Nutrient limitation and algal blooms in urbanizing tidal creeks

Tidal creeks are commonly found in low energy systems on the East and Gulf Coasts of the United States, and are often subject to intense watershed human development. Many of these creeks are receiving urban and suburban runoff containing nutrients, among other pollutants. During the period 1993-2001, we studied three tidal creeks located in southeastern North Carolina, a rapidly urbanizing area. All three creeks received anthropogenic nutrient loading. Oligohaline to mesohaline stations in upper tidal creek regions had much higher nutrient (especially nitrate-N) concentrations than lower creek areas, and hosted spring and summer phytoplankton blooms that at times exceeded 200 μg chlorophyll a l⁻¹. Phytoplankton biomass during winter was low at all stations in all three creeks. Spring and summer nutrient addition bioassay experiments were conducted to characterize the nutrients limiting phytoplankton growth. Water from high salinity stations in all three creeks always showed significant positive responses to nitrate-N inputs, even at concentrations as low as 50 μg N l⁻¹. Low salinity stations in upper creek areas often showed significant responses to nitrate-N inputs, but on occasion showed sensitivity to phosphorus inputs as well, indicating the influence of anthropogenic nitrate loading. During several experiments, one of the upper stations showed no positive response to nutrient inputs, indicating that these stretches were nutrient replete, and further phytoplankton growth appeared to be light-limited either by phytoplankton self-shading or turbidity. Water from upper creek areas yielded much higher chlorophyll a concentrations in bioassay experiments than did lower creek water. In general, these urbanizing tidal creeks were shown to be very sensitive to nitrogen loading, and provide a physical environment conducive to phytoplankton bloom formation in nutrient-enriched areas. Tidal creeks are important ecological resources in that they are considered to be nursery areas for many species of fish and shellfish. To protect the ecological function of these small, but very abundant estuarine systems, management efforts should recognize their susceptibility to algal blooms and focus on control of nonpoint source nutrient inputs, especially nitrogen.     

Seasonal and interannual variability in the taxonomic composition and production dynamics of phytoplankton assemblages in Crater Lake,Oregon

Taxonomic composition and production dynamics of phytoplankton assemblages in Crater Lake, Oregon, were examined during time periods between 1984 and 2000. The objectives of the study were (1) to investigate spatial and temporal patterns in species composition, chlorophyll concentration, and primary productivity relative to seasonal patterns of water circulation; (2) to explore relationships between water column chemistry and the taxonomic composition of the phytoplankton; and (3) to determine effects of primary and secondary consumers on the phytoplankton assemblage. An analysis of 690 samples obtained on 50 sampling dates from 14 depths in the water column found a total of 163 phytoplankton taxa, 134 of which were identified to genus and 101 were identified to the species or variety level of classification. Dominant species by density or biovolume included Nitzschia gracilis, Stephanodiscus hantzschii, Ankistrodesmus spiralis, Mougeotia parvula, Dinobryon sertularia, Tribonema affine, Aphanocapsa delicatissima, Synechocystis sp., Gymnodinium inversum, and Peridinium inconspicuum. When the lake was thermally stratified in late summer, some of these species exhibited a stratified vertical distribution in the water column. A cluster analysis of these data also revealed a vertical stratification of the flora from the middle of the summer through the early fall. Multivariate test statistics indicated that there was a significant relationship between the species composition of the phytoplankton and a corresponding set of chemical variables measured for samples from the water column. In this case, concentrations of total phosphorus, ammonia, total Kjeldahl nitrogen, and alkalinity were associated with interannual changes in the flora; whereas pH and concentrations of dissolved oxygen, orthophosphate, nitrate, and silicon were more closely related to spatial variation and thermal stratification. The maximum chlorophyll concentration when the lake was thermally stratified in August and September was usually between depths of 100 m and 120 m. In comparison, the depth of maximum primary production ranged from 60 m to 80 m at this time of year. Regression analysis detected a weak negative relationship between chlorophyll concentration and Secchi disk depth, a measure of lake transparency. However, interannual changes in chlorophyll concentration and the species composition of the phytoplankton could not be explained by the removal of the septic field near Rim Village or by patterns of upwelling from the deep lake. An alternative trophic hypothesis proposes that the productivity of Crater Lake is controlled primarily by long-term patterns of climatic change that regulate the supply of allochthonous nutrients.     

