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Summary: The interference of plant compounds with bacterial quorum sensing (QS) is a major mechanism through which plants and bacteria communicate. However, little is known about the modes of action and effects on signal integrity during this type of communication. We have recently shown that the plant compound rosmarinic acid (RA) specifically binds to the Pseudomonas aeruginosa RhlR QS receptor. To determine the effect of RA on expression patterns, we carried out global RNA‐seq analysis. The results show that RA induces the expression of 128 genes, amongst which many virulence factor genes. RA triggers a broad QS response because 88% of the induced genes are known to be controlled by QS, and because RA stimulated genes were found to be involved in all four QS signalling systems within P. aeruginosa. This finding was confirmed through the analysis of transcriptional fusions transferred to wt and a rhlI/lasI double mutant. RA did not induce gene expression in the rhlI/lasI/rhlR triple mutant indicating that the effects observed are due to the RA‐RhlR interaction. Furthermore, RA induced seven sRNAs that were all encoded in regions close to QS and/or RA induced genes. This work significantly enhances our understanding of plant bacteria interaction. [ABSTRACT FROM AUTHOR]

Transporters of the cation diffusion facilitator (CDF) family form dimers that export transitionmetals fromthe cytosol. The opportunistic pathogen Pseudomonas aeruginosa encodes three homologous CDF genes, czcD (PA0397), aitP (PA1297), and yiiP (PA3963). The three proteins are required for virulence in a plant host model. Disruption of the aitP gene leads to higher Fe2+ and Co2+ sensitivity together with an intracellular accumulation of these ions and to a decreased survival in presence of H2O2. Strains lacking czcD and yiiP showed low Zn2+ sensitivity. However, in iron-rich media and in the presence of Zn2+ these strains secreted higher levels of the iron chelator pyoverdine. Disruption of czcD and yiiP in a non-pyoverdine producer strain and lacking the Zn2+-transporting ATPase, increased the Zn2+ sensitivity and the accumulation of this ion. Most importantly, independent of the pyoverdine production strains lacking CzcD or YiiP, presented lower resistance to imipenem, ciprofloxacin, chloramphenicol, and gentamicin. These observations correlated with a lower survival rate upon EDTA-lysozyme treatment and overexpression of OprN and OprD porins. We hypothesize that while AitP is an Fe2+/Co2+ efflux transporter required for Fe2+ homeostasis, and ultimately redox stress handling, CzcD, and YiiP export Zn2+ to the periplasm for proper Zn2+-dependent signaling regulating outer membrane stability and therefore antibiotic tolerance. [ABSTRACT FROM AUTHOR]

Background The bioactive form of vitamin D3, i.e. 1,25-dihydroxyvitamin D3 (1,25(OH) 2 D 3 ) vitamin D has been shown to modulate monocytes/macrophages physiology and its response against bacterial infections. Pseudomonas aeruginosa ( P. aeruginosa ) is an opportunistic bacterial pathogen that can most frequently be fatal in immunocompromised infected people. Methods We investigated the ex vivo effect of 1,25(OH) 2 D 3 on monocyte-derived macrophages function against P. aeruginosa infection. Results Relative vitamin D receptor (VDR) mRNA expression was significantly increased in infected and 1,25(OH) 2 D 3 -treated macrophages compared to controls ( p < 0.01). Treatment with 1,25(OH) 2 D 3 markedly resulted in up-regulation of nitric oxide (NO) and IL-1β production and down-regulation of IL-10 levels (respectively, p = 0.029, p = 0.048 and p = 0.008). Additionally, 1,25(OH) 2 D 3 significantly increased M1/M2 macrophage ratio ( p < 0.05) and slightly reduced intracellular bacterial development. Furthermore, the arginase activity, P. aeruginosa phagocytosis and killing were significantly increased in cells that were both infected and 1,25(OH) 2 D 3 -treated compared to the infected, but not 1,25(OH) 2 D 3 -treated macrophages (respectively, p < 0.001, p < 0.01 and p < 0.001). Conclusions We show in this study that bioactive from of vitamin D [1,25-dihydroxyvitamin D3 (1,25D3)] can enhance M1 macrophage polarization and their bactericidal protective activity against P. aeruginosa . Future works would involve improving the treatment response through dose-dependent effect studies, both in ex vivo and in vivo models. [ABSTRACT FROM AUTHOR]

Objectives Manuka honey is a broad-spectrum antimicrobial agent that seems to affect different bacteria in many different ways. It has been shown to be bactericidal against Pseudomonas aeruginosa by destabilizing the cell wall, but we aimed to investigate whether there were further intracellular target sites. Methods In this study inhibitory effects of manuka honey on P. aeruginosa were investigated using hydrophobicity assays, two-dimensional electrophoresis, quantitative RT–PCR, transmission electron microscopy and motility assays. Results Exposure of P. aeruginosa to manuka honey reduced both swarming and swimming motility. Moreover, this was a consequence of de-flagellation of the bacterial cell, which was correlated with decreased expression of the major structural flagellin protein, FliC, and concurrent suppression of flagellin-associated genes, including fliA, fliC, flhF, fleN, fleQ and fleR. The differential expression of the flagellar regulon in the presence of manuka honey was mapped schematically. Flagella are integral to bacterial adhesion, the initiation of infection and biofilm formation, and swarming has been associated with increased virulence. Conclusions By limiting motility in vitro, we infer that manuka honey impacts on the virulence of P. aeruginosa. This deduction must now be tested in vivo. [ABSTRACT FROM PUBLISHER]

Objectives For fluoroquinolones, the area under the free plasma concentration–time curve divided by the MIC (fAUC/MIC) best predicts bacterial killing in mice and outcomes in patients. However, it is unknown whether the shape of the antibiotic concentration profile affects resistance emergence. Our objective was to compare killing and resistance between ciprofloxacin concentration profiles with different shapes at the same fAUC/MIC and identify the durations of ciprofloxacin exposure that minimize resistance emergence. Methods Static time–kill studies over 24 h using Pseudomonas aeruginosa ATCC 27853 assessed fAUC/MIC of 44 and 132 of ciprofloxacin (MICCIP = 0.25 mg/L) and fAUC/MIC of 22, 44 and 132 of ciprofloxacin plus an efflux pump inhibitor (MICCIP+EPI = 0.031 mg/L) at initial inocula of 104, 105 and 106 cfu/mL. Ciprofloxacin was added at 0 h and rapidly removed at 1, 4, 10, 16 or 24 h. Mutant frequencies and MICs were determined at 24 h. Results High ciprofloxacin concentrations over 1–10 h yielded more rapid and extensive initial killing compared with 16 and 24 h exposures at the same fAUC/MIC. No resistance emerged for 1–10 h exposures, although regrowth of susceptible bacteria was extensive. Ciprofloxacin exposure over 24 h yielded less regrowth, but ciprofloxacin-resistant bacteria at 5× MIC amplified by over 5 log10 and almost completely replaced the susceptible bacteria by 24 h; MICs increased 4- to 8-fold. Resistance also emerged on 3× MIC, but not 5× MIC, plates when efflux was inhibited. Conclusions Pre-existing resistant subpopulations amplified extensively with 24 and 16 h exposures, but not with shorter durations. The shape of the ciprofloxacin concentration profile was critical to minimize resistance emergence. [ABSTRACT FROM PUBLISHER]

