Biomass relationship to growth and phosphate uptake ofPseudomonas fluorescens,Escherichia coli andAcinetobacter radioresistens in mixed liquor medium

The ability ofPseudomonas fluorescens, Escherichia coli andAcinetobacter radioresistenns to remove phosphate during growth was related to the initial biomass as well as to growth stages and bacterial species. Phosphate was removed by these bacteria under favourable conditions as well as under unfavourable conditions of growth. Experiments showed a relationship between a high initial cell density and phosphate uptake. More phosphate was released than removed when low initial cell densities (10²–10⁵ cells ml^–1) were used. At a high initial biomass concentration (10⁸ cells ml^–1), phosphate was removed during the lag phase and during logarthmic growth byP. fluorescens. Escherichia coli. at high initial biomass concentrations (10⁷ cells ml^–1), accumulated most of the phosphate during the first hour of the lag phase and/or during logarithmic growth and in some cases removed a small quantily of phosphate during the stationary growth phase.Acinetobacter radioresistens, at high initial cell densities (10⁶, 10⁷ cells ml^–1) removed most of phosphate during the first hour of the lag phase and some phosphate during the stationary growth phase.Pseudomonas fluorescens removed phosphate more thanA. radioresistens andE. coli with specific average ranges from 3.00–28.50 mg L^–1 compared to average ranges of 4.92–17.14 mg L^–1 forA. radioresistens and to average ranges of 0.50–8.50 mg L^–1 forE. coli.     

Life table demography and population growth of Brachionus variabilis Hempel, 1896 in relation to Chlorella vulgaris densities

We studied the life history variables and population growth characteristics of Brachionus variabilis, which was recorded for the first time from Mexico. The animals were fed Chlorella, using five concentrations (0.25, 0.5, 1, 2 and 4 × 10⁶ cells ml^–1) at 25 °C. Food density was observed to have significant effect on life expectancy, average lifespan, gross reproductive rate, net reproductive rate, generation time and population growth rate. The average lifespan ranged from 3 to 6 days depending on the food density. The net reproductive rate ranged from 2 to 7 neonates female^–1 d^–1. The rate of population increase per day varied from 0.14 to 0.35. The highest net reproductive rate and average lifespan and life expectancy were recorded at Chlorella concentrations of 1 × 10⁶ and 2 × 10⁶ cells ml^–1.     

Effect of two microalgae concentrations on body size and egg size of the rotifer <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis>

The rotifer, Brachionus calyciflorus, was grown with two algae species (Chlorella sp. and Scenedesmus obliquus) at different concentrations (0.1, 1 and 10 × 10⁶ cells ml⁻¹). The body size (lorica biovolume) of individual rotifer and their egg size were measured when the populations were roughly in the exponential phase of population growth. The body size of the rotifers differed significantly (P < 0.05) among the two algae species used, however this effect was not observed for egg size. The body size of rotifers fed on higher densities of Chlorella sp. (10 × 10⁶ cells ml⁻¹) was significantly larger than for those fed on lower and medium densities (0.1 and 1 × 10⁶ cells ml⁻¹). Body size and egg size of rotifers fed with different amounts of Scenedesmus did not differ significantly. The egg size was significantly larger at higher food level of Chlorella. A significantly positive correlation was observed between the adult rotifer body size and their egg size.     

Ecological problems of Lake Ladoga: causes and solutions

We studied the outcome of competition between a large (Brachionus calyciflorus) and a small (Anuraeopsis fissa) rotifer species at five algal (Scenedesmus acutus) concentrations (0.5 × 10⁶ to 40.5 × 10⁶ cells ml^–1) and with varying initial densities in mixed populations (100 to 0% of B. calcyciflorus or A. fissa), the combined initial biomass being 0.2 µg ml^–1 in all test jars. Experiments were conducted at 28 ± 1 °C.Regardless of food concentration, B. calcyciflorus showed a greater increase in biomass than A. fissa, peak densities (mean ± standard error) at the lowest food concentration in the controls being 1.34 ± 0.31 µg dry weight ml^–1 and 0.82 ± 0.08 dry weight ml^–1, respectively. At the lower food concentrations, A. fissa displaced B. calyciflorus and vice versa at the higher food concentrations. At the intermediate food concentrations of 4.5 × 10⁶ cells ml^–1, B. calyciflorus outcompeted A. fissa only if its initial population density was three times higher. The rates of population growth in controls varied from 0.792 ± 0.06 d^–1 to 1.492 ± 0.13 d^–1 for B. calyciflorus and 0.445 ± 0.04 to 0.885 ± 0.01 for A. fissa depending on food level. When both species were introduced together, low food levels favoured higher abundance of A. fissa than B. calyciflorus, suggesting, in nature, it is likely that small Anuraeopsis colonize oligotrophic water bodies more successfully than larger Brachionus. The results also suggest that the outcome of competition depends not only on the size of the competing species and food availability but also on their colonizing density.     

