Effective isolation of bacterioplankton genus Polynucleobacter from freshwater environments grown on photochemically degraded dissolved organic matter

Effective isolation of freshwater bacterioplankton belonging to genus Polynucleobacter from a shallow eutrophic lake and its tributary was achieved by size-selective filtration with a 0.7-μm pore filter and cultivation on R2A agar medium. Partial 16S rRNA gene analysis showed that over 80% of all the strains were highly similar to the Polynucleobacter cluster. Essential medium components for effective cultivation are pyruvate, yeast extract and peptone, whereas soluble starch and glucose are not necessary. Isolate KF001 (affiliated with Polynucleobacter subcluster D) has a strict requirement for organic acids as carbon sources, and we hypothesize that the Polynucleobacter cluster of bacteria could utilize compounds formed via photochemically dissolved organic matter (DOM) degradation for growth. Because organic acids form from solar radiation of DOM in aquatic environments, carbon sources that are typical products of DOM photochemical degradation were added to the medium. These compounds were readily utilized by KF001 in this study. Finally, we observed the stimulation of strain KF001 activity by photochemical degradation of natural lake water. Our findings suggest a carbon flow of DOM photoproducts to Polynucleobacter in the freshwater microbial loop.     

Assessment of the microbial quality of irrigation water in a prairie watershed

Aims:  To assess levels of faecal contamination in the Qu'Appelle River (Saskatchewan, Canada) and its suitability for irrigation, by using the Colilert-18/Quanti-Tray technology.
Methods and Results:  Various sites located along the Qu'Appelle River were sampled weekly from May to August 2005–2007. A total of 594 freshwater samples were collected and analysed for enumeration of Escherichia coli using the Colilert-18. The false-positive rate for E. coli detection using Colilert-18 was at most 1·5%. Throughout the irrigation period (June to August), up to 85% of the water samples collected from one of the irrigation water-pumping sites exceeded the recommended limit of 100 CFU per 100 ml. Spikes in E. coli counts were generally concomitant with the sudden rise in river flows. A sub-sample of confirmed E. coli isolates were typed by randomly amplified polymorphic DNA (RAPD). RAPD analysis revealed a high degree of genetic diversity among E. coli isolates. A significant association between RAPD patterns and the month of E. coli isolation was demonstrated.
Conclusions:  Colilert-18 provides an effective means for assessing microbial quality of irrigation water.
Significance and Impact of the Study:  Qu'Appelle River is subject to variability of faecal contamination during irrigation times and monitoring throughout irrigation season is important for ensuring safe production practices.     

Influence of Conductivity Dissipation on Benthic Macroinvertebrates in the North Fork Holston River,Virginia Downstream of a Point Source Brine Discharge during Severe Low-Flow Conditions

The North Fork Holston River (NFHR) was examined in fall 2005 to determine the potential impact of a brine discharge on benthic macroinvertebrates. Conductivity of the discharge ranged from 5900 to 10,930 μ S/cm with a highest measurement of 18,000 μ S/cm. During normal flow, conductivity dissipates rapidly downstream of the discharge; however, low-flow conditions in 2005 hindered dissipation. Benthic macroinvertebrate surveys determined statistically significant differences (p = .05) in all metrices between sites excluding taxa richness. Hydropsychid caddisflies were the predominant taxa at the first two sites below the brine discharge, accounting for 74.3 and 68.8% of the organisms collected. Therefore, Ephemeroptera-Plecoptera-Trichoptera (EPT) abundance minus Hydropsychidae [EPT-H] was the most predictive parameter measured. When hydropsychids were removed from statistical analyses, mean EPT abundance was highest at upstream reference sites 1 and 2 (256 and 297, respectively) and reduced at all downstream sites, ranging from 24 to 52 EPT organisms. Virginia Stream Condition Index (SCI) scores indicated impairment downstream of the discharge with scores of 46.9, 41.8, and 55.8, well below the impairment threshold of 61. These results suggest subchronic, intermittent toxicity occurs under low-flow and may contribute to altered benthic macroinvertebrate assemblages downstream of the brine discharge for approximately 26 miles.     

