Attached and Unattached Bacterial Communities in a 120-Meter Corehole in an Acidic, Crystalline Rock Aquifer

The bacteria colonizing geologic core sections (attached) were contrasted with those found suspended in the groundwater (unattached) by examining the microbiology of 16 depth-paired core and groundwater samples using a suite of culture-independent and culture-dependent analyses. One hundred twenty-two meters was continuously cored from a buried chalcopyrite ore hosted in a biotite-quartz-monzonite porphyry at the Mineral Park Mine near Kingman, Ariz. Every fourth 1.5-m core was acquired using microbiologically defensible methods, and these core sections were aseptically processed for characterization of the attached bacteria. Groundwater samples containing unattached bacteria were collected from the uncased corehole at depth intervals corresponding to the individual cores using an inflatable straddle packer sampler. The groundwater was acidic (pH 2.8 to 5.0), with low levels of dissolved oxygen and high concentrations of sulfate and metals, including ferrous iron. Total numbers of attached cells were less than 10⁵ cells g of core material⁻¹ while unattached cells numbered about 10⁵ cells ml of groundwater⁻¹. Attached and unattached acidophilic heterotrophs were observed throughout the depth profile. In contrast, acidophilic chemolithotrophs were not found attached to the rock but were commonly observed in the groundwater. Attached communities were composed of low numbers (<40 CFU g⁻¹) of neutrophilic heterotrophs that exhibited a high degree of morphologic diversity, while unattached communities contained higher numbers (ca. 10³ CFU ml⁻¹) of neutrophilic heterotrophs of limited diversity. Sulfate-reducing bacteria were restricted to the deepest samples of both core and groundwater. 16S ribosomal DNA sequence analysis of attached, acidophilic isolates indicated that organisms closely related to heterotrophic, acidophilic mesophiles such as Acidiphilium organovorum and, surprisingly, to the moderately thermophilic Alicyclobacillus acidocaldarius were present. The results indicate that viable (but possibly inactive) microorganisms were present in the buried ore and that there was substantial distinction in biomass and physiological capabilities between attached and unattached populations.     

Ontogenetic Study of the Effects of Energetic Nutrients on Amino Acid Metabolism of Rat Cerebral Cortex

We studied the effect of various energetic nutrients on metabolism of l-[U-¹⁴C]leucine and [1–¹⁴C]glycine in cerebral cortex of rats at different ages. At gestational age, glucose and lactate stimulated protein synthesis from l-[U-¹⁴C]leucine and [1–¹⁴C]glycine and from l-[U-¹⁴C]leucine, respectively; glucose, -OH-butyrate and lactate stimulated lipid synthesis from l-[U-¹⁴C]leucine. At 10 days of age, glucose, mannose, and fructose stimulated protein synthesis, and glucose and mannose stimulated oxidation to CO₂ as well as lipid synthesis from l-[U-¹⁴C]leucine. In adult rats, glucose, mannose, and fructose stimulated protein synthesis from l-[U-¹⁴C]leucine and [1–¹⁴C]glycine; glutamine also markedly decreased the oxidation of l-[U-¹⁴C]leucine and [1–¹⁴C]glycine in 10–day-old and adult rats.     

Subsurface denitrification in a forest riparianzone: Interactions between hydrology and supplies ofnitrate and organic carbon

The influence of hydrology andpatterns of supply of electron donors and acceptors onsubsurface denitrification was studied in a forestriparian zone along the Boyne River in southernOntario that received high nitrogen inputs from a sandaquifer. Two hypotheses were tested: (1) subsurfacedenitrification is restricted to localized zones ofhigh activity; (2) denitrification zones occur atsites where groundwater flow paths transportNO₃ ^– to supplies of available organiccarbon. A plume of nitrate-rich groundwater withconcentrations of 10–30 mg N L^–1 flowed laterallyat depths of 1.5–5 m in sands beneath peat for ahorizontal distance of 100–140 m across the riparianzone to within 30–50 m of the river. In situ acetyleneinjections to piezometers revealed that significantdenitrification was restricted to a narrow zone ofsteep NO₃ ^– and N₂O decline at theplume margins. The location of these denitrificationsites in areas with steep gradients of groundwater DOCincrease supported hypothesis 2. Many of thesedenitrification hotspots occurred near interfacesbetween sands and either peats or buried river channeldeposits. Field experiments involving in situadditions of either glucose or NO₃ ^– topiezometers indicated that denitrification wasC-limited in a large subsurface area of the riparianzone, and became N-limited beyond the narrow zone ofNO₃ ^– consumption. These data suggest thatdenitrification may not effectively removeNO₃ ^– from groundwater transported at depththrough permeable riparian sediments unlessinteraction occurs with localized supplies of organicmatter.     

