Temporal stability of marine phytoplankton in a subtropical coastal environment

We investigated the temporal stability of phytoplankton at a subtropical coastal site for 9 months by conducting chlorophyll and flow cytometric measurements at relatively high frequency (roughly at 2–5 day interval). Phytoplankton cells were grouped based on their sizes obtained from flow cytometric signals. We also conducted dilution experiments to estimate the growth and grazing mortality rates of different phytoplankton groups to assess whether the temporal stability of phytoplankton abundances was related with phytoplankton growth/grazing rates. Based on size-fractionated chlorophyll measurements, there was some indication that smaller phytoplankton cells were more stable than larger ones. However, by cytometric counting, there was no evidence for greater stability in small cells. Synechococcus, which had the lowest stability and dominated the <1 μm size class, showed a strong seasonal cycle that was highly dependent on temperature whereas eukaryotes did not have evident seasonal cycles. In general, biomass of a phytoplankton group consisting of several size classes was more stable than that of its sub component, consistent with the hypothesis that higher diversity leads to higher stability, probably related with the effect of statistical averaging (portfolio effect). Stability of heterotrophic bacteria was much higher than that of phytoplankton, leading to the speculation that bacteria were more diverse than phytoplankton. Phytoplankton stability was not related with their growth or grazing mortality rates. Our study suggests that species diversity should be taken into account when considering the temporal stability of phytoplankton.     

Sources of Escherichia coli in a Coastal Subtropical Environment

Sources of Escherichia coli in a coastal waterway located in Ft. Lauderdale, Fla., were evaluated. The study consisted of an extensive program of field measurements designed to capture spatial and temporal variations in E. coli concentrations as well as experiments conducted under laboratory-controlled conditions. E. coli from environmental samples was enumerated by using a defined substrate technology (Colilert-18). Field sampling tasks included sampling the length of the North Fork to identify the river reach contributing high E. coli levels, autosampler experiments at two locations, and spatially intense sampling efforts at hot spots. Laboratory experiments were designed to simulate tidal conditions within the riverbank soils. The results showed that E. coli entered the river in a large pulse during storm conditions. After the storm, E. coli levels returned to baseline levels and varied in a cyclical pattern which correlated with tidal cycles. The highest concentrations were observed during high tide, whereas the lowest were observed at low tide. This peculiar pattern of E. coli concentrations between storm events was caused by the growth of E. coli within riverbank soils which were subsequently washed in during high tide. Laboratory analysis of soil collected from the riverbanks showed increases of several orders of magnitude in soil E. coli concentrations. The ability of E. coli to multiply in the soil was found to be a function of soil moisture content, presumably due to the ability of E. coli to outcompete predators in relatively dry soil. The importance of soil moisture in regulating the multiplication of E. coli was found to be critical in tidally influenced areas due to periodic wetting and drying of soils in contact with water bodies. Given the potential for growth in such systems, E. coli concentrations can be artificially elevated above that expected from fecal impacts alone. Such results challenge the use of E. coli as a suitable indicator of water quality in tidally influenced areas located within tropical and subtropical environments.     

Ubiquity and persistence of Escherichia coli in a Midwestern coastal stream

Dunes Creek, a small Lake Michigan coastal stream that drains sandy aquifers and wetlands of Indiana Dunes, has chronically elevated Escherichia coli levels along the bathing beach near its outfall. This study sought to understand the sources of E. coli in Dunes Creek's central branch. A systematic survey of random and fixed sampling points of water and sediment was conducted over 3 years. E. coli concentrations in Dunes Creek and beach water were significantly correlated. Weekly monitoring at 14 stations during 1999 and 2000 indicated chronic loading of E. coli throughout the stream. Significant correlations between E. coli numbers in stream water and stream sediment, submerged sediment and margin, and margin and 1 m from shore were found. Median E. coli counts were highest in stream sediments, followed by bank sediments, sediments along spring margins, stream water, and isolated pools; in forest soils, E. coli counts were more variable and relatively lower. Sediment moisture was significantly correlated with E. coli counts. Direct fecal input inadequately explains the widespread and consistent occurrence of E. coli in the Dunes Creek watershed; long-term survival or multiplication or both seem likely. The authors conclude that (i) E. coli is ubiquitous and persistent throughout the Dunes Creek basin, (ii) E. coli occurrence and distribution in riparian sediments help account for the continuous loading of the bacteria in Dunes Creek, and (iii) ditching of the stream, increased drainage, and subsequent loss of wetlands may account for the chronically high E. coli levels observed.     

