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Effect of macrophage secretory products on elaboration of virulence factors by planktonic and biofilm cells of Pseudomonas aeruginosa.

Macrophages, which constitute the first line of defense, pour their secretions in the mileu following stimulation with pathogens. These secretory products, referred to as macrophage secretory products (MSPs), can influence ultimate outcome of an infection. In the present investigation, it was observed that different strains of Pseudomonas aeruginosa vary in their ability to stimulate macrophages leading to variability in generation of macrophage secretory products. Cytokine levels, reactive nitrogen intermediates and protein content of macrophage secretory products generated with biofilm cells of P. aeruginosa was found to be more as compared to their planktonic counterparts. The effect of macrophage secretory products produced in response to interaction of macrophages with P. aeruginosa on elaboration of virulence factors produced by planktonic and biofilm cell forms of this pathogen was assessed. Significant enhancement in growth and elaboration of all the virulence determinants by both the cell forms was observed when P. aeruginosa was grown in presence of supernatants with macrophage secretory products. Implications of these findings in relation to urinary tract infections induced by P. aeruginosa have been discussed.

Alginates↗

In vitro biofilm characterization and activity of antifungal agents alone and in combination against sessile and planktonic clinical Candida albicans isolates.

Thirty clinical isolates of Candida albicans were collected from blood or other sterile site infections. Biofilm dry weight and metabolic activity were measured for each isolate. Planktonic and sessile antifungal susceptibilities of each isolate were determined for amphotericin B deoxycholate, caspofungin, and voriconazole. Sessile susceptibilities were determined for the combination of caspofungin/voriconazole. No significant differences in biofilm dry weight or metabolic activity were found between bloodstream and other invasive isolates. Planktonic MIC90 values and sessile MIC90 (SMIC90) values were 0.25 and 2, 0.06 and >256, and 0.5 and 2 microg/mL for amphotericin, voriconazole, and caspofungin, respectively. The SMIC90 of the combination of caspofungin/voriconazole against sessile isolates was 0.5/2 microg/mL. Therefore, the source of invasive C. albicans clinical isolates did not affect in vitro biofilm formation. Susceptibility to antifungal agents decreased when C. albicans was associated with biofilm, and the combination of caspofungin/voriconazole did not appear to provide enhanced activity compared with caspofungin alone.

Antifungal Agents↗

Activity of sodium metabisulfite against planktonic and biofilm Staphylococcus species.

Biofilm-forming staphylococci cause a majority of intravascular catheter-related infections. We evaluated the effect of sodium metabisulfite, a preservative commonly added to intravenously administered pharmaceuticals as an antioxidant and previously used as a catheter lock solution, on planktonic and biofilm staphylococci at clinically encountered concentrations. Sodium metabisulfite exhibited bactericidal activity against planktonic Staphylococcus aureus, Staphylococcus lugdunensis, and Staphylococcus epidermidis at concentrations of 512, 512, and 1024 microg/mL, respectively. A concentration of 720 microg/mL inhibited cell growth by all 3 species in a biofilm formation assay. However, established S. aureus and S. lugdunensis biofilms showed less than 1.5 log10 decreases in viable cell counts when treated with 720 microg/mL of sodium metabisulfite for 24 h. These in vitro results suggest that the use of sodium metabisulfite as a catheter lock may inhibit staphylococcal colonization of catheters, thereby preventing catheter-related infection.

Biofilms↗

A simple model of plankton population dynamics coupled with a LES of the surface mixed layer.

The concentration of phytoplankton in the sea is affected by biological processes, such as growth/mortality rates, predatory zooplankton concentrations and nutrient levels. Phytoplankton concentrations are also influenced by physical processes, in particular the mixing properties of the local fluid environment. On planktonic scales (approximately 10-1000 microm) one can assume the local turbulent flow is isotropic, with no distinction between horizontal and vertical mixing. However, agglomerations of phytoplankton into patches are observed on larger scales of up to hundreds of metres, whose formation will be influenced by the anisotropic advection/mixing properties and large-eddy structures prevalent in the surface mixed layer. This paper presents the results of the coupling of a large-eddy simulation (LES) model of the mixed layer with an advection-diffusion system of coupled equations for nitrate-phytoplankton-zooplankton (NPZ) concentration, incorporating sub-grid parameterizations of the biological processes. Typically these include phytoplankton growth due to light levels and ambient nitrate concentration, offset by grazing losses due to the presence of zooplankton. The primary goal of this work is to investigate how the characteristics of the mixed layer turbulence influence the observed distribution of phytoplankton. One novel feature is the incorporation of a 'vortex-force' term in the LES code in order to generate Langmuir circulations. It has been speculated that the enhanced mixing rates associated with 'Langmuir turbulence' play a significant role in regulating planktonic activity. Results derived from the coupled LES-NPZ model, run with and without the presence of Langmuir circulations, are presented in order to investigate these ideas.

