Comparative study of two meromictic basins of Lake Banyoles (Spain) with sulphur phototrophic bacteria

The annual limnological dynamics of two meromictic basins of Lake Banyoles (C-III and C-IV) have been studied and compared on the basis of their physical, chemical and biological characters. Stability values calculated for both basins gave 865 g cm cm⁻² and 495 g cm cm⁻² for C-III and C-IV respectively. These values are in agreement with the fact that C-IV was almost completely mixed during winter. In this basin, during stratification, the monimolimnion increased in thickness as the stability increased. Isolation of the respective monimolimnia resulted in the development of anoxic conditions and the accumulation of sulphide in both C-III and C-IV, which favoured the development of dense populations of sulfur phototrophic bacteria. The purple sulphur bacterium Chromatium minus and the green sulphur bacterium Chlorobium phaeobacteroides were identified as the main components of these photosynthetic populations. The different depths at which the O₂/H₂S boundary was situated in both basins (and consequently the different light intensity reaching this zone) determined the growth of these bacteria. Light intensities at the chemocline of C-IV reached values up to 5% of surface incident light. In contrast, in C-III this variable was sensibly lower, with values depending on season and seldom reaching 1%. Phototrophic bacteria were consequently found earlier in C-IV than in C-III, where no significant concentrations were found until August. Finally stability is discussed as an important factor controlling chemical and biological dynamics in meromictic lakes.     

Distribution of iron in Lake Banyoles in relation to the ecology of purple and green sulfur bacteria

The distribution of iron both in suspended sediment and in the water column has been studied during summer stratification in Lake Banyoles. In this lake, near bottom springs, a very fine material suspended sediment remains in suspension. Dissolved Fe²⁺ in interstitial water of this suspended sediment, is related to redox potential and to the bottom water inflow. In the water column, soluble iron is present in the hypolimnion of the six different basins forming Lake Banyoles. Under those conditions Fe²⁺ is partially removed by sulfide produced in the anoxic sediment. In addition, a peak of Fe²⁺ found at the density gradient level in basins C-III, C-IV and C-VI. A three compartment model on the dynamics of the processes involving iron in Lake Banyoles is proposed. The bottom springs supply oxygen to the anoxic hypolimnion affecting chemical processes of the iron cycle. The presence of phototrophic sulfur bacteria in the anoxic monimolimnion of basins C-III and C-IV can be related to the kinetics of Fe²⁺ and sulfide. In C-III sulfide concentration exceeds Fe²⁺ concentration whereas in C-IV sulfide is not detectable and iron reached values up to 60 mM. The presence of phototrophic sulfur bacteria in iron-containing environments with no detectable sulfide is explained by the ability of such microorganisms to use FeS as electron donor instead of H₂S.     

Rotifer vertical distributions in a meromictic basin of Lake Banyoles (Spain)

The vertical distribution of rotifer populations has been analyzed in a meromictic basin (III) of the karstic Lake Banyoles. This basin had, at the time of the study, an anaerobic monimolimnion from around 17 m in summer or 21 in winter to 25 m, its maximum depth. However, below this layer a warmer microaerophilic zone with suspended marls is found in a chimney to a depth of 130 m.The rotifer distributions of several years are compared. The presence of large populations of rotifers at the oxic-anoxic interface is constant through the years, but highly variable regarding the total abundance and relative proportions of the species, depending on the importance of the vertical mixing in winter and the posterior segregation of water layers as a function of production and decomposition processes. Filinia hofmanni, Filinia longiseta longiseta are species with a restricted occurrence at the oxycline and are not present in the other non meromictic basins of the lake. Filinia terminalis, more broadly distributed, also has dense populations in summer at the oxycline in basin III, but disappears from the plankton during this season in the other basins. Other species, such as Polyarthra dolichoptera, Anuraeopsis fissa, Keratella cochlearis and Kellicottia longispina, also showed maxima at the oxycline in summer, as well as other maxima in the metalimnion. Populations at the oxycline are probably different ecotypes adapted to low oxygen conditions, but having the advantage of high food availability.     

