The effects of initial population density on the competition for limiting nutrients in two freshwater algae

Eighteen long-term competition experiments were performed on two freshwater algae, a blue-green alga, Anabaena flos-aquae, and a diatom, Cyclotella sp., under controlled light and temperature conditions and various nutrient limitations. As predicted, Anabaena displaced Cyclotella when nitrate was in short supply to both species, whereas Cyclotella became dominant when both species were phosphate-limited. The two species stably coexisted when phosphate and silicate were limited. Anabaena either displaced or coexisted with Cyclotella when nitrate and phosphate or nitrate, phosphate and silicate were limited, depending on their initial density ratios. This study revealed strong effects of initial population densities on the outcomes of algal competition for limiting nutrients.     

Regulation of Expression of Nitrate and Dinitrogen Assimilation by Anabaena Species

Anabaena sp. strain 7120 appeared more responsive to nitrogen control than A. cylindrica. Growth in the presence of nitrate strongly repressed the differentiation of heterocysts and fixation of dinitrogen in Anabaena sp. strain 7120, but only weakly in A. cylindrica. Nitrate assimilation by ammonium-grown cultures was strongly repressed in Anabaena sp. strain 7120, but less so in A. cylindrica. The repressive effect of nitrate on dinitrogen assimilation in Anabaena sp. strain 7120, compared to A. cylindrica, did not correlate with a greater rate of nitrate transport, reduction to ammonium, assimilation into amino acids, or growth. Although both species grew at similar rates with dinitrogen, A. cylindrica grew faster with nitrate, incorporated more ¹³NO₃⁻ into amino acids, and assimilated (transported) nitrate at the same rate as Anabaena sp. strain 7120. Full expression of nitrate assimilation in the two species occurred within 2.5 h (10 to 14% of their generation times) after transfer to nitrate medium. The induction and continued expression of nitrate assimilation was dependent on protein synthesis. The half-saturation constants for nitrate assimilation and for nitrate and ammonium repression of dinitrogen assimilation have ecological significance with respect to nitrogen-dependent growth and competitiveness of the two Anabaena species.     

Blue-green algae and selection in rotifer populations

Summary Observation in natural populations of the rotifer Asplanchna girodi suggested blue-green algae may have toxic effects on rotifers. The blue-greens Anabaena flos-aquae and Lyngbya sp. were isolated from a pond in Central Florida along with three electrophoretically identified genotypes of A. girodi. When tested Anabaena had marked effect on the reproductive rate of A. girodi and the effect was different among genotypes. Likewise, Lyngbya also depressed rotifer reproduction, but required an order of magnitude higher concentration to produce the same effects as Anabaena. The toxicity of these algae was corroborated by mouse bioassays. Results clearly show these blue-greens have the capacity to differentially reduce reproductive rates of rotifer genotypes. Because of this capability blue-greens could play an important role in the ecology of natural selection in rotifer populations.     

Heterocyst formation and nitrogenase synthesis in Anabaena sp.

Summary When filaments from a culture of Anabaena sp. growing photoautotrophically with nitrate as a nitrogen source are placed in a nitrate-free mineral medium and incubated anaerobically in the light, the formation of heterocysts and the synthesis of nitrogenase both begin after a lag of about 24 hours. During the lag period, about 70% of the phycocyanin is destroyed. Under an atmosphere of N₂-CO₂, the nitrogenase activity rises to a peak value, and then falls markedly as growth at the expense of N₂ begins. Phycocyanin synthesis resumes concomitantly with growth. Under an atmosphere of Ar-CO₂, the formation of heterocysts and the synthesis of nitrogenase proceed to higher levels than those observed under N₂-CO₂, and the nitrogenase level is thereafter maintained. Under these conditions, neither growth nor resynthesis of phycocyanin occurs, and phycocyanin eventually falls to about 10% of its initial level in the filaments; however, growth can be promptly initiated if N₂ is admitted to the system. The implications of these findings are discussed.     

