Effect of Some Unicellular Algae on Escherichia coli Populations in Sea Water and Oysters

Acrylic acid was detected in sea water in which oysters had been maintained and was shown to be derived from unicellular algae containing dimethyl-β-propiothetin (DMPT). Anticoliform activity was noted in samples containing acrylic acid. Intensification of the effect was achieved in laboratory studies and Phaeodactylum tricornutum (DMPT present) caused a rapid decline of Escherichia coli populations in sea water and in oysters which had been fed this alga. Pavlova lutheri (DMPT absent) was ineffective against E. coli .     

Accumulation of Xanthosine by Escherichia coli in the Presence of Angustmycin C

Effect of angustmycin C, an adenosine analogue having an unusual sugar linked to adenine, on Escherichia coli was studied. It inhibited biosynthesis of RNA and DNA preferentially and xanthosine was excreted from the inhibited cells into the medium. During the course of its growth with addition of the antibiotic, the specific activity of inosine-5′-phosphate dehydrogenase of E. coli cells increased six times. These effects could be explained by the inhibitory effect of angustmycin C on xanthosine-5′-phosphate aminase which resulted in decreased level of IMP-dehydrogenase. Accumulation of xanthosine induced by the antibiotic reached the concentration of 940 μg/ml under an appropriate condition.     

Resistance to Coliphage Infection Induced in Escherichia coli by Growth in the Presence of a Surfactant

Heavy slime exudates surrounding Escherichia coli cells grown in media containing 1% sodium dodecybenzene sulfonate apparently block adsorption of coliphages to susceptible strains.     

Presence of Bacteriophage-Like Inhibitory Particles in Escherichia coli

Bacteriophage-like particles were found in the supernatant fluids of Escherichia coli O111a and O111:B(4). Caution is urged in the study of deoxyribonucleic acid synthesis and replication in these strains.     

The SecA Subunit of Escherichia coli Preprotein Translocase Is Exposed to the Periplasm

SecA undergoes conformational changes during translocation, inserting domains into and across the membrane or enhancing the protease resistance of these domains. We now show that some SecA bound at SecYEG is accessible from the periplasm to a membrane-impermeant probe in cells with a permeabilized outer membrane but an intact plasma membrane.     

Antibiotic Resistance in Salmonella enterica Serovar Typhimurium Exposed to Microcin-Producing Escherichia coli

Microcin 24 is an antimicrobial peptide secreted by uropathogenic Escherichia coli. Secretion of microcin 24 provides an antibacterial defense mechanism for E. coli. In a plasmid-based system using transformed Salmonella enterica, we found that resistance to microcin 24 could be seen in concert with a multiple-antibiotic resistance phenotype. This multidrug-resistant phenotype appeared when Salmonella was exposed to an E. coli strain expressing microcin 24. Therefore, it appears that multidrug-resistant Salmonella can arise as a result of an insult from other pathogenic bacteria.     

Fluctuations in Escherichia coli O-serotypes in Pigs Throughout Life in the Presence and Absence of Antibiotic Treatment

A detailed study, throughout life, of the coliform and Escherichia coli gut flora of two pigs, is presented. One pig was given oral tetracycline for 3 d on two separate occasions; the other pig was not treated and housed separately. The fluctuations in numbers of coliforms, O-serotypes of E. coli , the occurrence and persistence of antibiotic resistance, and their interrelations, are analysed for both animals. Oral tetracycline profoundly affected the proportion of antibiotic sensitive and resistant E. coli , and, by selecting for R-factor bearing O-serotypes, limited the number of O-serotypes isolated from individual faceal specimens. Specific O-serotypes exhibited marked differences in their ability to persist in the gut of the control pig. The R-factor bearing O-serotypes, selected by tetracycline treatment, also showed differences in ability to persist after treatment indicating that properties other than the possession of R-factors, decide colonizing ability. The size of the colony sample for serotype analysis is discussed.     

Nucleic Acid and Protein Synthesis in Reconstituted Lyophilized Escherichia coli Exposed to Air

S ummary : Escherichia coli strain BT^-, freeze dried from distilled water, loses viability by exposure to air in the dry state, and decay curves of such bacteria were determined. The amounts of tritiated thymidine incorporated in the DNA of such 'dead', rehydrated bacteria are compatible with an assumption that the synthesis of DNA proceeds till the completion of its replication cycle, but not beyond. Induction of β-galactosidase served as an indicator of RNA and protein synthesis in the dead bacteria. Such induction occurred after a short lag after which it proceeded at a rate similar to that observed in lyophilized, untreated bacteria. The loss of viability of these bacteria cannot be accounted for by defects in the production of DNA or RNA and protein synthesis; it is most probably due to damage to the DNA initiation point.     

