OPTIMAL GROWTH CONDITIONS AND ANTIOXIDATIVE ACTIVITIES OF CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)1

We isolated the unialgal strain of Cylindotheca closterium (Ehrenb.) Reimann et J. C. Lewin and produced an axenic strain using an antibiotic cocktail of enriched f/2 artificial seawater medium. The optimal growth conditions were estimated under 27 different combinations of temperature, salinity, and nutrients, and mass culture was performed based on the best specific growth conditions. Its antioxidant activities were determined from the extracts of methanol, water, and enzymes (proteases and carbohydrases). The maximum specific growth rate (μ_max) varied from 0.63 to 0.97 · d^?1. The maximum cell density was 7.20 × 10⁴ cells · mL^?1, while the μ_max was 0.82 · d^?1 in culture conditions of 20°C, 30 psu (practical salinity unit), and “F” nutrient concentrations on day 10 of the culture period. The scavenging rates for 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical were 72.5% and 69.4% from Viscozyme and methanol extracts, respectively. The enzymatic extracts of C. closterium prepared by the hydrolyses of Amyloglucosidase (AMG) and Viscozyme showed 45.8% and 45.5% nitric‐oxide‐scavenging rates, slightly lower than the activity of alpha‐tocopherol (α‐tocopherol) but similar to butylated hydroxytoluene (BHT). The extract from methanol and water showed 44.8% and 44.4% scavenging rates, statistically similar with BHT. The metal‐chelating activities of the Kojizyme, Alcalase, methanol, Viscozyme, and Neutrase extracts were 67.1, 53.9, 53.2, 52.1, and 50.2 %, respectively, five to six times higher than the commercial antioxidants. The AMG, Viscozyme, and Neutrase extracts showed a remarkable linoleic acid peroxidation inhibition, which was higher than BHT and statistically similar with α‐tocopherol.     

USING QUANTITATIVE PCR TO DETERMINE THE DISTRIBUTION OF A SEMICRYPTIC BENTHIC DIATOM,NAVICULA PHYLLEPTA (BACILLARIOPHYCEAE)1

Evidence has accumulated during the last decade showing that many established diatom morpho‐species actually consist of several semicryptic or truly cryptic species. As these species are difficult or even impossible to differentiate by microscopic analysis, there is virtually no information on how they behave in natural environments. In this study, we developed a quantitative real‐time PCR (qPCR) assay using TaqMan probes^® targeted to the internal transcribed spacer 1 (ITS1) to assess the spatial distribution and seasonal dynamics of an important component of the microphytobenthos of intertidal sediments. Navicula phyllepta Kützing is a brackish‐marine morpho‐species with a cosmopolitan distribution. Axenic clones of this species were isolated from natural assemblages of benthic diatoms at different intertidal stations in the Westerschelde estuary (The Netherlands). At least two distinct semicryptic species of N. phyllepta were present, as shown by differences in the quantity of DNA per cell, the ITS1 sequences and the copy number of ITS per cell. DNA and chl a concentrations extracted from sediment surface samples were closely correlated, showing that the DNA used for subsequent analysis mostly belonged to the microalgal community. The results of real‐time qPCR from sites throughout the estuary and over several seasons agreed well with microscopic counts. Additionally, the seasonal pattern of the two forms of N. phyllepta showed an overlapping, but unique distribution along the estuary.     