Cell to cell recognition between the ciliatePseudomicrothorax dubius and its food organisms: the role of surface charges

Summary The importance of charged groups during phagocytic recognition of filamentous Cyanobacteria (Oscillatoria formosa andAnabaena spp.) by the stenophagic ciliatePseudomicrothorax dubius has been studied. Anionic and cationic domains are evenly and randomly distributed over the cyanobacterial surface, as demonstrated with scanning electron microscopy following labeling with colloidal gold (–) and colloidal gold coupled with poly-L-lysine (+). The phagocytosis ofOscillatoria was inhibited when filaments were treated with cationic reagents such as poly-L-lysine (pLL), FeCl₃ and carbodiimide. In contrast elimination of cationic charges on theOscillatoria surface by treatment with poly-L-glutamic acid (pLGa) or colloidal gold did not affect phagocytosis. The effects of sequential treatment with pLL and pLGa demonstrated that pLL reduced phagocytosis of pLGa-pretreatedOscillatoria, whereas the pLGa restored phagocytosis of pLL-pretreated filaments. Scanning electron microscopy showed that pLL- or pLGa- treated filaments can still adsorb the oppositely charged colloidal gold particles on their surface. However, the treatment of filaments with pLL followed by pLGa prevented subsequent labeling with gold as well as with pLL-gold particles. Filaments ofAnabaena spp., which are not normally ingested byPseudomicrothorax, were also treated individually or sequentially with pLL and pLGa. None of these treatments, however, provoked phagocytosis ofAnabaena byPseudomicrothorax. We suggest that the surface charge alone does not play a crucial role in phagocytic recognition inPseudomicrothorax and that phagocytosis-specific molecules are implicated.     

A comparison of the influence of iron on the growth and nitrate metabolism of Anabaena and Scenedesmus

A comparative study of growth and nitrate metabolism of Anabaena flos-aquae (Lyng.) Bréb. and Scenedesmus bijugatus var. seriatus Chodat investigated possible mechanisms for the iron-stimulated increases in growth specific for blue-green algae in mixed algal communities. Algae were separately grown in an morganic medium with varying concentrations of iron and nitrate to determine the effects on each organism. Iron was found to be a limiting nutrient for cultures of both Anabaena and Scenedesmus as determined by chlorophyll a concentrations and cell enumeration. Both iron and nitrate stimulated the specific activity of nitrate reductase, nitrite reductase, and glutamine synthetase in Anabaena. Iron enrichment did not increase the activity of the enzymes in Scenedesmus, but inhibited the activity of nitrate reductase and glutamine synthetase. The stimulation of growth by iron in cells grown under iron limiting conditions was associated with increased nitrate metabolism in Anabaena but not in Scenedesmus.     

Benthic cyanobacterial bloom impacts the reefs of South Florida (Broward County, USA)

Benthic cyanobacteria of the genus Lyngbya can form prominent mats and blooms in tropical and subtropical coral reef and seagrass habitats worldwide. A Lyngbya bloom on the reef tract offshore of Broward County, Florida, was first noted in 2002, and although it is seasonally variable in its distribution and abundance, it has persisted and spread over the past 3 years. In this study, the most abundant species of Lyngbya found in the blooms have been identified and compared to other species of Lyngbya by morphological and molecular methods. The most common species of Lyngbya is consistent with the properties of Lyngbya confervoides C. Agardh. The 16S ribosomal DNA sequence shares 88–92% identity with other known Lyngbya sequences, suggesting that this bloom consists primarily of a new, previously unsequenced species of Lyngbya. The second most common Lyngbya in the bloom is consistent with Lyngbya polychroa. This persistent bloom is a concern because it smothers octocorals and other invertebrates and negatively impacts these southeastern Florida reefs.     

Effect of Anabaena flos-aquae on the abilities of Daphnia and Keratella to feed and reproduce on unicellular algae

SUMMARY.

