. Four cultures could not be assigned to a genus on the basis of the diagnostic criteria used. Only five isolates, in the genera Pseudomonas and Hormodendrum (Cladosporium), grew abundantly in a mineral salts solution with JP-4 fuel as the sole source of carbon. The presence of fuel utilizers in a fuel system may be a better index to potential problems that have been correlated with microbial contamination than the presence of aerobic sporeforming bacilli.     

Corrosion of aluminum alloy 2024 by microorganisms isolated from aircraft fuel tanks

Abstract

Microorganisms frequently contaminate jet fuel and cause corrosion of fuel tank metals. In the past, jet fuel contaminants included a diverse group of bacteria and fungi. The most common contaminant was the fungus Hormoconis resinae. However, the jet fuel community has been altered by changes in the composition of the fuel and is now dominated by bacterial contaminants. The purpose of this research was to determine the composition of the microbial community found in fuel tanks containing jet propellant-8 (JP-8) and to determine the potential of this community to cause corrosion of aluminum alloy 2024 (AA2024). Isolates cultured from fuel tanks containing JP-8 were closely related to the genus Bacillus and the fungi Aureobasidium and Penicillium. Biocidal activity of the fuel system icing inhibitor diethylene glycol monomethyl ether is the most likely cause of the prevalence of endospore forming bacteria. Electrochemical impedance spectroscopy and metallographic analysis of AA2024 exposed to the fuel tank environment indicated that the isolates caused corrosion of AA2024. Despite the limited taxonomic diversity of microorganisms recovered from jet fuel, the community has the potential to corrode fuel tanks.     

Microbial challenges for domestic heating oil storage tanks

Microbial growth on hydrocarbons is common in nature and used in bioremediation of contaminated sites, whereas in fuel storage tanks this phenomenon can affect the stability of the fuel and the tank. The impact of microbial growth and produced metabolites on materials, which are used in the construction of storage tanks, were analyzed. In contrast to metal tank components, polymeric materials did not affect or were influenced by microorganisms. Zinc was highly corroded by microbial growth, most likely due to the formation of organic acids that were produced during microbial growth on hydrocarbons. A contaminated water phase in a storage tank of a heating system was simulated with a self‐constructed pump test bench. Microbial growth began in the water phase of the storage tank and microbes were distributed throughout the tank system, through water‐in‐oil microemulsions. No microbial growth was observed in oil that was previously contaminated, indicating that essential nutrients had been depleted. The identification and removal of these essential nutrients from fuels could minimize or prevent microbial contamination. The results are discussed with regard to developing recommendations for the design and operation of domestic heating oil storage tanks to lower the risk of technical failure due to microbial contamination.     

Effects of polyurethane foams on microbial growth in fuel-water systems

Four, open-cell, ester-base polyurethane foams were examined for their effect on growth of fuel-utilizing organisms in jet fuel-water systems. Three foams contained a potential biocide, tetraethylthiuram E (0.66%), sodium omadine (0.07%), or zinc omadine (0.07%), all w/v. These were compared with a control foam which did not contain an additive. Each foam was examined in fuel-water systems containing JP-4 fuel, JP-4 fuel plus 0.1% anti-icing additive (AIA), or JP-5 fuel. Pure cultures of a fuel-grown bacterium, Pseudomonas aeruginosa, and of a fuel-grown fungus, Hormodendrum (Cladosporium) sp., served as test organisms. In control cultures without foam and in cultures containing control foam, P. aeruginosa achieved maximum stationary-phase populations of approximately 10⁸ viable cells per ml, and Hormodendrum sp. produced an extensive mycelial mat. In the three fuel systems examined, tetraethylthiuram E- and sodium omadine-containing foams had little effect on growth of the bacterium; foam with zinc omadine decreased the rate of bacterial growth but had little effect on total populations. Tetraethylthiuram E decreased the rate of fungal growth and showed its greatest effect in JP-4 plus AIA. Foam with sodium omadine or zinc omadine markedly decreased fungal growth in all three fuel systems. The data suggest that either sodium omadine or zinc omadine in polyurethane foam may be a useful antifungal agent; and that tetraethylthiuram E and AIA could exert a synergistic effect, particularly at AIA concentrations which have been reported to occur in some field situations.     

