Factors controlling bacterial production in marine and freshwater sediments

We collected benthic bacterial production data measured by ³H thymidine incorporation (TTI) (25 studies), frequency of dividing cells (FDC) (3 studies), dark-C0₂ assimilation (1 study) and ³H-adenine uptake (2 studies) from the literature, which included 18 marine, 6 river, and 2 lake studies. In all of the studies that used the TTI method, ³H-DNA was isolated and incubations were carried out at in situ temperatures. Most of the researchers also determined ³H-DNA extraction efficiencies and isotope dilution, thus interpretable estimates of bacterial production were used in the analysis. In marine sediments, bacterial production rates were linked to bacterial biomass, bacterial abundance, sediment organic matter, temperature, and sediment chlorophyll a, with these variables explaining between 40% and 68% of the variation in production rates. Simple relationships between production and bacterial biomass or bacterial abundance, or between production and sediment organic matter, were improved by also including temperature in the analysis of marine sediments. Sediment organic matter explained an appreciable fraction (58%) of the observed production in freshwater sediments. Temperature was the most powerful predictor of the observed variability in specific growth rates (r ² = 0.48 and r ² = 0.58) in marine and freshwater sediments, respectively. Thus, bacterial production and specific growth rates are most closely linked to substrate supply and temperature in marine and freshwater sediments. Offprint requests to: B. C. Sander.     

Importance of advective mass transfer and sediment surface area for streambed microbial communities

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Thermophilic anaerobes in Arctic marine sediments induced to mineralize complex organic matter at high temperature

Marine sediments harbour diverse populations of dormant thermophilic bacterial spores that become active in sediment incubation experiments at much higher than in situ temperature. This response was investigated in the presence of natural complex organic matter in sediments of two Arctic fjords, as well as with the addition of freeze‐dried Spirulina or individual high‐molecular‐weight polysaccharides. During 50°C incubation experiments, Arctic thermophiles catalysed extensive mineralization of the organic matter via extracellular enzymatic hydrolysis, fermentation and sulfate reduction. This high temperature‐induced food chain mirrors sediment microbial processes occurring at cold in situ temperatures (near 0°C), yet it is catalysed by a completely different set of microorganisms. Using sulfate reduction rates (SRR) as a proxy for organic matter mineralization showed that differences in organic matter reactivity determined the extent of the thermophilic response. Fjord sediments with higher in situ SRR also supported higher SRR at 50°C. Amendment with Spirulina significantly increased volatile fatty acids production and SRR relative to unamended sediment in 50°C incubations. Spirulina amendment also revealed temporally distinct sulfate reduction phases, consistent with 16S rRNA clone library detection of multiple thermophilic Desulfotomaculum spp. enriched at 50°C. Incubations with four different fluorescently labelled polysaccharides at 4°C and 50°C showed that the thermophilic population in Arctic sediments produce a different suite of polymer‐hydrolysing enzymes than those used in situ by the cold‐adapted microbial community. Over time, dormant marine microorganisms like these are buried in marine sediments and might eventually encounter warmer conditions that favour their activation. Distinct enzymatic capacities for organic polymer degradation could allow specific heterotrophic populations like these to play a role in sustaining microbial metabolism in the deep, warm, marine biosphere.     

Microbial communities from Arctic marine sediments respond slowly to methane addition during ex situ enrichments

Anaerobic methanotrophic archaea (ANME) consume methane in marine sediments, limiting its release to the water column, but their responses to changes in methane and sulfate supplies remain poorly constrained. To address how methane exposure may affect microbial communities and methane- and sulfur-cycling gene abundances in Arctic marine sediments, we collected sediments from offshore Svalbard that represent geochemical horizons where anaerobic methanotrophy is expected to be active, previously active, and long-inactive based on reaction-transport biogeochemical modelling of porewater sulfate profiles. Sediment slurries were incubated at in situ temperature and pressure with different added methane concentrations. Sediments from an active area of seepage began to reduce sulfate in a methane-dependent manner within months, preceding increased relative abundances of anaerobic methanotrophs ANME-1 within communities. In previously active and long-inactive sediments, sulfur-cycling Deltaproteobacteria became more dominant after 30 days, though these communities showed no evidence of methanotrophy after nearly 8 months of enrichment. Overall, enrichment conditions, but not methane, broadly altered microbial community structure across different enrichment times and sediment types. These results suggest that active ANME populations may require years to develop, and consequently microbial community composition may affect methanotrophic responses to potential large-scale seafloor methane releases in ways that provide insight for future modelling studies.     

