Cytoklepty in the plankton: A host strategy to optimize the bioenergetic machinery of endosymbiotic algae.

Endosymbioses have shaped the evolutionary trajectory of life and remain ecologically important. Investigating oceanic photosymbioses can illuminate how algal endosymbionts are energetically exploited by their heterotrophic hosts and inform on putative initial steps of plastid acquisition in eukaryotes. By combining three-dimensional subcellular imaging with photophysiology, carbon flux imaging, and transcriptomics, we show that cell division of endosymbionts (Phaeocystis) is blocked within hosts (Acantharia) and that their cellular architecture and bioenergetic machinery are radically altered. Transcriptional evidence indicates that a nutrient-independent mechanism prevents symbiont cell division and decouples nuclear and plastid division. As endosymbiont plastids proliferate, the volume of the photosynthetic machinery volume increases 100-fold in correlation with the expansion of a reticular mitochondrial network in close proximity to plastids. Photosynthetic efficiency tends to increase with cell size, and photon propagation modeling indicates that the networked mitochondrial architecture enhances light capture. This is accompanied by 150-fold higher carbon uptake and up-regulation of genes involved in photosynthesis and carbon fixation, which, in conjunction with a ca.15-fold size increase of pyrenoids demonstrates enhanced primary production in symbiosis. Mass spectrometry imaging revealed major carbon allocation to plastids and transfer to the host cell. As in most photosymbioses, microalgae are contained within a host phagosome (symbiosome), but here, the phagosome invaginates into enlarged microalgal cells, perhaps to optimize metabolic exchange. This observation adds evidence that the algal metamorphosis is irreversible. Hosts, therefore, trigger and benefit from major bioenergetic remodeling of symbiotic microalgae with potential consequences for the oceanic carbon cycle. Unlike other photosymbioses, this interaction represents a so-called cytoklepty, which is a putative initial step toward plastid acquisition.

Computational flow cytometry of planktonic populations for the evaluation of microbiological-control programs in district cooling plants.

Biofouling poses a serious concern for the district cooling (DC) industry. Current industry practises for monitoring biofouling continue to rely on culture-based methods for microbial enumeration, which are ultimately flawed. Computational flow cytometric (cFCM) analyses, which offer enhanced reproducibility and streamlined analytics versus conventional flow cytometry were applied to samples taken from 3 sites in each of 3 plants over a 5-week sampling program. We asked whether the application of cFCM to monitoring planktonic community dynamics in DC plants could be able to provide sufficient information to enhance microbiological-control strategies at site and inform about plant performance impacts. The use of cFCM enabled the evaluation of biocide dosing, deep cleaning treatment efficiencies and routes of microbial ingress into the studied systems. Additionally, inherent risks arising from the reintroduction of microbiological communities into recently cleaned WCT basins from contaminated cooling waters were identified. However, short-term dynamics did not relate with plant performance metrics. In summary, the insights offered by this approach can inform on plant status, enable evaluations of microbial loads during biofouling mitigation programs and, ultimately, enhance industry management of the biofouling process.

Bacterioplankton reveal years-long retention of Atlantic deep-ocean water by the Tropic Seamount.

Seamounts, often rising hundreds of metres above surrounding seafloor, obstruct the flow of deep-ocean water. While the retention of deep-water by seamounts is predicted from ocean circulation models, its empirical validation has been hampered by large scale and slow rate of the interaction. To overcome these limitations we use the growth of planktonic bacteria to assess the retention time of deep-ocean water by a seamount. The selected Tropic Seamount in the North-Eastern Atlantic is representative for the majority of isolated seamounts, which do not affect the surface ocean waters. We prove deep-water is retained by the seamount by measuring 2.4× higher bacterial concentrations in the seamount-associated or 'sheath'-water than in deep-ocean water unaffected by seamounts. Genomic analyses of flow-sorted, dominant sheath-water bacteria confirm their planktonic origin, whilst proteomic analyses of the sheath-water bacteria, isotopically labelled in situ, indicate their slow growth. According to our radiotracer experiments, it takes the sheath-water bacterioplankton 1.5 years to double their concentration. Therefore, the seamount should retain the deep-ocean water for 1.8 years for the deep-ocean bacterioplankton to grow to the 2.4× higher concentration in the sheath-water. We propose that turbulent mixing of the seamount sheath-water stimulates bacterioplankton growth by increasing cell encounter rate with ambient dissolved organic molecules.

