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Factors affecting catalase expression in Pseudomonas aeruginosa biofilms and planktonic cells.

Previous work with Pseudomonas aeruginosa showed that catalase activity in biofilms was significantly reduced relative to that in planktonic cells. To better understand biofilm physiology, we examined possible explanations for the differential expression of catalase in cells cultured in these two different conditions. For maximal catalase activity, biofilm cells required significantly more iron (25 microM as FeCl(3)) in the medium, whereas planktonic cultures required no addition of iron. However, iron-stimulated catalase activity in biofilms was still only about one-third that in planktonic cells. Oxygen effects on catalase activity were also investigated. Nitrate-respiring planktonic cultures produced approximately twice as much catalase activity as aerobic cultures grown in the presence of nitrate; the nitrate stimulation effect could also be demonstrated in biofilms. Cultures fermenting arginine had reduced catalase levels; however, catalase repression was also observed in aerobic cultures grown in the presence of arginine. It was concluded that iron availability, but not oxygen availability, is a major factor affecting catalase expression in biofilms.

Aerobiosis↗

Comparative in vitro activity of vancomycin and levofloxacin in combination with rifampin against planktonic versus sessile cells of Staphylococcus epidermidis.

STUDY OBJECTIVE: To evaluate the activity of vancomycin and levofloxacin alone and combined with rifampin against planktonic and sessile cells. INTERVENTION: Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of the three drugs were determined against a clinical isolate of methicillin-resistant Staphylococcus epidermidis (MRSE 23) and a reference strain of MRSE (ATCC 35984). MEASUREMENTS AND MAIN RESULTS: The MICs/MBCs of vancomycin, levofloxacin, and rifampin against MRSE 23 were 0.78/0.78 microg/ml, 0.19/0.19 microg/ml or below, and 0.19/0.19 microg/ml or below, and against ATCC 35984 were 0.78/1.56 microg/ml, 0.19/0.19 microg/ml or below, and 0.19/0.19 microg/ml or below, respectively. A 99.9% killing activity was achieved with vancomycin, levofloxacin, and vancomycin-levofloxacin against planktonic cells of MRSE 23 (18.9, 21.3, and 17.5 hrs, respectively) and only with levofloxacin against ATCC 35984 (21.5 hrs). No regimen achieved 99.9% killing activity against sessile cells. CONCLUSION: Adding rifampin was antagonistic against planktonic cells and had an additive effect against sessile cells. Activity typically reported using nutrient-rich, planktonic cells may not be applicable to sessile cells under environmental and growth restrictions.

Drug Therapy, Combination↗

[Effect of the rate of phytoplankton growth on the spaced-time dynamics of plankton communities in an homogeneous environment].

We use a conceptual mathematical reaction-diffusion model to investigate the mechanisms of spatial structure formation and complex temporal dynamics of plankton in a heterogeneous environment. We take into account basic trophic interactions, namely, "prey-predator" interactions between phytoplankton, zooplankton, and fish in upper layers of natural waters. We consider plankton as a passive contaminant in turbulent waters. We show that plankton structure formation can result from the difference in phytoplankton growth rate in neighboring habitats. Phytoplankton and zooplankton biomass is shown to undergo both regular and chaotic oscillations. The fish predation rate substantially affects the spatial and temporal dynamics of plankton in a heterogeneous environment.

Animals↗

High plankton densities reduce mercury biomagnification.

This study tests the hypothesis that differences in the densities of plankton across lakes lead to significantvariation in Hg accumulation by the biota. The study comprised (i) a field program to measure and compare total Hg concentrations in phytoplankton (0.4-45 microm), two size classes of zooplankton (45-202 and >202 microm), and fish to phytoplankton and zooplankton densities from 20 lakes in the northeastern United States and (ii) a statistical analysis of fish Hg burdens and plankton densities from 38 additional EMAP lakes in the northeastern United States. We found that phytoplankton density was negatively correlated with Hg concentrations within both the phytoplankton and their primary consumers (small zooplankton). We also found a negative correlation between zooplankton density and Hg concentrations in zooplankton and in both herbivorous and predatory fish. Zooplankton density alone explained more than 40% of the variation in predatory fish Hg levels across lakes. We discuss several mechanisms to explain the negative relationship of plankton density to Hg concentrations in lower trophic levels and to the biomagnification of Hg to fish. Our results suggest that measures of plankton density, which are relatively easy and inexpensive to estimate, may be effective predictors of Hg biomagnification potential across lakes.