Experimental degradation of plant materials in Hudson river sediments

We examined photopigment degradation and transformation in sediment microcosms that received different detrital source materials (planktonic, littoral, terrestrial) in the presence or absence of amphidpods (Gammarus sp.). Additions of realistic quantities of particulate organic matter resulted in detectable changes in pigment concentration and composition despite insignificant changes in total organic matter. The transformation of chlorophyll a to total phaeophorbide was significantly higher in all high quality (high nitrogen) detritus treatments containing amphipods. The highest production of phaeophorbide was in the higher quality detritus (blue-green algae, Anabaena cylindrica, and macrophyte, Vallisneria americana) when compared to red maple (Acer rubrum). Phaeophytin formation was not related to amphipod grazing and thus may be determined more by microbial heterotrophic processes. The degradation product of the carotenoid lutein, lutein 5,6 expoxide, was formed in all treatments. Phaeopigment composition can be used to infer differences in heterotrophic activity and will help in the interpretation of photopigment distribution in field samples.     

A bioassay to assess the potential effects of sediment resuspension on phytoplankton community composition

A laboratory assay (SAGA or Sediment Algal Growth Assay) was developed to assess the potential impact of sediment resuspension on the structure of phytoplankton communities, and to evaluate the effectiveness of various sediment treatments in decreasing the abundance of blue-green algae in the event of sediment resuspension during storms. In assays with sediment from eutrophic Akanoi Bay, Lake Biwa, Japan, 7–11 species of phytoplankton seeded from the sediments grew during the 3-week assay indicating that sediment resuspension has the potential to increase both phytoplankton biomass and species diversity. Treatment of sediments with Ca(NO3)2 substantially decreased phytoplankton biomass (measured as chlorophyll concentration) in assays with sediments from Akanoi Bay and the North Basin of Lake Biwa. Further, among various oxidation treatments of sediments, Ca(NO3)2 was most effective in decreasing or preventing filamentous blue-green algal growth in N- and P-replete media. In contrast, when sediments were added to P-limited phytoplankton dominated by green algae and diatoms, no growth of blue-green algae occurred regardless of sediment treatment. This revised version was published online in June 2006 with corrections to the Cover Date.     

Spatial and halophyte-associated microbial communities in intertidal coastal region of India

Microbial communities in intertidal coastal soils respond to a variety of environmental factors related to resources availability, habitat characteristics, and vegetation. These intertidal soils of India are dominated with Salicornia brachiata, Aeluropus lagopoides, and Suaeda maritima halophytes, which play a significant role in carbon sequestration, nutrient cycling, and improving microenvironment. However, the relative contribution of edaphic factors, halophytes, rhizosphere, and bulk sediments on microbial community composition is poorly understood in the intertidal sediments. Here, we sampled rhizosphere and bulk sediments of three dominant halophytes (Salicornia, Aeluropus, and Suaeda) from five geographical locations of intertidal region of Gujarat, India. Sediment microbial community structure was characterized using phospholipid fatty acid (PLFA) profiling. Microbial biomass was significantly influenced by the pH, electrical conductivity, organic carbon, nitrogen, and sodium and potassium concentrations. Multivariate analysis of PLFA profiles had significantly separated the sediment microbial community composition of regional sampling sites, halophytes, rhizosphere, and bulk sediments. Sediments from Suaeda plants were characterized by higher abundance of PLFA biomarkers of Gram-negative, total bacteria, and actinomycetes than other halophytes. Significantly highest abundance of Gram-positive and fungal PLFAs was observed in sediments of Aeluropus and Salicornia, respectively than in those of Suaeda. The rhizospheric sediment had significantly higher abundance of Gram-negative and fungal PLFAs biomarkers compared to bulk sediment. The results of the present study contribute to our understanding of the relative importance of different edaphic and spatial factors and halophyte vegetation on sediment microbial community of intertidal sediments of coastal ecosystem.     