Identification to species level of live single microalgal cells from plankton samples with matrix-free laser/desorption ionization mass spectrometry.

INTRODUCTION: Marine planktonic communities are complex microbial consortia often dominated by microscopic algae. The taxonomic identification of individual phytoplankton cells usually relies on their morphology and demands expert knowledge. Recently, a live single-cell mass spectrometry (LSC-MS) pipeline was developed to generate metabolic profiles of microalgae. OBJECTIVE: Taxonomic identification of diverse microalgal single cells from collection strains and plankton samples based on the metabolic fingerprints analyzed with matrix-free laser desorption/ionization high-resolution mass spectrometry. METHODS: Matrix-free atmospheric pressure laser-desorption ionization mass spectrometry was performed to acquire single-cell mass spectra from collection strains and prior identified environmental isolates. The computational identification of microalgal species was performed by spectral pattern matching (SPM). Three similarity scores and a bootstrap-derived confidence score were evaluated in terms of their classification performance. The effects of high and low-mass resolutions on the classification success were evaluated. RESULTS: Several hundred single-cell mass spectra from nine genera and nine species of marine microalgae were obtained. SPM enabled the identification of single cells at the genus and species level with high accuracies. The receiver operating characteristic (ROC) curves indicated a good performance of the similarity measures but were outperformed by the bootstrap-derived confidence scores. CONCLUSION: This is the first study to solve taxonomic identification of microalgae based on the metabolic fingerprints of the individual cell using an SPM approach.

Comparative proteomic and genomic analyses of Brucella abortus biofilm and planktonic cells.

The present study aimed to explore the differences in protein and gene expression of Brucella abortus cultured under biofilm and planktonic conditions. The proteins unique to biofilms and planktonic B. abortus were separated by two­dimensional (2­D) electrophoresis and then identified by matrix­assisted laser desorption/ionization­tandem time of flight­mass spectrometry (MALDI­TOF/TOF­MS). High­throughput sequencing and bioinformatic analyses were performed to identify differentially expressed genes between B. abortus cultured under biofilm and planktonic conditions. The proteins and genes identified by proteomic and genomic analyses were further evaluated via western blot and reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analyses. 2­D electrophoresis identified 20 differentially expressed protein spots between biofilms and planktonic cells, which corresponded to 18 individual proteins (12 downregulated and 6 upregulated) after MALDI­TOF/TOF­MS analysis, including elongation factor Tu and enolase. RT­qPCR analysis revealed that all of the 18 genes were downregulated in biofilms compared with planktonic cells. Western blot analysis identified 9 downregulated and 3 upregulated proteins. High­throughput sequencing and bioinformatic analyses identified 14 function and pathway­associated genes (e.g., BAbS19_I14970). RT­qPCR analysis of the 14 genes showed that they were upregulated in biofilm compared with in planktonic state. In conclusion, these differentially expressed genes may play important roles in bacterial defense, colonization, invasion, and virulence.

A novel application of Gini coefficient for the quantitative measurement of bacterial aggregation.

Non-surface attached bacterial aggregates are frequently found in clinical settings associated with chronic infections. Current methods quantifying the extent to which a suspended bacterial population is aggregated mainly rely on: (1) cell size distribution curves that are difficult to be compared numerically among large-scale samples; (2) the average size/proportion of aggregates in a population that do not specify the aggregation patterns. Here we introduce a novel application of Gini coefficient, herein named Aggregation Coefficient (AC), to quantify the aggregation levels of cystic fibrosis Pseudomonas aeruginosa (CF-PA) isolates in vitro using 3D micrographs, Fiji and MATLAB. Different aggregation patterns of five strains were compared statistically using the numerical AC indexes, which correlated well with the size distribution curves plotted by different biovolumes of aggregates. To test the sensitivity of AC, aggregates of the same strains were treated with nitric oxide (NO), a dispersal agent that reduces the biomass of surface attached biofilms. Strains unresponsive to NO were reflected by comparable AC indexes, while those undergoing dispersal showed a significant reduction in AC index, mirroring the changes in average aggregate sizes and proportions. Therefore, AC provides simpler and more descriptive numerical outputs for measuring different aggregation patterns compared to current approaches.