Rudd (Scardinius erythrophthalmus) introduced to the Iberian peninsula: feeding ecology in Lake Banyoles

The first data on the ecology of rudd (Scardinius erythrophthalmus) introduced to the Iberian peninsula are presented. The habitat and diet variation of rudd were studied in Lake Banyoles (Spain), an oligotrophic karstic lake dominated by exotic fish species. Rudd were strictly littoral and the diet was based on detritus and plant material. The most important animal prey were the cladocerans Daphnia longispinaand Scapholeberis rammneri, amphipods and several late stages of nematoceran dipterans. Rudd were more zooplanktivorous in spring and autumn and less in summer. There was also a size-dependent diet shift, from microcrustaceans to macroinvertebrates. The diet of rudd was also distinguished by the importance of plant material and various small neustonic invertebrates, particularly S. rammneriand late stages of nematocerans, showing a strong resource partitioning with other fish species. The degree of herbivory in Lake Banyoles was lower than usual.     

Sedimentary photosynthetic pigments of algae and phototrophic bacteria in Lake Hamana,Japan: temporal changes of anoxia in its five basins

We analyzed photosynthetic pigments of algae and bacteria (phototrophic sulfur bacteria: Chromatium and brown Chlorobium) in sediment cores and water samples obtained from five basins of Lake Hamana, a brackish, eutrophic, holomictic lake in Japan, and discussed our findings in relation to the distribution of the phototrophs. The four outer basins are connected to the central basin by narrow inlets. The prevalence of anoxia in Lake Hamana was demonstrated by the widespread presence of bacterial pigments in each core. The construction of training walls in 1954–1956 to direct tidal currents into the lake via Imagire-guchi Channel, the sole inlet for seawater, increased the lake water circulation, suppressed the development of anoxia, and caused Chromatium to disappear. Strong correlations (r ² 0.7) between total algal carotenoid (TAC) and total bacterial carotenoid (TBC) contents in each core were found in four basins. We ascribe this to the induction of anoxia by water stratification and algal proliferation, which precede the growth of phototrophic sulfur bacteria in the deeper layers of the water column. The slopes of the TBC–TAC correlations in the sediment cores, indicating the extent and stability of anoxia at each site, differed among basins (0.23–0.67) and were inversely related to the exchange rate of water by seawater intrusion in each basin.     

Pelodictyon phaeoclathratiforme sp. nov., a new brown-colored member of the Chlorobiaceae forming net-like colonies

A new strain of the green sulfur bacteria was isolated from the monimolimnion of Buchensee (near Radolfzell, Lake Constance region, FRG). Single cells were rod-shaped, nonmotile and contained gas vacuoles. Typical net-like colonies were formed by ternary fission of the cells. As photosynthetic pigments bacteriochlorophylls a, e, isorenieratene and -isorenieratene were present. Sulfide, sulfur and thiosulfate were used as electron donors during anaerobic phototrophic growth. Besides carbon dioxide, acetate and propionate could serve as carbon sources under mixotrophic conditions in the light. Like all other members of the green sulfur bacteria, the new bacterium is strictly anaerobic and obligately phototrophic. The possession of gas vacuoles and the formation of net-like colonies and the guanine plus cytosine content of the DNA (47.9 mol% G+C) are typical characteristics of the genus Pelodictyon. Because of its photosynthetic pigments which differ from those of Pelodictyon clathratiforme, strain BU 1 represents a new species, P. Phaeoclathratiforme sp. nov.     

Phototrophic bacterial production of oleic acid in an illuminated upflow anaerobic sludge blanket reactor

Phototrophic bacterial cells in the effluent from a lighted upflow anaerobic sludge blanket reactor supplied with a medium containing 142 mg S (as SO₄ ^2–) l^–1 accumulated a 6.8% w/w oleic acid content in cells and 19 mg cell-bound oleic acid l^–1 in the effluent. Pure cultures of Rhodopseudomonas palustris and Blastochloris sulfoviridis isolated from the effluent also accumulated 5.1 and 6.4% w/w oleic acid contents in cells, respectively. The oleic acid content in the cells recovered from the LUASB reactor effluent was related to the phototrophic bacterial population in the LUASB reactor. The inverse relationship was observed in the LUASB reactor between phototrophic bacterial growth and sulfate concentration in the influent.     