Investigation of enhancement of two processes, sedimentation and conjugation, when bacteria are concentrated in ultrasonic standing waves

Cells aggregate and can be recovered from suspension when exposed to an ultrasonic standing wave field. The acoustic force on individual cells in a standing wave decreases with particle volume. A plane ultrasonic field generated by a transducer driven at 3.3 MHz was used here to investigate the removal of Escherischia coli, cells with dimensions of the order of 1.0 m, from batch suspension by sedimentation over a range of concentrations (10³ to 10¹⁰ cells ml^–1). Cell removal efficiencies greater than 90% were achieved at initial concentrations of 10¹⁰ cells ml^–1. Removal efficiencies decreased gradually to zero, as initial bacterial concentration was reduced to 10⁷ cells ml^–1. It was found that, when low concentrations of E. coli (10³ to 10⁵ cells ml^–1) were added to suspensions of larger particles (i.e. yeast cells) that were of sufficient concentration to form aggregates in the sound field, E. coli could be harvested to an efficiency of 40%. The results imply that the E. coli became trapped and sediment with aggregates of larger particles. Some strains of bacteria are capable of DNA transfer by conjugation. The transfer rate of E. coli RP4 plasmid is order of magnitudes greater when conjugation occurs on solid medium rather than in liquid suspension. We have investigated whether the conjugation rate would also be higher in ultrasonically induced E. coli clumps than in free suspension. The donor strain was mixed with a recipient strain of E. coli, then sonicated in a capillary at 4.6 MHz in a tubular transducer for 5 min. The bacteria aggregated successfully. Results showed a three-fold increase in the rate of conjugation compared to a liquid mating control.     

Production and nutritional quality of the rotifer Brachionus plicatilis fed marine Chlorella sp. at different cell densities

The effect of different cell densities of marine Chlorella sp. on the growth rate, doubling time and production of the rotifer Brachionus plicatilis was investigated. A significant increase in rotifer production was achieved at a density of 50 × 10⁶ Chlorella cells ml^–1. The nutritional quality of rotifers grown at different concentrations of Chlorella is discussed.     

Utilization of cyanobacteria by Brachionus calyciflorus: Anabaena flos-aquae (NRC-44-1) as a sole or complementary food source

The rotifer Brachionus calyciflorus can utilize the cyanobacterium Anabaena flos-aquae as either a sole or supplementary food source in laboratory culture. Positive population growth rates accompany food densities of 10 or 100 µg dry weight ml^–1, but slightly negative rates are found at a lower density (1.0 µg ml^–1). These results are consistent for rotifers feeding on two strains of A. flos-aquae, UTEX-1444 and NRC-44-1, with slightly enhanced survivorship and reproduction with the latter food. A 1:1 mixture (by dry weight) of Euglena gracilis and A. flos-aquae (NRC-44-1) produces survivorship comparable to that of control rotifer cohorts fed E. gracilis alone, but elicits significantly greater fecundity and population growth rates than found with the control food suspension at the same biomass density.     