Metabolomic and proteomic changes induced by growth inhibitory concentrations of copper in the biofilm-forming marine bacterium Pseudoalteromonas lipolytica.

Copper is an essential element for living cells but this metal is present in some marine environments at such high concentrations that it can be toxic for numerous organisms. In polluted areas, marine organisms may develop specific adaptive responses to prevent cell damage. To investigate the influence of copper on the metabolism of a single organism, a dual approach combining metabolomics and proteomics was undertaken on the biofilm-forming bacterial strain Pseudoalteromonas lipolytica TC8. In order to highlight differential adaptation according to the phenotype, the response of P. lipolytica TC8 to copper stress was studied in planktonic and biofilm culture modes under growth inhibitory copper concentrations. As expected, copper exposure led to the induction of defense and detoxification mechanisms. Specific metabolite and protein profiles were thus observed in each condition (planktonic vs. biofilm and control vs. copper-treated cultures). Copper exposure seems to induce drastic changes in the lipid composition of the bacterial cell membrane and to modulate the abundance of proteins functionally known to be involved in copper cell homeostasis in both planktonic and biofilm culture modes. Much more proteins differentially expressed after copper treatment were observed in biofilms than in planktonic cells, which could indicate a more heterogeneous response of biofilm cells to this metallic stress.

Global distribution of Trebouxiophyceae diversity explored by high-throughput sequencing and phylogenetic approaches.

Trebouxiophyceae are a ubiquitous class of Chlorophyta encountered in aquatic and terrestrial environments. Most taxa are photosynthetic, and many acts as photobionts in symbiotic relationships, while others are free-living. Trebouxiophyceae have also been widely investigated for their use for biotechnological applications. In this work, we aimed at obtaining a comprehensive image of their diversity by compiling the information of 435 freshwater, soil and marine environmental DNA samples surveyed with Illumina sequencing technology in order to search for the most relevant environments for bioprospecting. Freshwater and soil were most diverse and shared more than half of all operational taxonomic units (OTUs), however, their communities were significantly distinct. Oceans hosted the highest genetic novelty, and did not share any OTUs with the other environments; also, marine samples host more diversity in warm waters. Symbiotic genera usually found in lichens such as Trebouxia, Myrmecia and Symbiochloris were also abundantly detected in the ocean, suggesting either free-living lifestyles or unknown symbiotic relationships with marine planktonic organisms. Altogether, our study opens the way to new prospection for trebouxiophycean strains, especially in understudied environments like the ocean.

Collective sinking promotes selective cell pairing in planktonic pennate diatoms.

Finding a partner in an inherently unsteady 3-dimensional system, such as the planktonic marine environment, is a difficult task for nonswimming organisms with poor control over their orientation. We experimentally investigate the process of cell pairing in pennate marine diatoms and present field evidence of its occurrence in the ocean. We describe the mechanism as a 3-step process in which pennate diatoms (i) vertically reorient while sinking from surface turbulent waters to a more stable environment (i.e., under the seasonal pycnocline), (ii) segregate from incompatible partners (e.g., dead or different sized cells), and (iii) pair with other partners as a result of the hydrodynamic instabilities generated by collective cell sinking. This is, eminently, a cell abundance-dependent process, therefore being more effective when population sinking is synchronized. We suggest that this selective process, enabling matching of size-compatible healthy partners, could be fundamental in understanding sexual reproduction in pennate diatoms.

Malachite green-conjugated multi-walled carbon nanotubes potentiate antimicrobial photodynamic inactivation of planktonic cells and biofilms of Pseudomonas aeruginosa and Staphylococcus aureus.