Filamentous Cells of Escherichia coli Formed in the Presence of Mitomycin

The effects of mitomycin C on cell elongation of Escherichia coli B were studied. Filament formation was most marked in cultures treated with a moderate level (1 mug/ml) of the antibiotic, becoming less obvious at higher levels (10 mug/ml). Cells treated with a bacteriostatic concentration (0.1 mug/ml or less) of mitomycin C were also significantly elongated. The filamentous or elongated cells appeared to lack septa, since their spheroplasts were considerably larger than those formed from normal cells. The appearance of empty spheres also indicated some defects in the surfaces of the filamentous cells. Electron micrographs of the filaments revealed a characteristic difference in the arrangement of the nuclei in the filaments formed in the presence of low (0.1 mug/ml) and high (5 mug/ml) concentrations of mitomycin C. The filaments formed by the low level of mitomycin C had normal well-defined nuclear bodies distributed along the long axis, whereas those formed by the elevated level of the antibiotic contained smaller nuclei. The latter were characteristically confined to the center of the cells and did not extend out to the tips of the filaments.     

Released products of pathogenic bacteria stimulate biofilm formation by Escherichia coli K-12 strains

It has recently been shown that pathogens with a limited capacity for sessile growth (like some Escherichia coli O157 strains) can benefit from the presence of other bacteria and form mixed biofilms with companion strains. This study addresses the question whether pathogens may influence attached growth of E. coli non-pathogenic strains via secreted factors. We compared the biofilm-modulating effects of sterile stationary-phase culture media of a biofilm non-producing strain of E. coli O157:H, a laboratory biofilm-producing E. coli K-12 strain and a biofilm-forming strain of the pathogen Yersina enterocolitica O:3. Sessile growth was monitored as biomass (crystal violet assay), exopolysaccharide (ELLA) and morphology (scanning electron and confocal laser microscopy). With two of the E. coli K-12 strains stimulation of biofilm formation by all supernatants was achieved, but only the pathogens' secreted products induced biomass increase in some 'biofilm-deficient' K-12 strains. Lectin-peroxidase labeling indicated changes in colanic acid and poly-N-acetylglucosamine amounts in extracellular matrices. The contribution of indole, protein and polysaccharide to the biofilm-modulating activities of the supernatants was compared. Indole, in concentrations equal to those established in the supernatants, suppressed sessile growth in one K-12 strain. Proteinase K significantly reduced the stimulatory effects of all supernatants, indicating a prominent role of protein/peptide factor(s) in biofilm promotion. The amount of released polysaccharides (rPS) in the supernatants was quantitated then comparable quantities of isolated rPS were applied during biofilm growth. The three rPS had notable strain-specific effects with regard to both the strain-source of the rPS and the E. coli K-12 target strain.     

Behavior of Enzyme Activities Exposed to Contamination by Heavy Metals and Dissolved Organic Carbon in Calcareous Agricultural Soils

To investigate the relevance of biochemical parameters in biogeochemical mechanisms of the soil, it is important to gather data related to different soil types under different pedogeoclimatic conditions. In this study, we investigated on the calcareous agricultural soils in the Saiss plain (North Morocco). Four agricultural soils exposed to multi-metal (Cr, Cu, Zn, and Ni) and organic matter (OM) contamination as a result of irrigation with Oued Fez and Oued Sebou waters that are affected by urban and industrial activities around the city of Fez were studied and compared to a reference site irrigated with uncontaminated water. The study concerned soil physicochemical properties and the activity of a range of enzymes [phosphatase (PHOS), arylsulfatase (SULF), urease (UREA), arylamidase (AMID), β-galactosidase (GALA), glucosidase (GLUC), and laccase (LACA)] related to nutrients cycles. Pearson's correlations between these parameters showed that soil enzymatic activities (PHOS, SULF, UREA, GALA, GLUC, and LACA) were correlated positively with heavy metals (Cu, Zn, and Cr) concentrations in the soil and also with dissolved organic carbon (DOC), and negatively with the aromaticity (AROM) of these compounds. Interestingly, analysis of intra-site correlations showed strong relationships among enzyme activities in the reference soil, while in contaminated soils, these activities were largely unrelated to each other. It was concluded that soil irrigation with heavy-metal- and OM-contaminated watercourses over decades has resulted in soils with high enzymatic activities function and nutrient turnover but altered relationships among geochemical cycles.     

Presence of Nuclear Bodies in Some Minicells of Escherichia coli

Electron microscopy of serial sections of Escherichia coli K-12 (P678-54) revealed that some "minicells" contain nuclear bodies.     