• 1 This study tests the hypothesis that the filamentous cyanobacterium, Anabaena flos-aquae, inhibits Daphnia's ability to feed and reproduce on unicellular algae more than it does Keratella's. Double-label radiotracer experiments using whole and sonieally disrupted Anabaena filaments showed that even low concentrations of Anabaena (5 × 10³ cells ml^?1) inhibited the abilities of Daphnia pulex and D. galeata mendotae to feed on Chlamydomonas and that this inhibition was due to both mechanical interference with feeding and increased food availability. A single-label radiotracer experiment showed that Anabaena (5 × 10³ and 2 × 10⁴ cells ml^?l) reduced the abilities of neonate, juvenile, and adult D. pulex to feed on Chlamydomonas but had a much more pronounced effect on adults.
• 2 Single-label radiotracer experiments using screened (25 μm mesh) and unscreened suspensions of whole Anabaena filaments showed that only very high concentrations of Anabaena (10⁵ cells ml^?1) reduced the ability of Keratella cochlearis to feed on Cryptomonas and that this inhibition probably was due to mechanical interference with feeding, since Keratella ate Anabaena extremely inefficiently - especially when single cells and short filaments were removed by screening.
• 3 Experiments comparing the survivorship and fecundity of individuals fed Anabaena, Chlamydomonas, or no food showed that the Anabaena was not toxic to either Keratella or D. galeata mendotae, was not utilized by Keratella, and was utilized by Daphnia, although not as well as Chlamydomonas was.
• 4 Anabaena (5 × 10³ or 10⁴ cells ml^?1) did not affect, or slightly increased, the population growth rate (r_m) of K. cochlearis on Cryptomonas, while it substantially increased that of D. pulex, Duphnia's ability to utilize Anabaena probably more than offset its reduced ability to feed on Cryptomonas. The presence of cyanobacterial filaments in natural plankton communities could adversely affect Daphnia and decrease its ability to competitively suppress Keratella and other rotifers if the filaments were of little nutritional value to the Daphnia, greatly interfered

     

Regulation of nitrogenase levels in Anabaena sp. ATCC 33047 and other filamentous cyanobacteria

Incubation in the dark of photoautotrophically grown N₂-fixing heterocystous cyanobacteria leads to a loss of nitrogenase activity. Original levels of nitrogenase activity are rapidly regained upon re-illumination of the filaments, in a process dependent on de novo protein synthesis. Ammonia, acting indirectly through some of its metabolic derivatives, inhibits the light-promoted development of nitrogenase activity in filaments of Anabaena sp. ATCC 33047 and several other cyanobacteria containing mature heterocysts. The ammonia-mediated control system is also operative in N₂-fixing filaments in the absence of any added source of combined nitrogen, with the ammonia resulting from N₂-fixation already partially inhibiting full expression of nitrogenase. High nitrogenase levels, about two-fold higher than those in normal N₂-fixing Anabaena sp. ATCC 33047, are found in cell suspensions which have been treated with the glutamine synthetase inhibitor l-methionine-d,l-sulfoximine or subjected to nitrogen starvation. Filaments treated in either way are insensitive to the ammonia-promoted inhibition of nitrogenase development, although this insensitivity is only transitory for the nitrogen-starved filaments, which become ammonia-sensitive once they regain their normal nitrogen status.Abbreviations Chl chlorophyll - EDTA ethylenediaminetetraacetic acid - MSX l-methionine-d,l-sulfoximine     

Decreased 5-HT2C receptor binding in the cerebral cortex and brain stem during pancreatic regeneration in rats

The purpose of this study was to investigate the role of central 5-HT_2C receptor binding in rat model of pancreatic regeneration using 60–70% pancreatectomy. The 5-HT and 5-HT_2C receptor kinetics were studied in cerebral cortex and brain stem of sham operated, 72 h pancreatectomised and 7 days pancreatectomised rats. Scatchard analysis with [³H] mesulergine in cerebral cortex showed a significant decrease (p < 0.05) in maximal binding (B_max) without any change in K_d in 72 h pancreatectomised rats compared with sham. The decreased B_max reversed to sham level by 7 days after pancreatectomy. In brain stem, Scatchard analysis showed a significant decrease (p < 0.01) in B_max with a significant increase (p < 0.01) in K_d. Competition analysis in brain stem showed a shift in affinity towards a low affinity. These parameters were reversed to sham level by 7 days after pancreatectomy. Thus the results suggest that 5-HT through the 5-HT_2C receptor in the brain has a functional regulatory role in the pancreatic regeneration.     