Purpose: Infections associated with medical devices that are caused by biofilms remain a considerable challenge for health care systems owing to their multidrug resistance patterns. Biofilms of Pseudomonas aeruginosa and Staphylococcus aureus can result in life-threatening situations which are tough to eliminate by traditional methods. Antimicrobial photodynamic inactivation (aPDT) constitutes an alternative method of killing deadly pathogens and their biofilms using reactive oxygen species (ROS). This study investigated the efficacy of enhanced in vitro aPDT of P. aeruginosa and S. aureus using malachite green conjugated to carboxyl-functionalized multi-walled carbon nanotubes (MGCNT). Both the planktonic cells and biofilms of test bacteria were demonstrated to be susceptible to the MGCNT conjugate. These MGCNT conjugates may thus be employed as a facile strategy for designing antibacterial and anti-biofilm coatings to prevent the infections associated with medical devices. Methods: Conjugation of the cationic dye malachite green to carbon nanotube was studied by UV-visible spectroscopy, high-resolution transmission electron microscopy, and Fourier transform infrared spectrometry. P. aeruginosa and S. aureus photodestruction were studied using MGCNT conjugate irradiated for 3 mins with a red laser of wavelength 660 nm and radiant exposure of 58.49 J cm-2. Results: Upon MGCNT treatment, S. aureus and P. aeruginosa were reduced by 5.16 and 5.55 log10 , respectively. Compared to free dye, treatment with MGCNT afforded improved phototoxicity against test bacteria, concomitant with greater ROS production. The results revealed improved biofilm inhibition, exopolysaccharide inhibition, and reduced cell viability in test bacteria treated with MGCNT conjugate. P. aeruginosa and S. aureus biofilms were considerably reduced to 60.20±2.48% and 67.59±3.53%, respectively. Enhanced relative MGCNT phototoxicity in test bacteria was confirmed using confocal laser scanning microscopy. Conclusion: The findings indicated that MGCNT conjugate could be useful to eliminate the biofilms formed on medical devices by S. aureus and P. aeruginosa.

Comparative in situ ROS mediated killing of bacteria with bulk analogue, Eucalyptus leaf extract (ELE)-capped and bare surface copper oxide nanoparticles.

This study demonstrates a simple one-pot green method for biosynthesis of terpenoids encapsulated copper oxide nanoparticles (CuONPs) using aqueous leaf extract of Eucalyptus globulus (ELE), as reducing, dispersing, and stabilizing agent. Indeed, the greater attachment and internalization of ELE-CuONPs in Gram-positive and -negative biofilm producing clinical bacterial isolates validated the hypothesis that terpenoids encapsulated CuONPs are more stable and effective antibacterial and antibiofilm agent vis-à-vis commercially available nano and micro sized analogues. Gas chromatography-mass spectroscopy (GC-MS) analysis of pristine ELE identified 17 types of terpenoids based on their mass-to-charge (m/z) ratios. Amongst them four bioactive terpenoids viz. terpineols, 2,6-octadienal-3,7-dimethyl, benzamidophenyl-4-benzoate and ß-eudesmol were found associated with the CuONPs as ELE-cap, and most likely involved in the nucleation and stabilization of ELE-CuONPs. Further, the Fourier transformed infrared (FTIR) analysis of ELE-CuONPs also implicated other functional biomolecules like proteins, sugars, alkenes, etc. with ELE terpenoids corona. Flow cytometric (FCM) data exhibited significantly enhanced intracellular uptake propensity of terpenoids encapsulated ELE-CuONPs and accumulation of intracellular reactive oxygen species (ROS), which ensued killing of planktonic cells of extended spectrum ß-lactamases (ESßL) producing Escherichia coli-336 (E. coli-336), Pseudomonas aeruginosa-621 (P. aeruginosa-621) and methicillin-resistant Staphylococcus aureus-1 (MRSA-1) clinical isolates compared to the bare surface commercial nano-CuO and bulk sized CuO. The study for the first-time demonstrated the (i) differential bio-nano interface activities due to ELE surface and varied cell wall composition of test bacterial isolates, (ii) antibacterial effect and biofilm inhibition due to disruption of proteins involved in adhesion and biofilm formation triggered by CuONPs induced intracellular oxidative stress, and (iii) indigenous terpenoids-capped bio-inspired CuONPs are more stable and effective antibacterial and antibiofilm agent as compared with commercially available nano-CuO and bulk-CuO.

Polyvinyl chloride biodegradation by Pseudomonas citronellolis and Bacillus flexus.