On the distribution patterns and population dynamics of sublittoral and profundal oligochaeta fauna from Lake Banyoles (Catalonia,NE Spain)

Oligochaeta assemblages from Lake Banyoles were composed of one species of Naididae and eight species of Tubificidae, Potamothrix heuscheri (8–1350 ind. m^–2) being the most abundant and widely distributed species, followed by Limnodrilus hoffmeisteri (25–858 ind. m^–2) and Psammoryctides barbatus (12–792 ind. m^–2). The remaining species were Potamothrix hammoniensis, Potamothrix bavaricus, Limnodrilus claparedeanus, Branchiura sowerbyi, Aulodrilus pigueti and Dero digitata, all of which occurred at lower densities and with restricted distributions.Regardless of sampling depth, maximum densities of oligochaetes were found in winter (up to 5,142 ind. m^–2 in December) and secondarily in June. Immature Tubificidae with hair setae and mature P. heuscheri were present and dominant throughout the year. Cestode parasites were recorded infesting L. hoffmeisteri and P. heuscheri populations.Oligochaeta densities increased with depth, but low oxygen concentrations (less than 1 mg l^–1) during a long period (4 months) in some basins of the lake acted as a key factor in reducing the density of worms and the species richness to one species, Potamothrix heuscheri.     

Vertical distributions of bacteria and algae in a steeply stratified humic lake under high grazing pressure from Daphnia longispina

The vertical distributions of bacteria and algae in a steeply stratified, highly humic lake were studied during three 24 h periods in summer. The highest bacterial and algal densities and biomasses were recorded in the anoxic hypolimnion. The bacterial biomass in the hypolimnion was composed mainly of photosynthetic green sulphur bacteria (Chlorobium) which occurred at very low light intensity (< 1.5 μmol m⁻² s⁻¹). The numbers and biomasses of bacteria, both in the epilimnion and hypolimnion, were mostly higher at night than during the day, indicating possible asynchrony between the production and loss of bacteria. Because of vertical migration, the diurnal vertical distribution of algae was more variable than that of bacteria. Particularly in July and August, when cryptomonads were abundant, the biomass of algae was much higher in the epilimnion during the day than at night. The flagellated chlorophytes, Chlamydomonas spp. and Scourfieldia cordiformis, stayed mainly in the upper hypolimnion close to the oxic-anoxic boundary zone where only a small proportion of Daphnia longispina was continuously present. Unpalatable Mallomonas chrysophytes with silicified plates and bristles, and small, presumably heterotrophic, flagellates stayed in the oxic epilimnion together with a dense (up to 300 ind l⁻¹) population of D. longispina. The results indicated that, besides the physical and chemical properties of the water column, grazing pressure by Daphnia longispina strongly affected the vertical distribution of microorganisms in this polyhumic lake.     

Distribution and breeding biology of offshore pelagic cyprinids and catfish in Lake Malawi/Niassa

Synopsis Lake Malawi/Niassa is a permanently stratified lake, consisting of an oxygenated layer of 230 m depth that overlies an anoxic zone that extends to the lake bottom at a maximum depth of over 700 m. Some fish are found throughout the oxygenated zone of the offshore waters, and lead a pelagic existence, although many of the species form part of the demersal community in shallower waters where the lake-bed is oxygenated. This paper reports on the distribution of cyprinids and catfish at six offshore locations, sampled with a mid-water trawl and gillnets, along the north-south axis of the lake. These two groups formed, respectively, 5% and 13% of the offshore fish biomass in the trawl catches. The offshore cyprinids comprised the small lake-spawningEngraulicypris sardella, the only species to have pelagic larvae, and the larger river-spawningOpsaridium microcephalum andO. microlepis. These cyprinids occurred mainly in the upper 100 m of the water column. The biomass of both larvae and adults ofE. sardella showed considerable fluctuations over the two years of study. The spawning site ofE. sardella remains a mystery, but is no longer believed to be in the offshore pelagic zone. The most abundant catfish was the mochokidSynodontis njassae, that exhibited diurnal movements from just above the oxic-anoxic boundary layer during the day to the surface layers at night. The clariid catfish of the genusBathyclarias were caught infrequently offshore, but owing to their large size, they could form a significant part of the biomass. Their abundance was difficult to quantify, as it was believed that the majority are active in the near-surface layers and were under-sampled by the trawl and gillnets. Information on some aspects of the breeding biology is included in the paper, although our knowledge in this area is very incomplete.     

