Screening of algal strains for metal removal capabilities

Eight algal species were tested for their ability to remove five toxic metalsduring 30-min exposures to single-metal (1 mg L^-1) solutions at pH7. Efficacy of metal bioremoval varied according to algal species and metal. Al⁺³ was best removed by the thermophilic blue-green alga(cyanobacterium) Mastigocladus laminosus, Hg⁺² and Zn⁺² by the thermophilic and acidophilic red alga Cyanidiumcaldarium, and Cd⁺² by C. caldarium and the green alga Scenedesmus quadricauda. All of these alga/metal combinations resultedin >90% metal removal. However, none of the eight algal speciesremoved more than 10% of Cr⁺⁶. Results indicate that some toxicmetals are more readily removed than others are by algae and that selectionof appropriate strains could potentially enhance bioremoval of specificmetals from wastewater at neutral pH.     

Studies on Nitrogen Fixation by Blue-green Algae: II. NITROGEN FIXATION BY MASTIGOCLADUS LAMINOSUS COHN

Mastigocladus laminosus Cohn, a blue-green alga of hot springs,has been obtained in pure bacteria-free culture and has beenshown to be capable of fixing elementary nitrogen. This appearsto be the first instance in which this capacity has been demonstratedin a blue-green alga belonging to a family other than the Nostocaceae.     

A study of several blue-green algae in the electron microscope

Summary All of the three blue-green algae, Anabaena cylindrica, Mastigocladus laminosus and Nostoc muscorum are characterized by the presence of multi-layered envelopes (sheath, wall and plasma membrane), photosynthetic lamellae and a variety of intracellular granules. Sections of heterocysts of Anabaena cylindrica showed the presence of an internal membrane system as well as lamellae. An unusual feature of the structure of Nostoc muscorum was the presence of densely stained intracellular membranes or lamellae. The results emphasize the variability in appearance of the internal structure of the blue-green algae and point to the need for detailed investigations of the influence of change in physiological environment on the anatomy of these organisms.     

Nitrogen Fixation by Thermophilic Blue-Green Algae (Cyanobacteria): Temperature Characteristics and Potential Use in Biophotolysis

Thermophilic, nitrogen-fixing, blue-green algae (cyanobacteria) were investigated for use in biophotolysis. Three strains of Mastigocladus laminosus were tested and were found to be equally effective in biophotolysis as judged by nitrogenase activity. The alga, M. laminosus NZ-86-m, which was chosen for further study, grew well in the temperature range from 35 to 50°C, with optimum growth at 45°C, at which temperature acetylene reduction activity was also greatest. The maximum tolerable temperature was 55°C. Acetylene reduction activity was saturated at a light intensity of 1 × 10⁴ ergs cm⁻² s⁻¹. Atmospheric oxygen tension was found to be slightly inhibitory to acetylene reduction of both slowly growing and exponentially growing cultures. Nonsterile continuous cultures, which were conducted to test problems of culture maintenance, could be operated for 2 months without any significant decrease in nitrogenase activity or contamination by other algae. Nitrogen-starved cultures of M. laminosus NZ-86-m produced hydrogen at comparable rates to Anabaena cylindrica. The conversion efficiency of light to hydrogen energy at maximum rates of hydrogen production was 2.7%.     

Hydrogen Production by the Thermophilic Alga Mastigocladus laminosus: Effects of Nitrogen, Temperature, and Inhibition of Photosynthesis

Hydrogen production by nitrogen-limited cultures of a thermophilic blue-green alga (cyanobacterium), Mastigocladus laminosus, was studied to develop the concept of a high-temperature biophotolysis system. Biophotolytic production of hydrogen by solar radiation was also demonstrated. Hydrogen consumption activity in these cultures was relatively high and is the present limiting factor on both the net rate and duration of hydrogen production.     