The accumulation of high amounts of petroleum-derived plastics in the environment has raised ecological and health concerns. The aim of this work was to study the biodegradative abilities of five bacterial strains, namely Pseudomonas chlororaphis, Pseudomonas citronellolis, Bacillus subtilis, Bacillus flexus and Chelatococcus daeguensis, towards polyethylene, polypropylene, polystyrene and polyvinyl chloride films under aerobic conditions. Preliminary screening resulted in the selection of P. citronellolis and B. flexus as potential PVC film degraders. Both strains were able to form a biofilm on the plastic film surface and to cause some modifications to the FTIR spectra of biomass-free PVC films. The two strains were then used to set up a PVC film biodegradation assay in 2-liter flasks. After 45 days incubation, fragmentation of the film was observed, suggesting that PVC biodegradative activity took place. Gel permeation chromatography analysis showed a reduction in average molecular weight of 10% for PVC incubated with P. citronellolis, with PVC polymer chains apparently attacked. Based on these results, the P. citronellolis strain was selected for biodegradation assays of two waste PVC films, used either nonsterile or subjected to ethanol sterilization. Chemical analyses on the incubated films confirmed the biodegradation of waste PVC plastics as shown by a gravimetric weight loss of up to about 19% after 30 days incubation. In summary, this work reports the biodegradation of PVC films by P. citronellolis and B. flexus. Both strains were shown to act mainly against PVC additives, exhibiting a low biodegradation rate of PVC polymer.

Tolerances of Deinococcus geothermalis Biofilms and Planktonic Cells Exposed to Space and Simulated Martian Conditions in Low Earth Orbit for Almost Two Years.

Fossilized biofilms represent one of the oldest known confirmations of life on the Earth. The success of microbes in biofilms results from properties that are inherent in the biofilm, including enhanced interaction, protection, and biodiversity. Given the diversity of microbes that live in biofilms in harsh environments on the Earth, it is logical to hypothesize that, if microbes inhabit other bodies in the Universe, there are also biofilms on those bodies. The Biofilm Organisms Surfing Space experiment was conducted as part of the EXPOSE-R2 mission on the International Space Station. The experiment was an international collaboration designed to perform a comparative study regarding the survival of biofilms versus planktonic cells of various microorganisms, exposed to space and Mars-like conditions. The objective was to determine whether there are lifestyle-dependent differences to cope with the unique mixture of stress factors, including desiccation, temperature oscillations, vacuum, or a Mars-like gas atmosphere and pressure in combination with extraterrestrial or Mars-like ultraviolet (UV) radiation residing during the long-term space mission. In this study, the outcome of the flight and mission ground reference analysis of Deinococcus geothermalis is presented. Cultural tests demonstrated that D. geothermalis remained viable in the desiccated state, being able to survive space and Mars-like conditions and tolerating high extraterrestrial UV radiation for more than 2 years. Culturability decreased, but was better preserved, in the biofilm consortium than in planktonic cells. These results are correlated to differences in genomic integrity after exposure, as visualized by random amplified polymorphic DNA-polymerase chain reaction. Interestingly, cultivation-independent viability markers such as membrane integrity, ATP content, and intracellular esterase activity remained nearly unaffected, indicating that subpopulations of the cells had survived in a viable but nonculturable state. These findings support the hypothesis of long-term survival of microorganisms under the harsh environmental conditions in space and on Mars to a higher degree if exposed as biofilm.

Virulence Factors Produced by Staphylococcus aureus Biofilms Have a Moonlighting Function Contributing to Biofilm Integrity.

Staphylococcus aureus is the causative agent of various biofilm-associated infections in humans causing major healthcare problems worldwide. This type of infection is inherently difficult to treat because of a reduced metabolic activity of biofilm-embedded cells and the protective nature of a surrounding extracellular matrix (ECM). However, little is known about S. aureus biofilm physiology and the proteinaceous composition of the ECM. Thus, we cultivated S. aureus biofilms in a flow system and comprehensively profiled intracellular and extracellular (ECM and flow-through (FT)) biofilm proteomes, as well as the extracellular metabolome compared with planktonic cultures. Our analyses revealed the expression of many pathogenicity factors within S. aureus biofilms as indicated by a high abundance of capsule biosynthesis proteins along with various secreted virulence factors, including hemolysins, leukotoxins, and lipases as a part of the ECM. The activity of ECM virulence factors was confirmed in a hemolysis assay and a Galleria mellonella pathogenicity model. In addition, we uncovered a so far unacknowledged moonlighting function of secreted virulence factors and ribosomal proteins trapped in the ECM: namely their contribution to biofilm integrity. Mechanistically, it was revealed that this stabilizing effect is mediated by the strong positive charge of alkaline virulence factors and ribosomal proteins in an acidic ECM environment, which is caused by the release of fermentation products like formate, lactate, and acetate because of oxygen limitation in biofilms. The strong positive charge of these proteins most likely mediates electrostatic interactions with anionic cell surface components, eDNA, and anionic metabolites. In consequence, this leads to strong cell aggregation and biofilm stabilization. Collectively, our study identified a new molecular mechanism during S. aureus biofilm formation and thus significantly widens the understanding of biofilm-associated S. aureus infections - an essential prerequisite for the development of novel antimicrobial therapies.