Computer Simulation↗

Identification of pectenotoxins in plankton, filter feeders, and isolated cells of a Dinophysis acuminata with an atypical toxin profile, from Chile.

A bloom of Dinophysis acuminata produced, in autumn of 2005, a closure of the scallop harvesting in Bahía Inglesa, in the Chilean III region. Isolated cells of this Dinophysis species were shown to contain 180 pg cell(-1) of pectenotoxin 2 but neither okadaic acid nor any of its analogs or derivatives (at least at a detectable level). Examination of plankton and filter-feeder samples covering an area of ca. 350 km, from the location where the toxicity was recorded to Bahía Tongoy, showed that the unique toxin profile found in the first bloom was widespread over that part of Chile and persisted for months. The analysis were carried out by HPLC-ESI-MS using positive ionization mode, with a detection limit below 2 ng of OA mL(-1) of methanolic extract. This is the first report of the presence of pectenotoxins in the plankton of the Pacific coast of America and in the studied filter feeders. This is also the first report of a Dinophysis species containing pectenotoxins and not any toxin of the okadaic acid group.

Animals↗

Large scale population structure and gene flow in the planktonic developing periwinkle, Littorina striata, in Macaronesia (Mollusca: Gastropoda).

Allozymes were used to investigate the genetic structure of 42 populations of the planktonic developing, Macaronesian periwinkle Littorina striata, throughout its entire geographic range (Azores, Madeira, Canary Islands and Cape Verde Islands). This periwinkle is presumed to have a high dispersal and gene flow potential, because it has a planktonic development. It is therefore expected to show little population genetic differentiation. Indeed, based on Wright's hierarchical F-statistics, no significant genetic differentiation could be detected among populations, at any of the specified hierarchical levels (i.e. population, island, and archipelago). Nevertheless, the Cape Verde Islands seemed genetically more diverse (highest mean number of alleles per locus). The number of loci revealing a significant genetic heterogeneity increased with increasing distance between populations, while private alleles based gene flow (Nm) estimates also revealed a tendency towards a geographical pattern. The distribution of rare and private alleles, might account for these observations which suggested some geographical effect. Because of the low frequency at which these alleles occur, their influence on the genetic population structure is negligible, and not picked up by F-statistics.

Journal Article↗

A comparison of plastic and plankton in the north Pacific central gyre.

The potential for ingestion of plastic particles by open ocean filter feeders was assessed by measuring the relative abundance and mass of neustonic plastic and zooplankton in surface waters under the central atmospheric high-pressure cells of the North Pacific Ocean. Neuston samples were collected at 11 random sites, using a manta trawl lined with 333 u mesh. The abundance and mass of neustonic plastic was the largest recorded anywhere in the Pacific Ocean at 334271 pieces km2 and 5114 g km2, respectively. Plankton abundance was approximately five times higher than that of plastic, but the mass of plastic was approximately six times that of plankton. The most frequently sampled types of identifiable plastic were thin films, polypropylene/monofilament line and unidentified plastic, most of which were miscellaneous fragments. Cumulatively, these three types accounted for 99% of the total number of plastic pieces.

Animals↗

A delay differential equations model of plankton allelopathy.

In this paper we have studied the dynamics of planktonic growth with special consideration on time dependent fluctuations in density of the species. We propose a modified delay differential equation model of the growth of two species of plankton having competitive and allelopathic effects on each other. The model system shows a stable limit cycle oscillation when the allelopathic effect is of a stimulatory nature.

Animals↗

Fatty acid profiles and their chemotaxonomy in planktonic species of Anabaena (Cyanobacteria) with straight trichomes.

