Regulation of planktonic bacterial growth rates: The effects of temperature and resources

We examined the potential limitation of bacterial growth by temperature and nutrients in a eutrophic lake. Dilution cultures from winter and summer were incubated at both high (>20°C) and low (4°C) temperatures and enriched with various combinations of organic carbon (C), inorganic nitrogen (N), and inorganic phosphorus (P). Bacterial abundance, ³H-thymidine incorporation, and ³H-leucine incorporation were measured over the growth cycle. For both winter and summer assemblages, low temperature limited growth even when resources (C, N, and P) were added. When temperature was adequate, bacterial growth in dilution cultures was co-limited by C, N, and P Additions of either C, P, or N and P alone provide little or only modest stimulation of growth, suggesting that under in situ conditions both nutrients and organic carbon limit bacterial growth. Our results provide little evidence of seasonal adaptation to low temperatures for bacterial communities in temperate lakes. Instead, bacterial growth appears to be temperature limited during winter and resource limited during summer. We propose that, in general, bacterial growth rates are temperature dependent up to a threshold, but that the patterns of change across temperature gradients are resource dependent, such that temperature has little effect on growth in resource-rich environments but a strong effect in resource-poor environments. Correspondence to: Marisol Felip     

Phytoplankton–bacterioplankton interactions in a neotropical floodplain lake (Laguna Bufeos,Bolivia)

Laguna Bufeos is a floodplain lake of the river Ichilo, a tributary of the Amazon basin situated in Bolivia. Nutrient addition assays involving whole water (<200 μm) as well as fractionated water (<0.8 μm) treatments were carried out in incubation tubes to test whether bacterial growth is limited by the availability of inorganic nutrients and to test whether bacteria are able to utilize inorganic nutrients directly or are stimulated by inorganic nutrients through increased production of phytoplankton. The assays were carried out during two extreme hydrological conditions, the high-water and the low-water period. During the high-water period experiment, neither N or P limited bacterial growth rates. During the low-water period, bacterial growth was P limited. Bacterial growth was stimulated in the fractionated as well as in the whole water treatments, indicating that bacterial growth was directly stimulated by P. Bacterial growth corrected for grazing losses (determined by means of dilution experiments) was significantly higher in the fractionated water containing only bacteria when compared to the whole water containing also grazers and phytoplankton. This suggests that bacterial growth was suppressed by competition with phytoplankton rather than stimulated through the production of dissolved organic matter by phytoplankton.     

Inadequacy of the Eucaryote Inhibitor Cycloheximide in Studies of Protozoan Grazing on Bacteria at the Freshwater-Sediment Interface

Four guilds from a lake sediment-water interface microbial community were isolated and tested for sensitivity to cycloheximide (0.1 to 200 mg liter⁻¹). Field experiments were conducted to compare the inhibition, dilution, and filtration methods for determining grazing rates. Cycloheximide inhibited anaerobic bacteria at 50 mg liter⁻¹, and inhibition of bacterial growth was observed in the grazing experiments. The results show that the assumption of selective inhibition of heterotrophic eucaryotes was violated and preclude the use of cycloheximide in grazing experiments.     

Bacterial diversity and activity along a salinity gradient in soda lakes of the Kulunda Steppe (Altai, Russia)

Here we describe the diversity and activity of sulfate reducing bacteria along a salinity gradient in four different soda lakes from the Kulunda Steppe (South East Siberia, Russia). For this purpose, a combination of culture-dependent and independent techniques was applied. The general bacterial and SRB diversity were analyzed by denaturing gradient gel electrophoresis (DGGE) targeting the 16S rDNA gene. DNA was used to detect the microbial populations that were present in the soda lake sediments, whereas ribosomal RNA was used as a template to obtain information on those that were active. Individual DGGE bands were sequenced and a phylogenetic analysis was performed. In addition, the overall activity of SRB was obtained by measuring the sulfate reduction rates (SRR) and their abundance was estimated by serial dilution. Our results showed the presence of minor, but highly active microbial populations, mostly represented by members of the Proteobacteria. Remarkably high SRR were measured at hypersaline conditions (200 g L⁻¹). A relatively high viable count indicated that sulfate reducing bacteria could be highly active in hypersaline soda lakes. Furthermore, the increase of sodium carbonate/bicarbonate seemed to affect the composition of the microbial community in soda lakes, but not the rate of sulfate reduction.     