In the arms race of bacterial pathogenesis, bacteria produce an array of toxins and virulence factors that disrupt core host processes. Hosts mitigate the ensuing damage by responding with immune countermeasures. The iron-binding siderophore pyoverdin is a key virulence mediator of the human pathogen Pseudomonas aeruginosa, but its pathogenic mechanism has not been established. Here we demonstrate that pyoverdin enters Caenorhabditis elegans and that it is sufficient to mediate host killing. Moreover, we show that iron chelation disrupts mitochondrial homeostasis and triggers mitophagy both in C. elegans and mammalian cells. Finally, we show that mitophagy provides protection both against the extracellular pathogen P. aeruginosa and to treatment with a xenobiotic chelator, phenanthroline, in C. elegans. Although autophagic machinery has been shown to target intracellular bacteria for degradation (a process known as xenophagy), our report establishes a role for authentic mitochondrial autophagy in the innate immune defense against P. aeruginosa. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is an opportunistic Gram-negative bacterial pathogen. A primary contributor to its ability to resist β-lactam antibiotics is the expression, following detection of the β-lactam, of the AmpC β-lactamase. As AmpC expression is directly linked to the recycling of the peptidoglycan of the bacterial cell wall, an important question is the identity of the signaling molecule(s) in this relationship. One mechanism used by clinical strains to elevate AmpC expression is loss of function of penicillin-binding protein 4 (PBP4). As the mechanism of the β-lactams is PBP inactivation, this result implies that the loss of the catalytic function of PBP4 ultimately leads to induction of antibiotic resistance. PBP4 is a bifunctional enzyme having both DD-carboxypeptidase and endopeptidase activities. Substrates for both the DD-carboxypeptidase and the 4,3-endopeptidase activities were prepared by multistep synthesis, and their turnover competence with respect to PBP4 was evaluated. The endopeptidase activity is specific to hydrolysis of 4,3-cross-linked peptidoglycan. PBP4 catalyzes both reactions equally well. When P. aeruginosa is grown in the presence of a strong inducer of AmpC, the quantities of both the stem pentapeptide (the substrate for the dd-carboxypeptidase activity) and the 4,3-cross-linked peptidoglycan (the substrate for the 4,3-endopeptidase activity) increase. In the presence of β-lactam antibiotics these altered cell-wall segments enter into the muropeptide recycling pathway, the conduit connecting the sensing event in the periplasm and the unleashing of resistance mechanisms in the cytoplasm. [ABSTRACT FROM AUTHOR]

In the management of multidrug-resistant infections in critically ill patients with multiorgan dysfunction, consideration must be given to the pharmacokinetics and pharmacodynamics of an antimicrobial agent to optimize dosing. We describe a 25-year-old woman who was undergoing thrice-weekly hemodialysis and developed multidrug-resistant Pseudomonas aeruginosa bacteremia secondary to infected left and right ventricular assist devices. After multiple courses of antibiotics, her blood cultures revealed that the infecting organism was becoming progressively more resistant to antibiotic options. Cefepime 2 g administered over 3 hours/day (in combination with colistimethate) provided adequate drug levels for multidrug-resistant, cefepime-intermediate P. aeruginosa bacteremia in this patient. We present the clinical case of this patient, followed by a discussion of possible therapeutic approaches to be considered, including illustration of the principles of using extended-infusion antimicrobial regimens, and present the patient's resulting clinical course. [ABSTRACT FROM AUTHOR]

Biofilm-forming marine bacterium Pseudomonas aeruginosa JP-11 was isolated from coastal marine sediment of Paradeep Port, Odisha, East Coast, India, which resisted up to 1,000 ppm of cadmium (Cd) as cadmium chloride in aerobic conditions with a minimal inhibitory concentration of 1,250 ppm. Biomass and extracellular polymeric substances (EPS) secreted by the cells effectively removed 58.760 ± 10.62 and 29.544 ± 8.02 % of Cd, respectively. The integrated density of the biofilm-EPS observed under fluorescence microscope changed significantly ( P ≤ 0.05) in the presence of 50, 250, 450, 650 and 850 ppm Cd. ATR-FTIR spectroscopy showed a peak at 2,365.09/cm in the presence of 50, 250, 450 and 650 ppm Cd which depicts the presence of sulphydryl group (-SH) within the EPS, whereas, a peak shift to 2,314.837/cm in the presence of 850 ppm Cd suggested the major role of this functional group in the binding with cadmium. On exposure to Cd at 100, 500 and 1,000 ppm, the expression profiles of cadmium resistance gene ( czcABC) in the isolate showed an up-regulation of 3.52-, 17- and 24-fold, respectively. On the other hand, down-regulation was observed with variation in the optimum pH (6) and salinity (20 g l) level. Thus, the cadmium resistance gene expression increases on Cd stress up to the tolerance level, but an optimum pH and salinity are the crucial factors for proper functioning of cadmium resistance gene. [ABSTRACT FROM AUTHOR]

The use of pesticides to increase agricultural production can result in the contamination of the environment, causing changes in the genetic structure of organisms and in the loss of biodiversity. This practice is also inducing changes in the rainforest ecosystem. In this work, a Pseudomonas aeruginosa isolated from a preservation soil area of the Brazilian Amazon Forest, without usage of any pesticide, was evaluated for its potential to degrade atrazine. This isolate presented all responsible genes ( atzA, atzB, atzC, atzD, atzE, and atzF) for atrazine mineralization and demonstrated capacity to use atrazine as a nitrogen source, having achieved a reduction of 44 % of the initial concentration of atrazine after 24 h. These results confirm gene dispersion and/or a possible contamination of the area with the herbicide, which reinforces global concern of the increase and intensive use of pesticides worldwide. [ABSTRACT FROM AUTHOR]

