Bacteria growing in biofilms often develop multicellular, three-dimensional structures known as microcolonies. Complex differentiation within biofilms of Pseudomonas aeruginosa occurs, leading to the creation of voids inside microcolonies and to the dispersal of cells from within these voids. However, key developmental processes regulating these events are poorly understood. A normal component of multicellular development is cell death. Here we report that a repeatable pattern of cell death and lysis occurs in biofilms of P. aeruginosa during the normal course of development. Cell death occurred with temporal and spatial organization within biofilms, inside microcolonies, when the biofilms were allowed to develop in continuous-culture flow cells. A subpopulation of viable cells was always observed in these regions. During the onset of biofilm killing and during biofilm development thereafter, a bacteriophage capable of superinfecting and lysing the P. aeruginosa parent strain was detected in the fluid effluent from the biofilm. The bacteriophage implicated in biofilm killing was closely related to the filamentous phage Pf1 and existed as a prophage within the genome of P. aeruginosa. We propose that prophage-mediated cell death is an important mechanism of differentiation inside microcolonies that facilitates dispersal of a subpopulation of surviving cells.

Our understanding of subsurface processes suffers from a profound observation bias: seismometers are sparse and clustered on continents. A new seismic recording approach, distributed acoustic sensing (DAS), transforms telecommunication fiber‐optic cables into sensor arrays enabling meter‐scale recording over tens of kilometers of linear fiber length. We analyze cataloged earthquake observations from three DAS arrays with different horizontal geometries to demonstrate some possibilities using this technology. In Fairbanks, Alaska, we find that stacking ground motion records along 20 m of fiber yield a waveform that shows a high degree of correlation in amplitude and phase with a colocated inertial seismometer record at 0.8–1.6 Hz. Using an L‐shaped DAS array in Northern California, we record the nearly vertically incident arrival of an earthquake from The Geysers Geothermal Field and estimate its backazimuth and slowness via beamforming for different phases of the seismic wavefield. Lastly, we install a fiber in existing telecommunications conduits below Stanford University and show that little cable‐to‐soil coupling is required for teleseismic P and S phase arrival detection.

A dark-green-pigmented marine bacterium, previously designated D2, which produces components that are inhibitory to common marine fouling organisms has been characterized and assessed for taxonomic assignment. Based on direct double-stranded sequencing of the 16s rRNA gene, D2T was found t o show the highest similarity (93%) to members of the genus Pseudoalteromonas. The G+C content of DZT is 42 molo/o, and it is a facultatively anaerobic rod and oxidase-positive. D Z T is motile by a sheathed polar flagellum, exhibited non-fermentative metabolism and required sodium ions for growth. The strain was not capable of using citrate, fructose, sucrose, sorbitol and glycerol but it utilizes mannose and maltose and hydrolyses gelatin. The molecular evidence, together with phenotypic characteristics, showed that this bacterium which produces an antifouling agent constitutes a new species of the genus Pseudoalteromonas. The name Pseudoalteromonas tunicata is proposed for this bacterium, and the type strain is DZT (= CCUG 2 6 7 5 7T).

Gymnodinium catenatum is an introduced toxic dinoflagellate that blooms intermittently and causes shellfish farm closure in the Huon Estuary, Tasmania, Australia. Seventy-five bacteria isolated from the estuary were tested for algicidal activity against this and other toxic and nontoxic algal species. Five isolates produced algicidal extracellular exudates. These algicidal species were a Pseudoalteromonas sp. (ACEM 4), a novel Zobellia sp. (ACEM 20), a strain of Cellulophaga lytica (ACEM 21) and 2 Firmicutes: a novel Planomicrobium sp. (ACEM 22) and a strain of Bacillus cereus (ACEM 32). This study is the first time Gram-positive bacteria have been associated with algicidal activities. Further data are presented on an algicidal Pseudoalteromonas species previously isolated from the Huon Estuary (Strain y). Supernatant produced by all 5 strains caused cell lysis and death in G. catenatum vegetative cells. No change or reversible ecdysis was noted for 2 other endemic dinoflagellate species. Algicidal or inhibitory activity was not activated via homoserine lactones, but bacterial quorum sensing for the isolates was shown by means of the AI-2 mechanism. Algicidal activity from field isolates was also influenced by strain or environmental variation. Bacteria were capable of losing or switching off their algicidal ability indicating that the presence of an algicidal species in the environment may not necessarily signify that they are currently algicidal. Concentrations of algicidal compounds required for algal lysis in laboratory experiments indicate that the 5 bacterial species can be effective against G. catenatum vegetative cells if they dominate the bacterial population in the estuary, particularly when attached to particles. KEY WORDS: Algicidal · Gymnodinium · Pseudoalteromonas · Cellulophaga · Flavobacteria · Harmful algal blooms Resale or republication not permitted without written consent of the publisher

