Heterotrophic prokaryotic production in ultraoligotrophic alpine karst aquifers and ecological implications

Spring waters from alpine karst aquifers are important drinking water resources. To investigate in situ heterotrophic prokaryotic production and its controlling factors, two different alpine karst springs were studied over two annual cycles. Heterotrophic production in spring water, as determined by [³H]leucine incorporation, was extremely low ranging from 0.06 to 6.83 pmol C L⁻¹ h⁻¹ (DKAS1, dolomitic-karst-spring) and from 0.50 to 75.6 pmol C L⁻¹ h⁻¹ (LKAS2, limestone-karst-spring). Microautoradiography combined with catalyzed reporter deposition-FISH showed that only about 7% of the picoplankton community took up [³H]leucine, resulting in generation times of 3–684 days. Principal component analysis, applying hydrological, chemical and biological parameters demonstrated that planktonic heterotrophic production in LKAS2 was governed by the respective hydrological conditions, whereas variations in DKAS1 changed seemingly independent from discharge. Measurements in sediments recovered from LKAS2, DKAS1 and similar alpine karst aquifers ( n =12) revealed a 10⁶-fold higher heterotrophic production (average 19 μmol C dm⁻³ h⁻¹) with significantly lower generation times as compared with the planktonic fraction, highlighting the potential of surface-associated communities to add to self-purification processes. Estimates of the microbially mediated CO₂ in this compartment indicated a possible contribution to karstification.     

The balance between photosynthesis and grazing in Antarctic mixotrophic cryptophytes during summer

SUMMARY 1. Grazing and photosynthetic contributions to the carbon balance of planktonic, mixotrophic cryptophytes in Lakes Fryxell and Hoare in the Taylor Valley, Antarctica were measured during November and December 2000.
2. The cryptophytes never became entirely photosynthetic, although carbon derived from grazing decreased in December. Individual grazing rates ranged between 5.28 and 10.08 bacteria cell⁻¹ day⁻¹ in Lake Fryxell and 0.36–11.76 bacteria cell⁻¹ day⁻¹ in Lake Hoare. Grazing rates varied temporally and with depth in the water column. In Lake Fryxell, which is a meromictic lake, highest grazing occurred just above the chemocline. Individual photosynthetic rates ranged from 0.23 to 1.35 pg C cell⁻¹ h⁻¹ in Lake Fryxell and 0.074 to 1.08 pg C cell⁻¹ h⁻¹ in Lake Hoare.
3. Carbon acquisition by the cryptophyte community gained through grazing ranged between 8 and 31% during November in Lake Fryxell, dropping to between 2 and 24% in December. In Lake Hoare grazing contributed 12–21% of the community carbon budget in November and 1–28% in December. Around 4% of the carbon acquired from grazing and photosynthesis was remineralised through respiration.
4. Mixotrophy is probably a major survival strategy for cryptophytes in the extreme lakes of the Dry Valleys, because perennial ice-cover severely limits light penetration to the water column, whereas these phytoflagellates are not normally mixotrophic in lower latitude lakes. The evidence suggests that mixotrophy may be a mechanism for supplementing the carbon budget, as well as a means of acquiring nutrients for growth.     

Mixotrophic cryptophytes and their predators in the Dry Valley lakes of Antarctica