VARIATION IN BENTHIC DIATOM (BACILLARIOPHYCEAE) IMMIGRATION WITH HABITAT CHARACTERISTICS AND CELL MORPHOLOGY1

We studied the hypothesis that diatom immigration abilities are related to their fitness for colonizing stream substrates. Diatom abundances on artificial substrates exposed for 24 h (the measure of immigration rate) and abundances of stream plankton were determined in six habitats. Diatom immigration varied among habitats from 50–2500 cells·cm^?2·d^?1. Immigration rates decreased 10-fold with increases in current from 10 to 30 cm·s^?1 but changed little during a 40-d summer period. Immigration abilities of diatom taxa were characterized as ratios of either their abundances or relative abundances in immigration assemblages versus in the plankton. Immigration abilities varied over 100-fold among different species. Immigration of some species could be characterized as slower than others in different streams; however, variation in immigration abilities of other species among streams indicated that environment also affected immigration. Diurnal variation in abundance and species composition of the immigration pool (stream plankton) can be important in assessing immigration abilities. Immigration ability may affect benthic diatom fitness. Monoraphid diatoms had a lower probability of immigrating from the plankton than araphid diatoms.     

THE MORPHOLOGY AND INTEGRATION OF VALVES AND BANDS IN NAVICULA MUTICA VAR. MUTICA (BACILLARIOPHYCEAE)1

Navicula mutica (Kütz.) var. mutica was isolated from the air, cloned on agar, cultured in soil-water bottle, and studied with transmission and scanning electron micros-ropy. The frustules were lanceolate to ovoid with rounded apices, with the apical axis 8.5 ± 3.2 μ and the trans-apical and the transapical axis 3.6 ± 0.6 μm. Striae were composed of two or three puncta, and the mantle bore a single row of puncta aligned with the striae. The ends of the raphe turned away from an isolated punctual in the central area of the valve. The mantle puncta and one or two of the valve-face puncta in each stria opened into a series of transapical grooves in the interior of the valve, the grooves contributing to the appearance of striae in the light microscope. The interior of the mantle also possessed a pair of longitudinal grooves, discontinuous at the apices of the valves. An undulate advalvar margin of the valvocopula likely articulates along the interior longitudinal groove of the mantle. The projections of the undulate margin are perhaps positioned between the transapical grooves and along the longitudinal groove between the dentiform structures formed by the intersection of the double-grooved system. The girdle bands each had two (occasionally three) rows of pores. The pleurae margins were straight and not undulate.     

MONITORING RAPID VALVE FORMATION IN THE PENNATE DIATOM NAVICULA SALINARUM (BACILLARIOPHYCEAE)1

After each division of a diatom cell, a new siliceous hypovalve is formed inside the silica deposition vesicle (SDV). We present the sequence of this early formation of the new valve in the pennate marine diatom Navicula salinarum (Grunow) Hustedt, visualized by using the fluorescent probe 2‐(4‐pyridyl)‐5‐((4‐(2‐dimethylaminoethylamino‐carbamoyl)methoxy)phenyl)oxazole (PDMPO). Our observations confirm that two‐dimensional expansion of the growing valve is a rapid process of no more than 15 min; three‐dimensional completion of the valve appears to be slower, lasting most of the time valve formation takes. The results are relevant to studies of the timing of molecular processes involved in valve formation (i.e. the bio‐ and morphogenesis of the SDV) in relation to uptake and transport of silicic acid. Use of this probe helps us to identify specific developmental stages for further detail analysis of diatom basilica formation, which eventually could lead to obtaining enriched SDV fractions.     