Grazing of attached bacteria by heterotrophic microflagellates

Four species of heterotrophic microflagellates were examined for their ability to graze attached and unattached bacteria. The species tested displayed pronounced differences in their ability to graze the bacteriumPseudomonas halodurans attached to chitin particles. Two species of microflagellates (Monas andCryptobia sp.) efficiently grazed unattached bacteria but showed little or no ability to graze attached or aggregated cells. In contrast,Rhynchomonas nasuta andBodo sp. showed marked preferences for attached and aggregated bacteria and a limited ability to graze unattached cells. The density of attached bacteria was reduced by an order of magnitude due to grazing byBodo andR. nasuta, even though the density of unattached bacteria was 5–90× the density of attached cells. The maximum densities attained by microflagellates in the cultures were related to the density of unattached bacteria forMonas andCryptobia but not forBodo andR. nasuta. Growth of the latter two species appeared to be related to the density of attached or aggregated bacteria. Based on the results of these experiments, it is concluded that the pelagic existence of microflagellates that graze attached bacteria may be strongly linked to the distribution of suspended particles and their associated bacteria. In addition, the removal of attached bacteria by microflagellates can significantly affect the density of bacteria attached to particles in the plankton. This activity may have important implications for the controversy concerning the relative importance of attached and free-living bacteria in the plankton.     

Activity Measurements of Planktonic Microbial and Microfouling Communities in a Eutrophic Estuary

[³H]thymidine incorporation, the rate of reduction of iodonitrotetrazolium violet (INT) to INT formazan normalized to DNA, and the ratio of ATP to DNA were adapted to measure the activity of attached and unattached microbial assemblages of Bayboro Harbor, Fla. Activity measurements by [³H]thymidine incorporation were made of cells attached to polystyrene culture dishes, in unfiltered water samples, and in the <1-μm-filtered fraction. In most cases, the activity of attached cells was greater than that of unattached cells either in unfiltered water samples or in the <1-μm fraction. The calculated thymidine incorporation rates for cells in the >1-μm fraction were higher than those for cells either in unfiltered water or in the <1-μm-filtered fraction. By the rate of reduction of INT to INT formazan normalized to DNA and by ATP-to-DNA ratios, attached cells were also more active than cells in unfiltered water samples. These results indicate that the microenvironment afforded by attachment is a more beneficial habitat for microbial growth. Reasons for greater activity by natural populations of attached bacteria are discussed.     

Uptake of15N labeled nitrate by root systems of sweet corn, carrot and white cabbage from 0.2–2.5 meters depth

Leaching of NO ₃ ^– from vegetable cropping systems can be very high compared to arable systems. This is a problem for vegetable growers in general as it decreases groundwater quality, and for organic growers in particular as the organic production is often limited by N. In a field experiment, we investigated the N uptake and root growth of three vegetables using minirhizotrons reaching 2.4 m with the purpose to study the relationship between vegetable root distribution and uptake of NO ₃ ^– from deep soil layers. NO ₃ ^– uptake was studied over a 6 d period at the end of September by injection of 15 NO ₃ ^– at four depths in the ranges: 0.2–0.8, 0.6–1.8, and 1–2.5 m under late sweet corn (Zea mays L. convar. Saccharata Koern.), carrot (Daucus carota L.), and autumn white cabbage (Brassica oleracea L. convar. capitata (L.) Alef. var. alba DC), respectively. The root depths of the three crops were 0.6, 1.3, and more than 2.4 m, respectively. Uptake of¹⁵N was close to zero from placements below root depth, and linear relationships were found between root density and¹⁵N uptake from different depths. N inflow rates (uptake per unit root length) were in the same range for all species and depths. This indicates that the very different N use efficiencies often found for vegetable crops depend on species specific differences in root development over time and space, more than on differences in N uptake ability of the single root. Thus deep rooting is important for deep N uptake. Knowledge about deep root growth enables design of crop rotations with improved N use efficiency based on re-cycling of deep soil NO ₃ ^– by vegetables.     