Real-time quantitative analysis of lipid disordering by aurein 1.2 during membrane adsorption, destabilisation and lysis

Effective antimicrobial peptides (AMPs) distinguish between the host and microbial cells, show selective antimicrobial activity and exhibit a fast killing mechanism. Although understanding the structure-function characteristics of AMPs is important, the impact of the peptides on the architecture of membranes with different lipid compositions is also critical in understanding the molecular mechanism and specificity of membrane destabilisation. In this study, the destabilisation of supported lipid bilayers (SLBs) by the AMP aurein 1.2 was quantitatively analysed by dual polarisation interferometry. The lipid bilayers were formed on a planar silicon oxynitride chip, and composed of mixed synthetic lipids, or Escherichiacoli lipid extract. The molecular events leading sequentially from peptide adsorption to membrane lysis were examined in real time by changes in bilayer birefringence (lipid molecular ordering) as a function of membrane-bound peptide mass. Aurein 1.2 bound weakly without any change in membrane ordering at low peptide concentration (5 μM), indicating a surface-associated state without significant perturbation in membrane structure. At 10 μM peptide, marked reversible changes in molecular ordering were observed for all membranes except DMPE/DMPG. However, at 20 μM aurein 1.2, removal of lipid molecules, as determined by mass loss with a concomitant decrease in birefringence during the association phase, was observed for DMPC and DMPC/DMPG SLBs, which indicates membrane lysis by aurein. The membrane destabilisation induced by aurein 1.2 showed cooperativity at a particular peptide/lipid ratio with a critical mass/molecular ordering value. Furthermore, the extent of membrane lysis for DMPC/DMPG was nearly double that for DMPC. However, no lysis was observed for DMPC/DMPG/cholesterol, DMPE/DMPG and E. coli SLBs. The extent of birefringence changes with peptide mass suggested that aurein 1.2 binds to the membrane without inserting through the bilayer and membrane lysis occurs through detergent-like micellisation above a critical P/L ratio. Real-time quantitative analysis of the structural properties of membrane organisation has allowed the membrane destabilisation process to be resolved into multiple steps and provides comprehensive information to determine the molecular mechanism of aurein 1.2 action.     

Regional invariance among microbial communities

Microbial ecology has focused much on causes of between-site variation in community composition. By analysing five data-sets each of aquatic bacteria and phytoplankton, we demonstrated that microbial communities show a large degree of similarity in community composition and that abundant taxa were widespread, a typical pattern for many metazoan metacommunities. The regional abundance of taxa explained on average 85 and 41% of variation in detection frequency and 58 and 31% of variation in local abundances for bacteria and phytoplankton, respectively. However, regional abundance explained less variation in local abundances with increasing environmental variation between sites within data-sets. These findings indicate that the studies of microbial assemblages need to consider similarities between communities to better understand the processes underlying the assembly of microbial communities. Finally, we propose that the degree of regional invariance can be linked to the evolution of microbes and the variation in ecosystem functions performed by microbial communities.
Ecology Letters (2010) 13: 118–127     

Iron‐mediated suppression of bloom‐forming cyanobacteria by oxine in a eutrophic lake