FRUSTULAR MORPHOLOGY AND TAXONOMIC AFFINITIES OF NAVICULA COMPLANATOIDES (BACILLARIOPHYCEAE)1

Live and prepared cells of the marine pennate diatom Navicula complanatoides Hust. were examined with light and electron microscopy. It has narrowly lanceolate valves (26–55 μm long, 4–5 μm wide) and girdles 10–24 μm in depth. Striae are parallel at the center of the valve (24–28 in 10 μm), becoming slightly convergent toward the apices. Electron microscopy revealed that the external valve surface presents a longitudinally ribbed appearance (20–28 parallel ribs at its maximum width), whereas internally, rectangular areolae are occluded by ricae. The raphe slit lies in a narrow axial area, and one side of the raphe sternum is deeper and folds over the other, obscuring the internal opening. Internally, the central virga on one side of the raphe and two virgae on the other are somewhat broader. A conspicuous pore (stigma) is present between the two broadened virgae. The girdle consists of valvocopulae, copulae, and pleurae. There are 16–20 bands per cingulum. The valvocopulae and copulae are hollow tube-like structures, with inner and outer portions contrsting in morphology. They decrease in diameter in an abvalvar direction. There are four pleurae. These are flat bands which facilitate overlap of the epicingulum and hypocingulum. Fundamental features of the valve and girdle reveal the distinctness of this species within Navicula. The areolae, external longitudinal ribs, and raphe structure suggest affinities with Pleurosigma, Gyrosigma, and Haslea. It is hypothesized that they share a derived state which indicates a recent common ancestor for these taxa. N. complanatoides and related species of the Naviculae microstigmatacae are distinctive enough to merit their own genus within the Naviculaceae.     

OBSERVATIONS OF A TUBE-DWELLING DIATOM: NAVICULA HAMULIFERA (BACILLARIOPHYCEAE)1

Colonies of the tube-dwelling diatom Navicula hamulifera Grunow living on mangrove prop roots in Indian River, Florida and at La Parguera, Puerto Rico, were studied using light and electron microscopy. Observations of the tube morphology and cell structure of this diatom from fresh samples and cultures are described, as well as the ultrastructural morphology of its frustule. The formation of tubes by this diatom is reported for the first time. Comparisons are made with the closest species; Navicula delognei V.H. and Navicula pseudoco moides Hendey.     

PERIZONIUM AND INITIAL VALVE FORMATION IN THE DIATOM NAVICULA CUSPIDATA (BACILLARIOPHYCEAE)1

This paper describes the perizonium and initial valve formation in Navicula cuspidata Kütz., based on light microscope (LM) and scanning electron microscope (SEM) observations. The perizonium consists of concentric over-lapping bands, laid down sequentially at the tips of the expanding biconical auxospore during its elongation. The central perizonial band has fimbriate edges and is considerably more rigid than the more distal bands. During auxospore elongation and the band secretion, the chloroplasts continuously oscillate between the two ends of the cell; this oscillation ceases once the elongation is complete. The initial valves, formed within the perizonium, are molded into the basically biconical shape of the perizonium except for a central flattening of each valve face. In contrast to the raphes in gametangial and vegetative valves which are surrounded by a smooth axial area, the raphes in initial valves lie within a raised ridge running along the apical axis of the valve. The regular pattern of apically oriented ridges on the outer surface of vegetative valves is also lacking on initial valves. Comparison of pore–pore spacing within striae of gametangial valves, initial values and post-initial valves (first division and vegetative cells) reveals that the pore–pore distance within striae is conserved at all sexual stages. However, the distance between striae is considerably larger in initial valves than in gametangial and post-initial valves. Vegetative interstriae spacing as well as the planar morphology of the valve face is regained at the first division of the initial cell. This suggests that the spacing between striae is dependent on the sexual stage of the cell during valve formation (i.e. not directly dependent on the cell size) and can be altered independently of the pore–pore spacing.     

OBSERVATIONS ON NAVICULA THALLODES (BACILLARIOPHYCEAE), A BLADE-FORMING DIATOM FROM THE BERING SEA1

A thallus-forming diatom, Navicula thallodes Proschkina-Lavrenko, previously known only from the original collection at Bering Island (U.S.S.R.), has been found at Amchitka Island in the Aleutians, Alaska. The most remarkable observation of the present report is that N. thallodes may form blades up to 50 cm long, which to our knowledge is the greatest length reported for a colonial diatom. SEM observations of this diatom are presented for the first time.     