Animals↗

Spirolide composition of micro-extracted pooled cells isolated from natural plankton assemblages and from cultures of the dinoflagellate Alexandrium ostenfeldii.

A novel micro-extraction technique was applied to the extraction of biologically active macrocyclic imines known as spirolides from pooled individual cells isolated from spirolide-rich plankton material. For comparison, this method was also applied to pooled individual cells isolated from a unialgal culture of the marine dinoflagellate Alexandrium ostenfeldii (Paulsen) Balech & Tangen, a species known to produce spirolides. Both athecate cells and motile forms of gonyaulacoid dinoflagellates derived from size-fractionated plankton material from Nova Scotia, Canada were sorted and pooled by the glass micropipette isolation technique and by flow cytometry. The development of a highly sensitive analytical method for spirolides (detection limit 2 ng ml(-1) for spirolide B) using liquid chromatography-mass spectrometry (LC-MS) and application to micro-extracted samples allowed the accurate determination of spirolide composition in as few as 50 cells. Total spirolide concentrations (fmol cell(-1)) calculated from pooled micropipette isolated cells were very consistent with those based upon bulk- or micro-extractions of A. ostenfeldii cells from unialgal batch cultures in exponential growth phase. The results of the pooled cell selection from field material from two sites in Nova Scotia confirmed the association of spirolides with vegetative cells of A. ostenfeldii and related athecate forms. Combining these techniques represents a highly sensitive method for the analysis of marine toxins within complex plankton matrices, even when the toxigenic species is in low abundance, by enrichment of the target organism.

Animals↗

Diatom/copepod interactions in plankton: the indirect chemical defense of unicellular algae.

Numerous coexisting species can be observed in the open oceans. This includes the complex community of the plankton, which comprises all free floating organisms in the sea. Traditionally, nutrient limitation, competition, predation, and abiotic factors have been assumed to shape the community structure in this environment. Only in recent years has the idea arisen that chemical signals and chemical defense can influence species interactions in the plankton as well. Key players at the base of the marine food web are diatoms (unicellular algae with silicified cell walls) and their main predators, the herbivorous copepods. It was assumed that diatoms represent a generally good food source for the grazers but recent work indicates that some species use chemical defenses. Secondary metabolites, released by these algae immediately after wounding, are targeted not against the predators themselves but rather at interfering with their reproductive success. This strategy allows diatoms to reduce the grazer population, thereby influencing the marine food web. This review addresses the chemical ecology of the defensive oxylipins formed by diatoms and the question of how these metabolites can act in such a dilute environment. Aspects of biosynthesis, bioassays, and the possible implications of such a chemical defense for the plankton community structure are also discussed.

Aldehydes↗

Chaos and regular dynamics in model multi-habitat plankton-fish communities.

This work is focused on the role of diffusive interaction between separate habitats in a patchy environment in plankton pattern formation. We demonstrate that conceptual reaction-diffusion mathematical models constitute an appropriate tool for searching and understanding basic mechanisms of plankton pattern formation and complex spatio-temporal plankton dynamics

Animals↗

Mercury levels in total suspended matter and in plankton of the Mediterranean basin.

Mercury levels in total suspended matter (TSM) and in plankton of sea water from some Mediterranean areas have been determined. The mercury concentration in TSM collected with a filter of 0.45 micron pore size ranges from 0.1 to 2.4 ng l-1 and from 0.1 to 4.7 ng mg-1, dry wt. These levels are 3-10 times higher than those observed in plankton (0.08-0.29 ng mg-1 dry wt.) collected with a 200 microns net mesh size. The larger amounts of mercury in TSM than in the matter collected by the plankton nets are probably due to the high abundance of smaller sized particles selected by the filter.

Animals↗

Fate and effects of 3,4-dichloroaniline (DCA) in marine plankton communities in experimental enclosures.