Exploring multiple effects of Zn0.15Mg0.85O nanoparticles on Bacillus subtilis and macrophages.

The increasing number of multidrug resistant bacteria raises a serious public-health concern, which is exacerbated by the lack of new antibiotics. Metal oxide nanoparticles are already applied as an antibacterial additive in various products used in everyday life but their modes of action have remained unclear. Moreover, their potential negative effects to human health are still under evaluation. We explored effects of mixed metal oxide Zn0.15Mg0.85O on Bacillus subtilis, as a model bacterial organism, and on murine macrophages. Zn0.15Mg0.85O killed planktonic bacterial cells and prevented biofilm formation by causing membrane damages, oxidative stress and metal ions release. When exposed to a sub-inhibitory amount of Zn0.15Mg0.85O, B. subtilis up-regulates proteins involved in metal ions export, oxidative stress response and maintain of redox homeostasis. Moreover, expression profiles of proteins associated with information processing, metabolism, cell envelope and cell division were prominently changed. Multimode of action of Zn0.15Mg0.85O suggests that no single strategy may provide bacterial resistance. Macrophages tolerated Zn0.15Mg0.85O to some extend by both the primary phagocytosis of nanoparticles and the secondary phagocytosis of damaged cells. Bacterial co-treatment with ciprofloxacin and non-toxic amount of Zn0.15Mg0.85O increased antibiotic activity towards B. subtilis and E. coli.

In vitro activity of antifungal combinations against planktonic and sessile cells of Candida albicans isolated from medical devices in an intensive care department.

BACKGROUND: Invasive fungal infections are an emerging health problem worldwide. They are responsible for a significant rate of morbidity and mortality. Infections caused by Candida albicans involve proliferation of biofilms on biotic or abiotic surface. These adherent communities exhibit characteristics distinct from planktonic cells such as the ability to tolerate high concentrations of antifungal. OBJECTIVE: The object of our study was focused on the determination of the susceptibility to amphotericin B, caspofungin, voriconazole and two antifungal combinations (amphotericin B/caspofungin and amphotericin B/voriconazole) of both planktonic and sessile cells of C. albicans, which were isolated from catheters. MATERIAL AND METHODS: The susceptibility of C. albicans to antifungals was determined using the broth microdilution method according to Clinical Laboratory Standards Institute CLSI (2008). A Checkerboard assay was employed to evaluate the efficacy of drugs combinations. Biofilm susceptibility was determined using a metabolic [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] (XTT) reduction assay. RESULTS: The minimal inhibitory concentrations of individual antifungal drugs determined against C. albicans biofilms (SMICs) were significantly higher (P<0.05) than planktonic ones (MICs). They went from 2 to 64µg/mL for amphotericin B, from 1 to 64µg/mL for caspofungin and from 2 to 128µg/mL for voriconazole. The combination of amphotericin B to caspofungin or to voriconazole decreased significantly the MIC values for planctonic (P<0.0001) and sessile cells (P=0.0016). Based on Fractional Inhibitory Concentration Index (FICI), no antagonistic interaction was observed. CONCLUSION: The obtained results showed that the combination of amphotericin B with either caspofungin or voriconazole can be used as a new strategy for management of systemic mycoses associated to medical devices.

A cell length-dependent transition in MinD-dynamics promotes a switch in division-site placement and preservation of proliferating elongated Vibrio parahaemolyticus swarmer cells.