Twenty-four axenic strains of planktonic Anabaena with straight trichomes, assigned to 7 species, were investigated by analyzing the pattern and content of their fatty acid composition and comparing their fatty acid composition with their morphological properties. In general, the fatty acids in planktonic Anabaena contained 14:0, 16:0, 16:1(cis-), 18:0, 18:1, 18:2, and 18:3(alpha) as their major components, and were classified as Type 2 according to the Kenyon-Murata system. These strains were further divided into 2 subtypes: 18 strains belonging to Type 2A, which contains 16:2 and 16:3, and 6 strains to Type 2B, which lacks 16:2 and 16:3. Fatty acid compositions of strains of A. solitaria, A. smithii, and A. kisseleviana closely corresponded to morphological properties; however, 10 strains of A. planctonica were divided into 4 clusters, and 3 strains of A. affinis into 2 clusters. These clusters should be taxonomically evaluated based on other aspects such as genetic characteristics.

Cyanobacteria↗

Are larvae of demersal fishes plankton or nekton?

A pelagic larval stage is found in nearly all demersal marine teleost fishes, and it is during this pelagic stage that the geographic scale of dispersal is determined. Marine biologists have long made a simplifying assumption that behaviour of larvae--with the possible exception of vertical distribution--has negligible influence on larval dispersal. Because advection by currents can take place over huge scales during a pelagic larval stage that typically lasts for several days to several weeks, this simplifying assumption leads to the conclusion that populations of marine demersal fishes operate over, and are connected over, similar huge scales. This conclusion has major implications for our perception of how marine fish populations operate and for our management of them. Recent (and some older) behavioural research-reviewed here-reveals that for a substantial portion of the pelagic larval stage of perciform fishes, the simplifying assumption is invalid. Near settlement, and for a considerable portion of the pelagic stage prior to that, larvae of many fish species are capable of swimming at speeds faster than mean ambient currents over long periods, travelling tens of kilometres. Only the smallest larvae of perciform fishes swim in an energetically costly viscous hydrodynamic environment (i.e., low Reynolds number). Vertical distribution is under strong behavioural control from the time of hatching, if not before, and can have a decisive, if indirect, influence on dispersal trajectories. Larvae of some species avoid currents by occupying the epibenthic boundary layer. Larvae are able to swim directionally in the pelagic environment, with some species apparently orientating relative to the sun and others to settlement sites. These abilities develop relatively early, and ontogenetic changes in orientation are seemingly common. Larvae of some species can use sound to navigate, and others can use odour to find settlement habitat, at least over small scales. Other senses may also be important to orientation. Larvae are highly aware of their environment and of potential predators, and some school during the pelagic larval stage. Larvae are selective about where they settle at both meso and micro scales, and settlement is strongly influenced by interactions with resident fishes. Most of these behaviours are flexible; for example, swimming speeds and depth may vary among locations, and speed may vary with swimming direction. In direct tests, these behaviours result in dispersal different from that predicted by currents alone. Work with both tropical and temperate species shows that these behaviours begin to be significant relatively early in larval development, but much more needs to be learned about the ontogeny of behaviour and sensory abilities in larvae of marine fishes. As a preliminary rule of thumb, behaviour must be taken into account in considerations of dispersal after the preflexion stage, and vertical distribution behaviour can influence dispersal from hatching. Larvae of perciform fishes are close to being planktonic at the start of the pelagic period and are clearly nektonic at its end, and for a substantial period prior to that. All these things differ among species. Larvae of clupeiform, gadiform and pleuronectiform fishes may be less capable behaviourally than perciform fishes, but this remains to be confirmed. Clearly, these behaviours, along with hydrography, must be included in modelling dispersal and retention and may provide explanations for recent demonstrations of self-recruitment in marine fish populations. Current work is directed at understanding the ontogeny of the gradual transition from planktonic to nektonic behaviour. Although it is clear that larvae of perciform fishes have the ability to strongly influence their dispersal trajectories, it is less clear whether or how these abilities are applied.

Animals↗

Phylogeography of the planktonic chaetognath Sagitta setosa reveals isolation in European seas.