Natural-Y Même polyurethane versus smooth silicone: analysis of the soft-tissue interaction from 3 days to 1 year in the rat animal model

The polyurethane foam-covered breast prosthesis is experiencing increased clinical use. The polyurethane is felt to be responsible for altering capsule formation and reducing the contracture rate. This study characterizes the soft-tissue response to the Natural-Y Même polyurethane foam versus smooth silicone in a rat model. Implants were fashioned from an unbacked polyurethane foam specimen used to cover the Natural-Y prosthesis, a silicone shell covered with the Natural-Y foam, and a smooth silicone control. Materials were placed subcutaneously into the backs of male Lew/SsN rats (n = 81) for 3, 7, 14, and 28 days and 3, 6, and 12 months. Implants were then harvested with their soft-tissue response and evaluated histologically. Analysis demonstrates that microstructuring of a surface, as opposed to a smooth material, will dramatically alter the early, intermediate, and late wound-healing events. The soft-tissue response was observed to be dependent on implant site, material chemistry, and morphology as characterized by exudate formation, macrophage invasion, multinucleated giant cell formation, collagen deposition, foam degradation, and angiogenesis.     

一株高效聚磷解淀粉芽孢杆菌的分离鉴定及其除磷条件优化

【背景】磷是引起水体富营养化的主要营养物质。生物除磷具有成本低、效率高且适用范围广等特点,成为近年来水处理研究领域的热点。目前虽然有一些聚磷菌被筛选获得,但它们的除磷效率不高,其除磷条件尚需优化。聚磷菌的固定化及回收利用也亟待研究。【目的】分离筛选并鉴定高效聚磷菌株,优化除磷环境条件,研究吸附材料对所筛菌株除磷的影响,为聚磷菌的开发利用提供理论依据。【方法】采用常规细菌分离筛选方法获得高效菌株,通过形态特征观察、生理生化实验和16S rRNA基因序列分析对菌株进行鉴定;结合单因素试验、Box-BehnkenDesign和响应面分析优化除磷条件;通过测定海绵、无纺布和聚氨酯泡沫对聚磷菌的吸附作用评估该材料的固定化效果。【结果】从连云港市某磷矿厂废水中分离筛选出一株高效聚磷细菌P49,经鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);优化后的最佳除磷条件为pH 6.8、温度31℃、装液量30.2%,在此条件下P49除磷率可达80.43%;应用聚氨酯泡沫的吸附效果优于海绵和无纺布。【结论】解淀粉芽孢杆菌P49具有良好的除磷效果,可为生物除磷提供微生物资源,用聚氨酯泡沫吸附可实现对该菌株的固定化和回收利用。     

Molecular characterization of a lipase-producing <Emphasis Type="Italic">Bacillus pumilus</Emphasis> strain (NMSN-1d) utilizing colloidal water-dispersible polyurethane

Bacillus pumilus strain NMSN-1d isolated from polyurethane-contaminated water was found to grow in high salt concentration (NaCl 10%, w/v) and degrade Impranil-DLN, water-dispersible polyurethane. The genetic relatedness of the isolate has been established by standard molecular biological techniques and the enzyme(s) involved in polyurethane degradation were also studied. A total of nine bacterial strains were isolated from polyurethane-polluted sites and characterized by conventional, microbiological and biochemical methods. These isolates were subjected to 16S ribosomal RNA gene amplification by PCR using specific primers. The genetic relatedness of the isolates was also ascertained by ribotyping and BLAST analysis of the 16S ribosomal RNA gene sequences. The bacterial isolates were grown in yeast extract-salts minimal broth medium supplemented with water-dispersible polyurethane (Impranil DLN) as a sole source of carbon. The promising isolate utilizing polyurethane and producing lipase was identified as Bacillus pumilus NMSN-1d. The polyurethane degradation has been studied in polyurethane-Rhodamine-B and Luria-Bertani-polyurethane plate assays. The activity of hydrolytic enzymes such as lipase and esterase was confirmed on 2xYT-olive oil and tributyrin-Tween 20 plate assay. The newly isolated Bacillus pumilus appears promising in the management of polyurethane waste and in production of industrially important enzymes.     