Distribution and abundance of zooplankton populations in Crater Lake,Oregon

The zooplankton assemblages in Crater Lake exhibited consistency in species richness and general taxonomic composition, but varied in density and biomass during the period between 1988 and 2000. Collectively, the assemblages included 2 cladoceran taxa and 10 rotifer taxa (excluding rare taxa). Vertical habitat partitioning of the water column to a depth of 200 m was observed for most species with similar food habits and/or feeding mechanisms. No congeneric replacement was observed. The dominant species in the assemblages were variable, switching primarily between periods of dominance of Polyarthra-Keratella cochlearis and Daphnia. The unexpected occurrence and dominance of Asplanchna in 1991 and 1992 resulted in a major change in this typical temporal shift between Polyarthra-K. cochlearis and Daphnia. Following a collapse of the zooplankton biomass in 1993 that was probably caused by predation from Asplanchna, Kellicottia dominated the zooplankton assemblage biomass between 1994 and 1997. The decline in biomass of Kellicottia by 1998 coincided with a dramatic increase in Daphnia biomass. When Daphnia biomass declined by 2000, Keratella biomass increased again. Thus, by 1998 the assemblage returned to the typical shift between Keratella-Polyarthra and Daphnia. Although these observations provided considerable insight about the interannual variability of the zooplankton assemblages in Crater Lake, little was discovered about mechanisms behind the variability. When abundant, kokanee salmon may have played an important role in the disappearance of Daphnia in 1990 and 2000 either through predation, inducing diapause, or both. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Thermal,chemical, and optical properties of Crater Lake,Oregon

Crater Lake covers the floor of the Mount Mazama caldera that formed 7700 years ago. The lake has a surface area of 53 km² and a maximum depth of 594 m. There is no outlet stream and surface inflow is limited to small streams and springs. Owing to its great volume and heat, the lake is not covered by snow and ice in winter unlike other lakes in the Cascade Range. The lake is isothermal in winter except for a slight increase in temperature in the deep lake from hyperadiabatic processes and inflow of hydrothermal fluids. During winter and spring the water column mixes to a depth of about 200–250 m from wind energy and convection. Circulation of the deep lake occurs periodically in winter and spring when cold, near-surface waters sink to the lake bottom; a process that results in the upwelling of nutrients, especially nitrate-N, into the upper strata of the lake. Thermal stratification occurs in late summer and fall. The maximum thickness of the epilimnion is about 20 m and the metalimnion extends to a depth of about 100 m. Thus, most of the lake volume is a cold hypolimnion. The year-round near-bottom temperature is about 3.5°C. Overall, hydrothermal fluids define and temporally maintain the basic water quality characteristics of the lake (e.g., pH, alkalinity and conductivity). Total phosphorus and orthophosphate-P concentrations are fairly uniform throughout the water column, where as total Kjeldahl-N and ammonia-N are highest in concentration in the upper lake. Concentrations of nitrate-N increase with depth below 200 m. No long-term changes in water quality have been detected. Secchi disk (20-cm) clarity varied seasonally and annually, but was typically highest in June and lowest in August. During the current study, August Secchi disk clarity readings averaged about 30 m. The maximum individual clarity reading was 41.5 m in June 1997. The lowest reading was 18.1 m in July 1995. From 1896 (white-dinner plate) to 2003, the average August Secchi disk reading was about 30 m. No long-term changes in the Secchi disk clarity were observed. Average turbidity of the water column (2–550 m) between June and September from 1991 to 2000 as measured by a transmissometer ranged between 88.8% and 90.7%. The depth of 1% of the incident solar radiation during thermal stratification varied annually between 80 m and 100 m. Both of these measurements provided additional evidence about the exceptional clarity of Crater Lake.     