Numerous planktonic species have disjunct distribution patterns in the world's oceans. However, it is unclear whether these are truly unconnected by gene flow, or whether they are composed of morphologically cryptic species. The marine planktonic chaetognath Sagitta setosa Müller has a discontinuous geographic distribution over the continental shelf in the northeastern Atlantic, Mediterranean Sea, and Black Sea. Morphological variation between these populations has been described, but overlaps and is therefore unsuitable to determine the degree of isolation between populations. To test whether disjunct populations are also genetically disjunct, we sequenced a 504-bp fragment of mitochondrial DNA comprising the cytochrome oxidase II region of 86 individuals. Sequences were highly variable; each represented a different haplotype. Within S. setosa, sequence divergence ranged from 0.2 to 8.1% and strong phylogeographic structure was found, with four main groups corresponding to the northeastern Atlantic, Mediterranean Sea (including Ligurian Sea, Tyrrhenian Sea and Gulf of Gabes), Adriatic Sea, and Black Sea. Two of these (Atlantic and Black Sea) were resolved as monophyletic clades, thus gene flow between disjunct populations of S. setosa has been extremely limited and lineage sorting has taken place. The deepest divergence was between Atlantic and Mediterranean/Black Sea populations followed by a split between Mediterranean and Black Sea populations. The Mediterranean/Black Sea clade comprised three groups, with the Adriatic Sea as the most likely sister clade of the Black Sea. These data are consistent with a colonization of the Black Sea from the Mediterranean. Furthermore, a possible cryptic species was found in the Black Sea with 23.1% sequence divergence from S. setosa. Two possibilities for the evolutionary origin of this species are proposed, namely, that it represents a relict species from the ancient Paratethys, or that it represents another chaetognath species that colonized the Black Sea more recently. Even though the exact timing of disjunction of S. setosa populations remains unclear, on the basis of the geological and paleoclimatic history of the European basins and our estimates of net nucleotide divergence, we suggest that disjunct populations arose through vicariance resulting from the cyclical changes in temperature and sea levels during the Pleistocene. We conclude that these populations have remained disjunct, not because of limited dispersal ability, but because of the inability to maintain viable populations in suboptimal, geographically intermediate areas.

Animals↗

Identification of peptide metabolites of Microcystis (Cyanobacteria) that inhibit trypsin-like activity in planktonic herbivorous Daphnia (Cladocera).

Cyanobacteria are recognized as producers of a broad variety of bioactive metabolites. Among these, the peptides synthesized by the non-ribosomal peptide synthetase pathway occur in high structural variability. One class of cyanobacterial peptides, the cyanopeptolins or micropeptins, have been shown to be strong inhibitors of vertebrate serine proteases, like trypsin. In the present study we screened extracts of ten strains of the unicellular cyanobacterium Microcystis sp. for their potential to inhibit trypsin-like activity in the planktonic crustacea Daphnia, the main herbivores in freshwater ecosystem. Respective standardized IC(50)'s varied for nearly two orders of magnitude. In HPLC fractions we could identify mainly cyanopeptolins as active compounds by MALDI-TOF mass spectrometry. Cyanopeptolins were found in 22 structural variants with 13 variants produced by one strain alone. Peptides of the microviridin class were moderately active while no activity was evident for microginins and microcystins. Among the cyanopeptolins only those were active that had an arginine or lysine residue N-terminal to the modified amino acid 3-amino-6-hydroxy-piperidone. Structural variants that had a tyrosine residue at this particular position did not inhibit trypsin-like activity. The highly variable composition of the side chain of cyanopeptolins had no marked effect on the activity. Among the six cyanobacterial strains we tested intensively two did not produce any cyanopeptolins and were accordingly less active as crude extracts. The present study underlines the potential importance of the biochemistry of cyanobacteria for the feeding ecology of a planktonic herbivore.

Animals↗

Biochemical prey recognition by planktonic protozoa.

Planktonic flagellates and ciliates are the major consumers of phytoplankton and bacterioplankton in aquatic environments, playing a pivotal role in carbon cycling and nutrient regeneration. Despite certain unicellular predators using chemosensory responses to locate and select their prey, the biochemical mechanisms behind prey reception and selection have not been elucidated. Here we identify a Ca(2+)-dependent, mannose-binding lectin on the marine dinoflagellate Oxyrrhis marina, which is used as a feeding receptor for recognizing prey. Blocking the receptor using 20 microM mannose-BSA inhibited ingestion of phytoplankton prey, Isochrysis galbana, by 60%. In prey selection studies, O. marina ingested twice as many 6 mum diameter beads coated with mannose-BSA as those coated with galNac-BSA. When pre-incubated with mannose-BSA, O. marina was no longer able to discriminate between different sugar-coated beads. Thus, these findings reveal molecular mechanisms of protozoan prey recognition. Our results also indicate the functional similarity between cellular recognition used by planktonic protozoa to discriminate between different prey items, and those used by metazoan phagocytic blood cells to recognize invading microorganisms.

Animals↗

Interstitial ciliates: benthic microaerophiles or planktonic anaerobes?

