Microdistribution of sulfate-reducing bacteria in sediments of a hypertrophic lake and their response to the addition of organic matter

To clarify the ecological significance of the association of sulfate-reducing bacteria (SRB) with sediment particle size, SRB utilizing lactate (l-SRB), propionate (p-SRB) and acetate (a-SRB) were examined with different sizes of sediment particles in a hypertrophic freshwater lake using the anaerobic plate count method. The numbers ofl-SRB anda-SRB were 10⁴–10⁵ colony forming units (CFU) per ml in the 0–3 cm layer and 10²–10³ CFU ml⁻¹ in the 10–13 cm layer while the numbers ofp-SRB were one or two orders lower than those ofl-SRB anda-SRB. A sediment suspension was fractionated into four fractions (<1, 1–10, 10–94 and >94 μm). The highest proportions ofl-SRB anda-SRB were found in the 10–94 μm fraction: 66–97% forl-SRB and 53–98% fora-SRB. The highest proportion ofp-SRB was found in the >94 μm fraction (70–74%). These results indicate that most SRB were associated with sediment particles. One isolate from an acetate-utilizing enrichment culture was similar toDesulfotomaculum acetoxidans, a spore-forming sulfate-reducing bacterium. When lactate and sulfate were added to sediment samples,l-SRB anda-SRB in the <10 μm-fraction grew more rapidly than those in whole sediment for the first 2 days. This result suggests that nutrients uptake by free-living and small particle-associated (<10 μm) SRB is higher than that by SRB associated with larger particles.     

Kinetics of sulfate respiration by free-living and particle-associated sulfate-reducing bacteria

Abstract Effects on sulfate respiration of association of sulfate-reducing bacteria (SRB) with solid particles (anion exchange resin and FeS-precipitate) were examined using Desulfovibrio desulfuricans . The rates of sulfide production by resin- and FeS-associated cells were 2–3% and 19–56% of that by free-living ones, respectively, under sulfate- and lactate-rich conditions. On the other hand, under sulfate-poor (less than 50 μM) and lactate-rich conditions the rate by FeS-associated cells was higher than that by free-living ones. The values of K _m (μM), half saturation constant of the Michaelis-Menten model, for sulfate were 244 for free-living cells, 8.96 for resin-associated ones and 8.42 for FeS-associated ones. Under lactate-poor and sulfate-rich conditions the rate by FeS-associated cells was similar to that by free-living ones. These results suggest that FeS-associated SRB are more advantageous than free-living ones under sulfate-poor environments such as freshwater sediments.     

Microbial consumption of dimethyl sulfide and methanethiol in coastal marine sediments

Abstract: Samples were taken from oxic and anoxic zones of three ecosystems: a cyanobacterial mat, a diatom film and a carbonate sediment. Dimethylsulfide (DMS) concentrations were determined by headspace analysis of sediment slurries; maximal amounts were in the upper 5–10 mm of the sediments of 20 μM (cyanobacterial mat), 8 μM (diatom film) and < 1 μM in the carbonate sediment. Dissolved DMS in the cyanobacterial mat, determined by centrifugation and cryogenic trapping, was about two orders of magnitude lower than from slurry estimations but its variation with depth was similar. CH₃SH concentrations in slurried samples, determined after treatment with tributylphosphine, ranged from 2 to 7 μM in the diatom mat and was below the limit of detection (< 0.1 μM) in the carbonate sediment. MPN counts of bacteria that grew on DMS under oxic and anoxic (nitrate added) conditions were determined at all three sites. Aerobic DMS utilizers peaked in the surface and decreased with depth, while the population of anaerobic DMS utilizers was relatively constant in the top 20 mm. Populations of DMS utilizers were highest in the cyanobacterial mat and lowest in the carbonate sediment. MPN's of thiosulfate utilizers, aerobic and anaerobic (nitrate added) were determined in the cyanobacterial mat. Populations of aerobic and anaerobic S₂O₃²⁻ utilizers were similar throughout the top 20 mm and comparable to those of DMS utilizers in the top 5 mm, but higher by about 100-fold below that zone. DMS and CH₃SH consumption rates were measured in slurries of sediments and aerobic rates were similar or only slightly higher than anaerobic rates; the latter were stimulated by nitrate.     