ECOLOGICAL DIFFERENTIATION BETWEEN SYMPATRIC PSEUDOCRYPTIC SPECIES IN THE ESTUARINE BENTHIC DIATOM NAVICULA PHYLLEPTA (BACILLARIOPHYCEAE)1

The occurrence of cryptic and pseudocryptic species, often living in sympatry, is widespread among microalgae. This phenomenon raises important questions about niche partitioning between these closely related species. To date, however, few studies have addressed the ecological mechanisms underlying sympatry in cryptic and pseudocryptic species. As a result, we have only a limited understanding of the factors that govern their distribution along environmental gradients. Here, we used the ribosomal internal transcribed spacer (ITS), 18S rRNA gene, and the RUBISCO LSU (rbcL) chloroplast gene sequence data together with cell wall morphology to show that estuarine populations of the widespread and common benthic diatom Navicula phyllepta Kütz. consist of pseudocryptic species. Growth rate measurements in function of salinity showed that N. phyllepta strains assigned to the different species differed in their tolerance to low salinities (<5 practical salinity units, psu), which was reflected by their different (but widely overlapping) distribution in the Westerschelde estuary (the Netherlands). Multiple regression analyses of the factors determining the abundance of the different species in field samples revealed that, in addition to salinity, sediment type and ammonium concentrations were probably equally important. Our results show that N. phyllepta sensu lato comprises different species with specialized ecophysiological characteristics rather than generalists with a broad adaptability to different environmental conditions.     

OBSERVATIONS OF A TUBE-DWELLING DIATOM: NAVICULA HAMULIFERA (BACILLARIOPHYCEAE)1

Colonies of the tube-dwelling diatom Navicula hamulifera Grunow living on mangrove prop roots in Indian River. Florida and at La Parguera, Puerto Rico, were studied using light and electron microscopy. Observations of the tube morphology and cell structure of this diatom from fresh samples and cultures are described, as well as the ultrastructural morphology of its frustule. The formation of tubes by this diatom is reported for the first time. Comparisons are made with the closest species; Navicula delognei V.H. and Navicula pseudocomoides Hendey.     

DAILY FLUCTUATIONS OF EXOPOLYMERS IN CULTURES OF THE BENTHIC DIATOMS CYLINDROTHECA CLOSTERIUM AND NITZSCHIA SP. (BACILLARIOPHYCEAE)1

Dynamics in the production of extracellular polymeric substances (EPS) were investigated for the benthic diatoms Cylindrotheca closterium (Ehrenberg) and Nitzschia sp. The effect of growth phase and light:dark conditions were examined using axenic cultures. Two EPS fractions were distinguished. Soluble EPS was recovered from the culture supernatant and represented polysaccharides that were only loosely associated with the cells. Bound EPS was extracted from the cells using warm (30° C) water and was more closely associated with the diatom aggregates. Concentrations of EPS exceeded internal concentrations of sugar throughout growth, indicating that EPS production is important in these organisms. Soluble and bound EPS revealed distinct differences in daily dynamics during the course of growth. Soluble EPS was produced continuously once cultures entered the stationary phase. During the stationary phase, chl a‐normalized EPS production rates equaled 6.4 and 3.4 d ^{? 1} for C. closterium and Nitzschia sp., respectively. In contrast, production of bound EPS occurred only in the light and was highest during the exponential phase. Up to 90% of the attached EPS that was produced in the light was degraded during the subsequent dark period. The monosaccharide distribution of EPS was constant during the course of the experiment. The soluble EPS consisted of high amounts of galactose and glucuronic acid, relative to rhamnose, glucose, xylose/mannose, and galacturonic acid. In contrast, glucose was the dominant monosaccharide present in the bound EPS. These differences suggest that the production of the two distinct EPS fractions is under different metabolic controls and probably serves different cellular functions.     