The fate and effects of single doses of 3,4-dichloroaniline (DCA) on North Sea coastal plankton communities enclosed by large plastic bags (contents 1.5 m3) were studied in two experiments lasting 5 and 6 weeks, respectively. The biodegradation of DCA was also studied in laboratory experiments, which were carried out simultaneously, using water from the enclosed model ecosystems. DCA was not degraded in the laboratory tests and probably also not degraded in the enclosed plankton communities, although concentrations in the water decreased during the experiments. This decrease appeared to be partly caused by diffusion of DCA through the walls of the enclosures. After the addition of single doses of 2, 10, and 25 microgram liter-1 DCA no effects on the enclosed plankton community could be detected. The addition of 0.1 mg liter-1 DCA had a clear influence on the species composition and the biomass development of the phytoplankton, changed the relative species composition of the zooplankton, and resulted in lower numbers of bacteria. In addition to these effects 1 mg liter-1 DCA limited the phytoplankton growth and resulted in mortality and inhibition of growth of the copepods.

Aniline Compounds↗

Nonylphenol induced changes in trophic web structure of plankton analysed by multivariate statistical approaches.

Microcosm studies unveil relevant information in ecotoxicology. However, statistical evaluation of data gained from the different parts of the ecosystem often remain on unsatisfying, discretely level, e.g. separately for different species communities. A combination of different multivariate statistical methods enables an ecological interpretation of the found changes in zoo- and phytoplankton abundances in connection with the physico-chemical variables. To investigate the effects of nonylphenol (NP) on plankton communities, NP was continuously applicated in microcosms by controlled release. Plankton taxa were identified and quantified, various physico-chemical variables were measured. Maximum NP concentration ranged between 29 and 129 microg/L. The most important abiotic variable explaining both zoo- and phytoplankton abundances was NP concentration. NP primarily inhibited Crustaceae, especially juveniles. Cladocera and Copepoda abundances decreased. Rotatoria tend to increase, probably due to a decline of competition and predator pressure. Shifts in phytoplankton structure occurred with a time lag. Aggregating phytoplankton on class level did not show NP effects, whereas aggregating them according to feeding protection strategies did. Thus NP influences phytoplankton at least prevailingly indirectly via zooplankton. Zooplankton shifts led to species composition with different feeding preferences and strategies. Thereby the feeding pressure on phytoplankton changed. NP reorganized the plankton interrelation. Whereas without NP phytoplankton composition was dominated by Rotatoria, in the NP treated microcosms the Crustaceae unveiled highest explanatory power. The found effects were reversible in all but the highest treated microcosm. There are indications that the found effects were caused by endocrine activity of the chemical. Iteratively combining multivariate statistical methods with ecological and toxicological knowledge proved to provide a deeper insight into the mode of ecosystem disturbance by toxic substances.

Animals↗

Antimicrobial effects of sanitizers against planktonic and sessile Listeria monocytogenes cells according to the growth phase.

This study was designed to investigate the individual or combined effects of sanitizers on survival of planktonic or sessile Listeria monocytogenes cells at different phase of growth. The sanitizers tested included: (i) acetic acid (pH 5.0), (ii) NaOH (pH 12.0), (iii) 10% Na2SO4, (iv) 10% Na2SO4 and acetic acid (pH 5.0), (v) 10% Na2SO4 and NaOH (pH 12.0), (vi) a quaternary ammonium (20 ppm) and (vii) glyceryl monolaurate (75 ppm). Results revealed a great efficacy of alkaline treatments on both sessile and planktonic cells with a slightly higher resistance of 6 h biofilms. Quaternary ammonium appeared very effective in killing more than 98% of cells, but a resistance of 7 days biofilm was observed. Other sanitizers did not succeed in inhibiting totally the pathogen but acted in a similar way on both sessile and planktonic cells. Renewing the medium or not do not seem to be the major cause of a resistance emergence.

Acetic Acid↗

Phage release from biofilm and planktonic Staphylococcus aureus cells.

The ability of pathogenic staphylococci to form biofilms facilitates colonization and the development of chronic infections. Therapy is hampered by the high tolerance of biofilms towards antibiotic treatment and the immune system. We found evidence that lysogenic Staphylococcus aureus cells in a biofilm and in planktonic cultures spontaneously release phages into their surroundings. Phages were detected over a much longer period in biofilm cultures than in planktonic supernatants because the latter were degraded by secreted proteases. Phage release in planktonic and biofilm cultures was artificially increased by adding mitomycin C. Two morphologically distinct phages in the S. aureus strain used in this work were observed by electron microscopy. We postulate that phage-release is a frequent event in biofilms. The resulting lysis of cells in a biofilm might promote the persistence and survival of the remaining cells, as they gain a nutrient reservoir from their dead and lysed neighboring cells. This might therefore be an early differentiation and apoptotic mechanism.