16S rRNA GENE HETEROGENEITY IN THE FILAMENTOUS MARINE CYANOBACTERIAL GENUS LYNGBYA1

The SSU (16S) rRNA gene was used to investigate the phylogeny of the cyanobacterial genus Lyngbya as well as examined for its capacity to discriminate between different marine species of Lyngbya. We show that Lyngbya forms a polyphyletic genus composed of a marine lineage and a halophilic/brackish/freshwater lineage. In addition, we found morphological and genetic evidence that Lyngbya spp. often grow in association with other microorganisms, in particular smaller filamentous cyanobacteria such as Oscillatoria, and propose that these associated microorganisms have led to extensive phylogenetic confusion in identification of Lyngbya spp. At the species level, the phylogenetic diversity obtained from the comparison of 16S rRNA genes exceeded morphological diversity in Lyngbya. However, the expectation that this improved phylogeny would be useful to species and subspecies identification was eliminated by the fact that phylogenetic species did not correlate in any respect with the species obtained from current taxonomic systems. In addition, phylogenetic identification was adversely affected by the presence of multiple gene copies within individual Lyngbya colonies. Analysis of clonal Lyngbya cultures and multiple displacement amplified (MDA) single‐cell genomes revealed that Lyngbya genomes contain two 16S rRNA gene copies, and that these typically are of variable sequence. Furthermore, intragenomic and interspecies 16S rRNA gene heterogeneity was approximately of the same magnitude. Hence, the intragenomic heterogeneity of the 16S rRNA gene overestimates the microdiversity of different strains and does not accurately reflect speciation within cyanobacteria, including the genus Lyngbya.     

Isolation and characterization of heterocysts from Anabaena sp. strain CA

A comparative study has been made on the pigment composition and nitrogenase activity of whole filaments and isolated beterocysts from a mutant strain of Anabaena CA. The whole cell absorption spectra of intact filaments and isolated heterocysts showed close resemblance especially between 550–700 nm region. On a quantitative basis the chlorophyll a content was found almost equal between the vegetative cell and heterocyst but the c-phycocyanin content in the heterocyst was about 1/2 that of the vegetative cell. The purification of the phycobiliprotein on DEAE-cellulose showed the presence of c-phycocyanin (_max 615 nm) and allophycocyanin (_max 645 nm, shoulder 620 nm). Isolated heterocysts under H₂ showed acetylene reduction rates of 57 nmol C₂H₄/mg dry wt·min (342 mol C₂H₄/mg chl a·h), whereas intact filaments reduced at the rate of 18 nmol C₂H₄/mg dry wt·min (108 mol C₂H₄/mg chl a·h). This rate accounts for 30% recovery of nitrogenase activity in isolated heterocysts compared to whole filaments. The activity was strictly light dependent and was linear under H₂ for more than 3 h. Addition of as little as 5% H₂ under argon stimulated the C₂H₂ reductionseveral fold. The acetylene reduction (nitrogenase activity) also showed tolerance to 5% added O₂ either under H₂ or argon. The results suggest that the heterocyst of Anabaena CA-V is different in some characteristics (viz., higher endogenous C₂H₂ reduction rate, prolonged activity and higher levels of phycobiliproteins) than those reported in other Anabaena species.     

Consequences of the iron-dependent formation of ferredoxin and flavodoxin on photosynthesis and nitrogen fixation on Anabaena strains

Iron-dependent formation of ferredoxin and flavodoxin was determined in Anabaena ATCC 29413 and ATCC 29211 by a FPLC procedure. In the first species ferredoxin is replaced by flavodoxin at low iron levels in the vegetative cells only. In the heterocysts from Anabaena ATCC 29151, however, flavodoxin is constitutively formed regardless of the iron supply.Replacement of ferredoxin by flavodoxin had no effect on photosynthetic electron transport, whereas nitrogen fixation was decreased under low iron conditions. As ferredoxin and flavodoxin exhibited the same K_m values as electron donors to nitrogenase, an iron-limited synthesis of active nitrogenase was assumed as the reason for inhibited nitrogen fixation. Anabaena ATCC 29211 generally lacks the potential to synthesize flavodoxin. Under iron-starvation conditions, ferredoxin synthesis is limited, with a negative effect on photosynthetic oxygen evolution.     