1. The ingestion rates of planktonic, mixotrophic cryptophytes in two perennially ice-covered Antarctic lakes in the McMurdo Dry Valleys, were investigated during the summer of 1997–1998.
2. In Lake Fryxell, which is meromictic, ingestion rates increased with depth in November and were highest in a cryptophyte maximum close to the chemocline. In Lake Hoare, which is unstratified and freshwater, there was no significant difference in ingestion rates with depth. In both lakes, the highest ingestion rates occurred in early summer, decreasing in December and January. Ingestion rates varied between 0.2 bacteria cell⁻¹ h⁻¹ and 3.6 bacteria cell⁻¹ h⁻¹.
3. During November, mixotrophic cryptophytes removed up to 13% of bacterial biomass day⁻¹ and had a greater grazing impact than heterotrophic nanoflagellates (HNAN). As summer progressed, the grazing impact of cryptophytes and HNAN became similar.
4. The maximum depth of cryptophytes in Lake Fryxell was predated by a population of the ciliate Plagiocampa. Plagiocampa had an ingestion rate of 0.13–0.19 cryptophytes cell⁻¹ h⁻¹. The grazing impact on the cryptophyte community was insignificant. However, the ciliate appeared to be indulging in temporary mixotrophy, sequestering the cryptophytes for a number of weeks before digesting them.
5. It is suggested that mixotrophy is an important survival strategy in the extreme lake ecosystems of the McMurdo Dry Valleys.     

Lactic acid production by Lactobacillus delbrueckii in a dual reactor system using packed bed biofilm reactor

Aims:  An integrated dual reactor system for continuous production of lactic acid by Lactobacillus delbrueckii using biofilms developed on reticulated polyurethane foam (PUF) is demonstrated.
Methods and Results:  Lactobacillus delbrueckii was immobilized on PUF, packed in a bioreactor and used in lactic acid fermentation. The rate of lactic acid production was significantly high with a volumetric productivity of 5 g l⁻¹ h⁻¹ over extended period of time. When coupled to a bioreactor, the system could be operated as dual reactor for over 1000 h continuously without augmentation of inoculum and no compromise on productivity.
Conclusions:  Polyurethane foams offer an excellent support for biofilm formation.
Significance and Impact of the Study:  The system was very robust and could be operated for prolonged period at a volumetric productivity of 4–6 g l⁻¹ h⁻¹.     

Antarctic stream ecosystems: physiological ecology of a blue-green algal epilithon

SUMMARY. 1. Several dozen summer meltwater streams are located in the McMurdo Sound region (c. 78°S 165°E) of southern Victoria Land. They are characterized by a highly variable flow regime at diel, seasonal and annual times caleis; wide fluctuations in temperature and nutrient content; and a very simple epilithic community of cyanophytes ( Nostoc spp., Oscillatoriaceae), bacteria, fungi and microherbivores.
2. The epilithon survives the dark Antarctic winter as dry, frozen mats which provide a large inoculum for growth the following summer. This overwintering assemblage retains a high metabolic capacity and responds rapidly to rehydration.
3. In a series of artificial substrate experiments, biomass accumulation rates were generally less than 0.1 In units d⁻¹. Colonization and growth on the substrates was inversely related to the suspended sediment load of the stream. There was also a visual correspondence between per cent algal cover of the natural streambed and the clarity of the streamwater. Sloughing losses may limit community biomass, particularly in the turbid flowing waters.
4. During running water conditions the mature communities had very low gross photosynthetic rates per unit chlorophyll (<0,1 μg C (μg chl a .h)⁻¹ and per unit carbon (<0,2 μg C (mg biomass C.h)⁻¹). Respiration was generally a high percentage (up to 92%) of gross photosynthesis, which probably reflected the high population densities of microheterotrophs in the community.
5. The floristically simple epilithic mats slowly accumulate to extreme biomass levels (>20 μg chl a cm ⁻², <20 mg C cm⁻²). Production rates per unit biomass are low, probably in response to the cold temperatures of the Antarctic stream environment, and the accumulated biomass represents several seasons of growth.     

Chlorella-bearing ciliates dominate in an oligotrophic North Patagonian lake (Lake Pirehueico, Chile): abundance, biomass and symbiotic photosynthesis