Identity and abundance of active sulfate-reducing bacteria in deep tidal flat sediments determined by directed cultivation and CARD-FISH analysis

The identity and abundance of potentially active sulfate-reducing bacteria (SRB) in several metre deep sediments of a tidal sand flat in the German Wadden Sea were assessed by directed cultivation and cultivation-independent CARD-FISH analysis (catalysed reporter deposition fluorescence in situ hybridization). Presumably abundant SRB from different sediment layers between 0.5 and 4 m depth were selectively enriched in up to million-fold diluted cultures supplemented with lactate, acetate or hydrogen. Partial 16S rRNA gene sequences obtained from highest dilution steps showing sulfide formation indicated growth of deltaproteobacterial SRB belonging to the Desulfobulbaceae and the Desulfobacteraceae as well as of members of the Firmicutes. Subsequent isolation resulted in 10 novel phylotypes of both litho- and organotrophic sulfate-reducing Deltaproteobacteria. Molecular pre-screening identified six isolates as members of the Desulfobulbaceae, sharing highest identities with either candidatus 'Desulfobacterium corrodens' (95-97%) or Desulfobacterium catecholicum (98%), and four isolates as members of Desulfobacteraceae, being related to either Desulfobacter psychrotolerans (98%) or Desulfobacula phenolica (95-97%). Relatives of D. phenolica were exlusively isolated from 50 and 100 cm deep sediments with 10 and 2 mM of pore water sulfate respectively. In contrast, relatives of D. corrodens, D. psychrotolerans and D. catecholicum were also obtained from layers deeper than 100 cm and with less than 2 mM sulfate. The high in situ abundance of members of both families in sediment layers beneath 50 cm could be confirmed via CARD-FISH analysis performed with a set of six SRB-specific oligonucleotide probes. Moreover, SRB represented a numerically significant fraction of the microbial community throughout the sediment (up to 7%) and reached even higher cell numbers in deep, sulfate-poor layers than in the sulfate-rich surface sediment. This relatively large community size of potentially active SRB in deep sandy sediments might on the one hand be a result of their syntrophic association with other anaerobes. Our results furthermore support the hypothesis that enhanced advective pore water transport might supply nutrients to microbial communities in deep sandy sediments and point to their so far unrecognized contribution to biogeochemical processes in Wadden Sea sediments.     

Detection of abundant sulphate-reducing bacteria in marine oxic sediment layers by a combined cultivation and molecular approach

The depth distribution and diversity of sulphate-reducing bacteria (SRB) was analysed in the upper intertidal zone of a sandy marine sediment of the Dutch island Schiermonnikoog. The upper centimetre of the sediment included the oxic-anoxic interface and was cut into five slices. With each slice, most probable number (MPN) dilution series were set up in microtitre plates using five different substrates. In the deeper sediment layers, up to 1 x 10(8) cm(-3) lactate-utilizing SRB were counted, corresponding to 23% of the total bacterial count. From the highest positive dilutions of the MPN series, 27 strains of SRB were isolated in pure culture. Sequencing of a 580 bp fragment of the 16S rDNA revealed that 21 isolates had identical sequences, also identical with that of the previously described species Desulfomicrobium apsheronum. However, the diversity of the isolates was higher with respect to their physiological properties: a total of 11 different phenotypes could be distinguished. Genomic fingerprinting by enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) revealed an even higher diversity of 22 different genotypes. A culture-independent analysis by PCR and denaturing-gradient gel electrophoresis (DGGE) revealed that the partial 16S rDNA sequence of the isolated D. apsheronum strains constituted a significant fraction of the Desulfovibrionaceae. The high subspecies diversity suggests that this abundant aggregate-forming species may have evolved adaptations to different ecological niches in the oxic sediment layers.     

Major substrates for microbial sulfate reduction in the sediments of Ise Bay, Japan

To clarify the anaerobic microbial interactions in the process of carbon mineralization in marine eutrophic environments, the microbial sulfate reduction and methane production rates were examined in coastal marine sediments of Ise Bay, Japan, in autumn 1990. Sulfate reduction rates (51–210 nmol ml⁻¹ day⁻¹ at 24°C) were much higher than the methane production ones (<1.78 nmol ml⁻¹ day⁻¹) in the surface sediments (top 2 cm) at the six stations surveyed (water depth: 10.7–23.3 m). Substrates for sulfate-reducing bacteria (SRB) were estimated after the addition of a specific inhibitor for SRB (20 mmol l⁻¹ molybdate) into the sediment slurry, from the substrate accumulation rates. In the presence of the inhibitor, sulfate reduction was completely stopped and volatile fatty acids (mainly acetate) were accumulated, although hydrogen was not. Methane production occurred markedly accompanied by consumption of the accumulated acetate from the third day after the addition of molybdate. The maximum rate of methane production was 1.2–1.9 μmol ml⁻¹ day⁻¹, which was similar to those in highly polluted freshwater sediments such as the Tama River, Tokyo, Japan. These results show that acetate is a common major substrate for sulfate reduction and methane production, and SRB competitively inhibit potential acetoclastic methanogenesis in coastal sediments. Methanogens may potentially inhabit the sediments at low levels of population density and activity.     

Sulfur Cycling in the Terrestrial Subsurface: Commensal Interactions, Spatial Scales, and Microbial Heterogeneity

Abstract Microbiological, geochemical, and isotopic analyses of sediment and water samples from the unconsolidated Yegua formation in east-central Texas were used to assess microbial processes in the terrestrial subsurface. Previous geochemical studies suggested that sulfide oxidation at shallow depths may provide sulfate for sulfate-reducing bacteria (SRB) in deeper aquifer formations. The present study further examines this possibility, and provides a more detailed evaluation of the relationship between microbial activity, lithology, and the geochemical environment on meter-to-millimeter scales. Sediment of varied lithology (sands, silts, clays, lignite) was collected from two boreholes, to depths of 30 m. Our findings suggest that pyrite oxidation strongly influences the geochemical environment in shallow sediments (∼5 m), and produces acidic waters (pH 3.8) that are rich in sulfate (28 mM) and ferrous iron (0.3 mM). Sulfur and iron-oxidizing bacteria are readily detected in shallow sediments; they likely play an indirect role in pyrite oxidation. In consistent fashion, there is a relative paucity of pyrite in shallow sediments and a low ³⁴S/³²S-sulfate ratio (0.2‰) (reflecting contributions from ³⁴S-depleted sulfides) in shallow regions. Pyrite oxidation likely provides a sulfate source for both oxic and anoxic aquifers in the region. A variety of assays and direct-imaging techniques of ³⁵S-sulfide production in sediment cores indicates that sulfate reduction occurs in both the oxidizing and reducing portions of the sediment profile, with a high degree of spatial variability. Narrow zones of activity were detected in sands that were juxtaposed to clay or lignite-rich sediments. The fermentation of organic matter in the lignite-rich laminae provides small molecular weight organic acids to support sulfate reduction in neighboring sands. Consequently, sulfur cycling in shallow sediments, and sulfate transport represent important mechanisms for commensal interaction among subsurface microorganisms by providing electron donors for chemoautotrophic bacteria and electron acceptors for SRB. The activity of SRB is linked to the availability of suitable electron donors from spatially distinct zones. Received: 10 November 1997; Accepted: 10 February 1998     

The distribution of chlorophylls, carotenoids and their breakdown products in Lake Kinneret (Israel) sediments

SUMMARY. 1. The sedimentary distribution of chlorophylls, carotenoids and their breakdown products following the winter-spring bloom of the dinoflagellate Peridinium gatunense in Lake Kinneret was determined both spatially and vertically within the sediments, using high-pressure liquid chromatography (HPLC).
2. All four stations (epilimnic, oxic St. J (depth = 8m) and D (12m) and hypolimnic, anoxic St. F (21 m) A (41 m) were characterized by the presence of chlorophyll a (chl-a) chl- b , chl- c , phaeophytins a (ph'tin- a ) and b , phaeophorbide a (ph'bide- a ) and lutein, fucoxanthin, diatoxanthin and β-carotene together with spectrally similar, but HPLC-resolved, breakdown products in the surface sediments. The central deep station A (41 m), which is seasonally anoxic, trapped and preserved significantly more pigments (e.g. chl- a = 16.35 μg g⁻¹ organic matter) than the more erosive, oxic littoral stations (chl- a = 1.10 and 0.76μg g^−l OM at St. D and J, respectively).
3. Whereas it was possible to ascribe the presence of lutein and chl- b to inputs of green algae and higher plants from the catchment, and fucoxanthin and chl- c ₁, c ₂ to winter diatom blooms (e.g. Melosira granulata ), there was a remarkable absence of peridinin and dinoxanthin from sediments, despite the dominance of dinoflagellates in the phytobiomass. This is consistent with the well-known oxic lability of this epoxy carotenoid in aquatic systems, and indicates efficient breakdown of this dinoflagellate organic matter in the water column, prior to sedimentation.
4. Although pigment concentrations all decreased with depths within sediments, lutein and β-carotene were selectively preserved compared to chl-a and fucoxanthin which, at 40m depth at St. A, decreased to < 1% of their surface concentrations.     

Detection and Enumeration of Sulphate-Reducing Bacteria in Estuarine Sediments by Competitive PCR

The distribution of sulphate-reducing bacteria (SRB) in the sediments of the Colne River estuary, Essex, UK covering different saline concentrations of sediment porewater was investigated by the use of quantitative competitive PCR. Here, we show that a new PCR primer set and a new quantitative method using PCR are useful tools for the detection and the enumeration of SRB in natural environments. A PCR primer set selective for the dissimilatory sulphite reductase gene (dsr) of SRB was designed. PCR amplification using the single set of dsr-specific primers resulted in PCR products of the expected size from all 27 SRB strains tested, including Gram-negative and positive species. Sixty clones derived from sediment DNA using the primers were sequenced and all were closely related with the predicted dsr of SRB. These results indicate that PCR using the newly designed primer set are useful for the selective detection of SRB from a natural sample. This primer set was used to estimate cell numbers by dsr selective competitive PCR using a competitor, which was about 20% shorter than the targeted region of dsr. This procedure was applied to sediment samples from the River Colne estuary, Essex, UK together with simultaneous measurement of in situ rates of sulphate reduction. High densities of SRB ranging from 0.2 ? 5.7 × 10⁸ cells ml^{? 1} wet sediment were estimated by the competitive PCR assuming that all SRB have a single copy of dsr. Using these estimates cell specific sulphate reduction rates of 10^{? 17} to 10^{? 15} mol of SO₄ ^{2 ?} cell^{? 1} day^{? 1} were calculated, which is within the range of, or lower than, those previously reported for pure cultures of SRB. Our results show that the newly developed competitive PCR technique targeted to dsr is a powerful tool for rapid and reproducible estimation of SRB numbers in situ and is superior to the use of culture-dependent techniques.     

Biochemical characteristics of surface sediments on the eastern Weddell Sea continental shelf, Antarctica: is there any evidence of seasonal patterns?

Biochemical characteristics of seafloor sediment off Austasen in the southeastern Weddell Sea were assayed in samples recovered in the early autumn and late spring of 2000 and 2003, respectively. Sediment was separated in the grain-size fractions >200 μm and <200 μm to distinguish biochemical characteristics in the fraction available for benthic suspension feeders (<200 μm). In the bulk sediment, the lipid (LPD) and carbohydrate (CHO) contents were significantly different between seasons with higher LPD content in the early autumn and higher CHO content in the late spring. In the grain-size fractions <200 μm, the LPD and protein (PRT) contents were significantly higher in the early autumn meaning that in this season the fraction available for benthic suspension feeders presented higher nutritive value. The relatively higher CHO concentrations observed in each fraction in the late spring were attributed to refractory matter, whereas the higher PRT and LPD concentrations found during the early autumn were associated with planktonic material settled after the summer phytoplankton bloom. Our results suggest that there is seasonal variation in the composition of organic matter in the sediment, with better nutritive quality in the early autumn, especially in the grain-size fraction available for benthic suspension feeders. These variations also suggest that the benthic community exploits the fresh organic matter accumulated after the summer throughout the Antarctic dark months leaving the sediment almost exhaust of LPD and with higher CHO contents, presumably of refractory nature, at the onset of the seasonal phytoplankton bloom of the following year.     

Microbial Ecosystem in Sunderban Mangrove Forest Sediment,North-East Coast of Bay of Bengal,India

This is the first documentation of seasonal and spatial fluctuation of the culturable microbial population collected from different zones in the sediment of the Sunderban mangrove forest. The population of cellulose degrading bacteria, [mean value of CFU 6.189 ± 1.025 × 10⁶ (g dry weight of sediment)^?1] was found to be maximum during post monsoon in the deep forest region, whereas, the fungal population [mean value of CFU 3.424 ± 0.886 × 10⁶ (g dry weight of sediment)^?1] was found to be maximum during pre-monsoon in the rooted region. The abundances of microbes, in decreasing order, studied from different zones are nitrifying bacteria [mean value of CFU 1.125 ± 0.359 × 10⁶ (g dry weight of sediment)^?1], phosphorous solubilizing bacteria (PSB) [mean value of CFU 0.805 ± 0.322 × 10⁶ (g dry weight of sediment)^?1], free living nitrogen fixing bacteria [mean value of CFU 0.417 ± 0.120 × 10⁶ (g dry weight of sediment)^?1] and sulfur reducing bacteria (SRB) [mean value of CFU 0.356 ± 0.125 × 10⁶ (g dry weight of sediment)^?1]. The content of organic carbon in the soil decreased from the deep forest region to the rooted and unrooted region but a reverse profile was found for soil salinity and soil silicate concentration. The results from the present study indicate that the monsoon cycle has a pronounced effect on the microbially dominated biogeochemistry in the sediment and consequently on the ecology of the Sundarban mangrove forest.     

Quantification of Sulfate-reducing Bacteria in Industrial Wastewater,by Real-time Polymerase Chain Reaction (PCR) Using <Emphasis Type="Italic">dsrA</Emphasis> and <Emphasis Type="Italic">apsA</Emphasis> Genes

Real-time polymerase chain reaction (PCR) is considered a highly sensitive method for the quantification of microbial organisms in environmental samples. This study was conducted to evaluate real-time PCR with SybrGreen detection as a quantification method for sulfate-reducing bacteria (SRB) in industrial wastewater produced by several chemical industries. We designed four sets of primers and developed standard curves based on genomic DNA of Desulfovibrio vulgaris from pure culture and on plasmids containing dissimilatory sulfate reductase (dsrA) or adenosine-5′-phosphosulfate reductase (apsA) genes of SRB. All the standard curves, two for dsrA and two for apsA genes, had a linear range between 0.95 × 10² and 9.5 × 10⁶ copies/μL and between 1.2 × 10³ and 1.2 × 10⁷ copies/μL, respectively. The theoretical copy numbers of the tenfold dilutions of D. vulgaris genomic DNA were best estimated (between 2.7 to 10.5 times higher than theoretical numbers) by the standard curve with DSR1F and RH3-dsr-R primers. To mimic the effect of foreign DNA in environmental samples, serial dilutions of D. vulgaris genomic DNA were mixed with Escherichia coli chromosomal DNA (40 ng per assay). This influenced neither PCR amplification nor the quantification of target DNA. Industrial wastewater was sampled during a 15-month period and analyzed for the presence of SRB, based on dsrA gene amplification. SRB displayed a higher abundance during the summer (about 10⁷–10⁸ targets mL⁻¹) and lower during the winter (about 10⁴–10⁵ targets mL⁻¹). The results indicate that our real-time PCR approach can be used for detection of uncultured SRB and will provide valuable information related to the abundance of SRB in durable environmental samples, such as complex and saline industrial wastewaters.     

Sulphate reduction in oxic and sub-oxic North-East Atlantic sediments

Abstract Oxic and sub-oxic N.-E. Atlantic sediments were examined for sulphate-reducing activity. Oxygen and/or nitrate reduction are probably the dominant mineralisation processes in the abyssal plain sediment studied. A low rate of sulphate reduction (0.1 nmol SO²⁻₄/ml/day) was recorded in the surface 5 cm of the continental slope sediment, together with the presence of a range of sulphate-reducing bacteria (SRB). A higher activity of sulphate reduction (2.2 nmol SO²⁻₄/ml/day) occurred in the continental shelf sediment which led to a small decrease in pore water sulphate and an increase in titration alkalinity. This sediment contained approx. 10²–10³ acetate, lactate and propionate oxidising SRB/ml. No low- M _r organic acids were detected in these sediments. However, amendment with 75 μM acetate stimulated sulphate-reducing activity in the shelf sediment.     

Successional development of sulfate-reducing bacterial populations and their activities in an activated sludge immobilized agar gel film

A combination of fluorescence in situ hybridization (FISH), microprofiles, and denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rDNA fragments followed by hybridization analysis with specific probes was applied to investigate successional development of sulfate-reducing bacteria (SRB) community structure and in situ sulfide production activity within an activated sludge immobilized agar gel film. In this model biofilm system, since biases arising from biofilm heterogeneity can be ignored, the population dynamics of SRB in the agar gel is directly related to physiological capability and in situ activity of SRB. Microelectrode measurements showed that an anoxic zone was already developed at the beginning (0 day), a first sulfide production of 0.054 mumol H2S m(-2) x s(-1) was detected during the first week, and the rate increased gradually to 0.221 mumol H2S m(-2) x s(-1) in the fifth week. The most active sulfide production zone moved upward to the chemocline and intensified with time to form a narrow zone with high volumetric sulfide production rates. This result coincided with the shift of the spatial distributions of SRB populations determined by FISH. In situ hybridization with probe SRB385 for mainly general SRB of the delta Proteobacteria plus some gram-positive bacteria and probe 660 for Desulfobulbus indicated that the most abundant populations of SRB were primarily restricted to near the oxic/anoxic interface (chemocline). A close observation of the development of the vertical distributions of SRB populations revealed that the cell numbers of Desulfobulbus tripled (from 0.5 x 10(8) to 1.5 x 10(8) cells cm(-3)) near the oxic/anoxic interface. Similar growth (from 1.0 x10(8) to 4.5 x 10(8) cells cm(-3)) of Desulfovibrio-like SRB that hybridized with probe SRB385 was observed. PCR-DGGE followed by hybridization analysis revealed that one Desulfobulbus strain was detected from the beginning, and another strain appeared after 1 week, coinciding with the first detected sulfide production. In addition, three strains hybridizing with probe 687 (possibly Desulfovibrio) were also dominant SRB in the agar gel.     

Analyses of Spatial Distributions of Sulfate-Reducing Bacteria and Their Activity in Aerobic Wastewater Biofilms

The vertical distribution of sulfate-reducing bacteria (SRB) in aerobic wastewater biofilms grown on rotating disk reactors was investigated by fluorescent in situ hybridization (FISH) with 16S rRNA-targeted oligonucleotide probes. To correlate the vertical distribution of SRB populations with their activity, the microprofiles of O₂, H₂S, NO₂⁻, NO₃⁻, NH₄⁺, and pH were measured with microelectrodes. In addition, a cross-evaluation of the FISH and microelectrode analyses was performed by comparing them with culture-based approaches and biogeochemical measurements. In situ hybridization revealed that a relatively high abundance of the probe SRB385-stained cells (approximately 10⁹ to 10¹⁰ cells per cm³ of biofilm) were evenly distributed throughout the biofilm, even in the oxic surface. The probe SRB660-stained Desulfobulbus spp. were found to be numerically important members of SRB populations (approximately 10⁸ to 10⁹ cells per cm³). The result of microelectrode measurements showed that a high sulfate-reducing activity was found in a narrow anaerobic zone located about 150 to 300 μm below the biofilm surface and above which an intensive sulfide oxidation zone was found. The biogeochemical measurements showed that elemental sulfur (S⁰) was an important intermediate of the sulfide reoxidation in such thin wastewater biofilms (approximately 1,500 μm), which accounted for about 75% of the total S pool in the biofilm. The contribution of an internal Fe-sulfur cycle to the overall sulfur cycle in aerobic wastewater biofilms was insignificant (less than 1%) due to the relatively high sulfate reduction rate.     

Growth characteristics of small and large free-living and attached bacteria in Lake Constance

The growth characteristics of small (0.2–1.0m) and large (1.0–3.0 (m) free-living and attached bacteria were studied in Lake Constance by comparing the spatial and seasonal dynamics of their biomass turnover time (ratio of biomass/production). The biomass of small free-living bacteria usually turned over significantly faster than that of large free-living bacteria throughout the water column. The turnover of attached bacterial biomass was characterized by large fluctuations. Occasionally, in aphotic water layers, it was as long as that of large free-living bacteria, but when large amounts of decaying organic particles were present, it was shorter than that of small free-living cells. Biomass turnover times of free-living bacteria were in the same range as their generation times, which were estimated from the increase in bacterial abundance in 3m prefiltered samples. The biomass turnover time of actively metabolizing bacteria was comparable to the generation time of actively metabolizing cells. These results indicate that the biomass turnover time is a useful indicator of the growth of different bacterial fractions, as it reflects their different amounts of participation in microbial processes of aquatic ecosystems.     

Phototrophic sulfur bacteria and sulfate-reducing bacteria causing red waters in a shallow brackish coastal lagoon (Prévost Lagoon, France)

Abstract In the shallow lagoon of Prévost (43°30'N, 3°54'E; French Mediterranean coast), red waters occurring periodically during warm summers are a consequence of a succession of ecological events beginning in the early spring with a bloom of algae ( Ulva lactuca ). In summer 1977, a red water was analyzed; in the early summer, the water turned anoxic and became rich in sulfide which originated from sulfate reduction in the first 10 cm of the sediment. Numbers of both phototrophic and sulfate-reducing bacteria (SRB) increased during spring and summer, and the genera in the prevailing populations did not change: Thiocapsa (80%) among the phototrophic bacteria and Desulfovibrio and Desulfobacter among the SRB. They were also dominant during the period of red waters. A few Chromatium and Thiocystis species were also identified. During red water periods, these bacteria grew very actively, removing all the sulfide produced by SRB, and accumulated in the whole water column. Consequently, the sulfate level increased to 5 mmol·1⁻¹ higher than the theoretical sulfate level calculated from salinity, showing the active oxidation of sulfide by phototrophic bacteria. After the dystrophic crisis, oxic conditions were reestablished and the phototrophic bacterial biomass was partly grazed by zoobenthos organisms which densely populated the sediment surface.     

Competition between anoxygenic phototrophic bacteria and colorless sulfur bacteria in a microbial mat

Abstract The populations of chemolithoautotrophic (colorless) sulfur bacteria and anoxygenic phototrophic bacteria were enumerated in a marine microbial mat. The highest population densities were found in the 0–5 mm layer of the mat: 2.0 × 10⁹ cells cm⁻³ sediment, and 4.0 × 10⁷ cells cm⁻³ sediment for the colorless sulfur bacteria and phototrophs, respectively. Kinetic parameters for thiosulfate-limited growth were assessed for Thiobacillus thioparus T5 and Thiocapsa roseopersicina M1, both isolated from microbial mats. For Thiobacillus T5, growing at a constant oxygen concentration of 43 μmol l⁻¹, μ_max was 0.336 h⁻¹ and K _s 0.8 μmol l⁻¹. Phototrophically grown Thiocapsa strain M1 displayed a μ_max of 0.080 h⁻¹ and a K _s of 8 μmol l⁻¹ when anoxically grown under thiosulfate limitation. In a competition experiment with thiosulfate as electron donor, Thiocapsa became dominant during a 10-h oxic/14-h anoxic regimen at continuous illumination, despite the higher affinity for thiosulfate of Thiobacillus .     

Microbial community structure and biomass estimates of a methanogenic Antarctic Lake ecosystem as determined by phospholipid analyses

Phospholipid analyses were performed on water column particulate and sediment samples from Ace Lake, a meromictic lake in the Vestfold Hills, Antarctica, to estimate the viable microbial biomass and community structure in the lake. In the water column, methanogenic bacterial phospholipids were present below 17 m in depth at concentrations which converted to a biomass of between 1 and 7×10⁸ cells/liter. Methanogenic biomass in the sediment ranged from 17.7×10⁹ cells/g dry weight of sediment at the surface to 0.1×10⁹ cells/g dry weight at 2 m in depth. This relatively high methanogenic biomass implies that current microbial degradation of organic carbon in Ace Lake sediments may occur at extremely slow rates. Total microbial biomass increased from 4.4×10⁸ cells/ liter at 2 m in depth to 19.4×10⁸ cells/liter at 23 m, near the bottom of the water column. Total nonarchaebacterial biomass decreased from 4.2 ×10⁹ cells/g dry weight in the surface sediment (1/4 the biomass of methanogens) to 0.06×10⁸ cells/g dry weight at 2 m in depth in the sediment. Phospholipid fatty acid profiles showed that microeukaryotes were the major microbial group present in the oxylimnion of the lake, while bacteria dominated the lower, anoxic zone. Sulfate-reducing bacteria (SRB) comprised 25% of the microbial population at 23 m in depth in the water column particulates and were present in the surface sediment but to a lesser extent. Biomass estimates and community structure of the Ace Lake eco-system are discussed in relation to previously measured metabolic rates for this and other antarctic and temperate ecosystems. This is the first instance, to our knowledge, in which the viable biomass of methanogenic and SRB have been estimated for an antarctic microbial community.     

Inhibiting sulfate-reducing bacteria in biofilms by expressing the antimicrobial peptides indolicidin and bactenecin

To identify novel, less-toxic compounds capable of inhibiting sulfate-reducing bacteria (SRB), Desulfovibrio vulgaris and Desulfovibrio gigas in suspension cultures were exposed to several antimicrobial peptides. The bacterial peptide antimicrobials gramicidin S, gramicidin D, and polymyxin B as well as the cationic peptides indolicidin and bactenecin from bovine neutrophils decreased the viability of both SRB by 90% after a 1-h exposure at concentrations of 25–100 μg ml⁻¹. To reduce corrosion by inhibiting SRB in biofilms, the genes for indolicidin and bactenecin were expressed in Bacillus subtilisBE1500 and B. subtilis WB600 under the control of the constitutive alkaline protease (apr) promoter, and the antimicrobials were secreted into the culture medium using the apr signal sequence. Bactenecin was also synthesized and expressed as a fusion to the pro-region of barnase from Bacillus amyloliquefaciens. Concentrated culture supernatants of B. subtilis BE1500 expressing bactenecin at 3 μg ml⁻¹ decreased the viability of Escherichia coli BK6 by 90% and the reference SRB D. vulgaris by 83% in suspension cultures. B. subtilis BE1500 and B. subtilis WB600 expressing bactenecin in biofilms also inhibited the SRB-induced corrosion of 304 stainless steel six to 12-fold in continuous reactors as evidenced by the lack of change in the impedance spectra (resistance polarization) upon addition of SRB and by the reduction in hydrogen sulfide and iron sulfide in batch fermentations with mild steel. A 36-fold decrease in the population of D. vulgaris in a B. subtilis BE1500 biofilm expressing bactenecin was also observed. This is the first report of an antimicrobial produced in a biofilm for in vivo applications and represents the first application of a beneficial, genetically-engineered biofilm for combating corrosion. Received 27 October 1998/ Accepted in revised form 21 February 1999