Nutrient limitation of phytoplankton in solar salt ponds in Shark Bay,Western Australia

The aim of this research was to examine nutrient limitation of phytoplankton in solar salt ponds of varying salinity at Useless Inlet in Western Australia. These ponds use solar energy to evaporate seawater for the purpose of commercial salt production. A combination of techniques involving water column nutrient ratios, comparisons of nutrient concentrations to concentration of magnesium ions and bioassays were used in the investigation. Comparisons of changes in dissolved inorganic nitrogen to phosphorus ratios and concentrations of dissolved inorganic nutrients against changes in concentrations of the conservative cation Mg²⁺ indicated that phytoplankton biomass was potentially nitrogen limited along the entire pond salinity gradient. Nutrient addition bioassays indicated that in low salinity ponds, phytoplankton was nitrogen limited but in high salinity ponds, phosphorus limited. This may be due to isolation of phytoplankton in bioassay bottles from in situ conditions as well as to changes in phytoplankton species composition between ponds, and the variable availability of inorganic and organic nutrient sources. The differences in limiting nutrient between methods indicate that phytoplankton cells may be proximally limited by nutrients that are not theoretically limiting at the pond scale. Dissolved organic nutrients constituted a large proportion of total nutrients, with concentrations increasing through the pond sequence of increasing salinity. From the change in nutrient concentrations in bioassay bottles, sufficient dissolved organic nitrogen may be available for phytoplankton uptake in low salinity ponds, potentially alleviating the dissolved inorganic nitrogen limitation of phytoplankton biomass. Guest Editors: J. John & B. Timms Salt Lake Research: Biodiversity and Conservation—Selected Papers from the 9th Conference of the International Society for Salt Lake Research     

Evaporation and the hydrologic budget of Crater Lake, Oregon

The hydrologic budget of Crater Lake, Oregon is investigated by taking advantage of its relatively simple geometry, climatic circumstances, and the concurrent availability of many years of traditional data. Buoy data are here utilized for the first time for this purpose. The lake gains water through precipitation and delayed runoff from the caldera sides and Wizard Island. The lake loses water through evaporation and seepage. Seepage can be estimated quite well from ice-covered precipitation-free intervals in 1985, and is 127 cm/year. Evaporation has previously been determined as a residual, but is here estimated directly from the floating buoy, with an approximate value of 76 cm/year, a downward revision from previous estimates. These losses are balanced by precipitation input, nearly all in the form of snow or snowmelt runoff. Factors contributing to the uncertainty in each of the water budget components are discussed in some detail. The buoy data corroborate previous findings based on studies of stage that evaporation is greatest on the coldest days. Seasonally, the greatest evaporation occurs in the autumn and the least in spring. Proxy records are used to extend the effective length of the buoy record. Monthly estimates of evaporation are calculated for 1950–1996 and used to deduce temporal characteristics. The standard deviation of water year precipitation is 4.6 times larger than that of evaporation. Thus the water budget is controlled more by variability of precipitation than evaporation. An additional 15 years of data since earlier studies confirm that the annual lake level variations from one September 30 to the next are highly correlated (r = 0.96) with Park Headquarters water year precipitation for the 42 years from 1961–2003. The lake rises 1.4 cm for every cm of measured precipitation over equilibrium value (168.6 cm) at Park Headquarters. Sources of this “magnification” are discussed.     