Macrozooplankton and the persistence of the deep chlorophyll maximum in a stratified lake

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Nutrient enrichment and nutrient regeneration stimulate bacterioplankton growth

Bacterial abundance results from predatory losses of individuals and replacement of losses through growth. Growth depends on sustained input of organic substrates and mineral nutrients. In this work we tested the hypothesis that bacterial growth in two oligotrophic Canadian shield lakes was limited by nitrogen (N) or phosphorus (P). We also determined whether consumer-regenerated resources contributed substantially to net bacterial growth. Two types of dilution assays were conducted to determine the response of bacteria to nutrient enrichment: diluted whole water (DWW, 1:9 whole/filtered with 0.2 m of filtered lake water) and diluted fractionated water (DFW, 1.0 m prefiltered then diluted as above). Replicate bottles in each dilution assay received either N (50 m), P (10 m), or both N and P enrichments. Controls received no nutrients. Resource-saturated growth rates and grazing rates were estimated from a standard dilution-growth approach. Bacterial growth was stimulated by addition of P alone and in combination with N. Consumers regenerated sufficient resources to support up to half the bacterial growth rate, but the benefit derived from consumers was minor when compared to mortality.     

Availability of dissolved organic carbon for planktonic bacteria in oligotrophic lakes of differing humic content

Bacterioplankton from 10 oligotrophic lakes, representing a gradient from clearwater to polyhumic, were grown in dilution cultures of sterile filtered lake water. The bacterial biomass achieved in the stationary phase of the dilution cultures was positively correlated with the amount of both humic matter and dissolved organic carbon (DOC) in the lakes. About the same fraction of the total DOC pool was consumed in the dilution cultures of all lakes (average 9.5%, coefficient of variation (CV) 24%), with approximately the same growth efficiency (average 26%, CV 28%). Thus, humic lakes could support a higher bacterial biomass than clearwater lakes due to their larger DOC pools. The relevance of the results to planktonic food webs of humic and clearwater lakes is discussed.     

Tests of the Critical Assumptions of the Dilution Method for Estimating Bacterivory by Microeucaryotes

The critical assumptions of the dilution method for estimating grazing rates of microzooplankton were tested by using a community from the sediment-water interface of Lake Anna, Va. Determination of the appropriate computational model was achieved by regression analysis; the exponential model was appropriate for bacterial growth at Lake Anna. The assumption that the change in grazing pressure is linearly proportional to the dilution factor was tested by analysis of variance with a lack-of-fit test. There was a significant (P < 0.0001) linear (P > 0.05) relationship between the dilution factor and time-dependent change in ln bacterial abundance. The assumption that bacterial growth is not altered by possible substrate enrichment in the dilution treatment was tested by amending diluted water with various amounts of dissolved organic carbon (either yeast extract or extracted carbon from lake sediments). Additions of carbon did not significantly alter bacterial growth rates during the incubation period (24 h). On the basis of these results, the assumptions of the dilution method proved to be valid for the system examined.     

Bacterioplankton growth, grazing mortality and quantitative relationship to primary production in a humic and a clearwater lake

Bacterial growth and grazing mortality were estimated from Mayto October in two south Swedish oligotrophic lakes, one beinga clearwater lake (water colour 5–10 mg Pt l^–1 DOC2.9–3.4 mg l^–1, Secchi disk depth 5.0–9.4m) and the other a humic, brownwater lake (water colour 105–165mg Pt l^–1, DOC 13.7–22.7mg l^–1, Secchi diskdepth 1.3–2.1 m). Specific rates of growth and grazingmortality were generally similar for both lakes. However, theabundance of bacteria was consistently 2–3 times higherin the water of the humic lake, suggesting that the total productionand consumption of bacterial cells were also higher than inthe dearwater lake. The ratio of bacterial secondary productionto primary production was higher in the humic lake than in theclearwater lake, indicating that the bacterioplankton of thehumic lake utilize allochthonous substrates, in addition tosubstrates originating from autochthonous primary production.Most of the bacterial loss in both lakes could be attributedto small protozoan grazers. This implies that allochthonousand autochthonous organic carbon fixed by bacterioplankton isless important in terms of carbon flow to higher trophic levelsthan would be expected if macrozooplankton were the dominantbacterivores, providing a more direct and efficient transferof carbon to larger organisms.     