CALCIUM RELEASE FROM INTRACELLULAR STORES IS NECESSARY FOR THE PHOTOPHOBIC RESPONSE IN THE BENTHIC DIATOM NAVICULA PERMINUTA (BACILLARIOPHYCEAE)1

Complex photoreceptor pathways exist in algae to exploit light as a sensory stimulus. Previous studies have implicated calcium in blue‐light signaling in plants and algae. A photophobic response to high‐intensity blue light was characterized in the marine benthic diatom Navicula perminuta (Grunow) in van Heurck. Calcium modulators were used to determine the involvement of calcium in the signaling of this response, and the fluorescent calcium indicator Calcium Crimson was used to image changes in intracellular [Ca²⁺] during a response. A localized, transient elevation of Calcium Crimson fluorescence was seen at the cell tip at the time of cell reversal. Intracellular calcium release inhibitors produced a significant decrease in the population photophobic response. Treatments known to decrease influx of extracellular calcium had no effect on the population photophobic response but did cause a significant decrease in average cell speed. As the increase in intracellular [Ca²⁺] at the cell tip corresponded to the time of direction change rather than the onset of the light stimulus, it would appear that Ca²⁺ constitutes a component of the switching mechanism that leads to reversal of the locomotion machinery. Our current evidence suggests that the source of this Ca²⁺ is intracellular.     

BENTHIC DIATOM AUTECOLOGY AND INFERENCE MODEL DEVELOPMENT FROM THE CANADIAN HIGH ARCTIC ARCHIPELAGO1

Diatom assemblages were analyzed from 64 lakes and ponds from Alert, Ellesmere Island and Mould Bay, Prince Patrick Island in the Canadian High Arctic Archipelago. Diverse water chemistry conditions and diatom communities were present in these sites. Small benthic taxa typically dominated diatom communities; however, assemblages were markedly different between Alert and Mould Bay sites in response to disparate water chemistry characteristics in the two regions. The most abundant taxa belonged to the genera Navicula, Cymbella, Achnanthes, Nitzschia, and Pinnularia. Canonical correspondence analysis indicated that pH, specific conductivity, dissolved organic carbon, and total phosphorus were the most important limnological variables in determining species composition. Diatom inference models were developed for pH, specific conductivity, and dissolved organic carbon using weighted averaging and weighted averaging partial least squares techniques; these had root mean square error of prediction/r²_boot values of 0.40/0.77, 0.28/0.70, and 0.24/0.55, respectively. These models are applicable to sites with large ranges of taxonomic and limnological variation and will allow the reconstruction of past changes of climate‐related limnological parameters from biostratigraphic records in future paleolimnological studies.     

TEMPERATURE EFFECTS ON SILICON LIMITED GROWTH OF THE LAKE MICHIGAN DIATOM STEPHANODISCUS MINUTUS (BACILLARIOPHYCEAE)1

The effect of temperature on the silicon limited growth and nutrient kinetics of Stephanodiscus minutus Grun. was examined using batch and semicontinuous culture methods. Short-term batch culture methods gave maximum growth rates which were essentially constant over the temperature range of 10° to 20°C (μ₃= 0.71–0.80 d^?1). The half-saturation constant for growth (K_s) was significantly lowest at 10°C (K_s= 0.31 μM Si; 0.22–0.41), and higher at both 15°C (K_s= 1.03 μM Si; 0.68–1.47) and 20°C (K_s= 0.88 μM Si; 0.60–1.22). Two methods were used to evaluate the semicontinuous experiments. The Droop relationship showed that the minimum cell quota was about 1.50 × 10^?7 nmol Si cell^?1, but there was much overlap in the results at all three temperatures. The Monod growth relationship for the semicontinuous experiments gave estimates of K_s which were lowest at 15°C (K_s= 0.12 μM Si), and higher at 10°C (K_s= 0.68 μM Si) and 20°C (K_s= 1.24 μM Si), although 95% confidence intervals overlapped. The maximum growth rate estimates for the semicontinuous experiments were similar at 10° and 15°, and higher at 20°C, but the number of points used in making the calculations makes the results less reliable than those from batch cultures. Generally, there were no consistent significant differences in the silicon limited growth of S. minutus over the temperature range studied. Our values of K_s for S. minutus are the lowest recorded for a freshwater diatom, and are consistent with the distribution of this species in nature. Generally, this species becomes abundant in areas with high phosphorus loading and very low silicon levels (low Si:P loading rates). Stephanodiscus species are also fossil indicators of eutrophication in north temperate lakes.     