Long-term survival of haptophyte and prasinophyte resting stages in marine sediment

Resting stages of marine phytoplankton have been shown to have potential for long-term survival and to remain viable in marine sediments for up to about a century. This study documents for the first time long-term survival in haptophytes and prasinophytes, by germination of resting stages of Isochrysis galbana and Mantoniella squamata from up to 40-year-old sediment layers. Germination was induced by setting up sediment slurries in L1 medium at 15°C. Cyst formation was induced in culture strains acquired from the germinations by keeping mixtures of strains in the light or dark at salinities of 20 or 30. The identity of the two species was confirmed by light and electron microscopy as well as LSU and SSU rDNA-based phylogenetic analyses.     

Coupling between benthic biomass of Microcystis and phosphorus release from the sediments of a highly eutrophic lake

Variations in microbial biomass and activity in the sediments of hypereutrophic Lake Vallentunasjön were followed during a period of 5 years. The data were compared to the calculated release of phosphorus from the sediments during the same period. A strong co-variation was found between biomass of Microcystis, heterotrophic bacterial activity in the sediments and internal phosphorus loading. These parameters exhibited mainly a declining trend during the investigation period. A pronounced stability of the sediment chemistry, including the fractional composition of the sediment phosphorus, during the studied period indicates that microbial activity affected the phosphorus release from the sediments. Calculations of the percentage of sediment bacteria that was associated to the mucilage of Microcystis colonies imply, together with the specific bacterial production, that Microcystis in the sediment stimulates bacterial production. In the highly phosphorus-saturated sediments of Lake Vallentunasjön this would ultimately lead to an increased release of phosphorus from the sediment. Lake Vallentunasjön does not follow the common pattern of recovery after reduction of external phosphorus loading. The large biomasses and long survival of Microcystis in the sediment are probably important reasons for the delayed recovery of the lake.     

Distribution of Aerobic Bacteria Which Contain Bacteriochlorophyll a

Sixteen strains of aerobic bacteria which contain bacteriochlorophyll a were isolated from the samples collected in aerobic marine environments: thalli of Enteromorpha linza, Porphyra sp., Sargussum horneri; beach sand; and the surface seawater from Aburatsubo Inlet. When they occurred, their proportions among the aerobic heterotrophic populations ranged from 0.9 to 1.1% in the seaweed samples and from 1.2 to 6.3% in the beach sand samples and were 0.9% in the seawater sample. The results suggested that the aerobic photopigmented bacteria widely inhabit aerobic marine environments.     

Temporal Variations in Heterotrophic Mesophilic Bacteria from a Marine Shallow Hydrothermal Vent off the Island of Vulcano (Eolian Islands, Italy)

Abstract The fluctuations of the total microbial abundance, the culturable heterotrophic bacterial population, and the composition of heterotrophic bacteria were investigated in relation to environmental parameters in a shallow, marine hydrothermal vent off the Island of Vulcano (Eolian Islands, Italy). Standing stock dynamics were studied by measuring the total population of picoplankton by direct count and the population of viable heterotrophic bacteria in water and sediment samples collected monthly. The environmental factors most strongly linked to the total microbial abundance and heterotrophic bacterial populations were pH and H₂S content in water and C/N ratio in sediment samples. The pattern of variation of microbial populations associated with water was different from those associated with sediment. Assessment of the qualitative composition of aerobic heterotrophic bacterial communities was based on 30 morphological and biochemical characteristics for each strain. Numerical analysis was used for an initial survey of the similarity among the isolates. The data were successively used to determine the structure and the metabolic potential of water and sediment bacteria. Metabolic properties varied between water- and sediment-isolated bacteria. Bacteria from water were structurally more diverse, and active in the use of carbohydrates, than those from sediment. Moreover, most of the sediment bacteria were able to grow at a high temperature (60 and 70°C). The fluctuations of bacterial characteristics in relation to environmental parameters present an evident temporal variation in water, but not in the sediment habitat. Received: 13 January 1997; Accepted: 7 August 1997     

