An alternative physiological role for the EmhABC efflux pump in Pseudomonas fluorescens cLP6a

<p>Abstract</p> <p>Background</p> <p>Efflux pumps belonging to the resistance-nodulation-division (RND) superfamily in bacteria are involved in antibiotic resistance and solvent tolerance but have an unknown physiological role. EmhABC, a RND-type efflux pump in <it>Pseudomonas fluorescens </it>strain cLP6a, extrudes hydrophobic antibiotics, dyes and polycyclic aromatic hydrocarbons including phenanthrene. The effects of physico-chemical factors such as temperature or antibiotics on the activity and expression of EmhABC were determined in order to deduce its physiological role(s) in strain cLP6a in comparison to the <it>emhB </it>disruptant strain, cLP6a-1.</p> <p>Results</p> <p>Efflux assays conducted with ¹⁴C-phenanthrene showed that EmhABC activity is affected by incubation temperature. Increased phenanthrene efflux was measured in cLP6a cells grown at 10°C and decreased efflux was observed at 35°C compared with cells grown at the optimum temperature of 28°C. Membrane fatty acids in cLP6a cells were substantially altered by changes in growth temperature and in the presence of tetracycline. Changed membrane fatty acids and increased membrane permeability were associated with ~30-fold increased expression <it>of emhABC </it>in cLP6a cells grown at 35°C, and with increased extracellular free fatty acids. Growth <it>of P. fluorescens </it>cLP6a at supra-optimal temperature was enhanced by the presence of EmhABC compared to strain cLP6a-1.</p> <p>Conclusions</p> <p>Combined, these observations suggest that the EmhABC efflux pump may be involved in the management of membrane stress effects such as those due to unfavourable incubation temperatures. Efflux of fatty acids replaced as a result of membrane damage or phospholipid turnover may be the primary physiological role of the EmhABC efflux pump in <it>P. fluorescens </it>cLP6a.</p

Genome Sequence of the Mesophilic Thermotogales Bacterium Mesotoga prima MesG1.Ag.4.2 Reveals the Largest Thermotogales Genome To Date

Here we describe the genome of Mesotoga prima MesG1.Ag4.2, the first genome of a mesophilic Thermotogales bacterium. Mesotoga prima was isolated from a polychlorinated biphenyl (PCB)-dechlorinating enrichment culture from Baltimore Harbor sediments. Its 2.97 Mb genome is considerably larger than any previously sequenced Thermotogales genomes, which range between 1.86 and 2.30 Mb. This larger size is due to both higher numbers of protein-coding genes and larger intergenic regions. In particular, the M. prima genome contains more genes for proteins involved in regulatory functions, for instance those involved in regulation of transcription. Together with its closest relative, Kosmotoga olearia, it also encodes different types of proteins involved in environmental and cell–cell interactions as compared with other Thermotogales bacteria. Amino acid composition analysis of M. prima proteins implies that this lineage has inhabited low-temperature environments for a long time. A large fraction of the M. prima genome has been acquired by lateral gene transfer (LGT): a DarkHorse analysis suggests that 766 (32%) of predicted protein-coding genes have been involved in LGT after Mesotogadiverged from the other Thermotogales lineages. A notable example of a lineage-specific LGT event is a reductive dehalogenase gene—a key enzyme in dehalorespiration, indicating M. prima may have a more active role in PCB dechlorination than was previously assumed

Degradation of a benzene–toluene mixture by hydrocarbon-adapted bacterial communities

We examined the rate of degradation of a benzene–toluene mixture in aerobic microcosms prepared with samples of an aquifer that lies below a petrochemical plant (SIReN, UK). Five samples exposed to different concentrations of benzene (from 0.6 to 317 mg l−1) were used. Fast degradation (approx. 1–6 mg l−1 day−1) of both contaminants was observed in all groundwater samples and complete degradation was recorded by the seventh day except for one sample. We also identified the microbial community in each of the samples by culture-independent techniques. Two of the less impacted samples harbour the aerobic benzene degrader Pseudomonas fluorescens, while Acidovorax and Arthrobacter spp. were found in the most polluted sample and are consistent with the population observed in situ. Hydrogenophaga was found in the deepest sample while Rhodoferax spp. were recovered in an alkaline sample (pH 8.4) and may also be implicated in benzene degradation. Time series analysis shows that each of the samples has a different community but they remain stable over the degradation period. This study provides new information on a well not previously studied (no. 309s) and confirms that adapted communities have the ability to degrade hydrocarbon mixtures and could be used in further bioaugmentation approaches in contaminated sites

Hydro-biogeochemical coupling beneath a large polythermal Arctic glacier: Implications for subice sheet biogeochemistry

