Specific uptake rates of amino acids by attached and free-living bacteria in a mesotrophic lake

Seasonal and spatial patterns of specific uptake rates of amino acids by bacteria in Lake Constance were studied. The total bacterial population was divided into small (0.2- to 1.0-micron) and large (1.0- to 3.0-micron) free-living bacteria and attached bacteria by fractionated filtration. Data for attached bacteria, received by retention on 3.0-micron-pore Nuclepore filters, were corrected for free-living bacteria in this fraction. Specific uptake rates based on autoradiography were also recorded. Specific uptake rates for attached bacteria ranged from 9.41 X 10(-11) to 6.11 X 10(-8) ng of C h-1 cell-1 and were therefore significantly greater than those for free-living bacteria during most time periods. However, they were not significantly different from those for cells proven to be active by autoradiography. Specific uptake rates for small free-living bacteria ranged between 7.68 X 10(-11) and 4.60 X 10(-9) ng of C h-1 cell-1. They were nearly in the same range of those for large free-living bacteria (5.10 X 10(-11) to 1.07 X 10(-8) ng of C h-1 cell-1), although both fractions exhibited pronounced differences in their seasonal and vertical distributions.     

Production of amino acids by free-living heterotrophic nitrogen-fixing bacteria

Summary Interest of microbial production of amino acids has been increased greatly since development of biotechnological methods. These methods represent a perspective way applied in a future large-scale manufacture of inexpensive amino acids. In this context, the isolation of producing organisms that may be exploited in the desing of alternative methods for the production of amino acids could be of primary importance.In this review we will describe the liberation of amino acids (methionine, lysine, arginine, tryptophane and glutamic acid) byAzotobacter andAzospirillum during growth in culture media with different carbon sources under diazotrophic and adiazotrophic conditions. These organisms may be useful in developing new methods for the industrial production of amino acids.     

Use of dissolved carbohydrates by planktonic bacteria in a mesotrophic lake

Dissolved carbohydrates comprise one of the largest pools of labile organic matter readily available for bacterial use in pelagic ecosystems. Despite this fact, very little is known about use of dissolved carbohydrates by planktonic bacteria. We studied use of total dissolved carbohydrates (TDCHO) by planktonic bacteria in mesotrophic Lake Constance, Germany, from April until August 1992. We examined the decrease of TDCHO over time together with the increase of bacterial numbers in 1-m filtered lake water incubated at in situ temperature in the dark. TDCHO analyses were done after hydrolysis by sulfuric acid as free monosaccharides and oxidation by periodate to formaldehyde with 3-methyl-2-benzothiazolinone-hydrazon-hypochloride (MBTH). Preliminary tests showed that hydrolysis by sulfuric acid gave higher yields than hydrolysis by HCl. Our results show that TDCHO are readily used by bacteria and compose substantial fractions of the C requirements for their growth. Concentrations of TDCHO varied between 1.7 and 5.5 m (glucose equivalents), and use rates varied between 0.47 and 3.43 g C liter^–1 h^–1. Highest rates of TDCHO use occurred during the phytoplankton spring bloom, during the clear-water phase in June, and during a phytoplankton bloom in August. Ratios of use of TDCHO/bacterial biomass production varied between 0.17 and 3.05. During the spring bloom, TDCHO and total dissolved amino acids (TDAA) were used in equal amounts. During the clear-water phase at chlorophyll a concentrations <3 g liter^–1, however, bacteria only consumed TDCHO and excreted amino acids. The growth efficiency based on the consumption of TDCHO and TDAA varied between 16 and 21% during the phytoplankton spring bloom and was 35% during the clear-water phase. Correspondence to: M. Simon     

Growth limitation of planktonic bacteria in a large mesotrophic lake

We studied nutrient-limitation of bacterioplankton growth in Lake Constance, a mesotrophic lake, between February and August in 1992. We amended 1-m filtrates with a single nutrient or nutrient combinations at 5 or 10 m final concentration, and the limiting nutrient or nutrient combination was inferred from the assay in which bacterial growth was most stimulated. The following nutrients were added individually or in combination: glucose, amino acids, peptone, and ammonium as C and N sources, and inorganic phosphate. From January until the beginning of the phytoplankton spring bloom in mid-April, C alone was growth-limiting. During the spring bloom a complex growth-limitation pattern occurred; first P was limiting, then for only 1 week C + N together, and thereafter P + C. During the clear-water phase with very low chlorophyll concentrations, P + C together limited bacterial growth again, interrupted by a period when C + N + P shortage caused a triple limitation. Later in the season, P + C were growth-limiting again. The growth efficiency (bacterial biomass produced/substrates used) on the basis of amino acid and carbohydrate used varied between 17 and 35%. The addition of various C and N sources indicated that the growth efficiency strongly depended on the quality of the substrates and the adaptation of the bacterial assemblages, for example, whether C and N originated from amino acids or glucose and ammonium.     

