Showing papers in "Microbial Ecology in 1996"
TL;DR: The utility and application of techniques suitable for determining microscale consortial activity, production, community composition, and interactions in the context of larger scale aquatic ecosystem structure and function are discussed.
Abstract: Molecular oxygen (O2) is a potent inhibitor of key microbial processes, including photosynthesis, N2 fixation, denitrification, sulfate reduction, methanogenesis, iron, and metal reduction reactions. Prokaryote survival and proliferation in aquatic environments is often controlled by the ability to tolerate exposure to oxic conditions. Many prokaryotes do not have subcellular organelles for isolating O2-producing from O2-consuming processes and have developed consortial associations with other prokaryotes and eukaryotes that alleviate metabolic constraints of high O2. Nutrient transformations often rely on appropriate cellular and microenvironmental, or microzonal, redox conditions. The spatial and temporal requirements for microenvironmental overlap among microbial groups involved in nutrient transformations necessitates close proximity and diffusional exchange with other biogeochemically distinct, yet complementary, microbial groups. Microbial consortia exist at different levels of community and metabolic complexity, as shown for detrital, microbial mat, biofilm, and planktonic microalgal-bacterial assemblages. To assess the macroscale impacts of consortial interactions, studies should focus on the range of relevant temporal (minutes to hours) and spatial (microns to centimeters) scales controlling microbial production, nutrient exchange, and cycling. In this review, we discuss the utility and application of techniques suitable for determining microscale consortial activity, production, community composition, and interactions in the context of larger scale aquatic ecosystem structure and function.
468 citations
TL;DR: The structures of bacterial communities were studied in activated sludge samples obtained from the aerobic and anaerobic zones of a wastewater treatment plant showing enhanced phosphorous removal by in situ hybridization with oligonucleotide probes complementary to selected regions of the 16S and 23S ribosomal RNA (rRNA) characteristic for defined phylogenetic entities.
Abstract: The structures of bacterial communities were studied in activated sludge samples obtained from the aerobic and anaerobic zones of a wastewater treatment plant showing enhanced phosphorous removal. Samples were analyzed by in situ hybridization with oligonucleotide probes complementary to selected regions of the 16S and 23S ribosomal RNA (rRNA) characteristic for defined phylogenetic entities (genera and larger groups). The microbial community structures revealed by molecular techniques were compared with the compositions of culturable bacterial communities, obtained from the characterization of 255 isolates from tryptone-soy (TS) agar and R2A agar. These isolates were characterized by 89 physiological tests and their cellular fatty acid patterns, and identified. Culture-dependent techniques indicated the following distribution: different Aeromonas spp. (2.7–8.3% on R2A agar; 45.0–63.7% on TS agar), Acinetobacter spp. (5.4–9.0% on R2A agar; 5.0–9.1% on TS agar), Pseudomonas spp. (up to 10% on R2A agar) and Shewanella putrefaciens (up to 3.0% on R2A agar), all members of the gamma subclass of Proteobacteria, were isolated most frequently. The relatively rare isolates of the beta subclass were identified as Acidovorax spp., Alcaligenes spp., and Comamonas spp., The Gram-positive bacteria (high DNA G+C) were assigned mainly to Arthrobacter spp., Microbacterium spp., and Mycobacterium phlei. In order to assess the in situ abundance of the most frequently isolated genus, Aeromonas, two rRNA-targeted oligonucleotide probes were developed. The two gamma proteobacterial genera Aeromonas and Acinetobacter constituted less than 5% of all bacteria. In situ, Proteobacteria belonging to the beta subclass and high G+C Gram-positive bacteria were dominant. From filamentous bacteria, Sphaerotilus spp. and Leptothrix spp. could be detected occasionally. In addition, one sample contained a high proportion of the morphologically distinct filaments of Microthrix parvicella.
220 citations
TL;DR: Methods designed to detect microorganisms involved in the biogeochemistry of nitrogen in the marine environment are rapidly being developed and deployed in ecological investigations and should complement the more familiar, but no less sophisticated methods that focus on quantification of in situ transformation rates.
Abstract: Methods designed to detect microorganisms involved in the biogeochemistry of nitrogen in the marine environment are rapidly being developed and deployed in ecological investigations. Probes based on phylogenetic sequences (usually rRNA) and those based on the sequences of functional genes or proteins have both been demonstrated in the nitrogen cycle. The most progress has been made for ammonia oxidizers; several sets of PCR primers have been described and their specificity may be optimized to allow detection of genetically and ecologically meaningful groups. For denitrifying bacteria, functional probes based on nitrite reductase show most promise. These approaches should complement the more familiar, but no less sophisticated, methods that focus on quantification of in situ transformation rates. Both approaches in combination will be useful in understanding regulation and environmental control of biogeochemical processes.
192 citations
TL;DR: A dramatic shift in functional microbial community structure was found during the 8-week composting process, and the data suggest that the Biolog assay may be a suitable approach to determine compost maturity.