Attached and unattached bacterial communities in a 120-meter corehole in an acidic, crystalline rock aquifer

The bacteria colonizing geologic core sections (attached) were contrasted with those found suspended in the groundwater (unattached) by examining the microbiology of 16 depth-paired core and groundwater samples using a suite of culture-independent and culture-dependent analyses. One hundred twenty-two meters was continuously cored from a buried chalcopyrite ore hosted in a biotite-quartz-monzonite porphyry at the Mineral Park Mine near Kingman, Ariz. Every fourth 1.5-m core was acquired using microbiologically defensible methods, and these core sections were aseptically processed for characterization of the attached bacteria. Groundwater samples containing unattached bacteria were collected from the uncased corehole at depth intervals corresponding to the individual cores using an inflatable straddle packer sampler. The groundwater was acidic (pH 2.8 to 5.0), with low levels of dissolved oxygen and high concentrations of sulfate and metals, including ferrous iron. Total numbers of attached cells were less than 10(5) cells g of core material(-1) while unattached cells numbered about 10(5) cells ml of groundwater(-1). Attached and unattached acidophilic heterotrophs were observed throughout the depth profile. In contrast, acidophilic chemolithotrophs were not found attached to the rock but were commonly observed in the groundwater. Attached communities were composed of low numbers (<40 CFU g(-1)) of neutrophilic heterotrophs that exhibited a high degree of morphologic diversity, while unattached communities contained higher numbers (ca. 10(3) CFU ml(-1)) of neutrophilic heterotrophs of limited diversity. Sulfate-reducing bacteria were restricted to the deepest samples of both core and groundwater. 16S ribosomal DNA sequence analysis of attached, acidophilic isolates indicated that organisms closely related to heterotrophic, acidophilic mesophiles such as Acidiphilium organovorum and, surprisingly, to the moderately thermophilic Alicyclobacillus acidocaldarius were present. The results indicate that viable (but possibly inactive) microorganisms were present in the buried ore and that there was substantial distinction in biomass and physiological capabilities between attached and unattached populations.     

Fluxes of carbohydrate metabolism in ripening bananas

The major fluxes of carbohydrate metabolism were estimated during starch breakdown by ripening bananas (Musa cavendishii Lamb ex Paxton). Hands of bananas, untreated with ethylene, were allowed to ripen in the dark at 21° C. Production of CO₂ and the contents of starch, sucrose, glucose and fructose of intact fruit were determined for a period of 10 d that included the climacteric. The detailed distribution of label was determined after supplying the following to cores of pulp from climacteric fruit: [U-¹⁴C]-, [1-¹⁴C]-, [3,4-¹⁴C]-and [6-¹⁴C]glucose, [U-¹⁴C]glycerol, ¹⁴CO₂. The data obtained were used to estimate the following fluxes, values given as mol hexose · (g FW)^–1 · h^–1 in parenthesis: starch to hexose monophosphates (5.9) and vice versa (0.4); hexose monophosphates to sucrose (7.7); sucrose to hexose (4.7); hexose to hexose monophosphate (3.8); glycolysis (0.5–1.6); triose phosphate to hexose monophosphates (0.14); oxidative pentose-phosphate pathway (0.48); CO₂ fixation in the dark (0.005). These estimates are related to our understanding of carbohydrate metabolism during ripening.We both thank Mr Richard Trethewey for his constructive criticism: S.A.H. thanks the Managers of the Broodbank Fund for a fellowship.     