Aerial release of bacteria from cot mattress materials and the sudden infant death syndrome

AIM: To investigate aerial release of bacteria from used cot mattresses and to assess factors that may influence this process. METHODS AND RESULTS: Movement on used mattresses, simulating that of an infant's head, significantly enhanced aerial release of naturally acquired bacteria from the polyurethane foams (total count data, P = 0.008; Staphylococcus aureus, P = 0.004) or from polyvinyl chloride covers (total count data, P = 0.001). Aerial release of naturally acquired bacteria from used cot mattresses showed high variability and was poorly correlated (R2 < or = 0.294) with bacterial cell density within the materials. In experiments involving inoculation of S. aureus and Escherichia coli onto the polyurethane of unused cot mattresses, aerial release of the species correlated well (R2 > or = 0.950) with inoculation density when simulated infant head movement was applied. Aerial release of these bacterial species from the material decreased with increase in width or aqueous content of the material, and was lower from polyurethane foam of a used cot mattress. CONCLUSIONS: Simulated infant movement and mattress related factors influence aerial release of bacteria from cot mattress materials. With simulated infant movement on cot mattress polyurethane foam, levels of airborne bacteria above the material are proportional to bacterial population levels inoculated onto the material. SIGNIFICANCE AND IMPACT OF THE STUDY: Cot mattresses harbouring relatively high levels of naturally acquired toxigenic bacteria, such as S. aureus, could pose a relatively high risk of infection to the infant's respiratory tract through increased aerial contamination. This has impact in the context of recent findings on cot mattress related risk factors for sudden infant death syndrome.     

The role of microbial biofilms in deterioration of space station candidate materials

Formation of microbial biofilms on surfaces of a wide range of materials being considered as candidates for use on the International Space Station was investigated. The materials included a fibre-reinforced polymeric composite, an adhesive sealant, a polyimide insulation foam, teflon cable insulation, titanium, and an aliphatic polyurethane coating. They were exposed to a natural mixed population of bacteria under controlled conditions of temperature and relative humidity (RH). Biofilms formed on the surfaces of the materials at a wide range of temperatures and RHs. The biofilm population was dominated by Pseudomonas aeruginosa, Ochrobactrum anthropi, Alcaligenes denitrificans, Xanthomonas maltophila, and Vibrio harveyi. The biocide, diiodomethyl-p-tolyl sulfone, impregnated in the polyurethane coating, was ineffective against microbial colonization and growth. Degradation of the polyurethane coatings was monitored with electrochemical impedance spectroscopy (EIS). The impedance spectra indicated that microbial degradation of the coating occurred in several stages. The initial decreases in impedance were due to the transport of water and solutes into the polymeric matrices. Further decreases were a result of polymer degradation by microorganisms. Our data showed that these candidate materials for space application are susceptible to biofilm formation and subsequent degradation. Our study suggests that candidate materials for use in space missions need to be carefully evaluated for their susceptibility to microbial biofilm formation and biodegradation.     

Candida keroseneae sp. nov., a novel contaminant of aviation kerosene

Aims: To characterize and identify a novel contaminant of aviation fuel. Methods and Results: Micro‐organisms (yeasts and bacteria) were isolated from samples of aviation fuel. A yeast that proved to have been unrecorded previously was isolated from more than one fuel sample. This novel yeast proved to be a new species of Candida and is described here. Ribosomal RNA gene sequence analyses of internal transcribed spacer (ITS) regions (including 5·8S subunit) plus the 26S D1/D2 domains showed the strains to cluster within the Candida membranifaciens clade nearest to, but distinct from, Candida tumulicola. Phenotypic tests were identical for both isolates. Physiological and biochemical tests supported their position as a separate taxon. The yeast was assessed for its effect on the main constituent hydrocarbons of aviation fuel. Conclusions: Two strains (IMI 395605^T and IMI 395606) belonging to the novel yeast species, Candida keroseneae, were isolated from samples of aircraft fuel (kerosene), characterized and described herein with reference to their potential as contaminants of aviation fuel. Significance and Impact of the Study: As a result of isolating a novel yeast from aviation fuel, the implications for microbial contamination of such fuel should be considered more widely than previously thought.     

Effect of the fuel system icing inhibitor diethylene glycol monomethyl ether on the biodegradation of jet fuel in soil