Assessment of Environmental Conditions That Favor Hepatotoxic and Neurotoxic Anabaena spp. Strains Cultured under Light Limitation at Different Temperatures

Abstract Toxic cyanobacterial mass occurrences have caused animal poisonings worldwide and may pose a health hazard for humans. Strains of the genus Anabaena are either non-toxic or produce hepatotoxins, microcystins (MCYST), or neurotoxins (such as anatoxin-a). In order to study which growth conditions favor hepatotoxic vs neurotoxic strains and how production of toxins varies, we compared the responses of two microcystin- and two anatoxin-a-producing Anabaena strains in continuous turbidostat cultures, at different temperatures, under growth-limiting light levels. Growth rates consistently remained <0.8 divisions per 24 h. Differences were strain-specific and not associated with hepatotoxicity or neurotoxicity. Thus, differential adaptation of strains to temperature and to growth-limiting light levels cannot explain why, in some cyanobacterial water blooms, hepatotoxic strains, and in others, neurotoxic ones become dominant. A statistical analysis of field data showed that the most significant discriminating factors between different types of blooms were the concentrations of dissolved PO₄-phosphorus and NO₃-nitrogen. Anabaena blooms with unknown neurotoxicity associated with low PO₄-phosphorus and high NO₃-nitrogen concentrations. Among other Anabaena blooms, the hepatotoxic ones associated with the lowest, and most of the non-toxic ones with higher concentrations of PO₄-phosphorus. Anabaena blooms that contained anatoxin-a and hepatotoxic Microcystis blooms showed tendencies towards the highest concentrations of PO₄-phosphorus. Non-toxic blooms dominated by genera other than Anabaena occurred over a wide range of growth conditions. In turbidostat cultures, maximal production of microcystins correlated with maximal growth rates. Light regulated the production of MCYST-LR variants, and temperature affected the production of MCYST-RR variants. Anatoxin-a seemed to be produced most under temperatures and light levels slightly suboptimal for growth. Under low light, considerable amounts of extracellular anatoxin-a were detected while microcystins consistently remained intracellular. Received: 25 August 1997; Accepted: 2 December 1997     

Effects of temperature, and availability of nitrogen and phosphorus on the abundance of Anabaena and Microcystis in Lake Biwa, Japan: an experimental approach

Under optimal nutrient conditions, both Microcystis sp. and Anabaena sp. isolated from Lake Biwa grew optimally at 28–32°C but differed in maximal growth rates, phosphate uptake kinetics, maximal phosphorus quotas, and growth responses to nitrogen and phosphorus limitation. The maximal growth rates of Microcystis and Anabaena were 1.6 and 1.25 divisions day⁻¹, respectively. With phosphate and nitrate in the growth-limiting range, the growth of Microcystis was optimal at an N : P ratio of 100 : 1 (by weight) and declined at lower (nitrogen limitation) and higher (phosphorus limitation) ratios. In contrast, Anabaena growth rates did not change at N : P ratios from 1000 : 1 to 10 : 1. Starting with cells containing the maximal phosphorus quota, Microcystis growth in minus-phosphorus medium ceased in 7–9 days, compared with 12–13 days for Anabaena. The phosphate turnover time in cultures starved to their minimum cell quotas was 7.9 min for Microcystis and 0.6 min for Anabaena. Microcystis had a higher K _s (0.12 μg P l⁻¹ 10⁻⁶ cells) and lower V _max (9.63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), than Anabaena (K _s 0.02 μg P l⁻¹ h⁻¹ 10⁻⁶ cells; V _max 46.25 63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), suggesting that Microcystis would not be able to grow well in phosphorus-limited waters. We conclude that in spite of the higher growth rate under ideal conditions, Microcystis does not usually bloom in the North Basin because of low availability of phosphorus and nitrogen. Although Anabaena has an efficient phosphorus-uptake system, its main strategy for growth in low-phosphorus environments may depend on storage of phosphorus during periods of abundant phosphorus supply, which are rare in the North Basin. Received: July 31, 2000 / Accepted: October 18, 2000     