1. Large mixotrophic ciliates ( Stentor araucanus , S. amethystinus and Ophrydium naumanni ) were a characteristic component of a temperate, oligotrophic lake in North Patagonia. During a 1-year study, the abundance, biomass and primary production of these large Chlorella -bearing ciliates were compared with those of the total plankton community.
2. Mixotrophic ciliates peaked in spring and from late summer to autumn, accounting for 1.6–43% (annual average: 16.3%) and 67–99% (annual average: 92%) of total ciliate abundance and biomass, respectively. Their contribution to total zooplankton biomass, including flagellates, rotifers, ciliates and crustaceans, was 14–76%, or 47% as an annual average. Endosymbiotic algae accounted for up to 25% of total autotrophic biomass (annual mean: 3.9%).
3. Maximum cell-specific photosynthetic rates of S. araucanus and S. amethystinus at light saturation varied between 80 and 4400 pg C ciliate^–1 h^–1 with high values during autumn and winter, and low values during summer. The depth-integrated rates of photosynthesis (0–40 m) of algal endosymbionts contributed 1–25% to total photosynthesis (annual mean: 6.5%).
4. A comparison of calculated ingestion rates with photosynthetic rates of Stentor indicates that photosynthate produced by endosymbionts generally exceeded heterotrophic food supply of Stentor during autumn and winter, but was much lower during summer, when food supply was high.
5. The mixotrophic ciliates represent an important 'link' between nanoplankton and higher trophic levels within the plankton community because of their high heterotrophic biomass and considerable contribution to total photosynthesis.     

Seasonal heterotrophic flagellate and bacterial plankton dynamics in a large oligotrophic lake– Loch Ness, Scotland

1. Bacterial production in the 0–30 m water column of Loch Ness was measured using a dual labelling procedure with [³H] thymidine and [¹⁴C] leucine between May 1993 and June 1994. In most cases the uptake of the two labels did not covary, suggesting unbalanced growth. Rates of bacterial production varied from undetectable to 46.2 μg C l^–1 day^–1. Highest production coincided with the period of highest primary production, but carbon derived from this source was insufficient to meet the bacterial carbon demand, which was met by allochthonous humic inputs to the system.
2. Heterotrophic flagellate (HNAN) grazing rates, measured using fluorescently labelled bacteria, ranged between 10.3 and 24.5 bacteria cell^–1 day^–1 at temperatures between 5 and 15 °C. They removed up to 27% of the bacterial production per day.
3. Heterotrophic flagellate specific growth rates ranged from 0.043 to 0.093 h^–1 between 5 and 15 °C, giving generation times of 7.4–16.1 h.
4. bacterial and HNAN abundances were not coupled, but the highest HNAN grazing impact related to a time of high bacterial productivity.     

Gas exchange in intact isolated chloroplasts from Chlamydomonas reinhardtii during starch degradation in the dark

During starch degradation in intact isolated chloroplasts from Chlamydomonas reinhardtii gas exchange was studied with a mass spectrometer. Oxygen uptake by intact chloroplasts in the dark never exceeded 1.5% of the starch degradation rate [maximum 15 nmol O₂ (mg Chl)⁻¹ h⁻¹ consumed. 1 000 nmol glucose (mg Chl)⁻¹h⁻¹ degraded]. Evolution of CO₂ under aerobic conditions [9.8–28 nmol (mg Chl)⁻¹ h⁻¹] was stimulated by addition of 0.1–0.5 m M oxaloacetate [393–425 nmol CO₂ (mg Chl)⁻¹ h⁻¹]. Pyridoxal phosphate (5 m M ) inhibited starch degradation by more than 80%, but had no effect on O₂ uptake. Starch degradation rates and CO₂ evolution did not differ under acrobic and anaerobic conditions. Increasing P_i in the reaction medium from 0.5 m M to 5.0 m M stimulated starch degradation by 230 and 260% under aerobic and anaerobic conditions, respectively. A rapid autooxidation of reduced ferredoxin was observed in a reconstituted system consisting of purified Chlamydomonas ferredoxin, purified Chlamydomonas NADP-ferredoxin oxidoreductase (EC 1.6.7.1) and NADPH. Addition of isolated thylakoids from C. reinhardtii did not affect the rate of O₂ uptake. Our results clearly indicate the absence of any oxygen requirement during starch degradation in isolated chloroplasts.     