Much of contemporary synthetic biology research relies on the use of bacterial chassis for plugging-in and plugging-out genetic circuits and new-to-nature functionalities. However, the microorganisms that are the easiest to manipulate in the laboratory are often suboptimal for downstream industrial applications, which can involve physicochemical stress and harsh operating conditions. In this Review, we advocate the use of environmental Pseudomonas strains as model organisms that are pre-endowed with the metabolic, physiological and stress-endurance traits that are demanded by current and future synthetic biology and biotechnological needs. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa displays tremendous metabolic diversity, controlled in part by the abundance of transcription regulators in the genome. We have been investigating P. aeruginosa's response to the host, particularly changes regulated by the host-derived quaternary amines choline and glycine betaine (GB). We previously identified GbdR as an AraC family transcription factor that directly regulates choline acquisition from host phospholipids (via binding to plcH and pchP promoters), is required for catabolism of the choline metabolite GB, and is an activator that induces transcription in response to GB or dimethylglycine. Our goal was to characterize the GbdR regulon in P. aeruginosa by using genetics and chemical biology in combination with transcriptomics and in vitro DNA-binding assays. Here we show that GbdR activation regulates transcription of 26 genes from 12 promoters, 11 of which have measureable binding to GbdR in vitro. The GbdR regulon includes the genes encoding GB, dimethylglycine, sarcosine, glycine, and serine catabolic enzymes and the BetX and CbcXWV quaternary amine transport proteins. We characterized the GbdR consensus binding site and used it to identify that the recently characterized acetylcholine esterase gene, choE (PA4921), is also regulated by GbdR. The regulon member not directly controlled by GbdR is the secreted lipase gene lipA, which was also the only regulon member repressed under GbdR-activating conditions. Determination of the GbdR regulon provides deeper understanding of how GbdR links bacterial metabolism and virulence. Additionally, identification of two uncharacterized regulon members suggests roles for these proteins in response to choline metabolites. [ABSTRACT FROM AUTHOR]

The aim of this work was to optimize the culture medium for biomass and PHA production by Pseudomonas aeruginosa 42A2, using a five-level-three-factor central composite rotatable design (CCRD) combined with response surface methodology (RSM). A total of 23 sets of experiments was developed to obtain the second-order polynomial equations that were used to predict biomass and PHA production in terms of three independent variables: carbon source, nitrate and phosphate concentrations. Canonical analysis of the response surface models showed that the optimum medium composition differed for the production of biomass and PHA. When the models were validated experimentally by culturing Pseudomonas aeruginosa 42A2 in the optimal media, similar values to the predicted ones were obtained for the biomass (18.73 g L-1) and PHA (4.52 g L -1) concentrations. Moreover, central composite rotatable design and canonical analysis of the response surfaces proved to be useful tools for determining the optimum composition of the culture medium. [ABSTRACT FROM AUTHOR]

Abstract: Background: Pseudomonas aeruginosa is one of the important causes of nosocomial infections that easily gains resistance to many antibiotics. This opportunistic pathogen is a major health hazard particularly in immunodeficient patients, patients in intensive care units (ICU) and burn units with life threatening outcome. The bacterium may be originated from different or common sources, and comprises a high colonization and transmission capacity. Objective: The aim of present study was to investigate the genotypic variation of Pseudomonas aeroginosa strains isolated from burn patients by using Random Amplified Polymorphic DNA (RAPD) method. Methods: Totally 70 clinical samples were collected from burn patients in Taleghani Burn Hospital of Ahvaz. Fifty out of total samples were positive for P. aeruginosa by application of conventional culture and biochemical identification tests. DNA was extracted from the isolates and the RAPD-PCR method was applied to the DNA extracts according to standard method using a short single primer of 272. The technique created repetitive electrophoresis patterns which was used for genotypic differentiation. Results: RAPD-PCR, created 9 genotypic profiles designated as I–IX with base pair length ranging from 180 to 2700. Each genotype showed between 3 and 6 different weight DNA bands. Genotype I was the most prevalent, identified in 10 bacterial isolates (20%). Genotypes I, II and VI were mostly common in patients with more severe burn, and were mainly isolated from wound and blood samples obtained from the same patients. Conclusion: In present study, we found RAPD-PCR technique as a useful tool for investigation of the genetic variation among P. aeruginosa strains. This is a rapid, low cost, genotypic method with high discriminatory power. The results could assist to screen for the original of infection caused by this organism with subsequent control of colonization and transmission. [Copyright &y& Elsevier]

Fire is a ubiquitous component of the Earth system that is poorly understood. To date, a global-scale understanding of fire is largely limited to the annual extent of burning as detected by satellites. This is problematic because fire is multidimensional, and focus on a single metric belies its complexity and importance within the Earth system. To address this, we identified five key characteristics of fire regimes—size, frequency, intensity, season, and extent-and combined new and existing global datasets to represent each. We assessed how these global fire regime characteristics are related to patterns of climate, vegetation (biomes), and human activity. Cross-correlations demonstrate that only certain combinations of fire characteristics are possible, reflecting fundamental constraints in the types of fire regimes that can exist. A Bayesian clustering algorithm identified five global syndromes of fire regimes, or pyromes. Four pyromes represent distinctions between crown, litter, and grassfueled fires, and the relationship of these to biomes and climate are not deterministic. Pyromes were partially discriminated on the basis of available moisture and rainfall seasonality. Human impacts also affected pyromes and are globally apparent as the driver of a fifth and unique pyrome that represents human-engineered modifications to fire characteristics. Differing biomes and climates may be represented within the same pyrome, implying that pathways of change in future fire regimes in response to changes in climate and human activity may be difficult to predict. [ABSTRACT FROM AUTHOR]

A chemically defined medium called KGm medium was used to isolate from a sample of sea water a bacterial strain, MW3A, capable of using N-3-oxohexanoyl- l-homoserine lactone as the sole carbon source. MW3A was clustered closely to Pseudomonas aeruginosa by 16S ribosomal DNA sequence analysis. It degraded both N-acylhomoserine lactones (AHLs) with a 3-oxo group substitution and, less preferably, AHLs with unsubstituted groups at C3 position in the acyl side chain, as determined by Rapid Resolution Liquid Chromatography. Its quiP and pvdQ homologue gene sequences showed high similarities to those of known acylases. Spent supernatant of MW3A harvested at 8-h post inoculation was shown to contain long-chain AHLs when assayed with the biosensor Escherichia coli [pSB1075], and specifically N-dodecanoyl- l-homoserine lactone and N-3-oxotetradecanoyl- l-homoserine lactone by high resolution mass spectrometry. Hence, we report here a novel marine P. aeruginosa strain MW3A possessing both quorum-quenching and quorum-sensing properties. [ABSTRACT FROM AUTHOR]