This article was published in the serial, Journal of Geophysical Research: Earth Surface [Wiley © American Geophysical Union]. It is also available at: http://dx.doi.org/10.1029/2009JF001602We analyze the interannual chemical and isotopic composition of runoff from a large, high Arctic valley glacier over a 5 year period, during which drainage evolved from a long-residence-time drainage system feeding an artesian subglacial upwelling (SGU) at the glacier terminus to a shorter-residence-time drainage system feeding an ice-marginal channel (IMC). Increased icemelt inputs to the SGU are thought to have triggered this evolution. This sequence of events provides a unique opportunity to identify coupling between subglacial hydrology and biogeochemical processes within drainage systems of differing residence time. The biogeochemistry of the SGU is consistent with prolonged contact between meltwaters and subglacial sediments, in which silicate dissolution is enhanced, anoxic processes (e.g., sulphate reduction) prevail, and microbially generated CO2 and sulphide oxidation drive mineral dissolution. Solute in the IMC was mainly derived from moraine pore waters which are added to the channel via extraglacial streams. These pore waters acquire solute predominantly via sulphide oxidation coupled to carbonate/silicate dissolution. We present the first evidence that microbially mediated processes may contribute a substantial proportion (80% in this case) of the total glacial solute flux, which includes coupling between microbial CO2-generation and silicate/carbonate dissolution. The latter suggests the presence of biofilms in subglacial/ice-marginal sediments, where local perturbation of the geochemical environment by release of protons, organic acids, and ligands stimulates mineral dissolution. These data enable inferences to be made regarding biogeochemical processes in longer-residence-time glacial systems, with implications for the future exploration of Antarctic subglacial lakes and other wet-based ice sheet environments

Global Distribution of Polaromonas Phylotypes - Evidence for a Highly Successful Dispersal Capacity

Bacteria from the genus Polaromonas are dominant phylotypes in clone libraries and culture collections from polar and high-elevation environments. Although Polaromonas has been found on six continents, we do not know if the same phylotypes exist in all locations or if they exhibit genetic isolation by distance patterns. To examine their biogeographic distribution, we analyzed all available, long-read 16S rRNA gene sequences of Polaromonas phylotypes from glacial and periglacial environments across the globe. Using genetic isolation by geographic distance analyses, including Mantel tests and Mantel correlograms, we found that Polaromonas phylotypes are globally distributed showing weak isolation by distance patterns at global scales. More focused analyses using discrete, equally sampled distances classes, revealed that only two distance classes (out of 12 total) showed significant spatial structuring. Overall, our analyses show that most Polaromonas phylotypes are truly globally distributed, but that some, as yet unknown, environmental variable may be selecting for unique phylotypes at a minority of our global sites. Analyses of aerobiological and genomic data suggest that Polaromonas phylotypes are globally distributed as dormant cells through high-elevation air currents; Polaromonas phylotypes are common in air and snow samples from high altitudes, and a glacial-ice metagenome and the two sequenced Polaromonas genomes contain the gene hipA, suggesting that Polaromonas can form dormant cells

Pseudomonas aeruginosa Population Structure Revisited

At present there are strong indications that Pseudomonas aeruginosa exhibits an epidemic population structure; clinical isolates are indistinguishable from environmental isolates, and they do not exhibit a specific (disease) habitat selection. However, some important issues, such as the worldwide emergence of highly transmissible P. aeruginosa clones among cystic fibrosis (CF) patients and the spread and persistence of multidrug resistant (MDR) strains in hospital wards with high antibiotic pressure, remain contentious. To further investigate the population structure of P. aeruginosa, eight parameters were analyzed and combined for 328 unrelated isolates, collected over the last 125 years from 69 localities in 30 countries on five continents, from diverse clinical (human and animal) and environmental habitats. The analysed parameters were: i) O serotype, ii) Fluorescent Amplified-Fragment Length Polymorphism (FALFP) pattern, nucleotide sequences of outer membrane protein genes, iii) oprI, iv) oprL, v) oprD, vi) pyoverdine receptor gene profile (fpvA type and fpvB prevalence), and prevalence of vii) exoenzyme genes exoS and exoU and viii) group I pilin glycosyltransferase gene tfpO. These traits were combined and analysed using biological data analysis software and visualized in the form of a minimum spanning tree (MST). We revealed a network of relationships between all analyzed parameters and non-congruence between experiments. At the same time we observed several conserved clones, characterized by an almost identical data set. These observations confirm the nonclonal epidemic population structure of P. aeruginosa, a superficially clonal structure with frequent recombinations, in which occasionally highly successful epidemic clones arise. One of these clones is the renown and widespread MDR serotype O12 clone. On the other hand, we found no evidence for a widespread CF transmissible clone. All but one of the 43 analysed CF strains belonged to a ubiquitous P. aeruginosa “core lineage” and typically exhibited the exoS+/exoU− genotype and group B oprL and oprD alleles. This is to our knowledge the first report of an MST analysis conducted on a polyphasic data set

Mycoremediation of petroleum contaminated soils: progress, prospects and perspectives

Mycoremediation, an aspect of bioremediation, has been investigated for some decades. However, there seems to be little progress on its commercial application to petroleum-contaminated soils despite some promising outcomes. In this review, mycoremediation is examined to identify development, limitations and perspectives for its optimal utilization on petroleum-contaminated soils. Mycoremediation agents and substrates that have been used for the treatment of petroleum contaminated soils have been identified, application methods discussed, recent advances highlighted and limitations for its applications accentuated. Possible solutions to the challenges in applying mycoremediation to petroleum-contaminated soils have also been discussed. From this review, we conclude that for optimal utilization of mycoremediation of petroleum-contaminated soils, ideal environmental, edaphic and climatic factors of a typical contaminated site must be incorporated into the approach from first principles. Development of application procedures that can easily translate laboratory results to field applications is also required