Enzymatic Combustion of Aromatic and Aliphatic Compounds by Manganese Peroxidase from Nematoloma frowardii

The direct involvement of manganese peroxidase (MnP) in the mineralization of natural and xenobiotic compounds was evaluated. A broad spectrum of aromatic substances were partially mineralized by the MnP system of the white rot fungus Nematoloma frowardii. The cell-free MnP system partially converted several aromatic compounds, including [U-¹⁴C]pentachlorophenol ([U-¹⁴C]PCP), [U-¹⁴C]catechol, [U-¹⁴C]tyrosine, [U-¹⁴C]tryptophan, [4,5,9,10-¹⁴C]pyrene, and [ring U-¹⁴C]2-amino-4,6-dinitrotoluene ([¹⁴C]2-AmDNT), to ¹⁴CO₂. Mineralization was dependent on the ratio of MnP activity to concentration of reduced glutathione (thiol-mediated oxidation), a finding which was demonstrated by using [¹⁴C]2-AmDNT as an example. At [¹⁴C]2-AmDNT concentrations ranging from 2 to 120 μM, the amount of released ¹⁴CO₂ was directly proportional to the concentration of [¹⁴C]2-AmDNT. The formation of highly polar products was also observed with [¹⁴C]2-AmDNT and [U-¹⁴C]PCP; these products were probably low-molecular-weight carboxylic acids. Among the aliphatic compounds tested, glyoxalate was mineralized to the greatest extent. Eighty-six percent of the ¹⁴COOH-glyoxalate and 9% of the ¹⁴CHO-glyoxalate were converted to ¹⁴CO₂, indicating that decarboxylation reactions may be the final step in MnP-catalyzed mineralization. The extracellular enzymatic combustion catalyzed by MnP could represent an important pathway for the formation of carbon dioxide from recalcitrant xenobiotic compounds and may also have general significance in the overall biodegradation of resistant natural macromolecules, such as lignins and humic substances.     

Responses of N fluxes and pools to elevated atmospheric CO2 in model forest ecosystems with acidic and calcareous soils

The objectives of this study were to estimate how soil type, elevated N deposition (0.7 vs. 7 g N m^–2y^–1) and tree species influence the potential effects of elevated CO₂ (370 vs. 570 mol CO₂ mol^–1) on N pools and fluxes in forest soils. Model spruce-beech forest ecosystems were established on a nutrient-rich calcareous sand and on a nutrient-poor acidic loam in large open-top chambers. In the fourth year of treatment, we measured N concentrations in the soil solution at different depths, estimated N accumulation by ion exchange resin (IER) bags, and quantified N export in drainage water, denitrification, and net N uptake by trees. Under elevated CO₂, concentrations of N in the soil solution were significantly reduced. In the nutrient-rich calcareous sand, CO₂ enrichment decreased N concentrations in the soil solution at all depths (–45 to –100%). In the nutrient-poor acidic loam, the negative CO₂ effect was restricted to the uppermost 5 cm of the soil. Increasing the N deposition stimulated the negative impact of CO₂ enrichment on soil solution N in the acidic loam at 5 cm depth from –20% at low N inputs to –70% at high N inputs. In the nutrient-rich calcareous sand, N additions did not influence the CO₂ effect on soil solution N. Accumulation of N by IER bags, which were installed under individual trees, was decreased at high CO₂ levels under spruce in both soil types. Under beech, this decrease occurred only in the calcareous sand. N accumulation by IER bags was negatively correlated with current-years foliage biomass, suggesting that the reduction of soil N availability indices was related to a CO₂-induced growth enhancement. However, the net N uptake by trees was not significantly increased by elevated CO₂. Thus, we suppose that the reduced N concentrations in the soil solution at elevated CO₂ concentrations were rather caused by an increased N immobilisation in the soil. Denitrification was not influenced by atmospheric CO₂ concentrations. CO₂ enrichment decreased nitrate leaching in drainage by 65%, which suggests that rising atmospheric CO₂ potentially increases the N retention capacity of forest ecosystems.     

Glucose and lactate utilization by the amygdala of male and female rats

Gonadal hormones appear to modulate brain energy metabolism, and morphological and functional sexual differences are found in the amygdaloid complex (AC) of rats. Our aim was to study the CO₂ production and lipid synthesis, measured by the rate of L-[U-¹⁴C]lactate or D-[U-¹⁴C]glucose utilization (in pmol.hr^–1.mg^–1), by AC slices in vitro of male and female rats. Lactate was more used than glucose as energy substrate (p < 0.01) but no sex-related difference was observed in glucose or lactate oxidation to CO₂ (p > 0.05) or on lipid synthesis obtained from both substrates (p > 0.05). In addition, there was no effect of the estrous cycle on lactate oxidation to CO₂ by the AC of females (p > 0.05). Based on the present data, it appears that the endogenous normal levels of gonadal hormones are not able to promote sex-related differences in the in vitro glucose or lactate utilization by the AC of rats.     

Soil and biomass carbon pools in model communities of tropical plants under elevated CO2

The experimental data presented here relate to the question of whether terrestrial ecosystems will sequester more C in their soils, litter and biomass as atmospheric CO₂ concentrations rise. Similar to our previous study with relatively fertile growth conditions (Körner and Arnone 1992), we constructed four rather nutrient-limited model communities of moist tropical plant species in greenhouses (approximately 7 m² each). Plant communities were composed of seven species (77 individuals per community) representing major taxonomic groups and various life forms found in the moist tropics. Two ecosystems were exposed to 340 l CO₂ l^–1 and two to 610 l l^–1 for 530 days of humid tropical growth conditions. In order to permit precise determination of C deposition in the soil, plant communities were initially established in C-free unwashed quartz sand. Soils were then amended with known amounts of organic matter (containing C and nutrients). Mineral nutrients were also supplied over the course of the experiment as timed-release full-balance fertilizer pellets. Soils represented by far the largest repositories for fixed C in all ecosystems. Almost 5 times more C (ca. 80% of net C fixation) was sequestered in the soil than in the biomass, but this did not differ between CO₂ treatments. In addition, at the whole-ecosystem level we found a remarkably small and statistically non-significant increase in C sequestration (+4%; the sum of C accretion in the soil, biomass, litter and necromass). Total community biomass more than quadrupled during the experiment, but at harvest was, on average, only 8% greater (i.e. 6% per year; n.s.) under elevated CO₂, mainly due to increased root biomass (+15%, P=0.12). Time courses of leaf area index of all ecosystems suggested that canopy expansion was approaching steady state by the time systems were harvested. Net primary productivity (NPP) of all ecosystems-i.e. annual accumulation of biomass, necromass, and leaf litter (but not plant-derived soil organic matter)-averaged 815 and 910 g m^–2 year^–1 at ambient and elevated CO₂, respectively. These NPPs are remarkably similar to those of many natural moist tropical forested ecosystems. At the same time net productivity of soil organic matter reached 7000 g dry matter equivalent per m² and year (i.e. 3500 g C m^–2 year^–1). Very slight yet statistically significant CO₂-induced shifts in the abundance of groups of species occurred by the end of the experiment, with one group of species (Elettaria cardamomum, Ficus benjamina, F. pumila, Epipremnum pinnatum) gaining slightly, and another group (Ctenanthe lubbersiana, Heliconia humilis, Cecropia peltata) losing. Our results show that: (1) enormous amounts of C can be deposited in the ground which are normally not accounted for in estimates of NPP and net ecosystem productivity; (2) any enhancement of C sequestration under elevated atmospheric CO₂ may be substantially smaller than is believed will occur (yet still very important), especially under growth conditions which permit close to natural NPP; and (3) species dominance in plant communities is likely to change under elevated CO₂, but that changes may occur rather slowly.     

Effects of solar radiation,humic substances and nutrients on phytoplankton biomass and distribution in Lake Solumsjö, Sweden

Impacts of solar radiation, humic substances and nutrients on phytoplankton abundance at different depths were investigated in a temperate dimictic lake, Lake Solumsjö. Penetration of solar radiation profiles at different depths, represented as light attenuation coefficient (K _d) were examined. Water sampling and downward irradiance of photosynthetically active radiation (PAR), ultraviolet-A (UV-A, 320–400 nm) and ultraviolet-B (UV-B, 280–320 nm) radiation were performed once a week and at three different times of the day (08.00, 12.00 and 16.00 hrs, local time) between September 13 and November 1, 1999. During the period of investigation, solar radiation above the water surface declined from 474 to 94 mol m^–2 s^–1 for PAR, from 1380 to 3.57 W m^–2 for UV-A and from 13.1 to 0.026 W m^–2 for UV-B, respectively. The attenuation coefficient (K _d) for UV-B radiation ranged from 3.7 to 31 m^–1 and UV-B radiation could not be detected at depths greater than 0.25 m. Humic substances measured at 440 nm ranged from 35.5 to 57.7 Pt mg l^–1. Mean values of biomass, estimated from chlorophyll a, in the whole water column (0–10 m) varied between 2.3 and 5.6 g l^–1 and a diel fluctuation was observed. During stratified conditions, high levels of iron (1.36 mg l^–1) and manganese (4.32 mg l^–1) were recorded in the hypolimnion, suggesting that the thermocline played a major role in the vertical distribution of phytoplankton communities in Lake Solumsjö. The high levels of iron and manganese stimulated the growth of Trachelomonas volvocinopsis in the hypolimnion at a depth of 10 m. Negative impacts of UV-B radiation on phytoplankton in lake Solumsjö are reduced due to the high levels of humic substances and the high degree of solar zenith angle at the latitude studied.     

Episodic sulphate export from wetlands in acidified headwater catchments: prediction at the landscape scale

Sulphate (SO ₄ ^–2 ) concentrations in 34 intensively measured Canadian Shield streams near the Dorset Research Centre, central Ontario, were used to test a hydrogeologic model that uses simple measures of wetland area and till depth to identify catchments that produce SO ₄ ^–2 pulses. Mean annual measured maximum SO ₄ ^–2 concentrations were significantly greater in shallow till (<1 m depth) catchments containing wetlands than catchments covered with deeper tills (>1 m depth) containing wetlands or catchments with no wetlands. Average maximum SO ₄ ^–2 concentrations in wetland catchments during years with dry summers were >20 mg/L in 19 of 20 catchments with average till depths of <1 m, whereas concentrations were <20 mg/L in 5 of 6 watersheds with average till depths of >1 m. Peaks in mean annual maximum SO ₄ ^–2 concentrations from wetland catchments with shallow till occurred during summers with rain fall 150–200 mm less than potential evaporation estimates. There were no significant differences in mean average annual SO ₄ ^–2 concentration among the different catchments during wet summers, with SO ₄ ^–2 concentrations ranging from 6 to 13 mg/L. These observations suggest that a large portion of the temporal and spatial variation in SO ₄ ^–2 chemistry and export can be predicted in headwater catchments of the Canadian Shield and perhaps in other landscapes where till depth influences upland-wetland hydrologic connections.     

Losses of inorganic carbon and nitrous oxide from a temperate freshwater wetland in relation to nitrate loading

We studied the export of inorganic carbon and nitrous oxide (N₂O) from a Danish freshwater wetland. The wetland is situated in an agricultural catchment area and is recharged by groundwater enriched with nitrate (NO₃ ^–) (1000 M). NO₃ ^– in recharging groundwater was reduced (57.5 mol NO₃ ^– m^–2 yr^–) within a narrow zone of the wetland. Congruently, the annual efflux of carbon dioxide (CO₂) from the sediment was 19.1 mol C m^–2 when estimated from monthly in situ measurements. In comparison the CO₂ efflux was 4.8 mol C m^–2 yr^–1 further out in the wetland, where no NO₃ ^– reduction occurred. Annual exports of inorganic carbon in groundwater and surface water was 78.4 mol C m^–2 and 6.1 mol C m^–2 at the two sites, respectively. N₂O efflux from the sedimenst was detectable on five out of twelve sampling dates and was significantly (P < 0.0001) higher in the NO₃ ^– reduction zone (0.35–9.40 mol m^–2 h^–1, range of monthly means) than in the zone without NO₃ ^– reduction (0.21–0.41 mol m^–2 h^–1). No loss of dissolved N₂O could be measured. Total annual export of N₂O was not estimated. The reduction of oxygen (O₂) in groundwater was minor throughout the wetland and did not exceed 0.2 mol 0₂ m^–2yr^–1. Sulfate (SO₄ ^––) was reduced in groundwater (2.1 mol SO₄ ^–– m^–2 yr^–1) in the zone without NO₃ ^– reduction. Although the NO₃ ^– in our wetland can be reduced along several pathways our results strongly suggest that NO₃ ^– loading of freshwater wetlands disturb the carbon balance of such areas, resulting in an accelerated loss of inorganic carbon in gaseous and dissolved forms.     

Viriobenthos Production and Virioplankton Sorptive Scavenging by Suspended Sediment Particles in Coastal and Pelagic Waters

Virus production in oxic surface sediments and virioplankton sorption to suspended particles was estimated across three stations in the Southern California region (33.4°N, 118.6°W). Viriobenthos production was estimated using a sterile sediment and filtered porewater dilution technique that targeted production from both attached bacteria and bacteria living free in the porewater, and attached bacteria alone. Potential virus production rates by bacteria free in the porewater ranged from 1.7 to 4.6 × 10⁸ VLP cm^–3 h^–1, while attached bacteria had slower potential production rates of between 0.4 and 1.1 × 10⁸ VLP cm^–3 h^–1, suggesting turnover rates of viruses in sediments (1–5 h) which are significantly higher than those of virioplankton (~24–48 h). Virioplankton adsorbed to small (<150 µm) suspended sediments at stations with high ambient suspended solid concentrations. Virioplankton scavenging rates combined with published sedimentation rates demonstrate that this mechanism of virus arrival could only account for 0.01% of daily benthic virus production. Calculated mortality rates of benthic bacteria (4–14% h^–1) suggest viruses may play an important role in sediment carbon cycling.     

The influence of sulphate on substrate utilization in a thermophilic sewage sludge digestor

Summary Bacterial sulphate reduction and the interaction between sulphate reduction and methane production was studied in an unadapted and sulphate-adapted thermophilic anaerobic sludge digestor. Addition of sulphate to a concentration of 5 mm (100 times the background level) did not influence gas production or volatile fatty acid concentration compared to the control digestor. When sulphate reduction was not limited by the sulphate concentration, the sulphate-adapted digestor had a sulphate reduction rate of 910 mol l^–1 day compared with 17 mol l^–1 day in the control digestor. The results indicate that the potential for sulphate reduction is low in a thermophilic sewage sludge digestor receiving a low sulphate concentration. Counts of sulphate-reducing bacteria and methanogens showed that sulphate-reducing bacteria were found only in significant numbers in the sulphate-adapted digestor and only with H₂/CO₂ as substrate. Only low numbers of acetate-utilizing sulphate-reducing bacteria were found in both digestors. When using radio-labelled acetate, the relative percentage of 2-labelled acetate converted to CO₂ was two to four times higher in the sulphate-adapted digestor compared to the control digestor. These results suggest that oxidation of acetate seems to play a larger role in the sulphate-adapted digestor.Offprint requests to: B. K. Ahring     

Dissolved organic carbon and its utilization in a riverine wetland ecosystem

Variations in dissolved organic carbon (DOC) concentrations of surface waters and subsurface interstitial groundwater of riparian and wetland soils to 1.2 m depth were evaluated in a riverine wetland ecosystem over one year. DOC was monitored at seven sites within the wetland pond, two sites on the inflow stream, and one site on the outflow stream. Surface concentrations in the inflow stream ranged from 0.74 to 11.6 mg C L^–1 and those of the outflow from 2.1 to 8.0 mg C L^–1 Average DOC from stream floodplain hydrosoils (3.1 to 32.1 mg C L^–1 was greater than DOC from the sediments below the stream channel (1.6 to 6.8 mg C L^–1 Surface DOC within the wetland varied seasonally, with greatest fluctuations in concentrations through the summer and autumn (range 4.8 to 32.6 mg C L^–1 ) during intensive macrophyte growth and bacterial production. DOC was less variable during the winter months (1.7 to 3.3 mg C L^–1 Within the wetland pond, average DOC concentrations (7.1 to 48.2 mg C L^–1) in the subsurface waters were significantly greater (p < 0.05) than average surface concentrations. The microbial availability of surface and subsurface DOC to bacteria was evaluated from losses of DOC by wetland bacteria grown on the DOC. Bacterial growth efficiencies ranged from 5 to 20% and were negatively correlated to the percentage of DOC removed by bacteria (r²=0.93). Throughout the ecosystem, DOC concentrations were greatest in the subsurface waters, but at most depths this DOC was a less suitable substrate than surface DOC for utilization by bacteria.     

Rates of Microbial Metabolism in Deep Coastal Plain Aquifers

Rates of microbial metabolism in deep anaerobic aquifers of the Atlantic coastal plain of South Carolina were investigated by both microbiological and geochemical techniques. Rates of [2-¹⁴C]acetate and [U-¹⁴C]glucose oxidation as well as geochemical evidence indicated that metabolic rates were faster in the sandy sediments composing the aquifers than in the clayey sediments of the confining layers. In the sandy aquifer sediments, estimates of the rates of CO₂ production (millimoles of CO₂ per liter per year) based on the oxidation of [2-¹⁴C] acetate were 9.4 × 10⁻³ to 2.4 × 10⁻¹ for the Black Creek aquifer, 1.1 × 10⁻² for the Middendorf aquifer, and <7 × 10⁻⁵ for the Cape Fear aquifer. These estimates were at least 2 orders of magnitude lower than previously published estimates that were based on the accumulation of CO₂ in laboratory incubations of similar deep subsurface sediments. In contrast, geochemical modeling of groundwater chemistry changes along aquifer flowpaths gave rate estimates that ranged from 10⁻⁴ to 10⁻⁶ mmol of CO₂ per liter per year. The age of these sediments (ca. 80 million years) and their organic carbon content suggest that average rates of CO₂ production could have been no more than 10⁻⁴ mmol per liter per year. Thus, laboratory incubations may greatly overestimate the in situ rates of microbial metabolism in deep subsurface environments. This has important implications for the use of laboratory incubations in attempts to estimate biorestoration capacities of deep aquifers. The rate estimates from geochemical modeling indicate that deep aquifers are among the most oligotrophic aquatic environments in which there is ongoing microbial metabolism.     

Detachment ofPseudomonas fluorescens from biofilms on glass surfaces in response to nutrient stress

The effects of glucose and nitrogen depletion on the colonization of glass Petri plates byPseudomonas fluorescens were studied in batch culture. Colonization of the surfaces was initiated before colonization of the bulk phase, and biofilm formation was observed. This resulted in an apparent lag in the batch growth curve for the cell suspension. The lag phase was an artifact caused by the partitioning of cells between the bulk and solid phase of the culture and was not due to a reduction in the growth rate of unattached cells. The specific growth rate of the unattached cells (0.331 hour^–1) was almost twice that determined for the total population (0.171 hour^–1). Consequently the growth rate of biofilm-forming bacteria cannot be determined in batch culture unless the growth of both attached and unattached cells is monitored, and batch growth curves may contain artifacts due to the formation and dispersion of biofilms. The depletion of either glucose or nitrogen led to the active detachment of cells from the biofilm. An increase in the hydrophobicity of unattached cells was noted on depletion of carbon. This increase was the result of emigration of cells from the surface into the bulk phase.Paper contribution number 128, Centre de Rechcrches Alimentaires de St Hyacinthe.