Triclosan is a broad-spectrum antimicrobial agent having low toxicity which facilitates its incorporation into numerous personal and health care products. Although triclosan acts against a wide range of gram-positive and gram-negative bacteria by affecting fatty acid biosynthesis, it is ineffective against the opportunistic pathogen Pseudomonas aeruginosa. Wild-type strain P. aeruginosa PAO1 was used as a model system to determine the effects of triclosan on fatty acid metabolism in resistant microorganisms. This was accomplished by cultivating P. aeruginosa PAO1 cultures in the presence of different concentrations of triclosan, monitoring growth rates turbidimetrically, and harvesting in stationary phase. Readily extractable lipids (RELs) were obtained from freeze-dried cells after washing and analyzed using gas chromatography coupled with mass spectrometry. Resultant data demonstrated that triclosan caused dose-dependent increases in the amounts of trans-C and trans-C fatty acids, with concomitant decreases in their respective cyclopropyl analogs. Triclosan did not affect the relative concentrations of saturated, cis unsaturated, or the overall ratios of combined C to C fatty acid species. The readily extractable lipid fractions contained triclosan proportional to triclosan concentrations in the growth media. The presence or absence of triclosan in either liquid or solid media did not affect the antimicrobial susceptibilities of P. aeruginosa PAO1 to a battery of unrelated antimicrobials. Triclosan decreased growth rate in a dose-dependant manner at soluble concentrations. Incorporation of triclosan into the REL fraction was accompanied by increased levels of trans unsaturated fatty acids, decreased levels of cyclopropyl fatty acids, and decrease in growth rate. These alterations may contribute to triclosan resistance in P. aeruginosa PAO1. [ABSTRACT FROM AUTHOR]

Increasing awareness of hazards associated with the use of antibiotic and chemical agents has accelerated investigations into plants and their extracts as new sources of antimicrobial agents. Therefore, this study is aimed at determining the antimicrobial activity of Poly Butylene Succinate extracts of Piper sarmentosum leaves and stems against six pathogenic strains associated with humans namely; Escherichia coli, Pseudomonas aeroginosa, Bacillus anthracis, Staphylocuccus aureus, Candida albicans and Candida neoformans by the disc diffusion method. The results revealed potent antimicrobial activity by both the leaves and stem extracts on P. aeroginosa at all the concentrations of the extracts used with an MIC values of 50mg/ml and 100 mg/ml obtained for with the stem and leaves extracts respectively by the micro broth method. The highest activity was exerted by the stem extract at 400mg/ml (20mm) and the least was by the leaves extract at 50mg/ml (9mm) on P. aeroginosa. However these extracts were unable to inhibit the growth of E. coli, B. anthracis, S. aureus, C. albicans and C. neoformans at all the tested concentrations. This confers utility of the extracts in treating infections associated with P. aeroginosa whereas they are unsuitable for infections associated with the other strains tested in this study. [ABSTRACT FROM AUTHOR]

Background: Otitis Media continues to be a major presentation in the ear, nose and throat clinic. Aim: This study aimed to isolate, characterize and identify the bacteriological and mycological etiologic agents of otitis media in Benin city. Patients and Methods: Ear discharge from 569 (299 males and 270 females) patients diagnosed clinically of otitis media between August 2009 and August 2010 were processed to recover the bacterial and fungal etiologic agents. Susceptibility test was performed on all bacterial isolate. Result: Pseudomonas aeruginosa (28.3%) was the predominant bacteria isolate causing otitis media followed by Staphylococcus aureus (21.0%), Klebsiella sp (8.9%), Proteus sp (8.2%), Alkaligenes spp (4.3%), Streptococcus pneumoniae (3.9%), Escherichia coli (3.0%) and Citrobacter freundi (1.7%). Fungi isolated were Aspergillus niger (9.2%), Candida albicans (5.4%), Candida tropicalis (3.0%), Aspergillus flavus (2.1%) and Candida parasilopsis (1.5%). 413 had a single organism isolated from the middle ear culture while twenty (3.51%) patients had mixed organisms isolated. Infection was highest among 0 - 5 years, and lowest among aged 18 - 23. All bacterial isolates were poorly susceptible to the antibacterial agents. Conclusion: The study uncovers a high frequency of bacteria associated otitis media with the finding of fungi too as a significant etiologic agent. [ABSTRACT FROM AUTHOR]

Proteus vulgaris strain named T6 which produced lipase (PVL) with nonpositional specificity had been isolated in our laboratory. To produce the lipase in large quantities, we cloned its gene, which had an opening reading frame of 864 base pairs and encoded a deduced 287-amino-acid protein. The PVL gene was inserted into the Escherichia coli expression vector pET-DsbA, and active lipase was expressed in E. coli BL21 cells. The secretive expression of PVL gene in Bacillus subtilis was examined. Three vectors, i.e., pMM1525 (xylose-inducible), pMMP43 (constitutive vector, derivative of pMM1525), and pHPQ (sucrose-inducible, constructed based on pHB201), were used to produce lipase in B. subtilis. Recombinant B. subtilis WB800 cells harboring the pHPQ-PVL plasmid could synthesize and secrete the PVL protein in high yield. The lipase activity reached 356.8 U/mL after induction with sucrose for 72 h in shake-flask culture, representing a 12-fold increase over the native lipase activity in P. vulgaris. The characteristics of the heterologously expressed lipase were identical to those of the native one. [ABSTRACT FROM AUTHOR]

Dispersal plays a crucial role in a range of evolutionary and ecological processes; hence there is strong motivation to understand its evolution. One key prediction is that the relative benefits of dispersal should be greater when dispersing away from close relatives, because in this case dispersal has the additional benefit of alleviating competition with individuals who share the same dispersal alleles. We tested this prediction for the first time using experimental populations of the opportunistic pathogen Pseudomonas aeruginosa. We measured the fitness of isogenic genotypes that differed only in their dispersal behaviors in both clonal and mixed populations. Consistent with theory, the benefit of dispersal was much higher in clonal populations, and this benefit decreased with increasing growth rate costs associated with dispersal. [ABSTRACT FROM AUTHOR]