Responses of Bacillus subtilis to hypotonic challenges: physiological contributions of mechanosensitive channels to cellular survival

Mechanosensitive channels are thought to function as safety valves for the release of cytoplasmic solutes from cells that have to manage a rapid transition from high- to low-osmolarity environments. Subsequent to an osmotic down-shock of cells grown at high osmolarity, Bacillus subtilis rapidly releases the previously accumulated compatible solute glycine betaine in accordance with the degree of the osmotic downshift. Database searches suggest that B. subtilis possesses one copy of a gene for a mechanosensitive channel of large conductance (mscL) and three copies of genes encoding proteins that putatively form mechanosensitive channels of small conductance (yhdY, yfkC, and ykuT). Detailed mutational analysis of all potential channel-forming genes revealed that a quadruple mutant (mscL yhdY yfkC ykuT) has no growth disadvantage in high-osmolarity media in comparison to the wild type. Osmotic down-shock experiments demonstrated that the MscL channel is the principal solute release system of B. subtilis, and strains with a gene disruption in mscL exhibited a severe survival defect upon an osmotic down-shock. We also detected a minor contribution of the SigB-controlled putative MscS-type channel-forming protein YkuT to cellular survival in an mscL mutant. Taken together, our data revealed that mechanosensitive channels of both the MscL and MscS types play pivotal roles in managing the transition of B. subtilis from hyper- to hypo-osmotic environments.     

Differential Growth Response of Colony-Forming α- and γ-Proteobacteria in Dilution Culture and Nutrient Addition Experiments from Lake Kinneret (Israel), the Eastern Mediterranean Sea, and the Gulf of Eilat

Even though it is widely accepted that bacterioplankton growth in lakes and marine ecosystems is determined by the trophic status of the systems, knowledge of the relationship between nutrient concentrations and growth of particular bacterial species is almost nonexistent. To address this question, we performed a series of culture experiments with water from Lake Kinneret (Israel), the eastern Mediterranean Sea, and the Gulf of Eilat (northern Red Sea). In the initial water samples, the proportion of CFU was typically <0.002% of the 4′,6′-diamidino-2-phenylindole (DAPI) counts. During incubation until the early stationary phase, the proportion of CFU increased to 20% of the DAPI counts and to 2 to 15% of the DAPI counts in unenriched lake water and seawater dilution cultures, respectively. Sequencing of the 16S ribosomal DNA of colony-forming bacteria in these cultures consistently revealed an abundance of α-proteobacteria, but notable phylogenetic differences were found at the genus level. Marine dilution cultures were dominated by bacteria in the Roseobacter clade, while lake dilution cultures were dominated by bacteria affiliated with the genera Sphingomonas and Caulobacter. In nutrient (glucose, ammonium, phosphate) addition experiments the CFU comprised 20 to 83% of the newly grown cells. In these incubation experiments fast-growing γ-proteobacteria dominated; in the marine experiments primarily different Vibrio and Alteromonas species appeared, while in the lake water experiments species of the genera Shewanella, Aeromonas, and Rheinheimera grew. These results suggest that major, but different, γ-proteobacterial genera in both freshwater and marine environments have a preference for elevated concentrations of nutrients and easily assimilated organic carbon sources but are selectively outcompeted by α-proteobacteria in the presence of low nutrient concentrations.     

Impact of fish predation on cladoceran body weight distribution and zooplankton grazing in lakes during winter