1. Published studies show that cyanobacteria have higher Fe requirements than eukaryotic algae. To test whether Fe availability can affect formation of a cyanobacterial bloom, a strong Fe chelator, oxine (8‐hydroxyquinoline, C₉H₇NO), was added to enclosures in eutrophic Lake 227 in the Experimental Lakes Area (ELA) (northwestern Ontario). 2. Aphanizomenon schindlerii growth was suppressed, and growth of eukaryotic chlorophytes significantly promoted in enclosures to which oxine had been added. Significant eukaryotic growth did not occur in enclosures treated with ammonium, suggesting that N supplied by degradation of oxine was not responsible for eukaryotic success in the oxine enclosures. 3. In situ Fe²⁺ measurements were unreliable because of interference from high concentrations of dissolved organic compounds. However, oxine rapidly promoted oxidation of Fe²⁺ to Fe³⁺ in deionised water, suggesting that rapid removal of Fe²⁺ also occurred in the oxine‐treated enclosures. 4. In batch cultures, 10 μm Fe and 10 μm oxine (a 1 : 1 ratio) completely inhibited the growth of the cyanobacteria Synechococcus sp. and Anabaena flos‐aquae and the chlorophytes Pseudokirchneriella subcapitata and Scenedesmus quadricauda. Increasing Fe 10‐fold to 100 μm Fe completely and partially reversed oxine inhibition in the two chlorophytes but could not overcome inhibition of the cyanobacteria, indicating that inhibition was Fe‐mediated at least in the eukaryotes. Since oxine binds Fe³⁺ in a 1 : 3 ratio (Fe : oxine), inhibition at a 1 : 1 ratio indicates that not all of the Fe is bound, and a mechanism involving Fe other than chelation was at least partly responsible for inhibition. 5. Collectively, the enclosure and laboratory results suggest that the outcome of competition between cyanobacteria and eukaryotic algae in the oxine‐treated enclosures in Lake 227 was likely a result of decreased availability of Fe, especially Fe²⁺. 6. The results suggest that remediation methods that dramatically restrict the supply rate of Fe²⁺ could reduce the relative abundance of cyanobacteria in eutrophic systems.     

Freshwater rock pools: a review of habitat characteristics,faunal diversity and conservation value

1. In the light of the recent surge of interest in small and often temporary wetlands as model systems for ecological and evolutionary research, this article reviews current knowledge on freshwater rock pools and their fauna. 2. Freshwater rock pools occur all over the globe in all major biomes and depend mainly on precipitation for filling. Rock pool clusters are some of the more persistent and oldest freshwater habitats worldwide. Interactions between climate and geology (e.g. limestone, sandstone, granite) generally determine the morphology and hydrology of rock pool habitats, with hydroperiods ranging from several days up to the whole year. Pool volume is usually small, resulting in strongly fluctuating environmental conditions, low conductivity and wide variations in pH (from 4.0 to 11.0) and temperature (from freezing point to 40 °C) often with well‐marked diel cycles. The highly variable environmental conditions, combined with the unpredictability of the flooding regime, require high stress tolerance of the inhabitants, with adaptations for surviving the dry phase such as the production of resistant stages and active emigration followed by recolonisation. 3. About 460 aquatic animal species have been recorded from freshwater rock pools around the world. Approximately 170 of these are passive dispersers, which mainly disperse as resting stages via wind and overflow of water between pools. Successful long distance dispersal seems limited. This group is composed mainly of rock pool specialists with a high degree of endemicity. The remaining taxa can be considered active dispersers, with migration usually restricted to the adult stage. Often these taxa are broadly distributed and occur in a wide range of temporary habitats in addition to rock pools. 4. The inherent characteristics of freshwater rock pools, such as their simple structure and occurrence on similar substrata all over the world, facilitate comparison of research results and promote rock pools as model systems for ecological and evolutionary research. Nevertheless, despite their potential as model systems, the unique fauna and their importance as sources of freshwater in dry countries, the ecology of freshwater rock pools remain virtually unexplored in large parts of the world.     