Enzymes Released from Escherichia coli With the Aid of a Servall Cell Fractionator

The release and stability of the enzymes S-adenosylhomocysteine nucleosidase, lysine decarboxylase, arginine decarboxylase, glutamic decarboxylase, formic hydrogenlyase, formic oxidase, and glucose oxidase from Escherichia coli during disruption of the organisms in a Servall-Ribi refrigerated cell fractionator were examined. With the possible exception of arginine decarboxylase, maximal activity was retained by all the enzymes reported here when the cell suspensions were processed at pressures necessary for rupture of all the organisms (15,000 to 25,000 psi). Considerable variation in the stability of different enzymes liberated by disruption at higher pressures (45,000 to 55,000 psi) was observed. It is reasonable to assume that mechanical forces rather than effects of temperature are responsible for inactivation of these enzymes.     

Growth of Escherichia coli in Model Distribution System Biofilms Exposed to Hypochlorous Acid or Monochloramine

Bacteria indigenous to water distribution systems were used to grow multispecies biofilms within continuous-flow slide chambers. Six flow chambers were also inoculated with an Escherichia coli isolate obtained from potable water. The effect of disinfectants on bacterial populations was determined after exposure of established biofilms to 1 ppm of hypochlorous acid (ClOH) for 67 min or 4 ppm of monochloramine (NH₂Cl) for 155 min. To test the ability of bacterial populations to initiate biofilm formation in the presence of disinfectants, we assessed the biofilms after 2 weeks of exposure to residual concentrations of 0.2 ppm of ClOH or 4 ppm of NH₂Cl. Lastly, to determine the effect of recommended residual concentrations on newly established biofilms, we treated systems with 0.2 ppm of ClOH after 5 days of growth in the absence of disinfectant. Whole-cell in situ hybridizations using fluorescently tagged, 16S rRNA-targeted oligonucleotide probes performed on cryosectioned biofilms permitted the direct observation of metabolically active bacterial populations, including certain phylogenetic groups and species. The results of these studies confirmed the resistance of established bacterial biofilms to treatment with recommended levels of disinfectants. Specifically, Legionella pneumophila, E. coli, and β and δ proteobacteria were identified within biofilms both before and after treatment. Furthermore, although it was undetected using routine monitoring techniques, the observation of rRNA-containing E. coli within biofilms demonstrated not only survival but also metabolic activity of this organism within the model distribution systems. The persistence of diverse bacterial species within disinfectant-treated biofilms suggests that current testing practices underestimate the risk to immunocompromised individuals of contracting waterborne disease.     

Identification and Quantification of Toxic Chemicals by Use of Escherichia coli Carrying lux Genes Fused to Stress Promoters

The luxCDABE bioluminescence genes of the Vibrio fischeri lux system have been used as a reporter system for different stress and regulatory promoters of Escherichia coli. Selected E. coli strains carrying lux genes fused to different promoters were exposed to various toxic chemicals, and the recorded luminescence was used for the characterization of the biologic signature of each compound. Analysis of these data with the aid of a proper algorithm allowed quantitative and qualitative assessment of toxic chemicals. Of the 25 tested chemicals, 23 were identified by this novel strategy in a 3-h procedure. This system can also be adapted for the identification of simple mixtures of toxic agents when the biologic signatures of the individual compounds are known. This biologic recognition strategy also provides a tool for evaluating the degree of similarity between the modes of action of different toxic agents.     

Presence of virulence and fitness gene modules of enterohemorrhagic Escherichia coli in atypical enteropathogenic Escherichia coli O26

Enterohemorrhagic Escherichia coli (EHEC) strains of serogroup O26 cause hemolytic-uremic syndrome (HUS) whereas atypical enteropathogenic E. coli (aEPEC) O26 typically cause uncomplicated diarrhea but have been also isolated from HUS patients. To gain insight into the virulence of aEPEC O26, we compared the presence of O island (OI) 122, which is associated with enhanced virulence in EHEC strains, among aEPEC O26 and EHEC O26 clinical isolates. We also tested these strains for the high pathogenicity island (HPI) which is a fitness island. All 20 aEPEC O26 and 20 EHEC O26 investigated contained virulence genes located within OI-122 (efa1/lifA, nleB, nleE, ent). In both aEPEC O26 and EHEC O26, OI-122 was linked to the locus for enterocyte effacement, forming a mosaic island which was integrated in pheU. Moreover, strains of these two pathotypes shared a conserved HPI. These data support a close relatedness between aEPEC O26 and EHEC O26 and have evolutionary implications. The presence of OI-122 in aEPEC O26 might contribute to their pathogenic potential.