Measurements of spectral optical properties and their relation to biogeochemical variables and processes in Crater Lake,Crater Lake National Park,OR

Spectral inherent optical properties (IOPs) have been measured at Crater Lake, OR, an extremely clear sub-alpine lake. Indeed Pure water IOPs are major contributors to the total IOPs, and thus to the color of the lake. Variations in the spatial distribution of IOPs were observed in June and September 2001, and reflect biogeochemical processes in the lake. Absorption by colored dissolved organic material increases with depth and between June and September in the upper 300 m. This pattern is consistent with a net release of dissolved organic materials from primary and secondary production through the summer and its photo-oxidation near the surface. Waters fed by a tributary near the lake’s rim exhibited low levels of absorption by dissolved organic materials. Scattering is mostly dominated by organic particulate material, though inorganic material is found to enter the lake from the rim following a rain storm. Several similarities to oceanic oligotrophic regions are observed: (a) The Beam attenuation correlates well with particulate organic material (POM) and the relationship is similar to that observed in the open ocean. (b) The specific absorption of colored dissolved organic material has a value similar to that of open ocean humic material. (c) The distribution of chlorophyll with depth does not follow the distribution of particulate organic material due to photo-acclimation resulting in a subsurface pigment maximum located about 50 m below the POM maximum.     

Nutrient limitation in a tropical saline lake: a microcosm experiment

There is increasing evidence that nitrogen limitation is of widespread occurrence in tropical lakes. Nonetheless, data on the deep tropical Lake Alchichica (Mexico) show that dissolved inorganic nitrogen (DIN) to soluble reactive phosphorus (SRP) ratio fluctuates widely. To elucidate further the role of nitrogen and phosphorus limitation on the phytoplankton growth in tropical saline lakes, we present the results of a series of nutrient enrichment experiments with natural assemblages of Lake Alchichica phytoplankton conducted monthly for a year. Our assays indicate that phosphorus and nitrogen alternate in limiting Lake Alchichica phytoplankton biomass. Phosphorous limited phytoplankton growth most (41.7%) of the time, followed by nitrogen (33.3% of the time), and both nutrients for the rest of the time (25.0%). This alternation in nitrogen and phosphorus responsible for phytoplankton growth limitation in Lake Alchichica is attributed to the combination of natural conditions (e.g., young volcanic terrain rich in phosphorus) that would favor nitrogen limitation and anthropogenic impacts (e.g., agricultural nitrogen fertilization) which would cause phosphorus limitation. Guest Editors: J. John & B. Timms Salt Lake Research: Biodiversity and Conservation—Selected papers from the 9th Conference of the International Society for Salt Lake Research     

Long-term observations of deepwater renewal in Crater Lake,Oregon

We examine observations of key limnological properties (primarily temperature, salinity, and dissolved oxygen), measured over a 14-year period in Crater Lake, Oregon, and discuss variability in the hypolimnion on time scales of days to a decade. During some years (e.g., 1994–1995), higher-than-average wintertime deep convection and ventilation led to the removal of significant amounts of heat and salt from the hypolimnion, while dissolved oxygen concentrations increase. In other years, such as the winter of 1996–1997, heat and salt concentrations increase throughout the year and dissolved oxygen levels drop, indicating conditions were dominated by the background geothermal inputs and dissolved oxygen consumption by bacteria (i.e., minimal deep convection). Over the entire 14 year period, no statistically significant trend was observed in the annual hypolimnetic heat and salt content. Measurements from several thermistors moored in the hypolimnion provide new insight into the time and space scales of the deep convection events. For some events, cool water intrusions are observed sequentially, from shallower depths to deeper depths, suggesting vertical mixing or advection from above. For other events, the cooling is observed first at the deepest sensors, suggesting a thin, cold water pulse that flows along the bottom and mixes more slowly upwards into the basin. In both cases, the source waters must originate from the epilimnion. Conditions during a strong ventilation year (1994–1995) and a weak ventilation year (1996–1997) were compared. The results suggest the major difference between these 2 years was the evolution of the stratification in the epilimnion during the first few weeks of reverse stratification such that thermobaric instabilities were easier to form during 1995 than␣1997. Thus, the details of surface cooling and wind-driven mixing during the early stages of␣reverse stratification may determine the net␣amount of ventilation possible during a particular year.     