Background: We have previously explored a therapeutic strategy for specifically targeting the profibrotic activity of IL-13 during experimental pulmonary fibrosis using a fusion protein comprised of human IL-13 and a mutated form of Pseudomonas aeruginosa exotoxin A (IL13-PE) and observed that the intranasal delivery of IL13-PE reduced bleomycininduced pulmonary fibrosis through its elimination of IL-13-responsive cells in the lung. The aim of the present study was to determine whether the presence of an immune response to P. aeruginosa and/or its exotoxin A (PE) would diminish the antifibrotic properties of IL13-PE. Methodology/Principal Findings: Fourteen days after P. aeruginosa infection, C57BL/6 mice were injected with bleomycin via the intratracheal route. Other groups of mice received 4 doses of saline or IL13-PE by either intranasal or intraperitoneal application, and were challenged i.t. with bleomycin 28 days later. At day 21 after bleomycin, all mice received either saline vehicle or IL13-PE by the intranasal route and histopatological analyses of whole lung samples were performed at day 28 after bleomycin. Intrapulmonary P. aeruginosa infection promoted a neutralizing IgG2A and IgA antibody response in BALF and serum. Surprisingly, histological analysis showed that a prior P. aeruginosa infection attenuated the development of bleomycin-induced pulmonary fibrosis, which was modestly further attenuated by the intranasal administration of IL13-PE. Although prior intranasal administration of IL13-PE failed to elicit an antibody response, the systemic administration of IL13- PE induced a strong neutralizing antibody response. However, the prior systemic sensitization of mice with IL13-PE did not inhibit the anti-fibrotic effect of IL13-PE in fibrotic mice. Conclusions: Thus, IL13-PE therapy in pulmonary fibrosis works regardless of the presence of a humoral immune response to Pseudomonas exotoxin A. Interestingly, a prior infection with P. aeruginosa markedly attenuated the pulmonary fibrotic response suggesting that the immune elicitation by this pathogen exerts anti-fibrotic effects. [ABSTRACT FROM AUTHOR]

Type VI secretion systems (T6SSs) contribute to interactions of bacterial pathogens and symbionts with their hosts. Previously, we showed that Pseudomonas aeruginosa T6S is posttranslationally activated upon phosphorylation of Fha1, an FHA domain protein, by PpkA, a membrane-spanning threonine kinase. Herein, additional structural, enzymatic and genetic requirements for PpkA-catalysed T6SS activation are identified. We found that PpkA plays an essential structural role in the T6SS, and that this role is intimately linked to its ability to promote secretion and phosphorylate Fha1. Protein localization and protein–protein interaction studies show that a complex containing Fha1 and the T6S ATPase, ClpV1 is recruited to the T6S apparatus in a phosphorylation-dependent manner. The mechanism of PpkA activation was also investigated. We identified critical PpkA autophosphorylation sites and showed that small molecule-induced dimerization of the extracellular domains of PpkA is sufficient to activate the T6SS. Finally, we discovered TagR, a component of the T6S posttranslational regulatory pathway that functions upstream of PpkA to promote kinase activity. We present a model whereby an unknown cue causes dimerization of the extracellular domains of PpkA, leading to its autophosphorylation, recruitment of the Fha1-ClpV1 complex, phosphorylation of Fha1, and T6SS activation. Our findings should facilitate approaches for identifying physiological activators of T6S. [ABSTRACT FROM AUTHOR]

The article presents a study which evaluates the use of virulence factor antagonists as adjunctive therapies for pseudomonas aeruginosa. It shows that these promising translational strategies may lead to adjunctive therapies development being able to improve results. It is concluded that these therapies have the ability to make improvement for the disease.

The article presents a study which shows the citrate-mediated iron uptake in Pseudomonas aeruginosa. It states that in an effort to name the components of a ferric citrate uptake system in Pseudomonas aeruginosa a mutant library of a siderophore-deificient strain (IA614) was constructed and screened for defects in citrate-promoted growth in an Fe-restricted medium. The report of the study indicate that the Fe delivered to Pseudomonas aeruginosa by citrate is released as Fe2+ prior to its transport into cells through Fe transport components.

Abstract: The virulent Pseudomonas aeruginosa bacteriophage LUZ24 (45,625 bp) was isolated from hospital sewage. It belongs to the family of the Podoviridae, and carries a bidirectionally transcribed dsDNA genome delineated by two direct terminal repeats of 184 bp. In vitro transcriptional analysis identified seven σ70 promoters, revealing a bias towards stronger promoter strength in the late genomic region. Reverse transcription demonstrated in vivo splicing of a 668 bp Group I intron embedded inside the DNA polymerase gene. Using mass spectrometry, nine structural proteins were identified as part of the phage particle. The lytic characteristics of LUZ24 are evaluated against its genomic content, which displays an overall 71% sequence similarity to the temperate phage PaP3. [Copyright &y& Elsevier]

The biofilm formation of Pseudomonas aeruginosa, an opportunistic human pathogen, is developed by cell-to-cell signaling, so-called quorum sensing (QS). To control the biofilm formation, we designed and synthesized new QS inhibitors of P. aeruginosa based on the structure of the previously known QS inhibitor, furanone. Newly synthesized compounds were a series of analogs of (5-oxo-2,5-dihydrofuran-3-yl)methyl alkanoate, and the structures of all six synthesized compounds was confirmed by NMR and GC/MS analyses. These new QS inhibitor candidates could remarkably inhibit both Pseudomonas QS signaling and biofilm formation, which were assayed by using the recombinant reporter system and flow cell confocal microscopy. The degree of QS inhibition by these new inhibitors varied from 20% to 90%. For the profound understanding about inhibition mechanism, we tried to estimate the binding energy between QS receptor, LasR, and our inhibitors from the in silico modeling system. The predicted binding pattern from the modeling system and our experimental data about QS inhibition were in good agreement. From these results, we suggest a new approach to develop the QS inhibitors and biofilm control agents based on structural modeling. [ABSTRACT FROM AUTHOR]

Abstract: Pseudomonas aeruginosa LBI produced surface active rhamnolipids when cultivated on waste from the sunflower-oil process under different conditions. These biosurfactants, which reduce the superficial and interfacial tensions between fluids, offer advantages over their chemical counterparts, especially because of their ecological acceptability. These molecules can be used in fields as diverse as chemical, pharmaceutical and petrochemical industries. In this work, we present the effect of C/N ratio on growth and production yield. The best production yields (Y P/S) were achieved for C/N ratios (in g/g) of 8/1 (0.22) and 6.4/1 (0.23). The product concentration was very satisfactory (7.3g/L) at C/N ratio of 8/1, especially when considering that the substrate was basically composed of wastes that would otherwise constitute an environmental disposal problem. [Copyright &y& Elsevier]

The article presents a research paper which illustrates the isolation and characterization of heavy metal resistant strain of Pseudomonas aeruginosa which were isolated from polluted sites in Assiut, Egypt. It mentions that in this research sixty-six isolates of Pseudomonas spp. were kept apart from wastewater of El-Malah canal situated in Assiut, Egypt, and were evaluated for their heavy metal tolerance. Moreover, it is mentioned that resistance to heavy metals, disinfectants, antiobiotics, detergents and other toxic substances were noted in a wide variety of bacteria, particularly in the genus Pseudomonas.