High density culture of the freshwater rotifer,Brachionus calyciflorus

The freshwater rotifer, Brachionus calyciflorus is one of the live food organisms used for the mass production of larval fish. In this study possibility of obtaining high density cultures of the freshwater rotifer B. calyciflorus were investigated. The two culture systems used differed in their air and dissolved oxygen supplies using three temperatures in each case: 24, 28 and 32 °C. Rotifers were batch-cultured using 5 l-vessels and fed with the freshwater Chlorella. The growth rate of rotifers significantly increased with an increase in temperature. The maximum density of the rotifers with air-supply at 24 °C, 6500 ind. ml^–1, was significantly lower than those cultured at 28 and 32 °C, i.e. 8600 and 8100 ind. ml^–1, respectively. Dissolved oxygen levels decreased with time and ranged from 0.8 to 1.4 mg l^–1 when the density of freshwater rotifer was the highest at each temperature. The highest density (19200 ind. ml^–1) of freshwater rotifer was obtained in cultures with a supply of oxygen at 28 °C. Densities of 13500 and 17200 ind. ml^–1 were found at 24 and 32 °C, respectively. Levels of NH₃-N increased with time and a dramatic increase of NH₃-N was observed at high temperatures. Levels of NH₃-N at 24, 28 and 32 °C were 13.2, 18.5 and 24.5 mg l^–1, respectively. These levels coincided with the highest rotifer density at each of the three temperatures. When rotifers were cultured with an oxygen-supply and pH was adjusted to 7, the maximum density of rotifer reached 33500 ind. ml^–1 at 32 °C . These results suggested that high density culture of freshwater rotifer, B. calyciflorus could be achieved under optimal conditions with DO value of exceeding 5 mg l^–1 and NH₃-N values of lower than 12.0 mg l^–1.     

Extracellular DNA in aquatic ecosystems may in part be due to phycovirus activity

In a eutrophic lake, a crash of the algal population was followed by a significant increase in the number of virus-like particles (from ca. 1 10⁶ ml^–1 to ca. 26 10⁶ ml^–1), and soon thereafter by an increase of the amount of extracellular DNA (from ca. 20 µg l^–1 to ca. 40 µg l^–1). The same pattern of correlation between decrease of algae and increase of viruses and extracellular DNA could be demonstrated by an in vitro experiment with a Chlorella-virus-system. Lysis of algae by viruses increased both the number of viruses and the amount of DNA in the culture medium. Extracellular DNA mainly consisted of material with a molecular weight below 500 bp.The Chlorella-virus-system is discussed. It could be used as a model-system for studying the dynamics of interaction of viruses and algae in aquatic ecosystems.     

Rotifers as predators on small ciliates

Clearance rates of Synchaeta pectinata, Brachionus calyciflorus and Asplanchna girodi on Tetrahymena pyriformis (46 µm in length) at a density of 10 cells ml^–1, in the presence of algal food, were 2.5 to 6.1 ml rot.^–1 day^–1. Clearance rates of these rotifers were, respectively, about 2, 3, and 13 times lower on Strobilidium gyrans (58 µm in length) than on T. pyriformis, indicating that the saltations of S. gyrans are an effective escape response. Clearance rates of S. pectinata were considerably lower on Colpidium striatum (81 µm) than on S. gyrans, suggesting that S. pectinata may not be able to ingest ciliates of this size. S. pectinata had a clearance rate of 19 ml rot.^–1 day^–1 on S. gyrans at a density of 1.2 cells ml^–1, in the absence of edible algal food. Rotifers may prey extensively on ciliates in natural plankton communities, ingesting 25 to 50 individuals in the 45–60 µm size range day^–1.     

Demographic parameters of Brachionus patulus Muller (Rotifera) exposed to sublethal DDT concentrations at low and high food levels

The combined effects of sublethal levels of DDT (0, 15, 30, 45, and 60 µg·l^–1) and food (Chlorella at 1 and 3 × 10⁶ cells ml^–1) on the demography of the rotifer Brachionus patulus were studied. The average lifespan, life expectancy, net reproductive rate (R_o) and intrinsic rate of increase (r) were significantly greater at the higher food level, but declined with increasing DDT concentration. A significant DDT food interaction was manifested in the toxicity-mitigating action of the higher food density at higher DDT levels. The r values were negative at DDT levels 30 µg · l^–1 and above, only when food density was low. The effective DDT concentration at which a given parameter value was reduced to 50% of that in the controls (EC50) was lower at lower food levels for both survivorship and reproduction, and lower for reproduction than for survivorship at either food level.     

Aerobic nitrate and nitrite reduction in continuous cultures of Escherichia coli E4