ENVIRONMENTAL CONTROL OF STALK LENGTH IN THE BLOOM‐FORMING,FRESHWATER BENTHIC DIATOM DIDYMOSPHENIA GEMINATA (BACILLARIOPHYCEAE)1

Blooms of the freshwater stalked diatom Didymosphenia geminata (Lyngb.) M. Schmidt in A. Schmidt typically occur in oligotrophic, unshaded streams and rivers. Observations that proliferations comprise primarily stalk material composed of extracellular polymeric substances (EPS) led us to ask whether or not the production of excessive EPS is favored under nutrient‐limited, high‐light conditions. We conducted experiments in outdoor flumes colonized by D. geminata using water from the oligotrophic, D. geminata–affected Waitaki River, South Island, New Zealand, to determine the relationship between D. geminata stalk length, cell division rates, and light intensity under ambient and nutrient‐enriched conditions. Stalk lengths were measured in situ, and cell division rates were estimated as the frequency of dividing cells (FDC). FDC responded positively to increasing light intensity and to nutrient additions (N+P and P). Under ambient conditions, stalk length increased as light level increased except at low ambient light levels and temperature. Nutrient enrichment resulted in decreased stalk length and negative correlations with FDC, with this effect most evident under high light. Our results are consistent with the hypothesis that extensive stalk production in D. geminata occurs when cell division rates are nutrient limited and light levels are high. Thus, photosynthetically driven EPS production in the form of stalks, under nutrient‐limited conditions, may explain the development of very high biomass in this species in oligotrophic rivers. The responses of FDC and stalk length under nutrient‐replete conditions are also consistent with occurrences of D. geminata as a nondominant component of mixed periphyton communities in high‐nutrient streams.     

BIOLOGICAL AND CHEMICAL CHARACTERISTICS OF THE GIANT DIATOM ETHMODISCUS (BACILLARIOPHYCEAE) IN THE CENTRAL NORTH PACIFIC GYRE

Cells of the giant diatom Ethmodiscus Castr. gathered from the upper 15 m were examined for O₂ evolution, nitrate reductase activity (NRA), C and N composition, internal NO concentrations, , and ¹⁵NO, ¹⁵NH , and ³²Si uptake in a series of cruises in the central N. Pacific gyre. The δ¹⁵N (2.56–5.09 ‰), internal NO concentrations (0.0– 11.5 mM NO⁻), and NRA (6.7 ± 4.7 × 10⁻⁴μM NO cell ⁻¹·h⁻¹) were consistent with recent exposure to elevated nitrate concentrations and utilization of deep NO as a primary N source. These results are similar to other diatoms that migrate vertically to the nutricline as part of their life cycle. Rate measures (Si[OH]₄ uptake, NRA, and O₂ evolution) indicated surface doubling times from 45 h to 75 h. Both NO and NH uptake in surface waters were low and inadequate to supply N needs at surface NO and NH concentrations. Our results suggest a partitioning in nutrient acquisition, with N acquired at depth and C and Si acquired at the surface. Doubling rates were two to three times higher than predicted from cell volume and C content models. These data are consistent with the observed elemental content being lower than expected because of the dominance of cell volume by the vacuole. Our calculations suggest that Ethmodiscus contributes little to the biogeochemistry of the upper water column via upward nutrient transport. Although reported as a paleo-upwelling indicator, thisevidence suggests that Ethmodiscus has adapted to the nutrient-poor open ocean by a vertical migration strategy and has biological characteristics inconsistent with a upwelling indicator.     