Temporal variability of attached and free-living bacteria in coastal waters

The temporal variability of the abundance and the incorporation of ³H-thymidine and ¹⁴C-glucose by attached and free-living bacteria, as well as their relation with environmental factors, were analyzed in a coastal marine ecosystem during a year. Both communities were quantitatively very different. Attached bacteria represented only 6.8% of the total bacterial abundance, whereas free-living bacteria represented 93.2%. The environmental factors most closely linked to the abundance and activity of free-living bacteria were temperature and the concentration of dissolved nutrients. Moreover, the free-living community showed similar temporal variations in abundance and in activity, with lower values in the cold months (from October to May). The attached community did not present the same pattern of variation as the free-living one. The abundance of the attached bacteria was mainly correlated to the concentration of particulate material, whereas their activity was correlated to temperature. We did not find a significant correlation between the abundance and the activity of the attached community. On the other hand, the activity per cell of the two communities did not present a clear temporal variation. Attached bacteria were more active than free-living ones in the incorporation of radiolabeled substrates on a per cell basis (five times more in the case of glucose incorporation and twice as active in thymidine incorporation). However, both communities showed similar specific growth rates. The results suggest that the two aquatic bacterial communities must not be considered as being independent of each other. There appears to be a dynamic equilibrium between the two communities, regulated by the concentrations of particulate matter and nutrients and by other environmental factors.Offprint requests to: M. Unanue.     

Bloom of filamentous bacteria in a mesotrophic lake: identity and potential controlling mechanism

Ephemeral blooms of filamentous bacteria are a common phenomenon in the water column of oligo- to mesotrophic lakes. It is assumed that the appearance of such morphotypes is favored by selective predation of bacterivorous protists and that filter-feeding zooplankton plays a major role in suppressing these bacteria. The phylogenetic affiliation of the important bloom-forming filamentous bacteria in freshwaters is presently unknown. Here we report the identification of dominant members of a filamentous bacterial assemblage during a bloom of such morphotypes in a mesotrophic lake. By molecular cloning and fluorescence in situ hybridization with specific oligonucleotide probes, up to 98% of filamentous cells in lake water could be assigned to a clade of almost identical (99% similarity) 16S rRNA gene sequence types, the cosmopolitan freshwater LD2 cluster. For a period of less than 1 week, members of the LD2 clade constituted >40% of the total bacterial biomass, potentially favored by high grazing of planktivorous protists. This is probably the most pronounced case of dominance by a single bacterioplankton species ever observed in natural freshwaters. In enclosures artificially stocked with the metazoan filter feeder Daphnia, bacteria related to the LD2 clade formed a significantly larger fraction of filaments than in enclosures where Daphnia had been removed. However, in the presence of higher numbers of Daphnia individuals, the LD2 bacteria, like other filaments, were eventually eliminated both in enclosures and in the lake. This points at the potential importance of filter-feeding zooplankton in controlling the occurrence and species composition of filamentous bacterial morphotypes in freshwater plankton.     

Variations in the uptake and metabolism of peptides and amino acids by mixed ruminal bacteria in vitro.

Mixed ruminal bacteria, isolated from sheep (Q and W) fed a concentrate and hay diet, were anaerobically incubated with either 14C-peptides or 14C-amino acids. Experiment 1 showed that uptake of both 14C-labeled substrates was rapid, but the rate for amino acids was twofold greater than for peptides (molecular weight, 1,000 to 200) initially but was similar after 10 min. Experiment 2 demonstrated that metabolism was also rapid; at least 90% of either 14C-labeled substrate was metabolized by 3 min. Of the radioactivity remaining in bacteria, approximately 30% was in the form of 14C-amino acids, but only in leucine, tyrosine, and phenylalanine. Supernatant radioactivity was contained only in tyrosine, phenylalanine, and mostly proline for incubations with 14C-amino acids but in up to 10 amino acids when 14C-peptides were the substrates. Short-term incubations (< 5 min; experiment 3) confirmed previous uptake patterns and showed that the experimental system was responsive to substrate competition. Experiment 4 demonstrated that bacteria from sheep Q possessed initial and maximum rates of 14C-amino acid uptake approximately fourfold greater (P < 0.01) than those of 14C-peptides, but with no significant differences (P > 0.1) between four 14C-peptide substrate groups with molecular weights of 2,000 to < 200. By contrast, bacteria from sheep W showed no such distinctions (P > 0.1) between rates for 14C-peptides and 14C-amino acids. Calculations suggested that peptides could supply from 11 to 35% and amino acids could supply from 36 to 68% of the N requirements of mixed ruminal bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)     