Bacteriolysis↗

Sensitivity of bacterial biofilms and planktonic cells to a new antimicrobial agent, Oxsil 320N.

The effective concentrations of disinfectants were determined for planktonic bacteria using the norms EN 1040 and NF T 72-150. This concentration corresponds to biocide efficacy after 5 min of contact, followed by neutralization. However, micro-organisms often colonize a substratum and form microcolonies or biofilms where they are enclosed in exopolymer matrices. Biofilms are commonly resistant to a broad range of antimicrobial agents, and resistance mechanisms involve exopolymer matrices, changes in gene expression and metabolic alterations. Due to these different resistance mechanisms, it is difficult to select and titrate antimicrobial agents to be effective against biofilms. In this context, SODIFRA developed a new disinfectant, Oxsil 320N (French patent 94 15 193). Oxsil 320N is an association of three active principles: hydrogen peroxide, acetic acid/peracetic acid and silver. This biocide was tested on planktonic bacteria and on 24-h biofilms formed on AISI 304 stainless steel surfaces. The effective concentration of Oxsil 320N was also determined on biofilms using SODIFRA recommendations (without neutralization of the biocide). Data showed that the antimicrobial efficacy measured on planktonic bacteria is not a reliable indicator of performance when biofilm is present. When biofilms were exposed to Oxsil 320N, the concentration needed to achieve a 10(5)-fold decrease in concentration was 10 times higher than that for bacterial suspensions (0.313% Oxsil 320N). An effective concentration of Oxsil 320N of 3.13% was required.

Acetic Acid↗

Some aspects of water quality in a polluted lowland river in relation to the intracellular chemical levels in planktonic and epilithic diatoms.

Changes in elemental concentrations of diatoms and river water from the river Erh-Jen were determined using scanning electron microscopy energy-dispersive X-ray microanalysis and inductively coupled plasma mass spectrometry. Relatively large amounts of copper and lead found in both planktonic and epilithic diatoms implied these algae might play an important role in biogeochemical cycles and in the transfer of those elements to higher trophic levels in the aquatic environment. Changes in elemental concentrations within diatom cells were found to vary with other elements within cells and the same or different elements in water. Planktonic and epilithic cells showed different correlation patterns. For epilithic diatoms, negative correlations were found between concentrations of total phosphorus and phosphate in water and those of phosphorus within cells, and between concentrations of lead in water and in cells. Concentrations of chromium and mercury within planktonic cells and those of phosphorus, manganese and lead within epilithic ones were found to be easily influenced by other elements in river water, indicating appearance of the competitive manner on uptake of such elements by algal cells. Relatively high concentration factors (CFs) for cadmium, mercury and lead by diatoms in this study suggested they are good accumulators for these heavy metals. Significant negative corrections were found between the CFs of diatoms and the concentrations of elements in river water.

Diatoms↗

Occurrence of planktonic blooms under environmental fluctuations and its possible control mechanism--mathematical models and experimental observations.

Planktonic blooms and its control is an intriguing problem in ecology. To investigate the oscillatory successions of blooms, three simple phytoplankton-zooplankton systems are proposed. It is observed that if the uptake function is linear and the process of toxin liberation is instantaneous, the oscillatory nature of blooms is not observed. On the other hand, periodic planktonic blooms are observed when toxin liberation process follows discrete time variation. The bloom phenomena described by this mechanism can be controlled through toxin producing phytoplankton (TPP). Introducing environmental fluctuation in the system, a critical value of time delay in terms of correlation time of the fluctuation is worked out. We observed from our mathematical analysis, numerical simulation and field observation that TPP and control of the rapidity of environmental fluctuation are key factors for the termination of planktonic blooms.

Animals↗

Planktonic or biofilm growth affects survival, hydrophobicity and protein expression patterns of a pathogenic Campylobacter jejuni strain.