We observed marine benthic interstitial ciliates Geleia sp. and Tracheloraphis sp. inhabiting the water column of a chemically stratified salt pond. This habitat is uncharacteristic for interstitial ciliates, yet they displayed active and abundant planktonic populations (up to 800 and 250 cells/liter, respectively) and a well-defined pattern of vertical distribution. Completely absent from the oxygenated epilimnion, they first appeared at the oxic/anoxic interface and were present throughout the anoxic hypolimnion. The data could not be explained by a passive removal (e.g. by currents) of these ciliates from their conventional habitat (soft sediments) to water column. The results suggest that 1) these ciliates favored an anoxic environment, and 2) they switched to a planktonic lifestyle as appropriate conditions (seasonal anoxia) developed in the water column. This sharply contrasts the classic view of these ciliates as specifically benthic and aerobic (albeit microaerophilic) organisms. We hypothesize that Geleia sp. and Tracheloraphis sp. can readily grow in either water column or benthos, but are typically found in sediments simply because they contain their preferred (anoxic) niche.

Anaerobiosis↗

A proteomic study of Escherichia coli O157:H7 NCTC 12900 cultivated in biofilm or in planktonic growth mode.

Escherichia coli 0157:H7 biofilms were studied by a new method of cultivation in order to identify some of the proteins involved in the biofilm phenotype. A proteomic analysis of sessile or planktonic bacteria of the same age was carried out by two-dimensional electrophoresis, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) and database searching. Comparison of two-dimensional gels showed clear differences between protein patterns of sessile and planktonic cells. Fourteen proteins increased in biofilms, whereas three decreased. From these 17 proteins, 10 were identified by MALDI-TOF-MS and could be classified into four categories according to their function: (1) general metabolism proteins (malate dehydrogenase, thiamine-phosphate pyrophosphorylase), (2) sugar and amino acid transporters (D-ribose-binding periplasmic protein, D-galactose-binding protein, YBEJ), (3) regulator proteins (DNA starvation protein and H-NS) and (4) three proteins with unknown function. The results of this study showed that E. coli O157:H7 modified the expression of several proteins involved in biofilm growth mode.

Amino Acid Sequence↗

In vitro effects of antimicrobial agents on planktonic and biofilm forms of Staphylococcus lugdunensis clinical isolates.

Staphylococcus lugdunensis is an atypically virulent coagulase-negative staphylococcal species associated with acute and destructive infections that often resemble Staphylococcus aureus infections. Several types of infection caused by S. lugdunensis (e.g., native valve endocarditis, prosthetic joint infection, and intravascular catheter infection) are associated with biofilm formation, which may lead to an inability to eradicate the infection due to the intrinsic nature of biofilms to resist high levels of antibiotics. In this study, planktonic MICs and MBCs and biofilm bactericidal concentrations of 10 antistaphylococcal antimicrobial agents were measured for 15 S. lugdunensis isolates collected from patients with endocarditis, medical device infections, or skin and soft tissue infections. Planktonic isolates were susceptible to all agents studied, but biofilms were resistant to high concentrations of most of the drugs. However, moxifloxacin was able to kill 73% of isolates growing in biofilms at </=0.5 mug/ml. Relative to the effect on cell density, subinhibitory concentrations of nafcillin substantially stimulated biofilm formation of most isolates, whereas tetracycline and linezolid significantly decreased biofilm formation in 93 and 80% of isolates, respectively. An unexpected outcome of MBC testing was the observation that vancomycin was not bactericidal against 93% of S. lugdunensis isolates, suggesting widespread vancomycin tolerance in this species. These data provide insights into the response of S. lugdunensis isolates when challenged with various levels of antimicrobial agents in clinical use.

Acetamides↗

Nitrate accumulation in aerobic hypolimnia: relative importance of benthic and planktonic nitrifiers in an oligotrophic lake.

Both nitrate and nitrous oxide accumulate in the hypolimnion of the oligotrophic Lake Taupo, New Zealand, throughout stratification. The two forms of oxidized nitrogen increase in concentration with increasing depth toward the sediments, where the dissolved concentrations of reduced nitrogen are two orders of magnitude higher than concentrations in the overlying water. Nitrification rates were measured by dark [C]CO(2) assays with and without the inhibitor nitrapyrin. The fastest rates were recorded for planktonic nitrifiers in the epilimnion and benthic species in the surficial 2.5 mm of the sediments. Nitrifying bacteria were least active in the deep hypolimnion. Deepwater accumulation of NO(3) in Lake Taupo must therefore be a product of benthic rather than planktonic nitrification.

Journal Article↗