ISOLATION,GROWTH, AND RESPIRATION OF A THERMOPHILIC BLUE-GREEN ALGA

Holton , Raymond W. (U. Texas, Austin.) Isolation, growth, and respiration of a thermophilic blue-green alga. Amer. Jour. Bot. 49(1): 1–6. Illus. 1962.—The isolation of a bacteria-free culture of Hapalosiphon laminosus (= Mastigocladus laminosus) from an algal mass obtained from Liard Hot Springs, British Columbia, is described. Seven different bacterial media at 2 incubation temperatures were used to test for bacteria in suspensions of partially broken up algae. The algal growth rate was obtained by calculating the slope of the straight line obtained in a plot of the cube root of the dry wt against time. In cultures aerated with air the growth was optimal between 35 and 50 C, with an upper limit at 55 and a lower limit at 25. Aeration with 1% CO₂ in air significantly increased growth rates and the optimum was at 45–50 C. Unaerated flasks grew more slowly than aerated ones. The endogenous Qo₂ (in μl/hr–mg dry wt) at 45 was 8.0 for growing cells and 2.8 for starved cells kept in darkness 18 hr. The Qo₂ was maximal between pH 7.0 and 8.5. The concentration of phosphate buffer affected the Qo₂ and rates in 0.02 M were slightly greater than those in distilled water and much greater than those in 0.4 M. The Qo₂ of growing cells was slightly stimulated by 2,4-dinitrophenol (DNP), whereas glucose or glucose plus DNP did not affect it. In contrast, the endogenous Qo₂ of starved cells was nearly doubled in glucose and tripled in DNP plus glucose.     

Morphological and physiological characteristics of a non-heterocystous strain of the cyanobacterium Mastigocladus laminosus Cohn from fumarolic soil on Mt Erebus,Antarctica

Summary An unusual strain of the thermophilic cyanobacterium Mastigocladus laminosus occurs on warm soils on the volcano Mt Erebus (77°32S, 167°8E). It differs morphologically from the two genetically distinct forms described from thermal habitats elsewhere. Heterocysts are lacking and true-branching is rare. Its photosynthetic rate, and the contrasting rates of two less thermotolerant algae from Mt Erebus soils, Phormidium fragile (Cyanobacteria) and Pseudococcomyxa simplex (Chlorophyta), were measured over the range -2° to 62°C. The optimum temperature range of M. laminosus was 35° to 50°C. Photosynthetic response to temperature of all three algae in the laboratory correlated well with distribution patterns in the field, confirming that zonation patterns were temperature controlled. M. laminosus retained viability following exposure to deep-freezing, freeze-thaw cycles and desiccation. Viability of the alga in culture was lost following exposure to 50°C in darkness for 42 days and following 42 days in the light at 0°C. Discussion suggests the alga would survive long distance airborne dispersal in the desiccated condition but would not survive the duration of overwinter darkness on moist soils at the warmer end of its range of occurrence in the field.     

Low molecular weight carbohydrates in Cyanidium caldarium and some related algae

Floridoside (2-O-glycerol-α-d-galactopyranoside) and a small amount of iso-floridoside (1-O-glycerol-α-d- galactopyranoside) were found in Cyanidium caldarium. Floridoside was also found in the red algae Porphyridium cruentum and Porphyra yezoensis, although in the latter iso-floridoside was the main component. Sucrose and glucose were found in the green algae Chlorella pyrenoidosa and Scenedesmus obliquus, and also in a blue-green alga, Anacystis nidulans. Another blue-green alga, Phormidium foveolarum, contains mostly trehalose. From these results and from morphological considerations, it is suggested that Cyanidium caldarium belongs to the primitive Rhodophyta.     

Studies with cyanidium caldarium,an anomalously pigmented chlorophyte

Summary Cyanidium caldarium, an alga found in acid hot springs troughout the world, has a morphology and developmental history resembling those of Chlorella, but contains C-phycocyanin and no chlorophyll other than chlorophyll a. The reasons for considering it to be a member of the Chlorophyta are reviewed. Cyanidium is also remarkable for its thermal and acid tolerance. It grows readily in the dark on sugar media. However, light is required for the formation of chlorophyll and phycocyanin except in occasional variant cells which can form limited amounts of these pigments in the dark. Light-grown Cyanidium carries out normal green plant photosynthesis but resembles the red and some of the blue-green algae in that chlorophyll-absorbed light is used with lower efficiency than that absorbed by phycocyanin.The possible significance of the unusual pigmentation of Cyanidium is discussed.Contribution no.23 from the Laboratory of Comparative Physiology and Morphology of The Kaiser Foundation.     