The effect of planktonic or biofilm modes of growth on survival, hydrophobicity and cellular protein expression patterns of a pathogenic Campylobacter jejuni strain were determined. This was achieved by growing the strain in brain heart infusion broth (with 1% yeast extract), or attached to glass beads in the same medium, at 37 degrees C for 48 h under microaerophilic conditions. Cells from the broth or the bead surfaces were stored at different temperatures (4, 10, 25 and 37 degrees C) for 28 days in phosphate buffered saline (PBS) and monitored at appropriate time intervals for culturable numbers and hydrophobicity by standard methods. In addition, cells were inoculated onto the surface of two processed meat products (a bologna and a summer sausage) vacuum packaged and stored at 4 degrees C for 28 days. Numbers of culturable cells were monitored at appropriate time intervals by standard methods. Cells from the broth or the bead surfaces were also examined for protein expression using two-dimensional protein electrophoresis. Results indicated that numbers of culturable cells in phosphate buffered saline decreased from approximately 6 log colony forming units (cfu) g(-1) to undetectable levels within 14-day storage in a temperature dependent manner. Hydrophobicity of broth grown cells decreased from 15% to 0% adherence to xylene over the same time in a temperature independent manner. Cells grown in a biofilm mode initially displayed a <0.3% adherence to xylene which was maintained during storage. Furthermore, cells grown in the biofilm mode decreased in number more rapidly on storage in buffer than their counterparts grown in broth. Numbers of culturable cells on meat decreased from approximately 5 log cfu g(-1) to undetectable levels within 14-day storage in a product dependent manner, with the most rapid decrease observed for the more acidic summer sausage. Cells grown in a biofilm mode decreased in number more rapidly on storage than broth grown cells. The protein expression patterns differed between planktonic and biofilm cells with seven unique and 12 up-regulated protein spots expressed in a growth mode specific manner. A number of the differentially expressed spots were tentatively identified, by comparison to existing literature, as surface- and stress-associated proteins. Despite the elicitation of some putative stress proteins, this study importantly indicates that biofilm cells of C. jejuni are less resistant to stress than their planktonic counterparts and may lack a sophisticated adaptive stress-resistance response. These findings have implication in determining the risks of infection associated with C. jejuni contamination on food.

Bacterial Adhesion↗

Coaggregation between freshwater bacteria within biofilm and planktonic communities.

The coaggregation ability of bacteria isolated from a freshwater biofilm was compared to those derived from the coexisting planktonic population. Twenty-nine morphologically distinct bacterial strains were isolated from a 6-month-old biofilm, established in a glass tank under high-shear conditions, and 15 distinct strains were isolated from the associated re-circulating water. All 44 strains were identified to genus or species level by 16S rDNA sequencing. The 29 biofilm strains belonged to 14 genera and 23.4% of all the possible pair-wise combinations coaggregated. The 15 planktonic strains belonged to seven genera and only 5.8% of all the possible pair-wise combinations coaggregated. Therefore, compared to the planktonic population, a greater proportion of the biofilm strains coaggregated. It is proposed that coaggregation influences biofilm formation and species diversity in freshwater under high shear.

Animals↗

Metabolism of Alcaligenes denitrificans in biofilm vs planktonic cells.

AIMS: To compare the effect of phosphorous concentration (200 mg P l(-1) and 20 mg P l(-1)) on the denitrifying efficiency of Alcaligenes denitrificans when in the form of planktonic cells or in the form of a biofilm, and to select the most adequate C/N ratio. METHODS AND RESULTS: Two types of assays were carried out: with planktonic cells and with cells in biofilm form. Anoxic bottles with the appropriate C/N and phosphorous concentration were incubated at 30 degrees C and submitted to orbital shaking at 150 rev min(-1). The specific activity of cells in biofilm form, in terms of substrate consumption, was significantly higher than cells in planktonic form. With regard to the effect of increasing phosphorous concentration, an increase in specific activity was also only evident when the cells were in biofilm form. CONCLUSIONS: The two forms showed different performances and phosphorous concentration only markedly affected the biofilm form. SIGNIFICANCE AND IMPACT OF THE STUDY: The importance of the C/N/P ratio in the denitrification process is demonstrated. As there was no report in the literature about the stoichiometric relationship of heterotrophic denitrification with citrate, its stoichiometry, including the requirement for cell synthesis, was determined.

Alcaligenes↗