Characterization of cytochrome c 6 from the cyanobacterium Anabaena PCC 7119

 A soluble monoheme c–type cytochrome c ₆ has been isolated from the cyanobacterium Anabaena PCC 7119. It is a basic protein, with a molecular mass of 9.7 kDa, which accepts electrons from Anabaena ferredoxin in the ferredoxin-NADP⁺reductase-dependent NADPH cytochrome c reductase activity assay. The turnover of the reaction has an optimum pH at 7.5. Flavodoxin can also replace ferredoxin in this assay, but with only 20% efficiency. Plastocyanin from Anabaena PCC 7119, as well as the c ₆ cytochromes from the green algae Chlorella fusca and Monoraphidium braunii are also shown to accept electrons from Anabaena ferredoxin. The reduction potential of cytochrome c ₆ at pH 6.7 was determined to be 338 mV and is pH dependent, with pK _a ^ox=8.4±0.1 and pK _a ^red≈9.5. The ferric and ferrous cytochrome forms and their pH equilibria have been studied using visible, EPR and ¹H-NMR spectroscopies. The amino acid sequence and the visible and NMR spectroscopic data indicate that the heme iron has a methionine-histidine axial coordination in the pH range 5–11. However, the EPR data for the ferricytochrome are complex and show that in this pH range five distinct forms are present. Between pH 5 and 9 the spectrum is dominated by two rhombic species, with g–values at 2.94, 2.29, 1.43 and at 2.84, 2.34, 1.56, which interconvert with a pK _a of 8.4. The NMR data also show a main interconversion between two cytochrome forms at this pH, which coincides with that determined from the pH dependence of the reduction potential. Both these forms were associated with a methionine-histidine heme-iron coordination by correlation with the visible and NMR spectral data, although having crystal field parameters atypical for this type of coordination. Anabaena cytochrome c ₆ is one more example of a heme protein for which the widely used crystal field analysis of the EPR data (truth diagram) fails to unequivocally determine the type of heme-iron ligation. Received: 17 May 1996 / Accepted: 13 January 1997     

Opheliidae (Polychaeta) from the Southwestern Atlantic ocean, with the description of Travisia amadoi n. sp., Ophelina gaucha n. sp. and Ophelina alata n. sp.

The diet of the brackish water tilapia Sarotherodon melanotheron is studied in various riverine and lacustrine systems of the Bia River basin (Côte d'Ivoire). Comparison of the diet between sampling sites (upper course, lower course and man-made Lake Ayame) shows significant differences. In the man-made lake, prey described as preferential include Asterionella (Bacillariophyceae) and Bosmina(Cladocera). In the lower course, the preferential preys are represented by Lyngbya(Cyanophyceae) while in the upper course Lyngbya and chironomid larvae are preferential. These differences are related to the altered environmental conditions generated by the construction of the dam on the Bia river main stream. In the man-made lake, there is no shift in diet either with the seasons (dry and rainy seasons) or with size.     

Sustained ammonia production by immobilized filaments of the nitrogen-fixing cyanobacterium Anabaena 27893

Summary Whole filaments of the N₂-fixing cyanobacterium Anabaena ATCC 27893 have been immobilized by entrapment in calcium alginate gel beads. In a continuous flow fluidized bed reactor sustained photosynthesis, N₂-fixation, and ammonia production have been achieved over a 130 hour period, the longest tested.     

Planktonic nitrogen fixation in Lake Malawi/Nyasa

Nitrogen (N₂) fixation has been identified as possibly an important source of “new” nitrogen (N) to the epilimnion of Lake Malawi but studies in 1999–2000 and 2002 (September–December) estimated that the contribution of N₂-fixation by heterocystous Anabaena filaments to the N budget of the lake’s epilimnion is only 3–4% of total N input. N₂-fixation rates in Lake Malawi, as estimated by the acetylene reduction assay were higher during the stratified season (September–March) than during the rest of the year. Planktonic N₂-fixation in Lake Malawi can be monitored by measuring heterocyst biovolume concentrations because a significant correlation (r ² = 0.945, P < 0.0001) was observed between the two parameters. Heterocyst density cannot be used to estimate N₂-fixation because heterocyst sizes in the lake change continuously as at least two Anabaena species with different heterocyst dimensions are present. During September–October 2002, a species similar to A. maxima, contained larger heterocysts (16.34 ± 2.46 and 13.25 ± 1.89 μm in cross and apical section dimensions, respectively). In November–December of 2002, A. discoidea dominated and had smaller sized heterocysts (8.92 ± 1.13 and 7.25 ± 0.95 μm in cross and apical section dimensions). Since planktonic N₂-fixation is higher near shore than offshore, its contribution to the N budget in the littoral zone where high densities of grazing benthic fish occur may be more critical to maintaining the high rates of benthic algal productivity observed.