Anoxygenic phototrophic bacteria of the high-altitude meromictic Lake Gek-Gel,Azerbaijan

The anoxygenic phototrophic bacterial community of the high-altitude meromictic Lake Gek-Gel (Azerbaijan) was investigated in September 2003. The highest concentration of bacteriochlorophyll e (48 μg/l) was detected at a depth of 30 m; the peak of bacteriochlorophyll a (4.5 μg/l) occurred at 29 m. Phylogenetic analysis revealed that brown-colored green sulfur bacteria Chlorobium phaeobacteroides predominated in the lake. Nonsulfur purple bacteria phylogenetically close to Blastochloris sulfoviridis were found in insignificant amounts; these organisms have not been previously reported in Lake Gek-Gel.     

On the significance of electron transport systems for growth of Rhodospirillum rubrum

The inhibitory effects of 2-hydroxybiphenyl on various electron transport reactions of isolated membranes and growth in the presence of malate of either phototrophic or chemotrophic cells of Rhodospirillum rubrum were studied. 50% inhibition of both oxygen uptake of whole cells and growth under chemotrophic conditions (i.e. aerobiosis in the dark) was achieved in the presence of 0.09 mM 2-hydroxybiphenyl. With isolated membranes the same effect on NADH oxidase was obtained with 0.08 mM of inhibitor. Succinate dependent respiratory reactions were inhibited by 50% at a concentration of 0.36 mM. Growth under phototrophic conditions (i.e. anaerobiosis in the light) was inhibited by 50% in the presence of 0.17 mM (wild type strain) or 0.21 mM (blue-green mutant, strain VI) of 2-hydroxybiphenyl. Photophosphorylation and light dependent NAD⁺ reduction by succinate were inhibited by 50% at concentrations of 0.21 mM and 0.03 mM of inhibitor, respectively. After phototrophic growth of the organisms for about five doublings of cell mass in the presence of 0.18 mM of 2-hydroxybiphenyl coloured carotenoids could no longer be detected. Membrane fractions of such cultures exhibited normal activities of succinate cytochrome c reductase but activities of NADH cytochrome c reductase were decreased by 80%. In comparison with a blue green mutant, strain VI, of R. rubrum light induced absorbance changes at 865 nm as well as activities of photophosphorylation were unaffected. However, no activity of light dependent NAD⁺ reduction with succinate could be detected. The data indicate that cellular respiration as well as chemotrophic growth depend largely on NADH dependent respiration. Phototrophic growth, on the other hand, is limited by photophosphorylation while energy dependent reversed electron flow to NAD⁺, if at all, is of rathe minor importance.Abbreviation BChl bacteriochlorophyll     

The regulation of cytochrome c oxidase of Rhodobacter capsulatus by light and oxygen

In order to distinguish between the regulatory effects of oxygen tension and light intensity on cytochrome c oxidase protein and enzymatic activity cells of Rhodobacter capsulatus were shifted from phototrophic (anaerobic, light) growth to aerobic-light, aerobic-dark and to anaerobic-dark conditions, respectively. During shift-experiments the formation of oxidase protein and regulation of oxidase activity was followed by immunological and enzymatic means. The results support the idea, that the formation of oxidase protein is regulated by oxygen tension and light intensity changes, whereas the regulation of oxidase activity seems only to be correlated to the oxygen tension. A DNA sequence involved in the oxygen-dependent regulation of cytochrome oxidase could be identified in the regulation-deficient oxidase mutant H41 of R. capsulatus. Immunological investigations of cytochrome c ₂ from mutant H41 demonstrated at the same time the participation of the c ₂-polypeptide in the regulation of cytochrome c oxidase.Abbreviations Bchl bacteriochlorophyll - CIE crossed immuno-electrophoresis - DMSO dimethyl sulfoxide     

Towards a phylogeny of phototrophic purple sulfur bacteria—16S rRNA oligonucleotide cataloguing of 11 species of Chromatiaceae

11 species from 7 genera of Chromatiaceae, depositing sulfur globules inside their cells, were analyzed by comparative oligonucleotide cataloguing of their 16S ribosomal RNA in order to determine the genealogical relationships to each other and to other Gram-negative phototrophic purple and non-phototrophic eubacteria. The rRNA data reveal that members of Chromatium, Amoebobacter, Lamprocystis, Thiocapsa, Thiocystis, Thiodictyon and Thiospirillum form a phylogenetically coherent grouping. The species investigated do not in each case cluster according to their present classification. Organisms storing sulfur inside their cells are moderately related to but clearly separated from members of Ectothiorhodospira, with which they form one of several sublines of group III of phototrophic purple bacteria as defined by Gibson et al. (1979).Dedicated to Professor H. G. Schlegel on the occasion of his 60th birthday.     