Vibrio parahaemolyticus exists as swimmer and swarmer cells, specialized for growth in liquid and on solid environments respectively. Swarmer cells are characteristically highly elongated due to an inhibition of cell division, but still need to divide in order to proliferate and expand the colony. It is unknown how long swarmer cells divide without diminishing the population of long cells required for swarming behavior. Here we show that swarmer cells divide but the placement of the division site is cell length-dependent; short swarmers divide at mid-cell, while long swarmers switch to a specific non-mid-cell placement of the division site. Transition to non-mid-cell positioning of the Z-ring is promoted by a cell length-dependent switch in the localization-dynamics of the division regulator MinD from a pole-to-pole oscillation in short swarmers to a multi-node standing-wave oscillation in long swarmers. Regulation of FtsZ levels restricts the number of divisions to one and SlmA ensures sufficient free FtsZ to sustain Z-ring formation by preventing sequestration of FtsZ into division deficient clusters. By limiting the number of division-events to one per cell at a specific non-mid-cell position, V. parahaemolyticus promotes the preservation of long swarmer cells and permits swarmer cell division without the need for dedifferentiation.

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles.

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those of suspended/sinking particles. Here, we present an all plastic (Polypropylene and Polycarbonate), multi-layer filtration system for collection of suspended particulate matter (SPM) at sea. This ultra-clean sampling device has been designed and developed specifically for trace element studies. Meticulous selection of all non-metallic materials and utilization of an in-line flow-through procedure minimizes any possible metal contamination during sampling. This system has been successfully tested and tweaked for determining trace metals (e.g., Fe, Al, Mn, Cd, Cu, Ni) on particles of varying size in coastal and open ocean waters. Results from the South China Sea at the South East Asia Time-Series (SEATS) station indicate that diurnal variations and spatial distribution of plankton in the euphotic zone can be easily resolved and recognized. Chemical analysis of size-fractionated particles in surface waters of the Taiwan Strait suggests that the larger particles (>153 µm) were mostly biologically derived, while the smaller particles (10 - 63 µm) were mostly composed of inorganic matter. Apart from Cd, the concentrations of metals (Fe, Al, Mn, Cu, Ni) decreased with increasing size.

The role of probiotic Lactobacillus acidophilus ATCC 4356 bacteriocin on effect of HBsu on planktonic cells and biofilm formation of Bacillus subtilis.

Bacillus subtilis is a Gram positive, aerobic and motile bacterium. Biofilm formation is an important feature of this bacterium which confers resistance to antimicrobial agents. The use of new antimicrobial reagents which eliminate biofilms are important and necessary. In this study, the effect of secondary metabolites (bacteriocin) from Lactobacillus acidophilus ATCC 4356 on Bacillus subtilis BM19 in the presence and absence of HBsu which is involved in the growth of planktonic cells and biofilm formation, is reported. HBsu nucleoprotein plays several roles in different processes of Bacillus subtilis cells such as replication, transcription, cell division, recombination and repair. In this study, for the first time, the effect of HBsu on biofilm formation is presented. RESULTS: In the absence of HBsu, purified bacteriocin from L. acidophilus ATCC 4356 was more effective in inhibiting growth of B. subtilis BM19 planktonic cells as well as biofilm formation. The presence of HBsu on the other hand led to increased biofilm formation.

In vitro antimicrobial effects and mechanisms of direct current air-liquid discharge plasma on planktonic Staphylococcus aureus and Escherichia coli in liquids.

The direct inactivation effects of an atmospheric pressure direct current (DC) air plasma against planktonic Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in aqueous solution are investigated in vitro. Upon plasma treatment, extensively analyses on cell culturability, metabolic capacity, membrane integrity, surface morphology, cellular proteins, nucleic acids and intracellular reactive oxygen species (ROS) for both bacterial species were carried out and significant antimicrobial effects observed. Compared with the cellular culturability, a sub-lethal viable but non-culturable (VBNC) state was induced while more S. aureus entered this state than E. coli. Damaged bacterial outer structures were observed and the total concentrations of cellular protein and nucleic acid decreased for both bacteria after plasma treatment. The plasma-induced aqueous reactive species (RS) and intracellular ROS might produce detrimental effects to the bacteria, while S. aureus was less susceptible to the discharge after a 20-min exposure compared to E. coli.