Abstract: The objective of this study was (a) to detect changes of the functional abilities of the microflora during composting of manure as a result of windrow turning frequency and (b) to detect differences between distinct zones within the windrows. Biolog GN microtiter plates containing 95 different carbon sources were inoculated with diluted suspensions of compost material containing 15,000 microorganisms per well (120 μl). We found a dramatic shift in functional microbial community structure during the 8-week composting process. The shift was more rapid when the compost windrows were turned. The substrate use pattern in the outer, well-aerated zone of the unturned windrow was similar to that of the turned windrows. Microbial biomass and respiration decreased more rapidly in the turned than in the unturned windrows, indicating a different pace of compost maturation. The data suggest that the Biolog assay may be a suitable approach to determine compost maturity.
169 citations
TL;DR: It is proposed that, in general, bacterial growth rates are temperature dependent up to a threshold, but that the patterns of change across temperature gradients are resource dependent, such that temperature has little effect on growth in resource-rich environments but a strong effect inresource-poor environments.
Abstract: We examined the potential limitation of bacterial growth by temperature and nutrients in a eutrophic lake. Dilution cultures from winter and summer were incubated at both high (>20°C) and low (4°C) temperatures and enriched with various combinations of organic carbon (C), inorganic nitrogen (N), and inorganic phosphorus (P). Bacterial abundance, (3)H-thymidine incorporation, and (3)H-leucine incorporation were measured over the growth cycle. For both winter and summer assemblages, low temperature limited growth even when resources (C, N, and P) were added. When temperature was adequate, bacterial growth in dilution cultures was co-limited by C, N, and P Additions of either C, P, or N and P alone provide little or only modest stimulation of growth, suggesting that under in situ conditions both nutrients and organic carbon limit bacterial growth. Our results provide little evidence of seasonal adaptation to low temperatures for bacterial communities in temperate lakes. Instead, bacterial growth appears to be temperature limited during winter and resource limited during summer. We propose that, in general, bacterial growth rates are temperature dependent up to a threshold, but that the patterns of change across temperature gradients are resource dependent, such that temperature has little effect on growth in resource-rich environments but a strong effect in resource-poor environments.
148 citations
TL;DR: The theoretical basis for the use of a molecular approach to N2 fixation is addressed, the strengths and weaknesses of the approach are highlighted, and case studies are provided that demonstrate the potential contribution of molecular biology approaches to studies of N2fixing organisms in the sea.
Abstract: Nitrogen fixation in the sea has attracted the attention of ecologists for decades. Much is known about the habitats in which it occurs and some of the factors that limit N2 fixation activity in different environments, but we still know little about the organisms that fix nitrogen, and what limits the growth and distribution of these organisms in marine environments. Molecular biology technological developments have provided tools for detecting and characterizing N2-fixing organisms in the environment. These techniques hold great promise for unraveling the mysteries and paradoxes of N2 fixation in the sea. In this review, we address the theoretical basis for the use of a molecular approach to N2 fixation, highlight the strengths and weaknesses of the approach, and provide case studies that demonstrate the potential contribution of molecular biology approaches to studies of N2 fixation in the sea.
118 citations
TL;DR: More than 34 species of Legionella have been identified, 20 of which have been isolated from both environmental and clinical sources as discussed by the authors, and the vast majority of Legionellosis is caused by the L. pneumophila species.
Abstract: The respiratory diseases produced by the Legionella genus of bacteria are collectively called Legionellosis. Presently more than 34 species of Legionella have been identified, 20 of which have been isolated from both environmental and clinical sources. The diseases produced by Legionella include the pneumonic form, Legionnaires' disease, and the flu-like form, Pontiac fever. Because the vast majority of Legionellosis is caused by the L. pneumophila species, this bacterium is the thrust of the discussion.
109 citations
TL;DR: The results indicated that even in low numbers, epibiotic protozoa may have a major grazing impact on free bacteria.
Abstract: Seasonal and depth variations of the abundance, biomass, and bacterivory of protozoa (heterotrophic and mixotrophic flagellates and ciliates) were determined during thermal stratification in an oligomesotrophic lake (Lake Pavin, France). Maximal densities of heterotrophic flagellates (1.9 × 103 cells ml−1) and ciliates (6.1 cells ml−1) were found in the metalimnion. Pigmented flagellates dominated the flagellate biomass in the euphotic zone. Community composition of ciliated protists varied greatly with depth, and both the abundance and biomass of ciliates was dominated by oligotrichs. Heterotrophic flagellates dominated grazing, accounting for 84% of total protistan bacterivory. Maximal grazing impact of heterotrophic flagellates was 18.9 × 106 bacteria 1−1h−1. On average, 62% of nonpigmented flagellates were found to ingest particles. Ciliates and mixotrophic flagellates averaged 13% and 3% of protistan bacterivory, respectively. Attached protozoa (ciliates and flagellates) were found to colonize the diatom Asterionella formosa. Attached bacterivores had higher ingestion rates than free bacterivorous protozoa and may account for 66% of total protozoa bacterivory. Our results indicated that even in low numbers, epibiotic protozoa may have a major grazing impact on free bacteria.
98 citations
TL;DR: Elevated viral abundance and their erratic distribution in the sediments suggest that viruses might play an important role in sediment microbial dynamics.