LOCATION AND EXPRESSION OF FRUSTULINS IN THE PENNATE DIATOMS CYLINDROTHECA FUSIFORMIS, NAVICULA PELLICULOSA, AND NAVICULA SALINARUM (BACILLARIOPHYCEAE)

A detailed immunocytochemical and biochemical study of the location and expression of frustulins, a family of proteins associated with the frustules of diatoms, has been performed for Cylindrotheca fusiformis Reimann et Lewin, Navicula pelliculosa (Brébisson et Kützing) Hilse, and Navicula salinarum (Grunow) Husted. Immunocytochemistry revealed that frustulins, which share homologous epitopes but are different in size, were predominantly located in the organic casing. Based on timed immunolocalization experiments and Western blotting analysis of cell extracts obtained sequentially after repleting silicate to Si-synchronized cells, the continuous presence of the frustulins in the mature and parental organic casing of the examined species was observed. The frustulins of N. pelliculosa appeared as proteins similar to those of C. fusiformis, sharing identical epitopes. The extractions, however, yielded a markedly lower abundance of frustulins in N. pelliculosa. Peak concentrations of extracted frustulins appeared to be expressed just ahead of the silicification process in C. fusiformis, whereas the level of expression in N. pelliculosa increased along with maturation of the new valves. For N. salinarum, the presence of the frustulins could not be confirmed properly by Western blotting, most probably because of the small sample volumes, inefficient extraction, and a lower amount of homologous frustulins in the casing of this species. It is concluded that the frustulins of these species are not associated with the silicalemma of the newly formed silica deposition vesicles and therefore do not seem to be involved in the silicification process itself. Overall, the results imply a structural role of the frustulins in the casing of diatoms rather than a regulation of the silicification process.     

DENSITY-DEPENDENT GROWTH,ECOLOGICAL STRATEGIES,AND EFFECTS OF NUTRIENTS AND SHADING ON BENTHIC DIATOM SUCCESSION IN STREAMS1

The importance of immigration, growth, and competition for nutrients and light in benthic diatom succession was studied in experimental channels in a low-nutrient stream. Diatom accumulation was greater in channels enriched with nitrate and phosphate (NP) than in control channels, reaching about 5 × 10⁶ and 2 × 10⁶ cells-cm^?2, respectively, after 30 d. Shading during late stages of community development reduced algal standing crop. Synedra ulna (Nitz.) Ehr. and Achnanthes minutissima Kütz. were codominant during early stages of community development in both habitats, but succession to an A. minutissima-dominated community was much faster in NP-enriched than in control conditions. Species dominating early stages tended to immigrate quickly, whereas species that increased in relative abundance during community development had either fast growth rates or fast immigration and average growth rates. Decreases in growth rates indicated resource supply became limiting during community development in control and enriched channels. Density-dependent competition was indicated because nutrient concentrations in the water column and light did not decrease during the 30-d study. Species autecologies were defined by effects of nutrient enrichment, shading, and community development on species growth rates. Differing autecologies of early and late succession species indicated that competition for nutrients was more important than competition for light. Species autecologies also indicated ecological strategies. The species most stimulated by nutrient enrichment were least able to maintain growth rates as algal abundances on substrata increased. In addition, these species that best sustained their growth rates during succession tended to have the highest immigration rates, indicating that drift and immigration may have been an important mechanism of persistence for some populations when resources become limiting within thick benthic mats.     

PROTOCOLS FOR THE REMOVAL OF BACTERIA FROM FRESHWATER BENTHIC DIATOM CULTURES1

In this study, we describe different combinations of physical separation and antibiotic treatment to remove associated bacteria from freshwater diatoms. Diatoms were purified either from natural epilithic biofilms or from unialgal cultures. We determined that for most strains, different purification procedures have to be combined individually. In a new approach, we show that for some diatom strains, the substitution of associated aquatic bacteria by an antibiotic‐sensitive Escherichia coli strain and subsequent treatment with antibiotics may be a successful strategy to obtain axenic diatom cultures. Axenic diatom cultures are essential to study the physiology and biochemistry of individual strains as well as their responses to environmental changes without interference of accompanying bacteria.