Partitioning of CO2 Incorporation Among Planktonic Microbial Guilds and Estimation of In Situ Specific Growth Rates

Partitioning of CO₂ incorporation into oxygenic phototrophic, anoxygenic phototrophic, and chemolithoautotrophic guilds was determined in a freshwater lake (Lake Cisó, Banyoles, Spain). CO₂ incorporation into the different types of microorganisms was studied at different depths, during diel cycles, and throughout the year. During winter holomixis, the whole lake became anoxic and both the anoxygenic and chemolithoautotrophic guilds were more active at the surface of the lake, whereas the activity of the oxygenic guild was negligible. During stratification, the latter guild was more active in the upper metalimnion, whereas the anoxygenic guild was more active in the lower metalimnion. Specific growth rates and doubling times were estimated for the most conspicuous phototrophic microorganisms. Doubling times for Cryptomonas phaseolus ranged between 0.5 and 192 days, whereas purple sulfur bacteria (Chromatiaceae-like) ranged between 1.5 and 238 days. These growth rates were similar to those calculated with a different approach in previous papers and indicate slow-growing populations with very large biomass. Overall, the annual total CO₂ incorporation in Lake Cisó was 220 g C m⁻². Most of the CO₂ incorporation, however, was due to the chemolithoautotrophic guild (61% during holomixis and 56% during stratification), followed by the anoxygenic phototrophic guild (35 and 19%, respectively) and the oxygenic phototrophs (4 and 25%, respectively), making dark carbon fixation the key process in the autotrophic metabolism of the lake.     

A Study of the Chromatiaceae in a Saline Meromictic Lake in Saskatchewan,Canada

Lamprocystis roseopersicina was found to be the dominant phototrophic bacterium in meromictic Deadmoose Lake. Using carbon-14 it was determined that phototrophic bacteria fixed 14.3 g C m⁻² year⁻¹, 17.1 % of the total limnetic primary production. Experiments indicated low light intensities (X = 8.87 × 10⁻² cal cm⁻²hr⁻¹) to be the most important factor limiting bacterial production. Lamprocystis was shown to be capable of photoorganoheterotrophic uptake of glucose.     

Photoinactivation of the detoxifying enzyme nitrophenol reductase from Rhodobacter capsulatus

The phototrophic bacterium Rhodobacter capsulatus E1F1 detoxifies 2,4-dinitrophenol by inducing an NAD(P)H-dependent iron flavoprotein that reduces this compound to the less toxic end product 2-amino-4-nitrophenol. This nitrophenol reductase was stable in crude extracts containing carotenes, but it became rapidly inactivated when purified protein was exposed to intense white light or moderate blue light intensities, especially in the presence of exogenous flavins. Red light irradiation had no effect on nitrophenol reductase activity. Photoinactivation of the enzyme was irreversible and increased under anoxic conditions. This photoinactivation was prevented by reductants such as NAD(P)H and EDTA and by the excited flavin quencher iodide. Addition of superoxide dismutase, catalase, tryptophan or histidine did not affect photoinactivation of nitrophenol reductase, thus excluding these reactive dioxygen species as the inactivating agent. Substantial protection by 2,4-dinitrophenol also took place when the enzyme was irradiated at a wavelength coinciding with one of the absorption peaks of this compound (365nm). These results suggest that the lability of nitrophenol reductase was due to the absorption of blue light by the flavin prosthetic group, thus producing an excited flavin that might irreversibly oxidize some functional group(s) necessary for enzyme catalysis. Nitrophenol reductase may be preserved in vivo from blue light photoinactivation by the high content of carotenes and excess of reducing equivalents in phototrophic growing cells.Abbreviations 2,4-DNP 2,4-dinitrophenol - ANP 2-amino-4-nitrophenol - EDTA ethylenediamine tetraacetic acid - MES 2-(N-Morpholino) ethanesulfonic acid - NPR nitrophenol reductase     