Characteristics of softwater streams in Rhode Island II. Composition and seasonal dynamics of macroalgal communities

Forty stream segments in Rhode Island, U.S.A., were examined seasonally from June 1979 to March 1982. Thirty-nine species of macroalgae were collected, respresenting 25 genera. The composition of the lotic flora was 54% green algae, 31% red algae, 5% blue-green algae, 5% xanthophytes, 3% chrysophytes and 3% diatoms. The majority of these taxa (85%) were filamentous. From a biweekly examination of five stream segments, macroalgal communities could be grouped according to light regime. Species in unshaded streams exhibited little seasonality, whereas in streams shaded by one or more layers of riparian canopy, maxima in species numbers and abundance occurred during colder seasons. The most widespread and abundant species were the blue-green alga Phormidium retzii, the green alga Draparnaldia acuta, and the diatom Eunotia pectinalis. P. retzii and E. pectinalis were aseasonal annuals, while D. acuta was primarily a winter-spring form. It appears that pH is a major factor affecting broad geographic distribution patterns of stream macroalgae, whereas the light regime established by overhanging canopy is an important factor which influences localized abundance and seasonality of lotic macroalgal communities. Niche pre-emption appears to be a common mode of resource space division among stream macroalgae in Rhode Island. E. pectinalis is the strongly developed dominant in this drainage system.     

Storage glucan and glucosyltransferase isozymes of cyanidioschyzon merolae: A primitive eukaryote

Cyanidioschyzon merolae, a primitive eukaryotic alga isolated from supposedly pure cultures of the thermoacidophilic alga, Cyanidium caldarium, has many of the characteristics of such prokaryotes as bacteria and the cyanobacteria. Cyanidioschyzon appears to have even more of these prokaryotic features than does Cyanidium. Cyanidioschyzon divides by binary fission as do most bacteria. Its thylakoids are arranged along the periphery of the cell, like the cyanobacteria. Its formation of storage glucan, as well as the type of sugar formed is more like that of the blue-green algae rather than that of the red algae. Cyanidioschyzon merolae may be much more primitive than Cyanidium caldarium, and could be the most primitive eukaryotic cell.     

Benthic algal populations respond to aluminum,acid, and aluminum-acid mixtures in artificial streams

Elevated aluminum (Al) concentrations are often associated with acid-stressed aquatic ecosystems, so it has been unclear whether acidic water or elevated Al is more responsible in changing community composition. Experiments were done to investigate effects of acidification and increased Al on the abundance of benthic algae in artificial streams supplied with natural water and nominal treatments of (a) pH 4.8, (b) 500 µg l^-1 Al, or (c) the mixture of pH 4.8 and 500 µg l^-1 Al compared to a control without added Al or acid. These treatments are referred to as Acid, Al-only, Acid + Al, and the control, respectively. In the Acid treatment the abundance of two diatoms, two green algae, dry weight biomass, and chlorophyll a decreased; one diatom and one filamentous blue-green alga increased. In the Al-only treatment, densities of two diatoms, one green alga, one blue-green alga, dry weight biomass, and chlorophyll a increased. In the Acid + Al treatment, abundances of one green alga, two blue-green algae, and concentrations of chlorophyll a decreased below the levels observed in the Acid treatment. Acid and Al concentrations were altered by each other and by chemical and biological processes in the stream system. Species of diatoms, green algae, and blue-green algae responded individually to treatments and mixtures of acid and Al. Shifts in the abundance of species may change food web relationships for higher-level consumers, and algae may be useful biomonitors of ecological stress.     

DISTRIBUTION AND PHYSIOLOGICAL DETERMINANTS OF BLUE-GREEN ALGAL NITROGEN FIXATION ALONG A THERMOGRADIENT1

The capacity of thermal algal-bacterial mats to fix nitrogen (N₂) was examined in an alkaline thermal stream, Rabbit Creek, of Yellowstone National Park. Nitrogenase activity and nitrogen-fixation rates of mat cores placed in serum bottles and incubated in situ were estimated by the acetylene-reduction technique. Active nitrogenase was not detected at 60 or 65 C in either the blue-green algal or bacterial undermat components of the mats. Acetylene was reduced by all mats ≤55 C along the thermogradient; mean fixation estimates for the mats ranged from 7 to 5,028 nmoles N₂ fixed · mg Chl a^?1· hr^?1. Maximum fixation occurred at 35 C in the stream; statistical comparison of mean rates ordered the thermogradient mats according to estimated activities: 35 > 40 > 30 > 50 ≥ 55 ≥ 45 C. Mats (≤40 C) dominated by species of Calothrix accounted for ca. 97% of the total nitrogen fixation observed in the stream; the remaining activity was associated with mats containing Mastigocladus laminosus Cohn. Light intensity significantly affected fixation rates of the Calothrix mats which responded in a linear fashion from 9–100% full sunlight (ca. 1,900 μEin · m^?2· sec^?1). Calothrix mats from 30 and 40 C had maximum nitrogenase activity at their growth temperature suggesting that nitrogen fixation along the thermogradient was optimally adapted to in situ temperatures.     

Cyanidium caldarium as a bridge alga between Cyanophyceae and Rhodophyceae: Evidence from immunodiffusion studies

The primer-independent phosphorylase isozyme, a₂, of Cyanidiumcaldarium was used for immunization of rabbits. The immune serumwas tested against pure a₂ isozymes from blue-green, red, andgreen algae. Double immunodiffusion in agar indicated that therewas structural similarity in the isozyme from Cyanidium caldarium,the blue-green algae, Oscillaloria princeps, Pleclonema nostocorumand the red alga, Rhodymenia pertusa. Complete fusion of theprecipitin lines was obtained with these algae. However, onlypartial fusion was observed with the a₂ isozymes from Chlorophyceaesuch as Chlorella pyrenoidosa and Spirogyra setiformis. Spurformation on the precipitin lines occurred when the isozymesfrom these algae were tested against the immuneserum. The results were interpreted as indicative of the possible transitionstatus of Cyanidium caldarium between prokaryotic blue-greenalgae and eukaryotic red algae. It would appear that the Chlorophyceaeevolved along different lines from Cyanophyceae than did theRhodophyceae. (Received November 25, 1975; )     

Composition and biosynthesis of thylakoid membrane polypeptides in the red alga Cyanidium caldarium: Comparison with the thylakoid polypeptide composition of higher plants and cyanobacteria

The polypeptide composition of thylakoid membranes of the red alga Cyanidium caldarium was studied by PAGE in the presence of lithium dodecyl sulfate. The thylakoid membranes were shown to contain 65 polypeptides with mol wt from 110 to 10 kDa. PS I isolated from C. caldarium cells is composed of at least 5 components, one of which is the chlorophyll-protein complex with mol wt of 110 kDa typical of higher plants. Cyt f, c ₅₅₂, b ₆ and b ₅₅₉ were identified. Inhibition of carotenoid biosynthesis with norflurazon caused no changes in the polypeptide composition of thylakoid membranes of the algae grown in dark. The suppression of the biosynthesis rate of some thylakoid polypeptides in the algae grown with norflurazon in light is a result of membrane photodestruction. Thylakoid membranes from C. caldarium cells are more similar in the number of protein components to thylakoid membranes from cells of the cyanobacterium Anacystis nidulans than to those of higher plants (Pisum sativum), which was proved by immune-blotting assays: Thylakoid membranes of the red alga and cyanobacteria contain 28 homologous polypeptides, while thylakoid membranes of the alga and pea, only 15.Abbreviations CD circular dichroism - CP chlorophyll-protein complex - LDS lithium dodecyl sulfate - NF norflurazon     

DEPRESSION OF MASTIGOCLADUS LAMINOSUS (CYANOPHYTA) NITROGENASE ACTIVITY UNDER NORMAL SUNLIGHT INTENSITIES1

The effect of light intensity (PAR) on the nitrogenase activity of Mastigocladus laminosus Cohn was studied by the acetylene reduction technique. Benthic mat from a thermal stream, Hot River, in Yellowstone National Park was used in both experimental and in situ incubations. This hot spring maintained a mean pH of 7.0, was essentially isothermal (ca. 50°C), and had virtually no upstream to downstream physicochemical gradients (P > 0.05). Two surveys of the stream showed that nitrogenase of the M. laminosus mat was significantly more active (P > 0.02) under low light intensities than under high intensities, 252 and 712 μE · m^?2· s^?1, respectively. Maximum activity of Hot River Mastigocladus (268 nmol C₂H₄· mg Chl a^?1· h^?1) occurred at 50% full midday light intensities; the rates at low light (mean = 247 nmol C₂H₄· mg Chl a^?1· h^?1) were significantly (P > 0.001) greater than those at high light (mean = 106). The results indicate that M. laminosus nitrogenase activity is low light adapted and suggest that the temporal pattern for nitrogen fixation might be significantly different from that of thermophilic Calothrix.     