Seasonal nutrient and plankton dynamics in a physical-biological model of Crater Lake

A coupled 1D physical-biological model of Crater Lake is presented. The model simulates the seasonal evolution of two functional phytoplankton groups, total chlorophyll, and zooplankton in good quantitative agreement with observations from a 10-year monitoring study. During the stratified period in summer and early fall the model displays a marked vertical structure: the phytoplankton biomass of the functional group 1, which represents diatoms and dinoflagellates, has its highest concentration in the upper 40 m; the phytoplankton biomass of group 2, which represents chlorophyta, chrysophyta, cryptomonads and cyanobacteria, has its highest concentrations between 50 and 80 m, and phytoplankton chlorophyll has its maximum at 120 m depth. A similar vertical structure is a reoccurring feature in the available data. In the model the key process allowing a vertical separation between biomass and chlorophyll is photoacclimation. Vertical light attenuation (i.e., water clarity) and the physiological ability of phytoplankton to increase their cellular chlorophyll-to-biomass ratio are ultimately determining the location of the chlorophyll maximum. The location of the particle maxima on the other hand is determined by the balance between growth and losses and occurs where growth and losses equal. The vertical particle flux simulated by our model agrees well with flux measurements from a sediment trap. This motivated us to revisit a previously published study by Dymond et al. (1996). Dymond et al. used a box model to estimate the vertical particle flux and found a discrepancy by a factor 2.5–10 between their model-derived flux and measured fluxes from a sediment trap. Their box model neglected the exchange flux of dissolved and suspended organic matter, which, as our model and available data suggests is significant for the vertical exchange of nitrogen. Adjustment of Dymond et al.’s assumptions to account for dissolved and suspended nitrogen yields a flux estimate that is consistent with sediment trap measurements and our model.     

Nutrient limitation of two saccharolytic clostridia; secretion, sporulation, and solventogenesis

Abstract Growth of the cellulolytic acidogen Clostridium strain C7 undergoing progressive nutrient limitation has been compared with that of the solventogen Clostridium beijerinckii . On the basis of cellulase secretion, differentiation of dissimilatory metabolism, and sporulation, different survival strategies by the two clostridia in progressive nutrient limitation can be discerned. In addition, the metabolic differentiation to butanol production in Clostridium beijerinckii can be specifically associated with the sporulation stage in which the forespore is enclosed by double membranes but not by a spore coat.     

Nutrient limitation of Microcystis aeruginosa in northern California Klamath River reservoirs

Nutrient limitations were investigated in Copco and Iron Gate Reservoirs, on the Klamath River in California, where blooms of the toxin-producing cyanobacterium Microcystis aeruginosa were first reported in 2005. Nutrient enrichment experiments conducted in situ in June and August, 2007 and 2008, determined responses in phytoplankton biomass, Microcystis abundance and microcystin concentration to additions of phosphorus and different forms of nitrogen (NH₄⁺, NO₃, and urea). Microcystis abundance was determined using quantitative PCR targeting the phycocyanin intergenic spacer cpcBA.Total phytoplankton biomass increased with additions of N both before and during Microcystis blooms, with no primary effects from P, suggesting overall N limitation for phytoplankton growth during the summer season. NH₄⁺ generally produced the greatest response in phytoplankton growth, while Microcystis abundance increased in response to all forms of N. Microcystis doubling time in the in situ experiments was 1.24–1.39 days when N was not limiting growth. The results from this study suggest availability of N during the summer is a key growth-limiting factor for the initiation and maintenance of toxic Microcystis blooms in Copco and Iron Gate Reservoirs in the Klamath River.     