Sucrose phosphorylase of the rumen bacterium Pseudobutyrivibrio ruminis strain A

Aims:  To verify the taxonomic affiliation of bacterium Butyrivibrio fibrisolvens strain A from our collection and to characterize its enzyme(s) responsible for digestion of sucrose.
Methods and Results:  Comparison of the 16S rRNA gene of the bacterium with GenBank showed over 99% sequence identity to the species Pseudobutyrivibrio ruminis . Molecular filtration, native electrophoresis on polyacrylamide gel, zymography and thin layer chromatography were used to identify and characterize the relevant enzyme. An intracellular sucrose phosphorylase with an approximate molecular mass of 52 kDa exhibiting maximum activity at pH 6·0 and temperature 45°C was identified. The enzyme was of inducible character and catalysed the reversible conversion of sucrose to fructose and glucose-1-P. The reaction required inorganic phosphate. The K _m for glucose-1-P formation and fructose release were 3·88 × 10⁻³ and 5·56 × 10⁻³ mol l⁻¹ sucrose, respectively – while the V _max of the reactions were −0·579 and 0·9  μ mol mg protein⁻¹ min⁻¹. The enzyme also released free glucose from glucose phosphate.
Conclusion:  Pseudobutyrivibrio ruminis strain A utilized sucrose by phosphorolytic cleavage.
Significance and Impact of the Study:  Bacterium P. ruminis strain A probably participates in the transfer of energy from dietetary sucrose to the host animal.     

Single-cell vs. bulk activity properties of coastal bacterioplankton over an annual cycle in a temperate ecosystem

The connections between single-cell activity properties of heterotrophic planktonic bacteria and whole community metabolism are still poorly understood. Here, we show flow cytometry single-cell analysis of membrane-intact (live), high nucleic acid (HNA) content and actively respiring (CTC+) bacteria with samples collected monthly during 2006 in northern Spain coastal waters. Bulk activity was assessed by measuring ³H-Leucine incorporation and specific growth rates. Consistently, different single-cell relative abundances were found, with 60–100% for live, 30–84% for HNA and 0.2–12% for CTC+ cells. Leucine incorporation rates (2–153 pmol L⁻¹ h⁻¹), specific growth rates (0.01–0.29 day⁻¹) and the total and relative abundances of the three single-cell groups showed marked seasonal patterns. Distinct depth distributions during summer stratification and different relations with temperature, chlorophyll and bacterial biovolume suggest the existence of different controlling factors on each single-cell property. Pooled leucine incorporation rates were similarly correlated with the abundance of all physiological groups, while specific growth rates were only substantially explained by the percentage of CTC+ cells. However, the ability to reduce CTC proved notably better than the other two single-cell properties at predicting bacterial bulk rates within seasons, suggesting a tight linkage between bacterial individual respiration and biomass production at the community level.     

Temporal dynamics in epipelic, pelagic and epiphytic algal production in a clear and a turbid shallow lake

SUMMARY 1. Pelagic and epipelic microalgal production were measured over a year in a pre-defined area (depth 0.5 m) in each of two lakes, one turbid and one with clear water. Further estimates of epiphytic production within reed stands were obtained by measuring production of periphyton developed on artificial substrata.
2. Total annual production of phytoplankton and epipelon was 34% greater in the turbid lake (190 g C m⁻² year⁻¹) than in the clearwater lake (141 g C m⁻² year⁻¹). However, the ratio of total production to mean water column TP concentration was two fold greater in the clearwater lake.
3. Phytoplankton accounted for the majority of the annual production (96%) in the turbid lake, while epipelic microalgal production dominated (77%) in the clear lake. The relative contribution of epipelic algae varied over the year, however, and in the turbid lake was higher in winter (11–25%), when the water was relatively clear, than during summer (0.7–1.7%), when the water was more turbid. In the clearwater lake, the relative contribution of epipelon was high both in winter, when the water was most clear, and in mid-summer, when phytoplankton production was constrained either by nutrients or grazing.
4. Compared with pelagic and epipelic primary production, epiphytic production within a reed stand was low and did not vary significantly between the lakes.
5. The study supports the theory of a competitive and compensatory trade-off between primary producers in lakes with contrasting nutrient concentrations, resulting in relatively small differences in overall production between clear and turbid lakes when integrating over the season and over different habitats.     

Spatial and temporal patterns of microcrustacean assemblage structure and secondary production in a wetland ecosystem

1. In contrast to extensive studies of zooplankton in lakes, the role of microcrustaceans in wetlands is not well studied. In this study, spatial and temporal patterns of microcrustacean assemblage structure and secondary production were quantified over a 2-year period in a southeastern U.S.A. wetland.
2. Thirty-two species, including 19 cladocerans, 10 copepods and three ostracods, generated different temporal patterns of density and production between vegetated ( Nymphaea ) and non-vegetated (open-water) zones reflecting species-specific differences in life histories.
3. Summer assemblages were dominated by small, planktonic filter-feeders, typified by high annual production/biomass ( P / B ) and daily production. In contrast, winter assemblages were dominated by larger, epibenthic detritivores with low P / B and high biomass. Seasonal shifts in the relative importance of planktonic species in the warmer months to benthic and epiphytic species in the cooler months suggest that energy flow pathways through microcrustaceans may vary seasonally.
4. Total annual production was higher during both years in the Nymphaea zone (13.0 g and 13.6 g DM m⁻² year⁻¹) than the open-water (8.2 and 6.3 g DM m⁻² year⁻¹), and was similar between years for the entire wetland pond (12.3 and 12.2 g DM m⁻² year⁻¹).
5. Although wetland ecosystems have been the subject of considerable ecological research in the past 20 years, our study is one of the few to demonstrate a highly diverse and relatively productive microcrustacean assemblage. Such comprehensive production studies can be used to quantify the ecological importance of microcrustaceans in freshwater wetland ecosystems.     

Holocene carbon burial by lakes in SW Greenland

The role of the Arctic in future global change processes is predicted to be important because of the large carbon (C) stocks contained in frozen soils and peatlands. Lakes are an important component of arctic landscapes although their role in storing C is not well prescribed. The area around Kangerlussuaq, SW Greenland (66–68°N, 49–54°W) has extremely high lake density, with ∼20 000 lakes that cover about 14% of the land area. C accumulation rates and standing stock (kg C m⁻²), representing late- to mid-Holocene C burial, were calculated from AMS ¹⁴C-dated sediment cores from 11 lakes. Lake ages range from ∼10 000 cal yr  bp to ∼5400 cal yr  bp , and reflect the withdrawal of the ice sheet from west to east. Total standing stock of C accumulated in the studied lakes for the last ∼8000 years ranged from 28 to 71 kg C m⁻², (mean: ∼42 kg C m⁻²). These standing stock determinations yield organic C accumulation rates of 3.5–11.5 g C m⁻² yr⁻¹ (mean: ∼6 g C m⁻² yr⁻¹) for the last 4500 years. Mean C accumulation rates are not different for the periods 8–4.5 and 4.5–0 ka, despite cooling trends associated with the neoglacial period after 4.5 ka. We used the mean C standing stock to estimate the total C pool in small lakes (<100 ha) of the Kangerlussuaq region to be ∼4.9 × 10¹³ g C. This C stock is about half of that estimated for the soil pool in this region (but in 5% of the land area) and indicates the importance of incorporating lakes into models of regional C balance at high latitudes.     

ADSORPTION OF FULVIC ACID ON ALGAL SURFACES AND ITS EFFECT ON CARBON UPTAKE

Adsorption of Suwannee River fulvic acid (SRFA) to algal surfaces of three green algae was studied at environmentally relevant pH values (4 –7) and SRFA concentrations (5–100 mg·L ^{− 1}). The influence of adsorbed SRFA on carbon uptake of Scenedesmus subspicatus Chodat was also examined. Although no adsorption was observed at neutral pH values (pH 6 and 7), at pH 4 up to 31 mg SRFA·m ^{− 2} and at pH 5 up to 4 mg SRFA·m ^{− 2} was adsorbed to the algal surfaces. Electrophoretic mobility measurements of S. subspicatus demonstrated an increase in the negative surface charge of the alga in the presence of SRFA at pH 4. The adsorbed SRFA also influenced ¹⁴C uptake in S. subspicatus; in this case, enhanced carbon uptake could be related to the amount of adsorbed SRFA. The binding of humic substances by algal surfaces was interpreted as the result of hydrogen bonding and hydrophobic interactions.     

Induction of nitrate reductase in needles of Scots pine seedlings by NOX and NO3−

The possibility to induce nitrate reductase (NR; EC 1.6.6.2) in needles of Scots pine ( Pinus sylvestris L.) seedlings was studied. The NR activity was measured by an in vivo assay. Although increased NR activities were found in the roots after application of NO₃⁻, no such increase could be detected in the needles. Detached seedlings placed in NO₃⁻ solution showed increasing NR activities with increasing NO₃⁻ concentrations. Exposure of seedlings to NO_x (70–80 ppb NO₂ and 8–12ppb NO) resulted in an increase of the NR activity from 10–20 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹ to about 400 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. This level was reached after 2–4 days of exposure, thereafter the NR activity decreased to about 200 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. Analyses of free amino acids showed low concentrations of arginine and glutamine in NO_x-fumigated seedlings compared to corresponding controls.     

Phytoplankton production and growth rate in Lake Tanganyika: evidence of a decline in primary productivity in recent decades

1. This study focused on phytoplankton production in Lake Tanganyika. We provide new estimates of daily and annual primary production, as well as growth rates of phytoplankton, and we compare them with values published in former studies.
2. Chlorophyll- a (chl- a ) in the mixed layer ranged from 5 to 120 mg chl- a  m⁻² and varied significantly between rainy and dry seasons. Particulate organic carbon concentrations were significantly higher in the south basin (with 196 and 166 mg C m⁻³ in the dry and the rainy season, respectively) than in the north basin (112 and 109 mg C m⁻³, respectively).
3. Carbon : phosphorus (C : P) ratios varied according to season. Phosphorus limitation seemed to occur more frequently than nitrogen limitation, especially during the rainy season. Severe P deficiencies were rare.
4. Measured particulate daily primary production ranged from 110 to 1410 mg C m⁻² day⁻¹; seasonal contrasts were well marked in the north basin, but less in the south basin, where primary production peaks occurred also in the rainy season. Estimates of annual primary production, based on daily primary production calculated from chl- a and water transparency, gave values lower than those reported in previous studies. Picophytoplankton accounted on average for 56% of total particulate production in the south basin during the wet season of 2003.
5. Phytoplankton growth rates, calculated from primary production, ranged from 0.055 to 0.282 day⁻¹; these are lower than previously published values for Lake Tanganyika.     

Oxygen uptake in developing eggs and larvae of the cod, Gadus morhua L.

Unfertilised cod eggs showed a mean oxygen uptake rate at 5°C of 0.089 μl O₂, dry wt.⁻¹ h⁻¹; this gradually rose to 0.768 μl O₂ mg dry wt.⁻¹ h⁻¹ in eggs about to hatch. From hatching to complete yolk absorption larvae respired at 1.6 μl O₂, mg dry wt.⁻¹ h⁻¹. During starvation following yolk absorption, uptake fell significantly to 1.1 μl O₂, mg dry ⁻¹ h⁻¹. Much of this decrease in oxygen consumption was shown to be caused by reduction in activity. Loss of weight during the embryo and larval phases could not easily be reconciled with total oxygen consumption; it is suggested that cod embryos and larvae may not rely solely upon endogenous energy reserves during development.     

Covariation of stream community structure and biomass of algae, invertebrates and fish with forest cover at multiple spatial scales

1. To evaluate the spatial extent of the effects of forest cover on stream ecosystems, we measured algae, invertebrate, and fish biomass and invertebrate and fish community structure in 38 small first- to third-order streams in the National Capital Region of Canada along with forest cover at different spatial scales.
2. We considered 55 spatial scales of forest cover including several buffer widths (doubling 10–320 m) and lengths (doubling 10–1280 m, entire riparian distance upstream from sampling area) and entire catchments to determine which spatial scale maximized the correlation with biomass and metrics of community structure.
3. The proportion of variability in biomass and structural metrics explained by forest cover generally increased with increasing scale, suggesting that catchment-wide disturbances are the most influential determinants of benthic and fish communities.
4. Catchment forest cover explained more variation in algal (adjusted r ²   =   0.54), invertebrate (adjusted r ²   =   0.51) and fish (adjusted r ²   =   0.33) biomass than structural metrics of invertebrates and fish (adjusted r ²   =   0.08–0.27).
5. Analyses of the partial effects of forest cover at three scales (reach, riparian and the entire catchment) on biomass and community structure metrics identified catchment and reach scales as being most influential and never detected a significant partial effect of forest cover at the riparian scale.
6. These results suggest that maintenance or protection of reach and riparian buffers alone will not sufficiently protect stream function and structure from catchment-wide impacts.     

Contribution of virus-induced lysis and protozoan grazing to benthic bacterial mortality estimated simultaneously in microcosms

In contrast to the water column, the fate of bacterial production in freshwater sediments is still a matter of debate. Thus, the importance of virus-induced lysis and protozoan grazing of bacteria was investigated for the first time simultaneously in a silty sediment layer of a mesotrophic oxbow lake. Microcosms were installed in the laboratory in order to study the dynamics of these processes over 15 days. All microbial and physicochemical parameters showed acceptable resemblance to field data observed during a concomitant in situ study, and similar conclusions can be drawn with respect to the quantitative impact of viruses and protozoa on the bacterial compartment. Viral decay rates ranged from undetectable to 0.078 h⁻¹ (average, 0.033 h⁻¹), and the control of bacterial production from below the detection limit to 36% (average, 12%). The contribution of virus-induced lysis of bacteria to the dissolved organic matter pool as well as to benthic bacterial nutrition was low. Ingestion rates of protozoan grazers ranged from undetectable to 24.7 bacteria per heterotrophic nanoflagellate (HNF) per hour (average, 4.8 bacteria HNF⁻¹ h⁻¹) and from undetectable to 73.3 bacteria per ciliate per hour (average, 11.2 bacteria ciliate⁻¹ h⁻¹). Heterotrophic nanoflagellate and ciliates together cropped up to 5% (average, 1%) of bacterial production. The viral impact on bacteria prevailed over protozoan grazing by a factor of 2.5–19.9 (average, 9.5). In sum, these factors together removed up to 36% (average, 12%) of bacterial production. The high number of correlations between viral and protozoan parameters is discussed in view of a possible relationship between virus removal and the presence of protozoan grazers.     

Generation of a reproducible nutrient-depleted biofilm of Escherichia coli and Burkholderia cepacia

An in vitro method of growing bacteria as a defined nutrient-depleted biofilm is proposed. The medium was defined nutritionally in terms of the quantitative composition and by the total amount of nutrient required to achieve a defined population size. Escherichia coli and Burkholderia cepacia were incubated on a filter support placed on a defined volume of solid medium. The change of biomass of the biofilm population was compared with the change in a planktonic culture. The size of the population in stationary phase was proportional to the concentration of limiting substrate up to 40 μmol cm⁻² glucose for E. coli and up to 2·7 × 10⁻⁹ mol cm⁻² iron for B. cepacia . Escherichia coli growing exponentially had a growth rate of μ = 0·30 h⁻¹ in a biofilm and μ = 0·96 h⁻¹ in planktonic culture. The growth rate, μ, for exponentially growing B. cepacia in a biofilm was 1·12 h⁻¹ and in planktonic culture 0·78 h⁻¹. This method allows the limitation of the size of a biofilm population to a chosen value.