The newly described green-pigmented bacterium Pseudoalteromonas tunicata (D2) produces target-specific inhibitory compounds against bacteria, algae, fungi, and invertebrate larvae and is frequently found in association with living surfaces in the marine environment. As part of our studies on the ecology of P. tunicata and its interaction with marine surfaces, we examined the ability of P. tunicata to form biofilms under continuous culture conditions within the laboratory. P. tunicata biofilms exhibited a characteristic architecture consisting of differentiated microcolonies surrounded by water channels. Remarkably, we observed a repeatable pattern of cell death during biofilm development of P. tunicata, similar to that recently reported for biofilms of Pseudomonas aeruginosa (J. S. Webb et al., J. Bacteriol. 185:4585-4595, 2003). Killing and lysis occurred inside microcolonies, apparently resulting in the formation of voids within these structures. A subpopulation of viable cells was always observed within the regions of killing in the biofilm. Moreover, extensive killing in mature biofilms appeared to result in detachment of the biofilm from the substratum. A novel 190-kDa autotoxic protein produced by P. tunicata, designated AlpP, was found to be involved in this biofilm killing and detachment. A ⌬alpP mutant derivative of P. tunicata was generated, and this mutant did not show cell death during biofilm development. We propose that AlpP-mediated cell death plays an important role in the multicellular biofilm development of P. tunicata and subsequent dispersal of surviving cells within the marine environment.

Pseudoalteromonas tunicata is a marine bacterium with the ability to prevent biofouling by the production of at least four target-specific compounds. In addition to these antifouling compounds, P. tunicata produces at least two pigments. These include a yellow and a purple pigment which, when combined, give the bacterium a dark green appearance. Transposon mutagenesis was used in this study to investigate the correlation between pigment production and the expression of specific antifouling phenotypes in P. tunicata. Four different categories of pigmentation mutants were isolated including yellow, dark-purple, light-purple and white mutants. The mutants were tested for their ability to inhibit the settlement of invertebrate larvae, algal spore germination, fungal growth and bacterial growth. The results showed that the yellow-pigmented mutants retained full antifouling activity, whereas the purple and white mutant strains had lost some, or all, of their ability to inhibit target organisms. This demonstrates that the loss of antifouling capabilities correlates with the loss of yellow pigment and not purple pigment. Sequencing and analysis of the genes disrupted by the transposons in these mutants identified a number of potential biosynthetic enzymes and transport systems involved in the synthesis and regulation of pigmentation and fouling inhibitors in this organism.

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A collection of 56 bacteria isolated from different surfaces in the marine environment were assayed for their effects on the germination of spores from the common green alga Ulva lactuca. Thirteen bacterial isolates were shown to inhibit spore germination. Of these bacteria, Pseudoalteromonas tunicata displayed the most pronounced effects against algal spores. Further characterisation of the anti-algal activity of P. tunicata was performed and it was found that this bacterium produces an extracellular component with specific activity toward algal spores that is heat-sensitive, polar and between 3 and 10 kDa in size. This biologically active compound was also found to prevent the germination of spores from the red alga Polysiphonia sp. and, given the widespread occurrence of P. tunicata in a range of marine habitats, this may suggest that it is effective against a variety of marine algae.