Photosynthetic regeneration of ATP using a strain of thermophilic blue-green algae

Photosynthetic ATP accumulation was shown in the presence of exogenous ADP plus orthophosphate on illumination to the intact cells of a strain of thermophilic blue-green algae isolated from Matsue hot springs, Mastigocladus sp. Kinetic studies of various effectors on the ATP accumulation proved that the ATP synthesis depends mainly on the cyclic photophosphorylation system around photosystem I (PS-I) in the algal cells. The temperature and pH optima for the accumulation were found at 45 degrees C and pH 7.5. Maximum yield was obtained with light intensity higher than 15 mW/cm(2). Borate ion exerted pronounced enhancement on the ATP synthesis. With a continuous reactor at a flow rate of 1 ml/hr at 45 degrees C and pH 7.5, efficient photoconversion of ADP (2mM, at substrate reservoir) to ATP (1mM, at product outlet) has been maintained for a period of 2.5 days, though the efficiency has decreased in a further 2-day period to the level of 0.5mM ATP/9.5 h of residence time.     

THE EFFECT OF SULFIDE ON THE BLUEGREEN ALGAE OF HOT SPRINGS. I. NEW ZEALAND AND ICELAND1

The concentration of soluble sulfide (H₂S, HS^?, S⁼)² in hot springs of New Zealand and Iceland has a “species-determining” effect which appears to override the effect of all other chemical factors excepting hydrogen ion concentration. The cosmopolitan high-temperature form (HTF) of Mastigocladus laminosus (Ag.) Cohn appears to be sensitive to concentrations of S⁼ over 0.15 mg·l^?1 in New Zealand and 0.25 mg·l^?1 in Iceland. It was absent in thermal streams where S⁼ level was high enough to exceed this concentration at a point below its optimal temperature range (i.e., below ca. 50 C). In low or non-S⁼ springs in these regions this alga formed mats to an upper temperature limit of 63–64 C. In contrast, one bluegreen alga (Oscillatoria amphigranulata van Goor) occurred abundantly only in moderate to high S⁼ springs of New Zealand; this was a species with an upper temperature limit of about 57 C. Field ¹⁴C-HCO₃^? incorporation experiments with various levels of added S⁼ confirmed the sensitivity of HTF Mastigocladus and the great tolerance of O. amphigranulata. In the latter, the photosynthetic incorporation of ¹⁴C-HCO₃^? was sustained or enhanced by the additions of S⁼ in the presence of 5 × 10^?5m DCMU, an inhibitor of photoelectron transport on the reducing side of photosystem II. It is possible, but not proven, that S⁼ may act as a photoreductant in this species.     

Morphological and genetic diversity of the thermophilic cyanobacterium,Mastigocladus laminosus (Stigonematales,Cyanobacteria) from Japan and Myanmar

The morphology and phylogeny of 13 strains of a thermophilic cyanobacterium, Mastigocladus laminosus Cohn, isolated from hot springs in Japan and Myanmar were analyzed to determine taxonomy and biogeography. From the morphological observations of cell size, there were significant differences among strains. Phylogenetic analyses based on 16S rRNA gene sequences revealed two lineages: Lineages I and II. Lineage I consisted of strains collected in Japan and reference strains from a previous study (CCMEE 5329 and 5331, Hakone, Japan); Lineage II included all of the Myanmar strains and one Japanese strain, and was a novel lineage in phylogeographic studies on M. laminosus. Since strains in the Lineage II tended to have larger cells than those in the Lineage I, the morphological and phylogenetic lineages corresponded well with each other.     

Evidence that electron-dense bodies in Cyanidium caldarium have an iron-storage role

The acidophilic and thermophilic unicellular red alga, Cyanidium caldarium (Tilden) Geitler, is widely distributed in acidic hot springs. Observation by transmission electron microscopy (TEM) showed that algae grown in Allen's medium contained electron-dense bodies with diameters from 100 to 200 nm. Electron dispersive x-ray analysis indicated that the electron-dense bodies contained high levels of iron, phosphorous, and oxygen; P/Fe ratios were from 1.3 to 2.0. The electron spin resonance (ESR) spectrum of the intact C. caldarium cells showed an isotropic signal at a g value of 2.00. Density-gradient centrifugation of the cell lysate yielded a fraction that included substances showing the isotropic ESR signal. EDTA treatment of this fraction reduced the ESR signal intensity, whereas it increased a signal that is typical of Fe(III)-EDTA. The fact that the isotropic signal dominates the ESR spectrum, together with a previous finding that iron is confined to the electron-dense bodies, led us to conclude that iron in the electron-dense bodies accounts for the isotropic ESR signal. Since the intensity of the ESR signal depends on the amount of iron in the cells, the electron-dense bodies are probably iron storage sites.