Sediment Resuspension Effects on the Benthic Microbial Loop in Experimental Microcosms

Sediment resuspension induced by anthropogenic disturbance is becoming a major threat of marine coastal ecosystems worldwide. The effects of sediment resuspension on the pelagic domain and on macro- and meiobenthos are well documented in the literature, whereas the effects on the benthic microbial components are nearly neglected. We have investigated the effects of sediment resuspension at two different disturbance levels on benthic bacterial abundance, biomass, and activities and on heterotrophic nanobenthos abundance in experimental microcosms. The results of our experiments pointed out that, independently from the amount of involved energy, sediment resuspension determined a general decrease of all benthic microbial components. The main effects consisted of a decreased abundance of the metabolically active bacterial fraction and of heterotrophic nanobenthos abundance. However, the amount of energy involved in sediment resuspension had differential effects on the structure and functioning of the benthic microbial loop, but only in the short term (i.e., within 36 h). Sediment resuspension had a stimulatory effect on activities of surviving bacterial cells, which, in turn, resulted in increased sediment organic C turnover rates. We hypothesize that such an effect, enhancing nutrient availability, might have relevant consequences on the trophic state of coastal marine ecosystem.     

Comparison of the Levels of Bacterial Diversity in Freshwater,Intertidal Wetland,and Marine Sediments by Using Millions of Illumina Tags

Sediment, a special realm in aquatic environments, has high microbial diversity. While there are numerous reports about the microbial community in marine sediment, freshwater and intertidal sediment communities have been overlooked. The present study determined millions of Illumina reads for a comparison of bacterial communities in freshwater, intertidal wetland, and marine sediments along Pearl River, China, using a technically consistent approach. Our results show that both taxon richness and evenness were the highest in freshwater sediment, medium in intertidal sediment, and lowest in marine sediment. The high number of sequences allowed the determination of a wide variety of bacterial lineages in all sediments for reliable statistical analyses. Principal component analysis showed that the three types of communities could be well separated from phylum to operational taxonomy unit (OTU) levels, and the OTUs from abundant to rare showed satisfactory resolutions. Statistical analysis (LEfSe) demonstrated that the freshwater sediment was enriched with Acidobacteria, Nitrospira, Verrucomicrobia, Alphaproteobacteria, and Betaproteobacteria. The intertidal sediment had a unique community with diverse primary producers (such as Chloroflexi, Bacillariophyta, Gammaproteobacteria, and Epsilonproteobacteria) as well as saprophytic microbes (such as Actinomycetales, Bacteroidetes, and Firmicutes). The marine sediment had a higher abundance of Gammaproteobacteria and Deltaproteobacteria, which were mainly involved in sulfate reduction in anaerobic conditions. These results are helpful for a systematic understanding of bacterial communities in natural sediment environments.     

Aerobic metabolic potential of microbial populations indigenous to deep subsurface environments

Subsurface sediment samples, collected from three boreholes ranging in depths from 0.1 to 260 m, were used in substrate mineralization studies to examine the aerobic metabolic potential of microbial populations indigenous to the deep subsurface. Mineralization was measured by quantifying the amount of ¹⁴CO₂ released from radiolabeled acetate, phenol, or 4‐methoxybenzoate added to subsurface sediments at 10 μg g^‐1. Mineralization of the three compounds was observed in all but a few of the subsurface samples and did not decrease with depth. In addition, mineralization data collected from similar geologic formations from the different boreholes indicated that there was significant lateral continuity of microbial activity. Regression analyses were performed to determine which environmental factors were related to microbial metabolic potential. Mineralization was positively correlated with heterotrophic abundance as measured by plate counts. Other parameters that appeared to influence metabolic potential included pH and clay content.     

Correlation of Direct Viable Counts with Heterotrophic Activity for Marine Bacteria

Viable-bacteria counts, heterotrophic activity, and substrate responsiveness of viable bacteria have been used to measure microbial activity. However, the relationship between these parameters is not clear. Thus, the direct viable count (DVC) method was used to analyze seawater samples collected from several different geographical locations. Samples collected from offshore waters of the South China Sea and western Pacific Ocean yielded DVC that indicated the presence of surface and subsurface peaks of viable, substrate-responsive bacteria which could be correlated with turnover rates of amino acids obtained by using uniformly ¹⁴C-labeled amino acids. DVC were always less than total viable counts (acridine orange direct counts), and the DVC subsurface peak occurred close to and within the chlorophyll a zone, suggesting algal-bacterial interactions within the layer. For comparison with the open-ocean samples, selected substrates were used to determine the response of viable bacteria present in seawater samples collected near an ocean outfall of the Barceloneta Regional Waste Treatment Plant, Barceloneta, Puerto Rico. The number of specific substrate-responsive bacteria at the outfall stations varied depending on the substrate used and the sampling location. Changes in the population size or physiological condition of the bacteria were detected and found to be associated with the presence of pharmaceutical waste.     

Growth and metabolism of the purple sulfur bacterium Thiocapsa roseopersicina under combined light/dark and oxic/anoxic regimens

The dominant purple sulfur bacterium of laminated sediment ecosystems in temperate environments, Thiocapsa roseopersicina, was cultivated in sulfide-limited continuous cultures (D=0.03 h^-1) subjected to various combined diel regimen of aeration and illumination in order to simulate environmental conditions in microbial mats. For comparison, cultures were grown under similar illumination regimens but continuously anoxic conditions.Bacteriochlorophyll a (BChla) and carotenoid synthesis was restricted to anoxic-dark periods and did not occur during oxic-light periods. An increase in the length of the oxic-light periods resulted in decreased pigment contents. However, phototrophic growth remained possible even at 20 h oxic-light/4 h anoxic-dark regimens. When anoxic conditions were maintained throughtout, BChla synthesis occurred both during light and dark periods.Glycogen was synthesized in the light and degraded in the dark. Calculations showed that degradation of 1/4–1/5 of the glycogen is sufficient to account for the BChla and carotenoid synthesis in the dark.The data showed that T. roseopersicina is very well adapted to cope with the combined oxygen and light regimes as they occur in microbial mats, which may explain the dominance of this bacterium in the purple layer of these sediment ecosystems.Non-standard abbreviations BChl bacteriochlorophyll - specific growth rate - D dilution rate - S_R concentration of limiting substrate in reservoir bottle     

Seasonal and spatial diversity of microbial communities in marine sediments of the South China Sea

This study was conducted to characterize the diversity of microbial communities in marine sediments of the South China Sea by means of 16S rRNA gene clone libraries. The results revealed that the sediment samples collected in summer harboured a more diverse microbial community than that collected in winter, Deltaproteobacteria dominated 16S rRNA gene clone libraries from both seasons, followed by Gammaproteobacteria, Acidobacteria, Nitrospirae, Planctomycetes, Firmicutes. Archaea phylotypes were also found. The majority of clone sequences shared greatest similarity to uncultured organisms, mainly from hydrothermal sediments and cold seep sediments. In addition, the sedimentary microbial communities in the coastal sea appears to be much more diverse than that of the open sea. A spatial pattern in the sediment samples was observed that the sediment samples collected from the coastal sea and the open sea clustered separately, a novel microbial community dominated the open sea. The data indicate that changes in environmental conditions are accompanied by significant variations in diversity of microbial communities at the South China Sea.     