Nitrate and nitrite was reduced by Escherichia coli E4 in a l-lactate (5 mM) limited culture in a chemostat operated at dissolved oxygen concentrations corresponding to 90–100% air saturation. Nitrate reductase and nitrite reductase activity was regulated by the growth rate, and oxygen and nitrate concentrations. At a low growth rate (0.11 h^–1) nitrate and nitrite reductase activities of 200 nmol · mg^–1 protein · min^–1 and 250 nmol · mg^–1 protein · min^–1 were measured, respectively. At a high growth rate (0.55 h^–1) both enzyme activities were considerably lower (25 and 12 nmol mg^–1 · protein · min^–1). The steady state nitrite concentration in the chemostat was controlled by the combined action of the nitrate and nitrite reductase. Both nitrate and nitrite reductase activity were inversely proportional to the growth rate. The nitrite reductase activity decreased faster with growth rate than the nitrate reductase. The chemostat biomass concentration of E. coli E4, with ammonium either solely or combined with nitrate as a source of nitrogen, remained constant throughout all growth rates and was not affected by nitrite concentrations. Contrary to batch, E. coli E4 was able to grow in continuous cultures on nitrate as the sole source of nitrogen. When cultivated with nitrate as the sole source of nitrogen the chemostat biomass concentration is related to the activity of nitrate and nitrite reductase and hence, inversely proportional to growth rate.     

Direct and indirect interactions for coexistence in a species-defined microcosm

Mechanisms for coexistence among micro-organisms were studied by using a species-defined microcosm, consisting of the bacterium Escherichia coli, the ciliate Tetrahymena thermophila and the alga Euglena gracilis. These organisms were chosen as representative of ecological functional groups i.e. decomposer, consumer and producer, respectively. Direct and indirect interactions among these organisms were evaluated by comparisons of their population dynamics in culture with different combinations of the three species. There was an E. coli cell density dependent predator–prey interaction between T. thermophila and E. coli which was only established when there were more than 10⁶ cells ml^–1 of E. coli. Indirect interactions were evaluated from the cultivation of each organism in media containing metabolites of the others. Metabolites from each population strongly accelerated the growth of their own populations and those of the others except for the self-toxicity effect of E. coli metabolites. These observations suggested that not only the cell–cell contact of direct interactions, but also metabolite-mediated indirect interactions supported the maintenance of the populations of each micro-organism and their coexistence. In natural ecosystems, there are many interactions and it is difficult to evaluate all those regulating community dynamics. The gnotobiotic microcosm used in this study was shown to be suitable for examining the specific, species–species microbial interactions.     

Effect of Aeolosoma sp. (Aphanoneura: Aeolosomatidae) on the Population Dynamics of Selected Cladoceran Species

Although oligochaete worms naturally coexist with cladocerans in many shallow freshwater ponds and lakes, their influence on the latter is not well established. In this work we studied the effect of Aeolosoma sp. on the population growth of Alona rectangula, Ceriodaphnia dubia, Daphnia pulex, Macrothrix triserialis and Moina macrocopa. Population growth studies were conducted at one algal food density (1 × 10⁶cells ml^–1 of Chlorella vulgaris). The experimental design was similar for all five cladoceran species, where we used 100 ml capacity transparent jars containing 50 ml of EPA medium with the desired algal density and three replicates for each treatment. The test medium was changed daily and fresh algal food was added. The initial density of each of the cladoceran species in the population growth studies was 0.4 ind ml^–1 while that of the worms 1.0 ind ml^–1. Following inoculation, we estimated daily the number of cladocerans and the worms for duration of 21 days. Regardless of the presence of worms, Moina macrocopa and Macrothrix triserialis showed rapid population growth while A. rectangula took more than 2 weeks to reach peak abundances. With the exception of M. triserialis, all the other our cladoceran species declined in the presence of Aeolosoma sp. The lowest peak population density (about 1 ind ml^–1) was observed for M. triserialisin controls. The remaining species had peak densities of about 3–5 ind ml^–1. The rates of population increase per day varied from 0.03 to 0.19 depending on the cladoceran taxa and the treatment. In general we found that pelagic taxa were more adversely affected by the presence of the worms than were the littoral cladocerans.     

Efficient production by Escherichia coli of recombinant protein using salting-out effect protecting against proteolytic degradation

To produce glucoamylase efficiently as a recombinant protein, E. coli was grown with 20 g (NH₄)₂SO₄ l^–1 which removed proteolytic activity but did not effect cell growth. Growth in M9 medium with 20 g (NH₄)₂SO₄ l^–1 produced 11 U glucoamylase ml^–1 compared to 7 U ml^–1 without addition. Furthermore, the glucoamylase activity was maintained at about 9 U ml^–1.     