Morphology and Notes on Morphogenesis during Cell Division of Deviata polycirrata n. sp. and of Deviata bacilliformis (Gelei, 1954) Eigner, 1995 (Ciliophora: Kahliellidae) from Argentina

ABSTRACT. Described herein are the morphology and certain morphogenetic stages of a new freshwater ciliate species, Deviata polycirrata n. sp., and of Deviata bacilliformis recorded in the soil of a dried temporary pond from Argentina. Ciliates were studied alive and after silver impregnation with Protargol. Deviata polycirrata n. sp. measures 130–180 × 45–70 μm in vivo. The species possesses 8–9 long cirral rows on the right and 9–13 on the left of the oral zone, and 3 dorsal rows of dikinetids. The adoral zone is composed of 39–48 membranelles. There are four macronuclear nodules and usually two micronuclei. A single contractile vacuole is located equatorially on the left body margin. This new species mainly differs from its congeners in having a higher number of cirral rows, the three long dorsal rows of dikinetids (vs. usually one to two dorsal rows of dikinetids), and a higher number of adoral membranelles. The other species reported here, D. bacilliformis, is recorded for the first time in Argentina. Unlike previous observations on this species, on the dorsal surface there are cirral rows that are preceded by cilia (combined cirral rows), and stomatogenesis begins with the proliferation of non‐ciliferous basal bodies some distance posterior to the buccal vertex.     

Behavior,metamorphosis, and muscular organization of the predatory rotifer Acyclus inquietus (Rotifera,Monogononta)

Abstract. The atrochid rotifer, Acyclus inquietus, is a sedentary predator that lives within the colonies of its prey, the rotifer Sinantherina socialis. After larvae infiltrate and become associated with the colony, they secrete a permanent gelatinous tube and undergo metamorphosis to the adult stage. We followed settlement and metamorphosis using bright-field microscopy to document specific larval behaviors after eclosion, and used epifluorescence and confocal microscopy of phalloidin-labeled specimens to visualize some of the morphological changes that occur during metamorphosis. Upon eclosion, larvae possess paired eyespots and a ciliated corona that functions strictly in locomotion. After leaving the parent's gelatinous tube, larvae eventually settle on unoccupied colonies of S. socialis or on other substrates if colonies are unavailable. Settlement involves a period of gliding among colony members before attachment with the foot and the secretion of a gelatinous tube. After settlement, there is a drastic reconfiguration of the corona that involves loss of the eyespots, loss of the coronal cilia, and the formation of the cup-shaped infundibulum, a deep depression in the anterior of the head that leads to the mouth. The development of the infundibulum involves the expansion of tissues around the mouth and is accompanied by a reorientation of the underlying musculature that supplies the infundibulum and allows its use in prey capture. The arrangement of the muscles in the trunk and foot regions, which contain outer circular (complete and incomplete) and inner longitudinal bands, remains unchanged between ontogenetic stages, and reflects the condition characteristic of other rotifers.     

Application of fast repetition rate fluorometry to phytoplankton photosynthetic parameters in freshwaters

Fast repetition rate fluorometry (FRRF) was successfully applied to various studies in modern oceanography. In this study, for the first time, the seasonality of phytoplankton photosynthetic parameters in a deep alpine lake was observed using FRRF in combination with the traditional ¹⁴C incubation technique. Special attention was given to the differences in photosynthetic behaviour during mixed and stratified conditions, characterised especially during summer by a deep chlorophyll maximum (DCM) dominated by the filamentous cyanobacterial species Planktothrix rubescens. Maximum light-utilisation efficiency (α*_14C) was in the range of 0.01–0.03 mgC (mg Chl-a)⁻¹ h⁻¹ (μmol phot. m⁻² s⁻¹)⁻¹, while maximum quantum yields for carbon fixation (Φ_C,max) varied from 0.01–0.07 molC (mol phot.)⁻¹. Higher values occurred during thermal stratification indicating acclimation of the phytoplankton assemblage. These findings were supported by FRRF-based estimates, although cyanobacterial blooms could not be characterised by FRRF-excitation due to methodological deficiencies. In general, however, instantaneous photosynthetic rates measured by FRRF-excitation correlate well at sub-saturating light-intensities with conventional ¹⁴C-uptake rates, although they operate on different time-scales.