A microautoradiographic study of the activity of attached and free-living bacteria

Microautoradiography, combined with epifluorescent microscopy, was used to evaluate the uptake of tritiated amino acids by a marine Pseudomonas sp. A comparison was made between the activity of bacteria free-living in the medium and bacteria which were attached to glass, polyethylene or polystyrene substrata. The proportion of active bacteria was lower for free-living cells (53–82%) and those attached to polystyrene (53–76%) than for those attached to glass (77–99%) or polyethylene (73–96%). For bacteria attached to glass, assimilated labelled substrate was retained within the cell over 3 h, whereas with polyethylene, labelled material was released from the cells and adsorbed on the surrounding substratum. Hence the physiological activity of attached bacteria depended on the chemical composition of the substratum.     

Anaerobic dechlorination of carbon tetrachloride by free-living and attached bacteria under various electron-donor conditions

The dechlorination of carbon tetrachloride (CCl₄) by free-living and attached bacteria under anaerobic conditions was studied to examine the relationship between porous media and electron donor. Two batch-type experiments, the free-living and attached bacterial systems, were conducted with and without addition of 0.5-mm glass beads. Glucose and acetate were selected as the primary electron donors because they are easily biodegradable. Direct epifluorescence technology, the DAPI (4′ 6-diamidino-2-phenylindole) method, was used for counting the microbial activities. Adding glass beads could accelerate the dechlorination rate of CCl₄. Removals of 44 %–57 % were observed in free-living bacterial system. Whereas a two- to fivefold increase in the CCl₄ dechlorination rate was observed in the attached system. Experimental results and thermodynamic calculations indicated that glucose is a better supplementary substrate than acetate for stimulating the dechlorinating capability of microorganisms because of its relatively high available free energy. A higher concentration of substrate provided more reducing power for attached bacteria to initiate the dechlorination reaction. The pseudo-first-order rate constants of CCl₄ dechlorination ranged from 0.007 day⁻¹ to 0.017 day⁻¹ and from 0.011 day⁻¹ to 0.0625 day⁻¹ for free-living and attached bacterial systems respectively. Microscopic observation revealed a three- to eightfold difference of microbial number between the free-living and attached bacterial systems. On the basis of the results in this study, we can conclude that the presence of porous media and an electron donor can change the dechlorination capabilities of the microorganisms. This work will be valuable in the design of in situ bioremediation as it discusses the specific area of the medium and supplementation with an electron donor to stimulate the indigenous microflora. Received: 21 June 1996 / Received revision: 2 September 1996 / Accepted: 29 September 1996     

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

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

Characterization of phytoplankton extracellular products (PDOC) and their subsequent uptake by heterotrophic organisms in a mesotrophic forest lake

The magnitude of phytoplankton release of photosyntheticallyproduced dissolved organic carbon (PDOC), the size distributionand chemical nature of the released substances, and the quantityand composition of PDOC that were immediately assimilated byheterotrophic bacteria were investigated in a meso- to oligotrophic,mesohumic forest lake by means of the ¹⁴CO₂ technique. The percentageextracellular release (PER) was comparatively high (up to {smalltilde}40%). The bulk of the released substances was of highmol. wt (>5000 dalton), with a minor contribution from lowmol. wt (<1000 dalton) compounds. Heterotrophic bacteriapreferentially assimilated the high molecular weight componentsof PDOC, and a polysaccharide of {small tilde}6000 dalton witha very short turnover time was a key substance in the carbonflow from autotrophic phytoplankton to heterotrophic bacteria.The shortcomings of the ¹⁴CO₂ method to assess phytoplanktonrelease of PDOC is critically evaluated and discussed.     

Importance of nutrients,organic matter and light availability on epilimnetic metabolic rates in a mesotrophic tropical lake

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Community structure of planktonic rotifers in a mesotrophic lake

Seasonal changes of species diversity (H_r) and average spatial overlap (ō_jk) of planktonic rotifers in a mesotrophic lake were examined with respect to their dependence on habitat diversity (H_h) and on a predator (Asplanchna herricki). H_r was positively correlated with the density of A. herricki and with H_h. In the absence of A. herricki there was no correlation between H_r and H_h. ō_jk was negatively correlated with the density of A. herricki and with H_h; the correlation between ō_jk and H_h was independent of the presence of the predator.     

Amino Acid assimilation and electron transport system activity in attached and free-living marine bacteria

[This corrects the article on p. 818 in vol. 45.].