Dynamics of Dimethyl Sulfide in a Marine Microbial Mat

Abstract The microbial mat was chosen as a model ecosystem to study dynamics of dimethyl sulfide (DMS) in marine sediments in order to gain insight into key processes and factors which determine emission rates. A practical advantage, compared to open ocean ecosystems, is that microbial mats contain high biomasses of different functional groups of bacteria involved in DMS dynamics, and that DMS concentrations are generally high enough to allow direct measurement of emission rates. Field data showed that, during the seasonal development of microbial mats, concentrations of chlorophyll a corresponded to dimethylsulfoniopropionate (DMSP). DMSP is an important precursor of DMS. It was demonstrated, with laboratory cultures, that various species of benthic diatoms produce substantial amounts of DMSP. The abundances of aerobic and anaerobic DMS- or DMSO-utilizing bacteria were estimated using the most-probable-number technique. Laboratory experiments with relatively undisturbed sediment cores showed that microbial mats act as a sink for DMS under oxic/light (day) conditions, and as a source of DMS under anoxic/dark (night) conditions. Axenic culture studies with Chromatium vinosum M2 and Thiocapsa pfennigii M8 (isolated from a microbial mat) showed that, under anoxic/light conditions, DMS was quantitatively converted to dimethylsulfoxide (DMSO). T. roseopersicina M11 converted DMSP to DMS and acrylate, apparently without use of either substrate. Received: 5 May 1997; Accepted: 21 August 1997     

Survival and Activity of Bacteria in a Deep,Aged Lake Sediment (Lake Constance)

Abstract Viable counts and potential activities of different bacteria were determined as a function of depth in the deep profundal sediment of Lake Constance, Germany. The sediment layer at the bottom of the lake had a total depth of about 7 m and was deposited in the time after the last ice age, i.e., over the past 13,000 years. The high clay content of the sediment prevents seepage. Below 25 cm all of the viable heterotrophic bacteria were present as heat-resistant spores. Numbers of viable spores of both aerobic and anaerobic heterotrophic bacteria decreased exponentially with sediment depth and were below the detection limit (5–55 cells ml⁻¹) at 4–6 m, i.e., in about 8,900-year-old sediment. Absence of viable heterotrophic bacteria in deeper sediment layers demonstrated that aseptic sampling conditions were achieved. The decrease of viable spores with depth may be interpreted as time-dependent death of spores resulting in a death rate of about 0.0013–0.0025 year⁻¹. Viable units of specific metabolic groups of bacteria were detected only in the upper sediment layers (0–50 cm). Nitrifying bacteria could not be detected below 30 cm. Methane-oxidizing bacteria were present in the sediment down to >30 cm, but were in a dormant state. Nitrate reduction activity decreased by a factor of 6 within the upper 25 cm of the sediment, but was still detected at 50 cm. Sulfate reduction, on the other hand, could not be detected at depths of 20 cm and below. By contrast, methanogenesis and methanogenic bacteria could be detected down to 50 cm. These observations indicate that bacteria eventually become nonviable in aged sediments. Received: 5 March 1996; Accepted: 12 March 1996     

Microbial biomass and activity in subsurface sediments from Vejen,Denmark

Subsurface sediment samples were collected from 4 to 31 m below landsurface in glacio-fluvial sediments from the Quaternary period. The samples were described in terms of pH, electrical conductivity, chloride concentration, organic matter content, and grain size distribution. Viable counts of bacteria varied from 0.5 to 1,203 x 103 colony forming units/g dry weight (gdw); total numbers of bacteria acridine orange direct counts (AODC) varied from 1.7 to 147 × 10⁷ cells/gdw; growth rates (incorporation of [³H]-thymidine) varied from 1.4 to 60.7 × 10⁴ cells/(gdw · day); and rate constants for mineralization of ¹⁴C-labelled compounds varied from 0.2 to 2.3 × 10^–3 ml/(dpm · day) for acetate, and from 0 to 2.0 × 10^–3 ml/(dpm · day) for phenol. Sediment texture influenced the total number of bacteria and potential for mineralization; with increasing content of clay and silt and decreasing content of sand, AODC increased and the mineralization rate declined. Intrinsic permeability calculated from grain size correlated positively with mineralization rate for acetate. Statistical correlation analysis showed high correlations between some of the abiotic parameters, but it was not possible to point out a single abiotic parameter that could explain the variation of size and activity of the microbial population. The microbial data obtained in these geologically young sediments were compared to literature data from older sediments, and this comparison showed that age and type of geological formation might be important for the size and activity of the microbial populations.Offprint requests to: H.-J. Albrechtsen.