Chaff piles of harvester ant (Messor spp.) nests in a desert ecosystem

Summary The plant species composition of the chaff piles of three species of harvester ant (Messor spp.) and the contribution of the chaff to the organic pool were studied from August 1985 to July 1987. There were distinct differences in the plant species composition of the chaff of the three species. We attribute this to the different diets of the three species, which reflect the relative sizes of their individuals and their foraging strategies. The amount of chaff accumulated varies greatly between the three species (Messor rugossus: 127–196 g · ha^–1 · y^–1;Messor ebeninus: 2823–4437 g · ha^–1 · y^–1;Messor arenarius: 2165–2535 g · ha^–1 · y^–1), although the number of nests per hectare is virtually the same. We found that the amount of chaff is related to the rate of activity and the size of the individuals of each of the three ant species. The total chaff accumulated during the study period was 19.2 kg · ha^–1, which is an important contribution to the organic matter in the soil in the Negev desert ecosystem.     

Oxygen availability for bacterial growth with a flow microcalorimeter

Summary The enthalpy change associated with aerobic growth of E. coli K₁₂ on minimal media with succinic acid as sole carbon and energy source, determined by flow microcalorimetry (with aerobic mixing cell) was 733.01±15.32 kJ·mol^–1. Molar growth yield was 39.6±1.2 g·mol^–1. When the microcalorimetric growth was limited by oxygen supply, the power-time curve was altered and the total heat evolved was less than the enthalpy change. The maximum thermal output corresponding to a fully aerobic growth in the calorimetric cell was 1.89×10^–3 W·ml^–1. Thus, the oxygen uptake rate was about 0.39 ml O₂·h^–1·ml^–1.     

Suspension feeding in Bithynia tentaculata (Prosobranchia,Bithyniidae), as affected by body size,food and temperature

The suspension feeding of Bithynia tentaculata was tested in laboratory experiments. The animals were fed in 1-1 aerated glass beakers, and filtration rates were calculated from changes in cell concentrations during the 6-h experiment. Temperature influenced the filtering rate, with minimum values of 5ml · ind^–1 · h^–1 at 5° C and maxima of 17.2 ml · ind^–1 · h^–1 at 18° C. Three food species of different size, motility and cell surface characteristics (Chlamydomonas reinhardii, Chlorella vulgaris and Chlorogonium elongatum) did not affect filtration rates. Suspension feeding increased with increasing food concentrations up to 12 nl · ml^–1, above which feeding rate was kept constant by lowering the filtering rates. Even the smallest animals tested (<4 mm body length) were found to be feeding on suspended food at a rate of 2.7 ml · ind^–1 · h^–1, and increasing rates up to 8.4 ml were found in the 6–7 mm size class. All size classes of Bithynia showed a circannual fluctuation of their filtration rates. The ecological consequences of Bithynia's ability to switch between two feeding modes, grazing and suspension feeding, are discussed.     

Overproduction and secretion of Bacillus circulans endo-β-1,3-1,4-glucanase gene (bglBC1) in B. subtilis and B. megaterium

A gene coding for endo--1,3-1,4-glucanase (lichenase) containing a recombinant plasmid, pLL200K, was transferred from Bacillus circulans into a new shuttle plasmid, pLLS920, by ligating linearized DNAs of pLL200K and pUB110. B. subtilis RM125 and B. megaterium ATCC14945 transformed with pLLS920 produced the endo--1,3-1,4-glucanase. The enzyme was produced during active growth with maximum activity. The B. subtilis (pLLS920) enzyme was 83 times (8522 mU ml^–1) more active than that of the gene donor cells (103 mU ml^–1). The B. megaterium (pLLS920) enzyme was 7 times (735 mU ml^–1) more active than that of the gene donor cells. While E. coli secreted only about 10% of the produced enzyme, B. subtilis excreted the enzyme completely into the medium and B. megaterium by about 98%. The plasmid pLLS920 was stable in B. megaterium (98%), and in B. subtilis (51%) but not in E. coli (29%).     

Segregation to non-dividing cells in recombinant Escherichia coli fed-batch fermentation processes

In Escherichia coli fermentation processes, a drastic drop in viable cell count as measured by the number of colony forming units per ml (c.f.u. ml^–1) is often observed. This phenomenon was investigated in a process for the production of the recombinant fusion protein, promegapoietin (PMP). After induction, the number of c.f.u. ml^–1 dropped to 10% of its maximum though the biomass concentration continued to increase. Flow cytometric analysis of viability and intracellular concentration of PMP showed that almost all cells were alive and contributed to the production. Thus, the drop in the number of c.f.u. ml^–1 probably reflects a loss of cell division capability rather than cell death.