Microbial degradation of jet fuel leads to the accumulation of sludge in fuel distribution systems and storage tanks. To prevent this phenomenon, the biocidal anti-icing inhibitor diethylene glycol monomethyl ether (DiEGME) is routinely added to the fuel. The fate of DiEGME in soil and its consequent effect on the biodegradation of jet fuel by indigenous soil microflora have not been investigated. The aim of this work was to study the kinetics of biodegradation of jet fuel in dark rendzina soil, as affected by the presence of DiEGME. Our data show that the degradability in soil of jet fuel amended with DiEGME was tenfold higher than that of non-amended fuel. Consequently, there was an increase in the jet-fuel-utilizing soil microbial populations during the 100 days of incubation of soil samples amended with jet fuel containing DiEGME. Gas chromatograms of distilled fractions of jet fuel extracted from the soil demonstrated that most of the light fractions' extracts could not be detected at the end of the 100-day incubation. The relative concentration of aromatic compounds in the soil contaminated with DiEGME-amended jet fuel increased during incubation, demonstrating the lower biodegradation rate of these components compared with other fuel components. DiEGME was partially degraded by the general microbial population of the soil. Maximal DiEGME degradation was obtained with specific jet-fuel-utilizing microbial strains – Pseudomonas aeruginosa and Cladosporium resinae – that were added to a carbon-free mineral medium. The degradation rate of DiEGME by specific strains or by soil mixed populations bore an inverse relationship to the DiEGME concentration. The finding that DiEGME can be degraded by indigenous soil microorganisms may have facilitated its utilization also by jet-fuel-degrading microorganisms.     

Microbial composition of the activated sludge of Moscow wastewater treatment plants

The contribution of the major technologically important microbial groups (ammonium- and nitrite-oxidizing, phosphate-accumulating, foam-inducing, and anammox bacteria, as well as planctomycetes and methanogenic archaea) was characterized for the aeration tanks of the Moscow wastewater treatment facilities. FISH investigation revealed that aerobic sludge were eubacterial communities; the metabolically active archaea contributed insignificantly. Stage II nitrifying microorganisms and planctomycetes were significant constituents of the bacterial component of activated sludges, with Nitrobacter spp. being the dominant nitrifiers. No metabolically active anammox bacteria were revealed in the sludge from aeration tanks. The sludge from the aeration tanks using different wastewater treatment technologies were found to have differing characteristics. Abundance of the nitrifying and phosphate-accumulating bacteria in the sludge generally correlated with microbial activity in microcosms and with efficiency of nitrogen and phosphorus removal from wastewater. The highest microbial numbers and activity were found in the sludge of the tanks operating according to the technologies developed in the universities of Hannover and Cape Town. The activated sludge from the Novokur’yanovo facilities, where abundant growth of filamentous bacteria resulted in foam formation, exhibited the lowest activity. The group of foaming bacteria included Gordonia spp. and Acinetobacter spp utilizing petroleum and motor oils, Sphaerotilus spp. utilizing unsaturated fatty acids, and Candidatus ‘Microthrix parvicella’. Thus, the data on abundance and composition of metabolically active microorganisms obtained by FISH may be used for the technological control of wastewater treatment.     

Anaerobic degradation of papermill sludge in a two-phase digester containing rumen microorganisms and colonized polyurethane foam

Summary The use of reticulated polyurethane foam as a support material for the immobilization of methanogenic associations and its application to the anaerobic treatment of fine particulate solid wastes was investigated. The colonization of polyurethane support particles in a continuous upflow reactor fed on a mixture of acetate, propionate and butyrate, was both rapid and dense. The combination of rumen microorganisms and colonized support particles in a two-phase digester resulted in an efficient anaerobic decomposition of papermill sludge.     

First use of two-dimensional polyacrylamide gel electrophoresis to determine phylogenetic relationships

Methods for microbial classification are not always capable of distinguishing between isolates at the species level. We have previously characterised four Ferroplasma isolates that were >98.9% similar at the 16S rDNA level, the isolates showed marked phenotypic differences, and one isolate was borderline on the 70% species boundary from DNA-DNA similarity data. In this study we have used statistical comparisons of two-dimensional polyacylamide gel electrophoresis gels for classification of closely related isolates. From the protein profile similarities an un-rooted tree was constructed that was congruent with a tree derived from DNA-DNA similarities.     

沉积型海底微生物燃料电池构建及其影响因素研究

海底微生物燃料电池具有底物丰富、可长期运行、维护成本低和环境友好等特点,具有很好的研究价值和广阔的发展前景。但由于其低的功率密度输出和长期运行的不稳定性,使海底微生物燃料电池尚未得到广泛地实际应用。选取海底沉积泥用于实验室构建的海底微生物燃料电池装置中,比较了在不同阳极材料、阴阳极面积比、阳极修饰、阳极泥下深度条件下海底微生物燃料电池的功率密度输出及其电化学性能,得出最佳的海底微生物燃料电池阳极材料为碳毡;阴极及电极最佳面积比为1∶1;最佳阳极修饰为氨水浸渍;最佳阳极泥下深度为2 cm。     