Abstract: Viruses were found to be very abundant in the top layer of the sediments of Lac Gilbert, Quebec. Viruses were extracted from the sediments using pyrophosphate buffer, and viruses from the diluted extracts were pelleted onto grids and enumerated using transmission electron microscopy. Viral abundance in the sediments ranged from 6.5 × 108 to 1.83 × 1010 ml−1, which is 10- to 1,000-fold greater than the number observed in the water column. This increase corresponds well with the 100- to 1,000-fold increase in bacterial abundance in the sediments. Viral abundance differed significantly among the surface sediment samples taken at different bottom depths and among samples taken at different depths of the water column. Viral abundance also varied significantly between the oxic and anoxic zones of the water column and the sediments. The virus-to-bacteria ratio varied greatly among the different sediment sites but not among depths in the water column. Viral abundance in the water column was related to bacterial abundance and chlorophyll concentration, whereas viruses in the sediments were most abundant in sediments with high organic matter content. Elevated viral abundance and their erratic distribution in the sediments suggest that viruses might play an important role in sediment microbial dynamics.
92 citations
TL;DR: An in situ field study of the motility patterns exhibited by Phormidium corallyticum and Beggiatoa spp.
Abstract: An in situ field study of the motility patterns exhibited by Phormidium corallyticum and Beggiatoa spp. in black-band disease of corals was conducted over a 5-day period. Measurements were made at a spatial resolution of 50 mm to document the horizontal migration of black-band across living coral tissue, while vertical migrations within the band were documented by observation and macrophotography of the black-band surface. It was determined that horizontal migration occurred both day and night, with the fastest movements by the front of the band during the day and the back of the band at night. Beggiatoa would rise to the band surface at night, and would often remain above the cyanobacterial population during extended periods of illumination the following day. The migration patterns are discussed in terms of motility cues and microbial physiology.
86 citations
TL;DR: Results from Signy Island and at continental sites on Alexander Island suggested that rates of microalgal colonization and community development might change significantly during current climate changes in Antarctica, and dominated by conspicuous autofluorescent indicator species with distinctive morphology.
Abstract: There is increasing evidence of climate change in Antarctica, especially elevated temperature and ultraviolet B (UVB) flux within the ozone “hole.” Its origins are debatable, but the effects on ice recession, water availability, and summer growth conditions are demonstrable. Light-dependent, temperature-sensitive, fast-growing organisms respond to these physical and biogeographical changes. Microalgae (cyanobacteria and eukaryotic algae), which are pioneer colonists of Antarctic mineral fellfield soils, are therefore highly suitable biological indicators of such changes. In frost-heaved soil polygons containing naturally sorted fine mineral particles, microalgal growth is restricted to a shallow zone of light penetration. By virtue of this light requirement, microalgae are exposed to extreme seasonal fluctuations in temperature (air and black-body radiation), photosynthetically active radiation, UV radiation, and desiccation. Dominance of conspicuous autofluorescent indicator species with distinctive morphology allowed quantification of responses using epifluorescence microscopy, and image analysis of undisturbed, unstained communities. However, the physical changes in climate, although significant in the long term, are gradual. The changes were therefore amplified experimentally by enclosing the communities at a fellfield site on Signy Island, maritime Antarctica, in cloches (small greenhouses). These were made of polystyrene of either UV transparent or UV opaque acrylic plastic, with or without walls. During a 6-year period, statistically significant changes were observed in microalgal colonization of the soil surface and in the morphology of filamentous populations. Evidence of community succession correlated with measured changes in local environment was found. Results from Signy Island and at continental sites on Alexander Island suggested that rates of microalgal colonization and community development might change significantly during current climate changes in Antarctica.
TL;DR: The results suggest that in this strain of S. marcescens caffeine is degraded to theobromine and/or paraxanthine (1,7-dimethylxanthine), and subsequently to 7-methylxanthines and xanthine.
Abstract: A strain of Serratia marcescens showing the ability to degrade caffeine and other methylxanthines was isolated from soil under coffee cultivation. Growth was observed only with xanthines methylated at the 7 position (caffeine, 1,3,7-dimethylxanthine; paraxanthine, 1,7-dimethylxanthine; theobromine, 3,7-dimethylxanthine and 7-methylxanthine). Paraxanthine and theobromine were released in liquid medium when caffeine was used as the sole source of carbon and nitrogen. When paraxanthine or theobromine were used, 3-methylxanthine, 7-methylxanthine, and xanthine were detected in the liquid medium. Serratia marcescens did not grow with theophylline (1,3-dimethylxanthine), 1-methylxanthine, and 3-methylxanthine, and poor growth was observed with xanthine. Methyluric acid formation from methylxanthines was tested in cell-free extracts by measuring dehydrogenase reduction of tetrazolium salt in native-polyacrylamide gel electrophoresis gel. Activity was observed for all methylxanthines, even those with which no bacterial growth was observed. Our results suggest that in this strain of S. marcescens caffeine is degraded to theobromine (3,7-dimethylxanthine) and/or paraxanthine (1,7-dimethylxanthine), and subsequently to 7-methylxanthine and xanthine. Methyluric acid formation could not be confirmed.
TL;DR: Low amounts of produced EPS and improvement of attachment indicated that mechanisms other than polysaccharide production are more important on substrata with low surface tensions (<25 mN m−1).