Escherichia coli mechanisms of copper homeostasis in a changing environment

Escherichia coli is equipped with multiple systems to ensure safe copper handling under varying environmental conditions. The Cu(I)-translocating P-type ATPase CopA, the central component in copper homeostasis, is responsible for removing excess Cu(I) from the cytoplasm. The multi-copper oxidase CueO and the multi-component copper transport system CusCFBA appear to safeguard the periplasmic space from copper-induced toxicity. Some strains of E. coli can survive in copper-rich environments that would normally overwhelm the chromosomally encoded copper homeostatic systems. Such strains possess additional plasmid-encoded genes that confer copper resistance. The pco determinant encodes genes that detoxify copper in the periplasm, although the mechanism is still unknown. Genes involved in copper homeostasis are regulated by MerR-like activators responsive to cytoplasmic Cu(I) or two-component systems sensing periplasmic Cu(I). Pathways of copper uptake and intracellular copper handling are still not identified in E. coli.     

Co-Detection of Virulent Escherichia coli Genes in Surface Water Sources

McNemar’s test and the Pearson Chi-square were used to assess the co-detection and observed frequency, respectively, for potentially virulent E. coli genes in river water. Conventional multiplex Polymerase Chain Reaction (PCR) assays confirmed the presence of the aggR gene (69%), ipaH gene (23%) and the stx gene (15%) carried by Enteroaggregative E. coli (EAEC), Enteroinvasive E. coli (EIEC) and Enterohermorrhagic E. coli (EHEC), respectively, in river water samples collected from the Berg River (Paarl, South Africa). Only the aggR gene was present in 23% of samples collected from the Plankenburg River system (Stellenbosch, South Africa). In a comparative study, real-time multiplex PCR assays confirmed the presence of aggR (EAEC) in 69%, stx (EHEC) in 15%, ipaH (EIEC) in 31% and eae (EPEC) in 8% of the river water samples collected from the Berg River. In the Plankenburg River, aggR (EAEC) was detected in 46% of the samples, while eae (EPEC) was present in 15% of the water samples analyzed using real-time multiplex PCR in the Plankenburg River. Pearson Chi-square showed that there was no statistical difference (p > 0.05) between the conventional and real-time multiplex PCRs for the detection of virulent E. coli genes in water samples. However, the McNemar’s test showed some variation in the co-detection of virulent E. coli genes, for example, there was no statistical difference in the misclassification of the discordant results for stx versus ipaH, which implies that the ipaH gene was frequently detected with the stx gene. This study thus highlights the presence of virulent E. coli genes in river water and while early detection is crucial, quantitative microbial risk analysis has to be performed to identify and estimate the risk to human health.     

Sources,impacts and trends of pharmaceuticals in the marine and coastal environment

There has been a significant investment in research to define exposures and potential hazards of pharmaceuticals in freshwater and terrestrial ecosystems. A substantial number of integrated environmental risk assessments have been developed in Europe, North America and many other regions for these situations. In contrast, comparatively few empirical studies have been conducted for human and veterinary pharmaceuticals that are likely to enter coastal and marine ecosystems. This is a critical knowledge gap given the significant increase in coastal human populations around the globe and the growth of coastal megacities, together with the increasing importance of coastal aquaculture around the world. There is increasing evidence that pharmaceuticals are present and are impacting on marine and coastal environments. This paper reviews the sources, impacts and concentrations of pharmaceuticals in marine and coastal environments to identify knowledge gaps and suggests focused case studies as a priority for future research.     

The survival response of Escherichia coli K12 in a natural environment

To verify the hypothesis of cryptic growth and viable but nonculturable (VBNC) state, survival responses of Escherichia coli cells were examined under oligotrophic microcosm conditions for an extended period. In the case of filtered distilled water at 4°C, E. coli cells definitely entered the VBNC state within 56 days. However, culturability and viability increased while the total number of cells declined after 110 days. This phenomenon can be explained by considering three possible states. The first is the existence of the VBNC state, the second is cryptic growth, and the third is the death of E. coli cells. In the case of artificial seawater at 4°C, VBNC E. coli cells confirmed the existence of two log units of elongated VBNC cells. Moreover, elongated VBNC cells showed the most significant change among all the other transformed cells. Also, E. coli cells in microcosms at 28°C indicated the entrance to the classical starvation survival state. In resuscitation tests, 1% diluted Luria-Bertani agar medium showed the highest level of resuscitation among amended agar media. To evaluate the survival ability of E. coli cells in the activated sludge samples, we used an E. colistrain XL-1 blue containing plasmids pQ2 including GFPcDNA (XL/GFP). In supernatant of activated sludge (SUP) at 28°C, XL/GFP cells entered the VBNC state after 10 days, whereas existence of VBNC cells was not detectable in resuspended activated sludge (ACT) at 28°C.     