Biofuel cells select for microbial consortia that self-mediate electron transfer

Microbial fuel cells hold great promise as a sustainable biotechnological solution to future energy needs. Current efforts to improve the efficiency of such fuel cells are limited by the lack of knowledge about the microbial ecology of these systems. The purposes of this study were (i) to elucidate whether a bacterial community, either suspended or attached to an electrode, can evolve in a microbial fuel cell to bring about higher power output, and (ii) to identify species responsible for the electricity generation. Enrichment by repeated transfer of a bacterial consortium harvested from the anode compartment of a biofuel cell in which glucose was used increased the output from an initial level of 0.6 W m(-2) of electrode surface to a maximal level of 4.31 W m(-2) (664 mV, 30.9 mA) when plain graphite electrodes were used. This result was obtained with an average loading rate of 1 g of glucose liter(-1) day(-1) and corresponded to 81% efficiency for electron transfer from glucose to electricity. Cyclic voltammetry indicated that the enhanced microbial consortium had either membrane-bound or excreted redox components that were not initially detected in the community. Dominant species of the enhanced culture were identified by denaturing gradient gel electrophoresis and culturing. The community consisted mainly of facultative anaerobic bacteria, such as Alcaligenes faecalis and Enterococcus gallinarum, which are capable of hydrogen production. Pseudomonas aeruginosa and other Pseudomonas species were also isolated. For several isolates, electrochemical activity was mainly due to excreted redox mediators, and one of these mediators, pyocyanin produced by P. aeruginosa, could be characterized. Overall, the enrichment procedure, irrespective of whether only attached or suspended bacteria were examined, selected for organisms capable of mediating the electron transfer either by direct bacterial transfer or by excretion of redox components.     

Spatial variation in frequency and intensity of antibiotic interactions among Streptomycetes from prairie soil

Antibiotic interactions are believed to be significant to microbial fitness in soil, yet little is known of the frequency, intensity, and diversity of antibiotic inhibition and resistance among indigenous microbes. To begin to address these issues, we studied the abilities of streptomycete isolates from prairie soil to inhibit growth and display resistance to antibiotics produced by a test collection of 10 streptomycete isolates. Wide variations in antibiotic inhibition and resistance for prairie isolates among three locations and four soil depths within a 1-m2 plot were revealed. Fewer than 10% of 153 prairie isolates inhibited all 10 test isolates, while more than 40% of the isolates did not inhibit any of the test isolates. No field isolate was resistant to all of the test isolates, nor was any isolate susceptible to all of the test isolates. No correlation between inhibition and resistance phenotypes was found, suggesting that inhibition and resistance are under independent selection. The significant spatial variation in the frequency and intensity of antibiotic inhibition implies that the fitness benefits of antibiotic production are not the same among locations in soil. In contrast, the consistency of resistance over space indicates that its significance to fitness across locations is stable or the costs of maintaining resistance in the absence of selection are small or nonexistent. The spatial clustering of antibiotic inhibitory activity suggests a variable matrix of selection pressures and microbial responses across the soil landscape.     

Development of a simple cultivation method for isolating hitherto-uncultured cellulase-producing microbes

Although enrichment culture is typically employed to isolate cellulolytic microbes, this approach tends to favor fast-growing species and discriminates against all others. Therefore, efforts to prevent the overgrowth of fast-growing species are necessary to isolate novel cellulase-producing strains. In this study, we developed a simple culture method for isolating hitherto-uncultured microbes that possess cellulase activity, particularly exocellulase. In this method, the microbial source (a forest soil) was suspended in sterilized water and inoculated onto a mineral salts agar medium, which was then overlaid with filter paper to sandwich the microbial suspension between the agar surface and paper. The filter paper fibers served to immobilize the microbial cells and were the dominant carbon source. Following cultivation at 30°C for 2 weeks, emerging colonies were isolated based on their morphology and were then subjected to phylogenetic and enzyme analyses. Using this method, 2,150 CFUs/g dry soil were obtained, and the ratio of fungal to bacterial isolates was approximately 4:1. Phylogenetic analyses revealed that most fungal and bacterial isolates belong to ten and two genera, respectively. Notably, all isolates possessed exocellulase activity, and several strains showed strong activity that was comparable to Trichoderma cellulase. Many isolates also exhibited cellulase and xylanase activity, and several strains possessed laccase activity. It is expected that the culture method described here will be useful for the isolation of hitherto-uncultured cellulolytic microbes and the identification of novel cellulases.