Abstract: Attachment strength and exopolysaccharide (EPS) production of Pseudomonas sp. (bacteria) and the diatom Amphora coffaeformis were studied on six different substrata with surface tensions between 19 and 64.5 mN m−1. Test panels of the materials were exposed to bacterial cultures between 3 and 120 hours, and to diatom cultures between 48 and 72 hours. Exopolysaccharide production by surface-associated cells was measured using the phenol sulfuric acid method. Attachment studies were run by exposing test panels to laminar flow pressure using a radial flow chamber. Highest EPS production by bacteria and diatoms was recorded on substrata with surface tensions above 30 mN m−1. Lowest EPS production occurred on substrata between 20 and 25 mN m−1. Highest EPS production and strongest adhesion was found on polycarbonate (33.5 mN m−1). Both test organisms improved their attachment strength with exposure time on most materials. However, amounts of produced EPS and improvement of attachment indicated that mechanisms other than polysaccharide production are more important on substrata with low surface tensions (<25 mN m−1). Simply producing more polysaccharides is not sufficient to overcome weak attachment on materials with low surface tensions. For example, adhesion of Pseudomonas sp. and A. coffaeformis on polytetrafluorethylene/perfluor-copolymer (PFA; 22 mN m−1). and glass (64.5 mN m−1. was equally strong although EPS production was much higher on glass than on PFA. This is somewhat surprising for A. coffaeformis because polysaccharide production has been considered the most important attachment mechanism of A. coffaeformis.
TL;DR: Phenanthrene-degrading bacteria were isolated from enrichment cultures of soils contaminated with creosote and jet fuel and it was inferred that the half-saturation constants were less than the lowest phenanthrene concentration tested.
Abstract: Phenanthrene-degrading bacteria were isolated from enrichment cultures of soils contaminated with creosote and jet fuel. The isolates from the creosote enrichments were classified by fatty acid methyl ester profiles as Acidovorax delafieldii and Sphingomonas paucimobilis; the bacterium from the jet fuel-contaminated soil was not identified and was designated strain JFD 11. All three isolates used phenanthrene as a sole carbon and energy source, and two of the isolates used fluoranthene as a sole carbon and energy source. Anthracene and fluorene were cometabolized by all three strains, but pyrene was not transformed. Naphthalene inhibited all of the strains, and 28-h cultures of A. delafieldii were inhibited by naphthalene concentrations as low as 5 ppm. Short-term degradation experiments were undertaken with center-well flasks and concentrations of phenanthrene ranging from 1.2 to 12.0 μm. Since initial degradation rates were not a function of phenanthrene concentration, it was inferred that the half-saturation constants were less than the lowest phenanthrene concentration tested.
TL;DR: In this paper, the authors compared molecular biological methods using rRNA probes to geochemical measurements at two different sites and found that sulfate reduction rates were low and 16S-like rRNAs comprised about 5% of the total rRNA extracted from the sediment.
Abstract: Past studies of microbial communities responsible for geochemical transformations have been limited by an inability to representatively cultivate, and then identify, the constituent members. Ribosomal RNA sequences, particularly 16S-like rRNAs, provide a measure of phylogenetic relationship that can now be used to examine the structure and diversity of microbial communities. Sulfate-reducing bacteria (SRB) play an important role in the sulfur cycle and the terminal mineralization of organic matter in estuarine and marine environments. Because the Gram-negative mesophilic SRB comprise a phylogenetically coherent assemblage, their communities are well suited to explorations through rRNA sequence-based methodologies. In this study we related molecular biological methods using rRNA probes to geochemical measurements at two different sites. At an unvegetated site in northwest Florida, rates of sulfate reduction were low and SRB rRNA comprised about 5% of the total rRNA extracted from the sediment. The other site, a salt marsh in New Hampshire, had higher rates of sulfate-reduction with SRB rRNA accounting for up to 30% of the total rRNA extracted from the sediment. SRB community structure differed dramatically between the two sites with Desulfobulbus rRNA much less abundant in the unvegetated site than in the salt marsh. The differences in these SRB communities reflect differences in the ecology of their habitats.
TL;DR: This work has shown that Rhizobium is the best-studied rhizobacterium, which forms a symbiosis with a restricted host range, and Azospirillum is another plant-growth-promoting rhizOBacterium which forms rhizocoenoses with a wide range of plants.
Abstract: Plant growth promotion by rhizobacteria is a widely spread phenomenon. However only a few rhizobacteria have been studied thoroughly. Rhizobium is the best-studied rhizobacterium. It forms a symbiosis with a restricted host range. Azospirillum is another plant-growth-promoting rhizobacterium which forms rhizocoenoses with a wide range of plants. In both bacteria, the interaction with the plant involves the attraction toward the host plant and the attachment to the surface of the root. Both bacteria are attracted to plant roots, but differ in specificity. Attachment to plant roots occurs in two steps for both bacteria: a quick, reversible adsorption, and a slow, irreversible anchoring to the plant root surface. However, for the two systems under study, the bacterial surface molecules involved in plant root attachment are not necessarily the same.
TL;DR: The results indicate that under natural conditions there is denitrification and hence a loss in nitrogen from the Amazon floodplain to the atmosphere, and the nitrogen removal of exposed sediments exceeded that of undisturbed wetland soils of temperate regions.