Sympatric metabolic diversification of experimentally evolved Escherichia coli in a complex environment

Sympatric diversification in bacteria has been found to contravene initial evolutionary theories affirming the selection of the fittest type by competition for the same resource. Studies in unstructured (well-mixed) environments have discovered divergence of an ancestor strain into genomically and phenotypically divergent types growing both on single and mixed energy sources. This study addresses the metabolic diversification in an Escherichia coli population that evolved over ~1,000 generations under aerobic conditions in the nutritional complexity offered by Luria–Bertani (LB) broth. The medium lacked glucose but contained a variety of other resources. Two distinct metabolically-diverged types, coinciding with colony morphologies, were found to dominate the populations. One type was an avid carbohydrate consumer, which could quickly utilize the available (alternative) substrates feeding into glycolysis. The second type was a slow grower, which was able to specifically consume acetate. The capacity to utilize acetate might be providing an advantage to this second type, suggesting an increased capability to deal with adverse conditions that occur in the later stages of growth. The diverged metabolic preferences of the two forms suggested differential and interactive ecological roles within the population. We postulate that these types used different alternative metabolic strategies occupying different niches in a sympatric manner as an outcome of adaptation to the complex environment.     

Escherichia coli genes expressed preferentially in an aquatic environment

Enteric bacteria are frequently found in aquatic environments, where they may pose a risk to human health. Although bacterial survival and persistence in such habitats has been studied extensively, there is almost no information about bacterial adaptation to these conditions at the level of changes in gene expression. As a first exploration of this field, we have carried out a screen designed to identify Escherichia coli genes that show increased expression in an aquatic environment. The screen was performed by subtractive hybridization on a genomic library and led to the identification of several RNA species more abundant in cells inoculated in this medium than in stationary- phase cultures after growth in rich medium. The genes identified include specific tRNA operons and a gene of unknown function, gapC , with similarities to glyceraldehyde-3-phosphate dehydrogenases. E. coli K-12 strains appear to have accumulated mutations in gapC , which may impede its translation, whereas natural isolates have an intact gapC gene. Sequence comparison of gapC with related genes suggests its acquisition by horizontal gene transfer from Gram-positive bacteria.     

Growth rhythms and productivity of a coastal subtropical eucalypt forest

Growth of plant parts in different species in Eucalyptus signata dominated forest in Queensland occurs at different times. Measurement of increments in root basal diameter combined with previously obtained regressions of root weight on root diameter allows separate estimation of the growth of woody root parts on a seasonal basis for the first time. The growth of lateral and of tap roots as well as of stems and leafy twigs, were separately measured on three tree species every 6 weeks for 2 years. Net primary productivity for the forest is estimated at 26 t ha^-1 yr^-1, of which 66% is aboveground production, and 78% is contributed by a single species. This productivity rate is unexpectedly high, and may reflect a growth stimulus due to release of nutrients following a recent fire.     

Statistical Considerations in the Sampling of Escherichia coli from Intestinal Sources for Serotyping

A theoretical study of the expected distribution of Escherichia coli serotypes among samples randomly drawn from populations containing different numbers, ranging from 2 to 40, was compared with experimental data based on the examination of 10 and 100 coiony samples from the same specimens. From these studies it was possible to lay down guide lines for the planning of efficient sampling programmes for determining most or all serotypes present in various animal species.     

Development of a software tool for in silico simulation of Escherichia coli using a visual programming environment

This study describes the development of a software tool, EcoSim, to assist users in implementing quantitative in silico simulation easily. It consists of four parts: extracellular environment and constraints setting mode, table for optimal metabolic flux distribution and chart for changes of substrate concentration, dynamic flux distribution viewer and dynamic hierarchical regulatory network viewer. Representation of a hierarchical regulatory network was constructed with defined modeling symbols and weight in the central Escherichia coli metabolism. All programming procedures for EcoSim were accomplished in a visual programming environment (LabVIEW). To illustrate quantitative in silico simulation with EcoSim, this program was performed on E. coli using glucose and acetate as carbon sources. The simulation results were in agreement with the experimental data obtained from the literature. EcoSim can be used to assist biologists and engineers in predicting and interpreting dynamic behaviors of E. coli under a variety of environmental conditions.     

Genetic richness of vibriophages isolated in a coastal environment

The purpose of this study was to characterize Vibrio parahaemolyticus viruses (VpVs) isolated from different environments within and adjacent to the Strait of Georgia, and to examine the relative influences of distance and environment on host-range and genetic richness. Nearly all seawater enrichment cultures (29/31) generated isolates, implying that VpVs were widespread in the virioplankton, yet at low abundances (< 1 l(-1)). Viruses were not detected in sediments (n = 99). Fourteen of the 16 viruses characterized were siphoviruses, with genome sizes ranging from approximately 45-106 kb, and half were capable of infecting other Vibrio species. The VpVs infected bacteria isolated from oysters and sediments fairly well (55% and 46% of the host-virus combinations, respectively), but were unable to infect many of the bacteria isolated from the water column (< 13% of 112 combinations). When compared with VpVs from oysters, it was clear that the major determinant of phenotypic (host-range) and genetic richness (by the DP-RAPD assay) was not geography, but the source environment from which the VpVs originated. Therefore, the VpV population within the Strait of Georgia is a highly diverse mixture of phenotypes and genotypes.     

Meteorological factors and ambient bacterial levels in a subtropical urban environment

We conducted a study to investigate the characteristics and determinants of ambient bacteria in Taipei, Taiwan from August 2004 to March 2005. We monitored ambient culturable bacteria in Shin-Jhuang City, an urban area in the Taipei metropolitan areas, using duplicate Burkard Portable Air Samplers with R2A agar. The average concentration of total bacteria was 1,986 colony-forming units per cubic meter of air (CFU/m³) (median?=?780 CFU/m³) over the study period, with the highest level in autumn. Most bacterial taxa had similar seasonal variation, with higher concentrations in autumn and winter. During the study period, Gram negative rods and cocci were predominant. Multivariate analyses indicated that wind speed and wind direction significantly influenced ambient bacterial distribution. Temperature and relative humidity were also important environmental factors positively associated with ambient bacterial concentrations. We observed statistically significant relationships between ambient bacteria and air pollutants, including sulfur dioxide (SO₂), ozone, particulate matter (aerodynamic diameter ≤10 μm (PM₁₀)), methane and total hydrocarbons. The concentrations of methane and total hydrocarbons during the previous day were positively associated with total bacteria and Gram negative rods, respectively. Ozone level on the previous day had a negative relationship with Gram negative cocci. SO₂ level with a 3-day lag was positively correlated with concentrations of both total bacteria and Gram negative cocci. In the future, more longitudinal studies are needed to confirm the relationships and possible mechanisms between ambient bacteria and meteorological factors, as well as to evaluate the ecological and health impacts of ambient bacteria.     

Increased susceptibility to repeated freeze-thaw cycles in Escherichia coli following long-term evolution in a benign environment

Background  

In order to study the dynamics of evolutionary change, 12 populations of E. coli B were serially propagated for 20,000 generations in minimal glucose medium at constant 37°C. Correlated changes in various other traits have been previously associated with the improvement in competitive fitness in the selective environment. This study examines whether these evolved lines changed in their ability to tolerate the stresses of prolonged freezing and repeated freeze-thaw cycles during adaptation to a benign environment.     

Fish and limnology of a thermal water environment in subtropical South America

Two thermal sources with water temperatures from 51 to 59°C flow into a stream of 2 to 5 m width and about 0.5 m depth at Agua Caliente (23° 44′, 64° 38′) in Jujuy province, Argentina. Data from 3 years sampling show that the influence of the thermal sources maintains the water temperature of the stream section at a high and constant level (from 24 to 35°C), different from the thermal regime of other streams in the area. Composition of water (N=13) has the following mean values: pH 8.36, conductivity 1591 μS cm^-1, dominant ions (in mg l^-1) CO₃ ^-- 12.77, CO₃H^- 140.27, Cl^- 246.86, SO₄ ^-- 460.14, Na⁺ 400.45, K⁺ 2.18, Ca⁺⁺ 27.68 and Mg⁺⁺ 2.14. Mean total dissolved solids: 1.3 g l^-1. Large amounts of SO₄ ^--, Na⁺, and Cl^- Sixteen fish species (2460 specimens) were captured in the warmed reach. Dominant families were Characidae, Cichlidae and Loricariidae. New geographic distribution information is provided for eight species, some of them with restricted northwestern Argentina distributions. Most abundant species were the eurytopic characid Astyanax bimaculatus, followed by the cichlid Bujurquina vittata. These species have the highest critical thermal maximum according to field experiments. Temperature of acclimatization is closer to lethal than in fishes from ‘normal’ habitats. Agua Caliente differs from other thermal habitats in the lack of isolation, its placement in a rain forest area, a high number of species, and the lack of cyprinodontoids. The fish fauna here represents an opportunistic invasion of a habitat with water parameters strongly different from those in the area, particularly temperature and salinity. Both faunistic and limnological traits make of Agua Caliente a new type of environment within the subtropics. This revised version was published online in August 2006 with corrections to the Cover Date.