Phytoplankton, bacterial production and protozoan bacterivory in stratified, brackish-water Lake Shira (Khakasia, Siberia)

Rates of oxygenic and anoxygenic photosynthesis, chemoautotrophic and heterotrophic bacterial production and protozoan bacterivory were measured in the pelagic zone of the stratified brackish-water lake with the purpose to determine the vertical distribution of these processes and to estimate their significance in the functioning of planktonic community of the lake. In midsummer, total daily primary productivity was about 1.3 g C m^–2, of which 72% was produced by the phytoplankton, 24% by the chemoautotrophic bacteria, and only 4% by the phototrophic sulphur bacteria. Thus anoxygenic photosynthesis is a negligible source of organic matter in the lake. The production of heterotrophic bacteria averaged 1.5 g C m^–2 d^–1 and exceeded the total photosynthesis of phytoplankton and photosynthetic bacteria by a factor of 1.5. The estimated total primary production was too low to sustain the bacterial production. Probably the carbon cycle in the lake is dependent on the input of allochthonous organic matter. As a rule, the maximal rates of primary production and heterotrophic bacterial production were found in the chemocline or at the upper boundary of the chemocline. Heterotrophic flagellates dominated among the protozoan populations and were the major consumers of the bacterioplankton production in the lake. They showed maximal ingestion rates from 2.3 to 2.9 mg C m^–3 h^–1 at the upper boundary of the chemocline, where they consumed from 50 to 54% of the production of heterotrophic bacteria. Data obtained indicate that in Lake Shira the oxic-anoxic interface is the site of the most intensive production and mineralization of organic matter.     

Effects of light quality on the physiology and the ecology of planktonic green sulfur bacteria in lakes

The effect of light quality on the selection of natural populations of Green Sulfur Bacteria (Chlorobiaceae) is considered to be a classic factor in the determination of their ecological niches. From the comparison among phototrophic bacterial populations of lakes, it is shown that brown and green pigmented groups of Chlorobiaceae have a differential distribution depending on depth. Statistical analyses prove that green species, which dominate at shallow oxic/anoxic boundaries, are correlated to light spectra enriched in long wavelengths, while brown ones are found when light spectra are enriched in the central region of the spectrum, as in deeper lake layers. Physiological experiments have been made withChlorobium limicola andC. phaeobacteroides cultures placed under different light quality conditions, in order to verify these hypotheses made on a field data basis. Results show that red and white light has more positive effects on the green bacterium than on the brown. Blue and green light illuminations have opposite consequences. Therefore, the effect of shallow depths and Chromatiaceae shading—which also increases the proportion of long wavelengths in light spectra—benefits the bacteriochlorophyll-based strategies of green species. On the other hand, the carotenoid-based strategies of brown ones are favored by the light climates usually dominant at greater depths. Thus, brown species are considered to be singular adaptations of Chlorobiaceae to depth, where bacteriochlorophyll light-harvesting is strongly limited by light quality.     

Expression of luciferase genes from different origins in Bacillus subtilis

Summary A group of vectors for luciferase expression in Bacillus subtilis was constructed. So far, only bacterial luciferases have been expressed in Bacillus, but in this study we wanted also to express genes encoding eukaryotic luciferases to perform direct comparisons of the light levels produced by the two different systems in B. subtilis. The vectors constructed can replicate both in Escherichia coli and B. subtilis, and the luciferase expression is strictly regulated due to the dual plasmid system used. Nearly a 100-fold increase in light production compared to previous results was achieved when genes encoding bacterial luciferase were inserted into the constructs and transformed into B. subtilis. An additional tenfold increase in light production was obtained when luciferase genes from the North American firefly (Photinus pyralis) or a click beetle (Pyrophorus plagiophtalamus) were introduced in a similar fashion into B. subtilis. Measurement of the light emission was performed without disruption of bacterial cells in a real-time manner, which is a common feature when working with all of these constructions. Structures of the shuttle vector constructs and results from light emission measurements are presented.