Abstract: Denitrification processes were measured by the acetylene-blockage technique under changing flood conditions along the aquatic/terrestrial transition zone on the Amazon floodplain at Lago Camaleao, near Manaus, Brazil. In flooded sediments, denitrification was recorded after the amendment with NO
3
−
(100 μmol liter−1) throughout the whole study period from August 1992 to February 1993. It ranged from 192.3 to 640.7 μmol N m−2 h−1 in the 0- to 5-cm sediment layer. Without substrate amendment, denitrification was detected only during low water in November and December 1992, when it occurred at a rate of up to 12.2 μmol N m−2 h−1 Higher rates of denitrification at an average rate of 73.3 μmol N m−2 h−1 were measured in sediments from the shallow lake basin that were exposed to air at low water. N2O evolution was never detected in flooded sediments, but in exposed sediments, it was detected at an average rate of 28.3 μmol N m−2 h−1 during the low-water period. The results indicate that under natural conditions there is denitrification and hence a loss in nitrogen from the Amazon floodplain to the atmosphere. Rates of denitrification in flooded sediments were one to two orders of magnitude smaller than in temperate regions. However, the nitrogen removal of exposed sediments exceeded that of undisturbed wetland soils of temperate regions, indicating a considerable impact of the flood pulse on the gaseous turnover of nitrogen in the Amazon floodplain.
TL;DR: A community hybridization technique was applied to determine the similarity and relative diversity of two samples by cross hybridization to determine broad-scale differences in soil microbial community structure, demonstrating that single bacterial species and a mixture of cultivable bacteria were less complex and only 5% similar to whole-community DNA or bacteria directly extracted from the soil.
Abstract: Broad-scale approaches seek to integrate information on whole microbial communities. It is widely recognized that culture techniques are too selective and unrepresentative to allow a realistic assessment of the overall structure of microbial communities. Techniques based on fatty acid or metabolic profiles determine the phenotypic composition of the community. Complementary information about the genotypic structure of soil microbial communities necessitates analysis of community DNA. To determine broad-scale differences in soil microbial community structure (i.e., differences at the whole community level, rather than specific differences in species composition), we have applied a community hybridization technique to determine the similarity and relative diversity of two samples by cross hybridization. In previous studies this assay failed with whole-soil community DNA. Usable hybridization signals were obtained using whole-soil DNA, in this study, by digesting the DNA with restriction enzymes before the labeling with a random-primer reaction. The community hybridization technique was tested using a graded series of microbial fractions, increasing in complexity, all isolated from the same soil sample. This demonstrated that single bacterial species and a mixture of cultivable bacteria were less complex and only 5% similar to whole-community DNA or bacteria directly extracted from the soil. Extracted bacterial and whole-community DNA were 75% similar to each other and equally complex. When DNA was extracted from four different agricultural soils, their similarities ranged from 35 to 75%. The potential usefulness of community hybridization applied to soil microbial communities is discussed.
TL;DR: It is demonstrated that determining conversion factors strongly depends on the selected calculation method (modified derivative, integrative, and cumulative) and these different estimates are reconciled when the very low proportion of active cells is accounted for.
Abstract: This study compares three independent me,-ods usec for estimat- ing bacterioplankton production in waters from the lagoon (mesotrophic) and the surrounding ocean (oligotrophic) of two atolls from the Tuamotu archipel- ago (French Polynesia). Thymidine and leucine incorporation were calibrated in dilution cultures and gave consistent results when the first was calibrated against cell multiplica- tion and the second against protein synthesis. This study demonstrates that determining conversion factors strongly depends on the selected calculation method (modified derivative, integrative, and cumulative). These different estimates are reconciled when the very low proportion of active cells is ac- counted for. Frequency of dividing-divided cells (FDDC) calibrated using the same dilu- tion cultures led to unrealistically high estimates of bacterial production. How- ever, highly significant correlations between FDDC and either thymidine- or leucine-specific incorporation per cell were found in lagoon waters in situ. These correlations became more positive when oceanic data were added. This suggests that the FDDC method is also potentially valid to determine bacteri- oplankton growth rates after cross calibration with thymidine or leucine meth- ods. If recommended precautions are observed, the three methods tested in the present study would give reliable production estimates.
TL;DR: Levels of merA-specific transcripts and Hg(II) volatilization were influenced more by microbial activity (as manipulated by nutrient additions) than by the concentration of total mercury, suggesting that rates of mercury vol atilization in the environment may not always be proportional to merA transcription.
Abstract: Bacterial transformation of mercury in the environment has received much attention owing to the toxicity of both the ionic form and organomercurial compounds. Bacterial resistance to mercury and the role of bacteria in mercury cycling have been widely studied. The genes specifying the required functions for resistance to mercury are organized on the mer operon. Gene probing methodologies have been used for several years to detect specific gene sequences in the environment that are homologous to cloned mer genes. While mer genes have been detected in a wide variety of environments, less is known about the expression of these genes under environmental conditions. We combined new methodologies for recovering specific gene mRNA transcripts and mercury detection with a previously described method for determining biological potential for mercury volatilization to examine the effect of mercury concentrations and nutrient availability on rates of mercury volatilization and merA transcription. Levels of merA-specific transcripts and Hg(II) volatilization were influenced more by microbial activity (as manipulated by nutrient additions) than by the concentration of total mercury. The detection of merA-specific transcripts in some samples that did not reduce Hg(II) suggests that rates of mercury volatilization in the environment may not always be proportional to merA transcription.
TL;DR: It appears that the bioavailability of various forms of adsorbed phosphate was determined primarily by the stability of the phosphate-surface complexes that they formed, rather than by the total amount of phosphate Adsorbed.
Abstract: A nonreductive community-level study of P availability was conducted using various forms of adsorbed P. Orthophosphate (Pi), inositol hexaphosphate (IHP), and glucose 6-phosphate (G6P) were adsorbed to a short-range ordered Al precipitate. These bound phosphates provided a P source sufficient to support the growth of microbial communities from acidic Brazilian soils (oxisols). Adsorbed IHP, the most abundant form of organic phosphate in most soils, had the lowest bioavailability among the three phosphates studied. Adsorbed G6P and Pi were almost equally available. The amount of adsorbed Pi (1 cmol P kg−1) required to support microbial growth was at least 30 times less than that of IHP (30 cmol P kg−1). With increased surface coverage, adsorbed IHP became more bioavailable. This availability was attributed to a change in the structure of surface complexes and presumably resulted from the decreased number of high-affinity surface sites remaining at high levels of coverage. It thus appears that the bioavailability of various forms of adsorbed phosphate was determined primarily by the stability of the phosphate-surface complexes that they formed, rather than by the total amount of phosphate adsorbed. IHP, having the potential to form stable multiple-ring complexes, had the highest surface affinity and the lowest bioavailability. Bioaggregates consisting of bacteria and Al precipitate were observed and may be necessary for effective release of adsorbed P. Bacteria in the genera Enterobacter and Pseudomonas were the predominate organisms selected during these P-limited enrichments.
TL;DR: Experiments in which host cells and phage ΦR2f were introduced into two soils of different texture revealed that host cells were primarily lysed in the presence of added nutrients, and phages reached highest titres in these nutrient-amended soils.
Abstract: The introduction of bacteriophages could potentially be used as a control method to limit the population size of engineered bacteria that have been introduced into soil. Hence, the ability of a species-specific phage, ΦR2f, to infect and lyse its host, a Pseudomonas fluorescens R2f transposon Tn5 derivative, in soil, was studied. Control experiments in liquid media revealed that productive lysis of host cells by phage ϕR2f occurred when cells were freely suspended, whereas cells present in alginate beads resisted lysis. The presence of nutrients enhanced the degree of lysis as well as the production of phage progeny, both with the suspended cells and with cells escaped from the alginate beads. Experiments in which host cells and phage ΦR2f were introduced into two soils of different texture revealed that host cells were primarily lysed in the presence of added nutrients, and phage reached highest titres in these nutrient-amended soils. Encapsulation of the host cells in alginate beads inhibited lysis by the phage in soil. Populations of free host cells introduced into soil that colonized the rhizosphere of wheat were not substantially lysed by phage ΦR2f. However, P. fluorescens R2f populations colonizing the rhizosphere after introduction in alginate beads were reduced in size by a factor of 1,000. Cells migrating from the alginate beads towards the roots may have been in a state of enhanced metabolic activity, allowing for phage ΦR2f infection and cell lysis.
TL;DR: The stability of the virulence following passage through insect-host and stability of molecular structure for the fusion products FP 8 and FP 25 suggest that asexual genetic recombination by protoplast fusion may provide an attractive method for the genetic improvement of biocontrol efficiency in entomopathogenic fungi.
Abstract: Genetic improvement of two different strains of the entomopathogenic fungus Beauveria bassiana for more effective control of Ostrinia nubilalis and Leptinotarsa decemlineata was obtained by crosses with the insecticidal toxin-producing strain Beauveria sulfurescens. Protoplast fusion between diauxotrophic mutants resulted in the recovery of some stable prototrophic fusion products. The low levels of virulence of the wild type strain B. bassiana 28 isolated originally from L. decemlineata were enhanced both on L. decemlineata and O. nubilalis for one of the hybrids obtained (FP 8) from the cross B. bassiana 28 × B. sulfurescens 2. Fusion product 25 obtained from the cross between B. sulfurescens and the highly pathogenic strain B. bassiana 147 showed a three-day reduction in the LT50 towards O. nubilalis. Southern blot hybridization with nine probe-enzyme combinations were conducted on genomic DNAs from the original wild strains, parental mutant strains, and fusion products. Additive banding patterns or unique banding pattern of either parental strain was observed in five hybrids, indicating their status as recombinant and/or partially diploid. Combination of RFLP markers indicative of both parental genomes was never observed with fusion product FP 25. The stability of the virulence following passage through insect-host and stability of molecular structure for the fusion products FP 8 and FP 25 suggest that asexual genetic recombination by protoplast fusion may provide an attractive method for the genetic improvement of biocontrol efficiency in entomopathogenic fungi.
TL;DR: Spatial and temporal variations in the concentrations of dissolved gases (CH4, CO2, and O2) in peat cores were studied using membrane inlet mass spectrometry (MIMS) and variations in vertical gas profiles were observed between randompeat cores taken from hollows on the same peat bog.
Abstract: Spatial and temporal variations in the concentrations of dissolved gases (CH4, CO2, and O2) in peat cores were studied using membrane inlet mass spectrometry (MIMS). Variations in vertical gas profiles were observed between random peat cores taken from hollows on the same peat bog. Methane concentrations in profiles (0–30 cm) generally increased with depth and reached maximum values in the range of 200–450 μm CH4 below about 13-cm depth. In some profiles, a peak of dissolved methane was observed at 7-cm depth. Oxygen penetrated to approximately 2-cm depth in the hollows. The sampling probe was used to continuously monitor CH4, CO2, and O2 concentrations at fixed depths in peat cores over periods of several days. The concentration of dissolved CO2 and O2 at 1-cm depth oscillated over a 24-h period with the maximum of CO2 concentration corresponding with the minimum of 02. Diurnal variations in CO2 but not CH4 were measured at 15-cm depth; dissolved CO2 levels decreased during daylight hours to a constant minimum concentration of 4.85 mm. This report also describes the application of MIMS for the measurement of gaseous diffusion rates in peat using an inert gas (argon); the value of D, the diffusion coefficient, was 2.07 × 10−8 m2 s−1.
TL;DR: In this paper, the authors studied the use of total dissolved carbohydrates (TDCHO) by planktonic bacteria in mesotrophic Lake Constance, Germany, from April until August 1992, and examined the decrease of TDCHO over time together with the increase of bacterial numbers in 1-μm filtered lake water incubated at in situ temperature in the dark.
Abstract: Dissolved carbohydrates comprise one of the largest pools of labile organic matter readily available for bacterial use in pelagic ecosystems. Despite this fact, very little is known about use of dissolved carbohydrates by planktonic bacteria. We studied use of total dissolved carbohydrates (TDCHO) by planktonic bacteria in mesotrophic Lake Constance, Germany, from April until August 1992. We examined the decrease of TDCHO over time together with the increase of bacterial numbers in 1-μm filtered lake water incubated at in situ temperature in the dark. TDCHO analyses were done after hydrolysis by sulfuric acid as free monosaccharides and oxidation by periodate to formaldehyde with 3-methyl-2-benzothiazolinone-hydrazon-hypochloride (MBTH). Preliminary tests showed that hydrolysis by sulfuric acid gave higher yields than hydrolysis by HCl. Our results show that TDCHO are readily used by bacteria and compose substantial fractions of the C requirements for their growth. Concentrations of TDCHO varied between 1.7 and 5.5 μm (glucose equivalents), and use rates varied between 0.47 and 3.43 μg C liter−1 h−1. Highest rates of TDCHO use occurred during the phytoplankton spring bloom, during the clear-water phase in June, and during a phytoplankton bloom in August. Ratios of use of TDCHO/bacterial biomass production varied between 0.17 and 3.05. During the spring bloom, TDCHO and total dissolved amino acids (TDAA) were used in equal amounts. During the clear-water phase at chlorophyll a concentrations <3 μg liter−1, however, bacteria only consumed TDCHO and excreted amino acids. The growth efficiency based on the consumption of TDCHO and TDAA varied between 16 and 21% during the phytoplankton spring bloom and was 35% during the clear-water phase.
TL;DR: Correlation analysis showed that cyanob bacterial abundance in the two phases (water and sediment) was influenced more by water than by soil properties, and dissolved inorganic nitrogen (DIN) and the ratio DIN: SRP correlated negatively with cyanobacterial abundance.
Abstract: In order to estimate the potential utilization of N2-fixing (heterocystous) cyanobacteria as natural biofertilizers in the Valencian rice fields (Spain), the distribution and seasonal variation of these microorganisms in water and sediment samples were evaluated, and the relationships among cyanobacterial abundance and physical and chemical characteristics of soil and water were investigated. N2-fixing cyanobacteria were present in all the samples analyzed (25 sampling points sampled three times per year during two years). The relative cyanobacterial abundance in soil and water followed contrasting patterns, maximum presence in soil coincided with minimum abundance in water. Correlation analysis showed that cyanobacterial abundance in the two phases (water and sediment) was influenced more by water than by soil properties. Salinity,mineralization variables, and soluble reactive phosphate (SRP) correlated positively with heterocystous cyanobacteria presence. Furthermore, dissolved inorganic nitrogen (DIN) and the ratio DIN:SRP correlated negatively with cyanobacterial abundance. However DIN:SRP ratio better described the cyanobacterial distribution, with a threshold effect: below the Redfield ratio value (7.2 in mass units) cyanobacterial abundance was clearly higher.
TL;DR: This work characterize bacterial isolates for 40 physiological attributes, and grouped the isolates hierarchically using two-stage density-linkage cluster analysis and used rarefied estimates of richness as a measure of community diversity in the various habitats.
Abstract: We present an approach to comparing the diversity and composition of bacterial communities from different habitats and for identifying which members of a community are most affected by an introduced bacterium. We use this method to explore both previously published and new data from field and growth chamber experiments in which we isolated heterotrophic bacteria from samples of root-free soil, roots of nontreated soybean seedlings, and from the roots of soybean seedlings grown from Bacillus cereus UW85nl—treated seeds. We characterize bacterial isolates for 40 physiological attributes, and grouped the isolates hierarchically using two-stage density-linkage cluster analysis. Multivariate analysis of variance and discriminant analysis of the relative frequencies of the clusters in the soil and rhizosphere habitats were then used to determine whether there were differences among the bacterial communities from the various habitats, and which of the clusters were most useful in discriminating among the communities. We used rarefied estimates of richness as a measure of community diversity in the various habitats. Introduction of UW85n 1 affected the composition and/or diversity of rhizosphere communities in three of four experiments.