Abstract: The transcription factor QscR is a regulator of quorum sensing in Pseudomonas aeruginosa and plays a role in controlling virulence in this prevalent opportunistic pathogen. This study outlines the discovery of a set of synthetic N-acylated homoserine lactones that are capable of either activating or strongly inhibiting QscR in a cell-based reporter gene assay. We demonstrate that the synthetic antagonists inhibit ligand-dependent QscR binding to DNA. Several of these ligands can selectively modulate QscR instead of LasR, or modulate the activity of both receptors, and represent new chemical tools to study the hierarchy of quorum-sensing signaling in P. aeruginosa. [Copyright &y& Elsevier]

Abstract: A series of 4-oxo-4H-pyrido[1,2-a]pyrimidine derivatives, substituted at the 2-position with piperidines bearing quaternary ammonium salt side chains, were synthesized and evaluated for their ability to potentiate the activity of the fluoroquinolone levofloxacin (LVFX) and the β-lactam aztreonam (AZT) in Pseudomonas aeruginosa. Attachment of the charged entity using an N-ethylcarbamoyloxy linker led to the discovery of the highly soluble compound 22 (D13-9001), which maintained good potency in vitro and displayed excellent activity in vivo in a rat pneumonia model of P. aeruginosa. [Copyright &y& Elsevier]

Abstract: Three new briarane-type diterpenoids, designated as robustolides A–C (1–3), were obtained from the female gorgonian coral Ellisella robusta. The structures of briaranes 1–3 were elucidated by the interpretation of spectral data analysis and the structure of 1 was further confirmed by a single-crystal X-ray diffraction analysis. Briaranes 1 and 2 were found to show weak activity against the bacteria Pseudomonas aeruginosa and Staphylococcus aureus. [Copyright &y& Elsevier]

Abstract: An efficient process for the discovery of inhibitors of DDAH enzymes, without the requirement for high throughput screening, is described. Physicochemical filtering of a 308,000-compound library according to drug likeness followed by reciprocal nearest neighbour selection produced a representative subset of 35,000 compounds. Virtual screening on a dual processor PC using FlexX, followed by biological screening, identified two hit series. Similarity searches of commercial databases and chemical re-synthesis of pure compounds resulted in SR445 as an inhibitor of Pseudomonas aeruginosa DDAH at 2μM. [Copyright &y& Elsevier]

Background. Pseudomonas aeruginosa is an opportunistic bacterium that can cause severe infection in susceptible patients. During the winter of 2001-2002, we investigated an outbreak of P. aeruginosa infection among patients in several hospitals across Norway. Methods. A nationwide outbreak investigation was performed with case finding, questionnaires, and product sampling. All available clinical and environmental P. aeruginosa strains were genotyped. Detailed information was collected from patients with the outbreak strain or with any P. aeruginosa in blood or cerebrospinal fluid samples. To identify risk factors, we conducted a case-control study among patients with P. aeruginosa isolated from blood or cerebrospinal fluid samples during October 2001-December 2002. Case patients were patients infected with the outbreak genotype, and control subjects were patients infected with other genotypes. Results. A total of 231 patients from 24 hospitals were identified as having the outbreak strain; 39 of these patients had positive blood culture results. Seventy-one patients (31%) died while hospitalized; all of the patients who died had severe underlying disease. Among 39 case patients and 159 control subjects, use of the moist mouth swab (adjusted odds ratio, 5.3; 95% confidence interval, 2.0-13.6) and receipt of mechanical ventilation (adjusted odds ratio, 6.4; 95% confidence interval, 2.3-17.2) were associated with infection due to the outbreak strain. Genotypically identical strains of P. aeruginosa were identified in 76 mouth swabs from 12 different batches and from the production line. Conclusions. Contamination of mouth swabs during production caused the largest-ever outbreak of P. aeruginosa infection in Norway. Susceptible patient groups should use only documented quality-controlled, high-level-disinfected products and items in the oropharynx. [ABSTRACT FROM AUTHOR]

Abstract: Rhomboids are ubiquitous integral membrane proteases that release cellular signals from membrane-bound substrates through a general signal transduction mechanism known as regulated intramembrane proteolysis (RIP). We present the NMR structure of the cytosolic N-terminal domain (NRho) of P. aeruginosa Rhomboid. NRho consists of a novel α/β fold and represents the first detailed structural insight into this class of intramembrane proteases. We find evidence that NRho is capable of strong and specific association with detergent micelles that mimic the membrane/water interface. Relaxation measurements on NRho reveal structural fluctuations on the microseconds–milliseconds timescale in regions including and contiguous to those implicated in membrane interaction. This structural plasticity may facilitate the ability of NRho to recognize and associate with membranes. We suggest that NRho plays a role in scissile peptide bond selectivity by optimally positioning the Rhomboid active site relative to the membrane plane. [Copyright &y& Elsevier]

Abstract: This article examines the activity of meropenem and imipenem against Pseudomonas aeruginosa isolates from the Meropenem Yearly Susceptibility Test Information Collection program between 1997 and 2005. In particular, it examines the activity of meropenem against imipenem-resistant strains and vice versa. Meropenem proved to be active against up to a 3rd of imipenem-resistant strains. [Copyright &y& Elsevier]

The availability of high-intensity synchrotron facilities, technological advances in data-collection techniques and improved data-reduction and crystallographic software have ushered in a new era in high-throughput macromolecular crystallography. Here, the de novo automated crystal structure determination at 1.28 Å resolution of an NAD(P)H-dependent FMN reductase flavoprotein from Pseudomonas aeruginosa PA01-derived protein Q9I4D4 using the anomalous signal from an unusually small number of S atoms is reported. Although this protein lacks the flavodoxin key fingerprint motif [(T/S) XTG XT], it has been confirmed to bind flavin mononucleotide and the binding site was identified via X-ray crystallography. This protein contains a novel flavin mononucleotide-binding site GSLRSGSYN, which has not been previously reported. Detailed statistics pertaining to sulfur phasing and other factors contributing to structure determination are discussed. Structural comparisons of the apoenzyme and the protein complexed with flavin mononucleotide show conformational changes on cofactor binding. NADPH-dependent activity has been confirmed with biochemical assays. [ABSTRACT FROM AUTHOR]

Abstract: Cytochrome c-551 from the mesophile Pseudomonas aeruginosa is an electronic transfer protein that contains 82 amino-acid residues and a c-type heme as the prosthetic group with low spin Fe(II) in the reduced form and low spin Fe(III) in the oxidized form of cytochrome c-551. We have studied the electronic properties of ferricytochrome c-551 from P. aeruginosa at high pH (9–11.4) by means of paramagnetic 1H NMR spectra and the T 1 and T 2 values of isotropically shifted proton resonances. We have also analyzed the temperature dependence of the hyperfine-shifts. Resonance assignment of some signals was based on 2D saturation transfer experiments, EXSY. These results indicate the existence of high-spin iron(III) cytochrome c-551 (S =5/2, 6A1) with penta or hexacoordinated symmetry at high pH. A spin transition from low-spin iron(III) to high-spin iron(III) has been produced. This transition can be associated with the deprotonation of the axially coordinated amino-acid to iron, His-16. We also discuss the spin crossover between different ferricytochromes c. [Copyright &y& Elsevier]

Abstract: The synergistic potential of ceftazidime plus amikacin or levofloxacin was assessed against 61 Pseudomonas aeruginosa isolates with variable susceptibility patterns to the 3 antibiotics. A checkerboard broth method and a disk diffusion method were used and compared. The latter, also easy to perform as a triple-disk assay, could be a helpful laboratory screening tool for drug synergism to drive possible combination treatments. [Copyright &y& Elsevier]

It is generally reasoned that lethal infections caused by opportunistic pathogens develop permissively by invading a host that is both physiologically stressed and immunologically compromised. However, an alternative hypothesis might be that opportunistic pathogens actively sense alterations in host immune function and respond by enhancing their virulence phenotype. We demonstrate that interferon-γ binds to an outer membrane protein in Pseudomonas aeruginosa, OprF, resulting in the expression of a quorum-sensing dependent virulence determinant, the PA-1 lectin. These observations provide details of the mechanisms by which prokaryotic organisms are directly signaled by immune activation in their eukaryotic host. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is an opportunistic pathogen which causes a variety of diseases, including respiratory tract infections in patients suffering from cystic fibrosis. Therapeutic treatment of P. aeruginosa infections is still very difficult because the bacteria exhibit high intrinsic resistance against a variety of different antibiotics and, in addition, form stable biofilms, e.g. in the human lung. Several virulence factors are produced by P. aeruginosa, among them the two lectins LecA and LecB, which exert different cytotoxic effects on respiratory epithelial cells and presumably facilitate bacterial adhesion to the airway mucosa. Here, the physiology has been studied of the lectin LecB, which binds specifically to L-fucose. A LecB-deficient P. aeruginosa mutant was shown to be impaired in biofilm formation when compared with the wild-type strain, suggesting an important role for LecB in this process. This result prompted an investigation of the subcellular localization of LecB by cell fractionation and subsequent immunoblotting. The results show that LecB is abundantly present in the bacterial outer-membrane fraction. It is further demonstrated that LecB could be released specifically by treatment of the outer-membrane fraction with p-nitrophenyl α-L-fucose, whereas treatment with D-galactose had no effect. In contrast, a LecB protein carrying the mutation D104A, which results in a defective sugar-binding site, was no longer detectable in the membrane fraction, suggesting that LecB binds to specific carbohydrate ligands located at the bacterial cell surface. Staining of biofilm cells using fluorescently labelled LecB confirmed the presence of these ligands. [ABSTRACT FROM AUTHOR]

The involvement of the chemotaxis gene cluster 1 (cheYZABW) and cheR in repellent responses of Pseudomonas aeruginosa to trichloroethylene (TCE) is described and three methyl-accepting chemotaxis proteins (MCPs) for TCE are identified. TCE chemotaxis assays of a number of deletion-insertion mutants of P. aeruginosa PAO1 revealed that the chemotaxis gene cluster 1 and cheR are required for negative chemotaxis to TCE. Mutant strains which contained deletions in pctA, pctB and pctC showed decreased responses to TCE. The pctA, pctB and pctC genes have been reported to encode MCPs for amino acids [K. Taguchi et al., Microbiology, 143, 3223–3229 (2000)]. The pctA mutation more severely impaired chemotactic responses to TCE than did those of pctB and pctC, suggesting that PctA is the major MCP for TCE among the three MCPs. The pctA, pctB and pctC mutant strains showed decreased responses to chloroform and methylthiocyanate. This result demonstrates that PctA, PctB and PctC are also involved in repellent responses to chloroform and methylthiocyanate. [Copyright &y& Elsevier]

Abstract: Two interlinked quorum sensing circuits, las and rhl, which control pathogenesis of Pseudomonas aeruginosa are further modulated by numerous regulators, including VqsR (virulence and quorum sensing regulator). High-density oligonucleotide microarrays were used to compare the global expression profile of a wild-type and VqsR mutant in ABC minimal medium. The expression of a large group of metabolic genes, ECF sigma factors as well as of many quorum-sensing genes previously not assigned to VqsR-regulon was found to be affected by the disruption of vqsR. [Copyright &y& Elsevier]

Biochemical oxygen demand (BOD) is an international regulatory environmental index for monitoring organic pollutants in wastewater and the current legislated standard test for BOD monitoring requires 5 days to complete (BOD5 test). We are developing a rapid microbial technique, MICREDOX®, for measuring BOD by eliminating oxygen and, instead, quantifying an equivalent biochemical co-substrate demand, the co-substrate being a redox mediator. Elevated concentrations of Proteus vulgaris, either as free cells or immobilised in Lentikat® disks, were incubated with an excess of redox mediator (potassium hexacyanoferrate(III)) and organic substrate for 1 h at 37 °C without oxygen. The addition of substrate increased the catabolic activity of the microorganisms and the accumulation of reduced mediator, which was subsequently re-oxidised at a working electrode generating a current quantifiable by a coulometric transducer. The recorded currents were converted to their BOD5 equivalent with the only assumption being a fixed conversion of substrate and known stoichiometry. Measurements are reported both for the BOD5 calibration standard solution (150 mg l-1 glucose, 150 mg l-1 glutamic acid) and for filtered effluent sampled from a wastewater treatment plant. The inclusion of a highly soluble mediator in place of oxygen facilitated a high ferricyanide concentration in the incubation, which in turn permitted increased concentrations of microorganisms to be used. This substantially reduced the incubation time, from 5 days to 1 h, for the biological oxidation of substrates equivalent to those observed using the standard BOD5 test. Stoichiometric conversion efficiencies for the oxidation of the standard substrate by P. vulgaris were typically 60% for free cells and 35–50% for immobilised cells. [Copyright &y& Elsevier]

Four hundred and sixty-four Pseudomonas aeruginosa strains were isolated in northern Lebanon at the Islami Hospital Microbiology department, in Tripoli. The purpose of this study was to evaluate the susceptibility of these strains to antibiotics, to compare this susceptibility according to the nature of the sample and the year of sampling. The results show that urinary samples were the most frequent (39.3%), followed by wound samples (21.2%), and ear samples (16.5%). The average rate of susceptible strains was 39.8% to ticarcillin, 56.9% to piperacillin, 58.2% to piperacillin + tazobactam, 74.1% to imipenem, 63.3% to ceftazidime, 60.4% to cefepime, 62.1% to aztreonam, 60.3% to netilmicin, 57.5% to gentamicin, 62.2% to tobramycin, 69% to amikacin, 100% to colistin, 45.4% to pefloxacin and ofloxacin, 57.7% to ciprofloxacin and 1.3% to rifampicin. The study showed that the strains isolated from pulmonary secretions were the most resistant to antibiotics. [Copyright &y& Elsevier]

An elastin-like polypeptide (ELP) composed of a polyhistidine tail (ELPH12) was exploited as a tunable, metal-binding biopolymer with high affinity toward cadmium. By taking advantage of the property of ELPH12 to undergo a reversible thermal precipitation, easy recovery of the sequestered cadmium from contaminated water was demonstrated as the result of a simple temperature change. In this study, batch soil washing experiments were performed to evaluate the feasibility of using ELPH12 as an environmentally benign strategy for removing cadmium from contaminated soil. The stability constant (log KL) for the cadmium-ELPH12 complex was determined to be 6.8, a value similar to that reported for the biosurfactant rhamnolipid. Two washings with 1.25 mg/mL of ELPH12 were able to remove more than 55% of the bound cadmium as compared to only 8% removal with ELP containing no histidine tail or 21% removal using the same concentration of EDTA. Unlike rhamnolipid from Pseudomonas aeruginosa ATCC 9027, which adsorbs extensively to soil, less than 10% of ELPH12 was adsorbed under all soil washing conditions. As a result a significantly lower concentration of ELPH12 (0.036 mM as compared to 5-10 mM of biosurfactants) was required to achieve similar extraction efficiencies. However, cadmium recovery by simple precipitation was incomplete due to the displacement of bound cadmium by zinc ions present in soil. Owing to its benign nature, ease of production, and selective tailoring of the metal binding domain toward any target metals of interest ELP biopolymers may find utility as an effective extractant for heavy metal removal from contaminated soil or ore processing. [ABSTRACT FROM AUTHOR]

g.a. o'may, d.g. allison and p. gilbert. 2004. To develop a method for studying the microbial spoilage of water-in-oil emulsions and to use this to investigate (i) the intrinsic stability of water-in-oil formulations and (ii) Pseudomonas aeruginosa SP1-induced spoilage of a proprietary emulsion. Aliquots of test emulsion were placed into wells of a microtitre plate and the opacity (492 nm) monitored at 120-min intervals over several hours. Cracking of the emulsion was associated with marked reductions in opacity. Rate and extent of change in O.D. could be used as indicators of spoilage. Spoilage of a laboratory emulsion formulation was investigated where microorganisms with demonstrated spoilage potential were incorporated either into the water phase prior to emulsification or where the proportion of contaminated water droplets was varied by dilution of contaminated emulsion with a sterile formulation. Results suggested that the route of introduction was a critical determinant of the probability of gross spoilage. Ps. aeruginosa SP1-induced spoilage of a proprietary formulation was found to be independent of growth in the formulation; rather it was attributed to the presence of a heat-labile extracellular spoilage-factor that was protease labile and possessed both lipase and polysorbate hydrolytic activity. Such spoilage potential was unique to one Ps. aeruginosa culture filtrate amongst five pseudomonads tested. The method is both rapid and reproducible, enables evaluation of the effects of route of contamination upon emulsion spoilage and has potential application in formulation development for cosmetic, pharmaceutical and food products. [ABSTRACT FROM AUTHOR]

Faropenem is a new oral ß-lactam antibiotic unique from carbapenems and other available ß-lactams. Determinants of the in vitro activity of ß-lactam antibiotics include affinity to penicillin-binding proteins (PBPs) and ß-lactamase stability. In this study, the binding affinity of faropenem to various PBPs and its impact on the morphology of Staphylococcus aureus and Escherichia coli were evaluated. In general, faropenem demonstrated high binding affinity to high-molecular-weight PBPs but low affinity to low-molecular-weight PBPs. In S. aureus and Streptococcus pneumoniae, faropenem exhibited high binding affinity to PBP1, followed by PBP3 and PBP2. In E. coli, faropenem showed the highest affinity for PBP2, followed by PBP1A, PBP1B, PBP3 and PBP4. In Proteus vulgaris, binding was highest to PBP4, followed by PBP1A, PBP2 and PBP3. In Serratia marcescens, faropenem bound preferentially to PBP2 and PBP4. Exposure of S. aureus to faropenem at minimum inhibitory concentrations (MICs) of 1/8 or 1/4 resulted in irregular septum formation. At 1 x MIC or higher, a larger number of lysed cells were observed. Exposure of E. coli to 1/8 x MIC or 1/4 x MIC also induced changes in cellular shape; the normal rod-shaped form changed to a spherical form in a time-dependent manner. After exposure of E. coli to 1 x MIC for 2 h, bulging-shaped E. coli cells were observed and after 4 h of exposure cell lysis was demonstrated. In the presence of 4 x MIC, spheroplast-like forms and cell lysis were observed. The morphological changes triggered by faropenem are in agreement with the PBP binding affinities reported. Thus, the high binding affinities of faropenem to PBPs from gram-negative and gram-positive bacteria are mirrored by its pronounced and concentration-dependent bactericidal effect. [ABSTRACT FROM AUTHOR]

Bacteria in nature frequently grow as biofilms, yet little is known regarding how biofilm bacteria morphologically adapt to low nutrient availability, which is common in unsaturated environments such as the terrestrial subsurface or on plant leaves. For unsaturated biofilms, in which the substratum may provide all nutrients, what are the relationships between nutrition and cell size and shape—the simplest metrics of cellular morphology? To address this question, we cultured Pseudomonas aeruginosa, a ubiquitous gram-negative bacterium that is environmentally and medically important, on membranes overlaying solid media, and then measured cellular dimensions using atomic force microscopy (AFM). Nutrition was controlled chemically by media composition and physically by stacking membranes to increase the path length for nutrient diffusion. Under conditions of carbon–nitrogen imbalance, low carbon bioavailability, or increased nutrient diffusional path length, cells elongated while maintaining constant width. A mathematical relationship suggests that, by elongating, biofilm bacteria strategically enlarge their nutrient collection surface without substantially changing the ratio of surface area to volume (SA/V). We conclude that P. aeruginosa growing as unsaturated biofilm with a planar nutrient source morphologically adapt to starvation by elongating. This adaptation, if generalizable, differs from a better-understood starvation response (i.e., cell size decreases; thus SA/V increases) for planktonic bacteria in well-mixed environments. [ABSTRACT FROM AUTHOR]