Ultraviolet radiation and bio-optics in Crater Lake,Oregon

Crater Lake, Oregon, is a mid-latitude caldera lake famous for its depth (594 m) and blue color. Recent underwater spectral measurements of solar radiation (300–800 nm) support earlier observations of unusual transparency and extend these to UV-B wavelengths. New data suggest that penetration of solar UVR into Crater Lake has a significant ecological impact. Evidence includes a correlation between water column chlorophyll-a and stratospheric ozone since 1984, the scarcity of organisms in the upper water column, and apparent UV screening pigments in phytoplankton that vary with depth. The lowest UV-B diffuse attenuation coefficients (K _d,320) were similar to those reported for the clearest natural waters elsewhere, and were lower than estimates for pure water published in 1981. Optical proxies for UVR attenuation were correlated with chlorophyll-a concentration (0–30 m) during typical dry summer months from 1984 to 2002. Using all proxies and measurements of UV transparency, decadal and longer cycles were apparent but no long-term trend since the first optical measurement in 1896.     

Nutrient limitation and plant species composition in temperate salt marshes

Addition of inorganic nitrogen, phosphorus and potassium in a factorial design in two ungrazed Wadden-Sea salt marshes at low and high elevations showed that nitrogen was the limiting nutrient. No effects of nutrient addition were detected in the 1st year, probably due to a considerable rainfall deficit during the growing season. In the 2nd year, which was more humid, only nitrogen addition caused significant effects in both the low salt marsh dominated by Puccinellia maritima and the high marsh dominated by Festuca rubra. No two-way or three-way interactions with phosphorus or potassium were found. In the low marsh, nitrogen addition had a negative effect on the biomass of Puccinellia, but a positive effect on the biomass of Suaeda maritima and on the total above-ground biomass. Puccinellia was replaced by Suaeda after nitrogen addition, due to shading. In the high salt marsh, no significant effects of fertilizer application on total above-ground biomass were found, due to the weak response of the dominant species Festuca rubra, which accounted for 95% of total biomass. The biomass of Spergularia maritima increased, however, as a response to nitrogen addition.The shoot length of Festuca was positively affected by nitrogen fertilization. It is suggested that stands of Festuca reached maximal biomass at the study site without fertilization and that its growth was probably limited by self-shading. Received: 22 September 1996 / Accepted: 5 April 1997     

High-elevation mountain hemlock growth as a surrogate for cool-season precipitation in Crater Lake National Park,USA

Snow dominates the hydrology and climate of the United States’ central Pacific Coast, but because local measurements of snowpack and winter precipitation often extend back only a few decades, observations by themselves are not adequate to describe potential amplitude of wintertime conditions. Here we present a set of updated and extended mountain hemlock (Tsuga mertensiana [Bong.] Carr.) tree-ring width records from Crater Lake National Park, Oregon, and use these data to make inferences about snowpack prior to the start of instrumental monitoring. In July and August 2013, we collected cores from 228 trees at seven high-elevation hemlock stands that surround the crater’s rim. The oldest tree had an inner ring date of CE 1474, and the longest ring-width chronology maintained a satisfactory common signal back to the middle of the 16th century. The growth of high-elevation mountain hemlock is strongly and inversely related to cool-season precipitation, making these records some of the most southerly examples of a robust inverse cool-season moisture signal in North American tree rings. The growth of these snow-limited forests does not appear to have been affected by the substantial decline in spring snowpack observed in the past two decades across the broader Cascade Range, and we did not find any indication of changing relationships between tree growth and either monthly or seasonal winter precipitation since the early 1990s. The exceptional three-year sequence in Crater Lake tree rings between CE 1809 and 1811, which includes the narrowest ring since CE 1540 and anatomical abnormalities produced by cold weather, leads us to conclude that 1809–1810 was the most snowy and severe winter to affect south-central Oregon during the past four and a half centuries.     