1. It is well accepted that fish, if abundant, can have a major impact on the zooplankton community structure during summer, which, particularly in eutrophic lakes, may cascade to phytoplankton and ultimately influence water clarity. Fish predation affects mean size of cladocerans and the zooplankton grazing pressure on phytoplankton. Little is, however, known about the role of fish during winter. 2. We analysed data from 34 lakes studied for 8–9 years divided into three seasons: summer, autumn/spring and winter, and four lake classes: all lakes, shallow lakes without submerged plants, shallow lakes with submerged plants and deep lakes. We recorded how body weight of Daphnia and then cladocerans varied among the three seasons. For all lake types there was a significant positive correlation in the mean body weight of Daphnia and all cladocerans between the different seasons, and only in lakes with macrophytes did the slope differ significantly from one (winter versus summer for Daphnia). 3. These results suggest that the fish predation pressure during autumn/spring and winter is as high as during summer, and maybe even higher during winter in macrophyte‐rich lakes. It could be argued that the winter zooplankton community structure resembles that of the summer community because of low specimen turnover during winter mediated by low fecundity, which, in turn, reflects food shortage, low temperatures and low winter hatching from resting eggs. However, we found frequent major changes in mean body weight of Daphnia and cladocerans in three fish‐biomanipulated lakes during the winter season. 4. The seasonal pattern of zooplankton : phytoplankton biomass ratio showed no correlation between summer and winter for shallow lakes with abundant vegetation or for deep lakes. For the shallow lakes, the ratio was substantially higher during summer than in winter and autumn/spring, suggesting a higher zooplankton grazing potential during summer, while the ratio was often higher in winter in deep lakes. Direct and indirect effects of macrophytes, and internal P loading and mixing, all varying over the season, might weaken the fish signal on this ratio. 5. Overall, our data indicate that release of fish predation may have strong cascading effects on zooplankton grazing on phytoplankton and water clarity in temperate, coastal situated eutrophic lakes, not only during summer but also during winter.     

Nitrogen deposition and warming – effects on phytoplankton nutrient limitation in subarctic lakes

The aim of this study was to predict the combined effects of enhanced nitrogen (N) deposition and warming on phytoplankton development in high latitude and mountain lakes. Consequently, we assessed, in a series of enclosure experiments, how lake water nutrient stoichiometry and phytoplankton nutrient limitation varied over the growing season in 11 lakes situated along an altitudinal/climate gradient with low N‐deposition (<1 kg N ha^?1 yr^?1) in northern subarctic Sweden. Short‐term bioassay experiments with N‐ and P‐additions revealed that phytoplankton in high‐alpine lakes were more prone to P‐limitation, and with decreasing altitude became increasingly N‐ and NP‐colimited. Nutrient limitation was additionally most obvious in midsummer. There was also a strong positive correlation between phytoplankton growth and water temperature in the bioassays. Although excess nutrients were available in spring and autumn, on these occasions growth was likely constrained by low water temperatures. These results imply that enhanced N‐deposition over the Swedish mountain areas will, with the exception of high‐alpine lakes, enhance biomass and drive phytoplankton from N‐ to P‐limitation. However, if not accompanied by warming, N‐input from deposition will stimulate limited phytoplankton growth due to low water temperatures during large parts of the growing season. Direct effects of warming, allowing increased metabolic rates and an extension of the growing season, seem equally crucial to synergistically enhance phytoplankton development in these lakes.     

Influence of hydrological seasonality on bacterioplankton in two neotropical floodplain lakes

Seasonal fluctuation in river stage strongly affects the ecological functioning of tropical floodplain lakes. This study was conducted to assess the influence of hydrological seasonality on bacterial production and abundance in two floodplain lakes of the Autana River, a blackwater river in the Middle Orinoco basin, Venezuela. Water samples for nutrient chemistry, chlorophyll a, and microbiological determinations were collected in two floodplain lakes and in the mainstem of the river during 1997–98. DOC and chlorophyll a concentrations were similar between mainstem and lake sites during high water when river and lakes were well connected but became different during the period of low water when the interaction was minimal. Higher values of bacterial production were observed in the floodplain lakes (0.62–1.03 g C l^–1 h^–1) compared to the mainstem sites (0.17–0.19 g C l^–1 h^–1) during the period of low water, while during the period of high water river and lake sites showed similar levels (0.04 g C l^–1 h^–1). Bacterial numbers followed bacterial production in the floodplain lakes, reaching higher numbers during the period of low water (1.41–2.40 × 10⁶ cells ml^–1). Availability of substrate and inorganic nutrients, pH, and inputs and losses of bacterial cells could be determining the observed seasonal patterns in bacterial production and abundance. The Autana lakes exhibited a strong seasonal pattern in the chemical and biological conditions, showing higher productivity during the lentic phase that lasted between 5 and 6 months.     

Bacterial Growth on Allochthonous Carbon in Humic and Nutrient-enriched Lakes: Results from Whole-Lake 13C Addition Experiments

Organic carbon (C) in lakes originates from two distinct sources—primary production from within the lake itself (autochthonous supply) and importation of organic matter from the terrestrial watershed (allochthonous supply). By manipulating the ¹³C of dissolved inorganic C, thereby labeling within-lake primary production, we examined the relative importance of autochthonous and allochthonous C in supporting bacterial production. For 35 days, NaH¹³CO₃ was added daily to two small, forested lakes. One of the lakes (Peter) was fertilized so that primary production exceeded total respiration in the epilimnion. The other lake (Tuesday), in contrast, was low in productivity and had high levels of colored dissolved organic C (DOC). To obtain bacterial C isotopes, bacteria were regrown in situ in particle-free lake water in dialysis tubes. The contribution of allochthonous C to bacterial biomass was calculated by applying a two-member mixing model. In the absence of a direct measurement, the isotopic signature of the autochthonous end-member was estimated indirectly by three different approaches. Although there was excess primary production in Peter Lake, bacterial biomass consisted of 43–46% allochthonous C. In Tuesday Lake more than 75% of bacterial growth was supported by allochthonous C. Although bacteria used autochthonous C preferentially over allochthonous C, DOC from the watershed contributed significantly to bacterial production. In combination with results from similar experiments in different lakes, our findings suggest that the contribution of allochthonous C to bacterial production can be predicted from ratios of chromophoric dissolved organic matter (a surrogate for allochthonous supply) and chlorophyll a (a surrogate for autochthonous supply).     

Acidified lakes: sediment treatment with sodium carbonate — a remedy?

Until now, additions of lime have been used to restore the buffering capacity of acidified lakes, but an alternative method which is more effective in the treatment of lakes with organogenic sediments has recently been applied in a full-scale experiment. The method, called CONTRACID, is based on the cation exchange properties of lake sediment. A sodium carbonate (soda ash) solution is injected into the sediment (by a harrow), so that the sediment becomes sodium stocked. A reverse exchange occurs during subsequent acidification. Liming has a limited effect on humic lakes, since Ca-humates have a reduced reverse exchange ability and also the lime, which remains undissolved, is rendered inactive. Ionic exchange processes and nutrient transport were studied in water/sediment cores andin situ enclosures after additions of soda ash-, lye- and lime solutions with subsequent re-acidification. Sodium carbonate additions in laboratory systems resulted in a sorption to the sediment of 42–62% of the added sodium ions (5 eq m⁻²) and a release of 14–78 mg Pm⁻² sediment. Similar results were obtained in the enclosures where phosphorus release stimulated algal growth. Sediment pH, elevated by the sodium base addition, was lowered by re-acidification. Limed systems released no phosphorus and only about 25% of the added lime remained active for future neutralization. With the injection of the sodium carbonate solution into the sediment, only about 12% of the added sodium was recovered in lake water by spring circulation. Lake water alkalinity was then 0.12 meq l⁻¹ and pH 6.7. Total phosphorus had been raised by 0.007 mg P l⁻¹ causing an increase in phytoplankton biomass. Observations indicate that manipulations of acidic lake sediment according to the CONTRACID method create a long-lasting neutralizing capacity and a biological stimulation (through phosphorus release), which makes the method an attractive alternative to frequent liming.     

Bottom–up carbon subsidies and top–down predation pressure interact to affect aquatic food web structure

Human impacts such as eutrophication, overexploitation and climate change currently threaten future global food and drinking water supplies. Consequently, it is important that we understand how anthropogenic resource (bottom–up) and consumer (top–down) manipulations affect aquatic food web structure and production. Future climate changes are predicted to increase the inputs of terrestrial dissolved organic carbon to lakes. These carbon subsidies can either increase or decrease total basal production in aquatic food webs, depending on bacterial competition with phytoplankton for nutrients. This study examines the effects of carbon subsidies (bottom–up) on a pelagic community exposed to different levels of top–down predation. We conducted a large scale mesocosm experiment in an oligotrophic clear water lake in northern Sweden, using a natural plankton community exposed to three levels of glucose addition (0, 420 and 2100 μg C l^?1 total added glucose) and three levels of young‐of‐the‐year perch Perca fluviatilis density (0, 0.56 and 2 individuals m^?3). Bacterioplankton production doubled with glucose addition, but phytoplankton production was unaffected, in contrast to previous studies that have manipulated carbon, nutrients or light simultaneously. This suggests that carbon addition alone is not sufficient to reduce autotrophic production, at least in an oligotrophic lake dominated by mixotrophic phytoplankton. Larval perch grazing did not produce a classical trophic cascade, but substantially altered the species composition of crustacean zooplankton and ciliate trophic levels. Glucose addition increased the biomass of rotifers, thus potentially increasing energy transfer through the heterotrophic pathway, but only when fish were absent. This study illustrates that changes in community structure due to selective feeding by top‐predators can determine the influence of bottom–up carbon subsidies.     

Seasonal changes in temperature and nutrient control of photosynthesis, respiration and growth of natural phytoplankton communities

1. To investigate the influence of elevated temperatures and nutrients on photosynthesis, respiration and growth of natural phytoplankton assemblages, water was collected from a eutrophic lake in spring, summer, autumn, winter and the following spring and exposed to ambient temperature and ambient +2, +4 and +6 °C for 2 weeks with and without addition of extra inorganic nutrients. 2. Rates of photosynthesis, respiration and growth generally increased with temperature, but this effect was strongly enhanced by high nutrient availability, and therefore was most evident for nutrient amended cultures in seasons of low ambient nutrient availability. 3. Temperature stimulation of growth and metabolism was higher at low than high ambient temperature showing that long‐term temperature acclimation of the phytoplankton community before the experiments was of great importance for the measured rates. 4. Although we found distinct responses to relatively small temperature increases, the interaction between nutrient availability, time of the year and, thus, ambient temperature was responsible for most of the observed variability in phytoplankton growth, photosynthesis and respiration. 5. Although an increase in global temperature will influence production and degradation of organic material in lakes, the documented importance of ambient temperatures and nutrient conditions suggests that effects will be most pronounced during winter and early spring, while the remaining part of the growth season will be practically unaffected by increasing temperatures.     

Experimental Evaluation of Conversion Factors for the [3H]Thymidine Incorporation Assay of Bacterial Secondary Productivity

The relationship between bacterial growth and incorporation of [methyl-³H]thymidine in oligotrophic lake water cultures was investigated. Prescreening, dilution, and addition of organic and inorganic nutrients were treatments used to prevent bacterivory and stimulate bacterial growth. Growth in unmanipulated samples was estimated through separate measurements of grazing losses. Both bacterial number and biovolume growth responses were measured, and incorporation of [³H]thymidine in both total macromolecules and nucleic acids was assayed. The treatments had significant effects on conversion factors used to relate thymidine incorporation to bacterial growth. Cell number-based factors ranged from 1.1 × 10¹⁸ to 38 × 10¹⁸ cells mol of total thymidine incorporation⁻¹ and varied with treatment up to 10-fold for the same initial bacterial assemblage. In contrast, cell biovolume-based conversion factors were similar for two treatment groups across a 16-fold range of [³H]thymidine incorporation rates: 5.54 × 10¹⁷ μm³ mol of total thymidine incorporation⁻¹ and 15.2 × 10¹⁷ μm³ mol of nucleic acid incorporation⁻¹. Much of the variation in cell number-based conversion factors was related to changes in apparent mean cell volume of produced bacteria. Phosphorus addition stimulated [³H]thymidine incorporation more than it increased bacterial growth, which resulted in low conversion factors.     

Experimental evaluation of conversion factors for the [h]thymidine incorporation assay of bacterial secondary productivity

The relationship between bacterial growth and incorporation of [methyl-H]thymidine in oligotrophic lake water cultures was investigated. Prescreening, dilution, and addition of organic and inorganic nutrients were treatments used to prevent bacterivory and stimulate bacterial growth. Growth in unmanipulated samples was estimated through separate measurements of grazing losses. Both bacterial number and biovolume growth responses were measured, and incorporation of [H]thymidine in both total macromolecules and nucleic acids was assayed. The treatments had significant effects on conversion factors used to relate thymidine incorporation to bacterial growth. Cell number-based factors ranged from 1.1 x 10 to 38 x 10 cells mol of total thymidine incorporation and varied with treatment up to 10-fold for the same initial bacterial assemblage. In contrast, cell biovolume-based conversion factors were similar for two treatment groups across a 16-fold range of [H]thymidine incorporation rates: 5.54 x 10 mum mol of total thymidine incorporation and 15.2 x 10 mum mol of nucleic acid incorporation. Much of the variation in cell number-based conversion factors was related to changes in apparent mean cell volume of produced bacteria. Phosphorus addition stimulated [H]thymidine incorporation more than it increased bacterial growth, which resulted in low conversion factors.