TL;DR: The ability of attack phase bdellovibrios to survive in an anoxic environment for up to nine days and their growth or survival under microaerobic conditions greatly expands the possible ecological niches in which the predators may be active members of the microbial community.
Abstract: The ability of intraperiplasmic and attack phase bdellovibrios to survive and/or grow under anoxic and microaerobic conditions was examined. Both halotolerant and nonhalotolerant bdellovibrio strains were examined. In all instances, the bdellovibrio strains were unable to grow under anoxic conditions, but were able to survive for periods of time in both the extracellular and intraperiplasmic forms. However, the intraperiplasmic organisms were observed to survive longer. Increased temperature hastened the loss of viability of both forms of the predatory bacteria in oxic and anoxic environments. Under microaerobic conditions, halotolerant bdellovibrios were observed to grow, although at a slightly reduced rate than in atmospheric oxygen, while two nonhalotolerant isolates survived but did not grow. The ability of attack phase bdellovibrios to survive in an anoxic environment for up to nine days and their growth or survival under microaerobic conditions greatly expands the possible ecological niches in which the predators may be active members of the microbial community.
TL;DR: Assimilation of 35S-precursors into microbial proteins was used to investigate toxicity and adaptational responses that occur in nutrient enriched and natural freshwater samples experimentally contaminated with benzene, toluene, trichloroethylene (TCE), or xylene, and distinct banding patterns of polypeptides differ with each chemical pollutant.
Abstract: Assimilation of 35S-precursors into microbial proteins was used to investigate toxicity and adaptational responses that occur in nutrient enriched and natural freshwater samples experimentally contaminated with benzene, toluene, trichloroethylene (TCE), or xylene. Experiments were conducted to analyze (1) the potential of using microbial community protein profiles for responsive identification of chemical pollutant exposure, (2) the inhibition of microbial productivity through reduction in rate of protein synthesis caused by specific chemical pollutants, and (3) whether selection of subpopulations in freshwater microbial communities challenged with chemical pollutants leads to adaptive strategies mediated by production of particular polypeptides. The results show that distinct banding patterns of polypeptides in the range of 30 to 100 kilodaltons that were obtained following collective cultivation of freshwater microorganisms differ with each chemical pollutant. Protein yield and radioisotope incorporation were reduced within ten minutes of microbial exposure to chemical pollutants in the following order: xylene < toluene < benzene < TCE. Adaptation of the freshwater microbial community to chemical pollutants prior to radioisotope incorporation produced differences in polypeptide profiles, in the banding patterns of radioactive polypeptides, and in the rate of radioisotope incorporation. The rate of radioisotope incorporation by freshwater microorganisms pre-adapted to chemical pollutants was lowest with xylene (88.1% reduction), followed by TCE (84.0% reduction), toluene (67.3% reduction), and benzene (43.5% reduction). In long-term radioisotope incorporation experiments, protein yield and polypeptide radioactivity was higher in the presence of chemical pollutants than in uncontaminated control samples, suggesting increased metabolic productivity attributable to the chemical pollutants.
TL;DR: The results of factorial analysis of variance suggested that Ni toxicity to photosynthesis is a consequence of a strong interaction among strain, Ni dosage, and duration of exposure.
Abstract: Nickel (Ni) toxicity to growth and photosynthesis was studied in four strains of Scenedesmus acutus f. alternans. Effects of Ni dosage and duration of exposure on growth and photosynthesis were strain specific. Large differences in responses of both growth and photosynthesis to Ni were detected between three resistant strains (B4, Cu-Tol, and Ni-Tol) and one sensitive strain (UTEX 72). Growth of UTEX 72 was ≥ 18 times more sensitive to Ni than those of the three resistant strains. The order of Ni dosages (fmol Ni/pg cell dry weight) causing 50% inhibition (D150) of growth rates in the four strains was Ni-Tol (10.5) > B4 (8.19) > Cu-Tol (4.60) > UTEX 72 (0.25). The effect of Ni dosage on photosynthetic rate as percentage of control corresponded to a saturation curve and was a strong function of duration of exposure. The DI50s of photosynthetic rates were ≥3.5 times lower in UTEX 72 than in the three resistant strains, and in all four strains they decreased sharply with the increase in duration of exposure. The order of the four strains in DI50s of photosynthetic rate was B4 (58.2) > Cu-Tol (38.0) > Ni-Tol (28.9) > UTEX 72 (8.24) for 6-h exposure and Ni-Tol (2.88) > Cu-Tol (1.30) > B4 (1.01) > UTEX 72 (0.15) for 24-h exposure. The DI50s of photosynthetic rate for 6-h exposure were higher than those of growth rate in all four strains, and for 24-h exposure they were lower, except in UTEX 72. Thus, the relative Ni sensitivity of growth and photosynthesis of the four strains depends on the duration of exposure. The results of factorial analysis of variance suggested that Ni toxicity to photosynthesis is a consequence of a strong interaction among strain, Ni dosage, and duration of exposure.