Nutrient pulsing as a regulator of phytoplankton abundance and community composition in Galveston Bay, Texas

Galveston Bay, Texas, is a large shallow estuary with a watershed that includes 60% of the major industrial facilities of Texas. However, the system exhibits low to moderate (2-20 μg l⁻¹) microalgal biomass with sporadic phytoplankton blooms. Both nitrogen (N) and phosphate (P) limitation of phytoplankton growth have been proposed for the estuary. However, shifts between N and P limitation of algae growth may occur due to annual fluctuations in nutrient concentrations. The primary goal of this work was to determine the primary limiting nutrient for phytoplankton in Galveston Bay. Nutrient addition bioassays were used to assess short-term (1-2 days) phytoplankton responses (both biomass and community composition) to potentially limiting nutrients. The experimental bioassays were conducted over an annual cycle using natural water collected from the center to lower part of the estuary. Total phytoplankton biomass increased in the nitrate (10 μM) additions in 11 of the 13 bioassays, but no significant increases were detected in the phosphate (3 μM)-only additions. Bioassay results suggest that the phytoplankton community was usually not phosphate limited. All major groups increased in biomass following nitrate additions but diatoms increased in biomass at a faster rate than other groups, shifting the community composition toward higher relative abundance of diatoms. The results of this study suggest that pulsed N input events preferentially favor increases in diatom biomass in this estuary. The broader implications of this study are that N pulsing events, primarily due to river discharge, play an important role in structuring the phytoplankton community in the Galveston Bay estuary.     

黄海和东海营养盐分布及其对浮游植物的限制

根据1997～1999年黄海和东海4个季节的现场调查资料,分析探讨了黄海、东海的营养盐分布特征及其对浮游植物生长的限制状况．结果表明,在长江口以东及其东北部海域终年存在一个范围很大的营养盐高值区．分析表明,这些营养盐主要来自长江冲淡水的扩展及苏北沿岸流的输送．此外,还获得了1998年长江流域特大洪水期间,迄今被观测到的长江冲淡水中营养盐的最大扩展范围,计算并研究了黄海、东海上层水中Si／N／P比值,结果表明,黄东海上层水中Si／N比值较高,Si不会成为黄东海浮游植物生长的限制因子;但在南黄海南部尤其是西南部、东海近岸及长江口以东海域,N／P比值很高(>30),说明与一般海洋环境的情况不同,黄海、东海有很大一部分海域浮游植物的生长受磷限制,而不是受氮限制．     

Representative freshwater bacterioplankton isolated from Crater Lake, Oregon

High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat.     

Lake Kinneret phytoplankton: Response to N and P enrichments in experiments and in nature

A series of water samples from Lake Kinneret was supplemented with 100 µM N (as NH₄ or as NO₃ and/or 10 µM orthophosphate-P. The yield of phytoplankton both as chlorophyll and in cell numbers of major species was determined after a two-week incubation. During these experiments, some of the algae present initially never multiplied (e.g. Peridinium and Peridiniopsis spp. Cryptomonas spp., Rhodomonas spp. and Crysochromulina parva); others e.g. Anomoeoneis exilis, Synedra sp., Chlamydomonas sp., Elakatothrix gelatinosa), undetected in the original sample, grew out during the incubation. Chlorophyte species (the majority of commonly observed forms in the lake) responded most readily to added nutrients.The results of these enrichment experiments were related to the long-term record of phytoplankton populations observed in the lake and suggest that through summer and fall, when ambient levels of both P and N are minimal, P was generally, but not always, the most limiting nutrient for algal growth. In the spring, after the decline of the Peridinium bloom, P appeared to be limiting the growth of Chlorophyta. Although most algal species grew equally well on NH₄ or NO₃, some species appeared to respond preferentially either to the former (Coelastrum, Chodatella) or to the latter (Chroococcus, Anomoeoneis) source of N.     

Representative Freshwater Bacterioplankton Isolated from Crater Lake, Oregon

High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat.