Daniel A. Kreeger

COMBO : a defined freshwater culture medium for algae and zooplankton

In order to conduct experiments on interactions between animals and food organisms, it is necessary to develop a medium that adequately supports the growth of both algae and zooplankton without the need to alter the medium to accommodate either the algae or the animals. We devised a freshwater medium, named COMBO, that supports excellent growth of both algae and zooplankton. Two types of algae, Ankistrodesmus falcatus and Stephanodiscus hantzschii, were reared in COMBO and their growth rates were not significantly different from those of algae grown in a reference medium (WC). One of these algae, A. falcatus, was then fed to a cladoceran, Daphnia pulicaria, which was also cultured in COMBO, and the resulting fecundities of D. pulicaria were compared to those of animals reared in natural surface water. We also determined whether the value of COMBO as a medium for D. pulicaria was affected by modifications in nitrogen or phosphorus concentration to evaluate whether the new medium will be useful in nutritional research. Lowering the N or P content of COMBO did not affect the reproductive performance of D. pulicaria. Other researchers have also reported excellent growth and reproduction by numerous algae and zooplankton reared in COMBO. Our results suggest that COMBO is an effective artificial, defined culture medium capable of supporting robust growth and reproduction of both freshwater algae and zooplankton.

Dam Removal: Challenges and Opportunities for Ecological Research and River Restoration

W flow is a “master variable” (sensu Power et al. 1995) that governs the fundamental nature of streams and rivers (Poff et al. 1997, Hart and Finelli 1999), so it should come as no surprise that the modification of flow caused by dams alters the structure and function of river ecosystems. Much has been learned during the last several decades about the adverse effects of dams on the physical, chemical, and biological characteristics of rivers (Ward and Stanford 1979, Petts 1984, Poff et al. 1997, Poff and Hart 2002). Increasing concerns about these impacts, together with related social and economic forces, have led to a growing call for the restoration of rivers by removing dams (AR/FE/TU 1999, Pejchar and Warner 2001). For the purposes of this paper, we define restoration broadly as an effort to compensate for the negative effects of human activities on ecological systems by facilitating the establishment of natural components and regenerative processes, although we acknowledge that these efforts rarely eliminate all human impacts (see Williams et al. 1997 for alternative definitions). Interest in dam removal as a means of river restoration has focused attention on important new challenges for watershed management and simultaneously created opportunities for advancing the science of ecology. One challenge lies in determining the magnitude, timing, and range of physical, chemical, and biological responses that can be expected following dam removal. This information is needed to decide whether and how dam removals should be performed to achieve specific restoration objectives (Babbitt 2002). Opportunities for advancing ecological research also exist because dam removal represents a major, but partially controllable, perturbation that can help scientists test and refine models of complex ecosystems. In contrast to the small-scale experiments that traditionally have been employed in stream and river ecology, the unusually large magnitude and spatial extent of dam removal WE DEVELOP A RISK ASSESSMENT FRAME-

EFFECT OF NUTRIENT AVAILABILITY ON THE BIOCHEMICAL AND ELEMENTAL STOICHIOMETRY IN THE FRESHWATER DIATOM STEPHANODISCUS MINUTULUS (BACILLARIOPHYCEAE)

The objective of this study was to examine the differences in the biochemical and elemental stoichiometry of a freshwater centric diatom, Stephanodiscus minutulus (Grun.), under various nutrient regimes. Stephanodiscus minutulus was grown at μmax or 22% of μmax under limitation by silicon, nitrogen, or phosphorus. Cell sizes for nutrient‐limited cultures were significantly smaller than the non‐limited cell sizes, with N‐limited cells being significantly smaller than all other treatments. Compared with the nutrient‐replete treatment, both carbohydrates and lipids increased in Si‐ and P‐limited cells, whereas carbohydrates increased but proteins decreased in N‐limited cells. All of the growth‐limited cells showed an increase of carbohydrate and triglyceride, and a decrease of cell size and polar lipids as a percentage of total lipids. The non‐limited cells also had a significantly higher chl a concentration and galactolipids as a percentage of total lipids than any of the limited treatments, and the low‐Si and low‐P cells had significantly higher values than the low‐N cells. The particulate C concentrations showed significant differences between treatments, with the Si‐ and P‐limited treatments being significantly higher than the N‐ and non‐limited treatments. Particulate Si did not show a strong relationship with any of the parameters measured, and it was the only parameter with no differences between treatments. The low‐Si cells had a significantly higher P content (about two times more) than any other treatment, presumably owing to the luxury consumption of P, and a correspondingly high phospholipid concentration. The elemental data showed that S. minutulus had a high P demand with low optimum N:P (4) and Si:P (10) ratios and a C:N:P ratio of 109:16:2.3. The particulate C showed a positive relationship with POM (r = 0.93), dry weight (r = 0.88), lipid (r = 0.87) and protein (r = 0.84, all P < 0.0001). Particulate N showed a positive relationship with galactolipids (r = 0.95), protein (r = 0.90), dry weight (r = 0.78), lipid (r = 0.75), and cell volume (r = 0.64, all P < 0.0001). It is evident that nutrient limitation in the freshwater diatom S. minutulus has pronounced effects on its biochemical and elemental stoichiometry.

Biochemical composition of three algal species proposed as food for captive freshwater mussels

To identify potential diets for rearing captive freshwater mussels, the protein, carbohydrate (CHO), and lipid contents of two green algae, Neochloris oleoabundans, Bracteacoccus grandis, and one diatom, Phaeodactylum tricornutum, were compared at different growth stages. The fatty acid and sterol composition were also identified. Protein was greatest (55–70%) for all species at late log growth stage (LL), and declined in late stationary (LS) growth. CHO was greatest at LS stage for all species (33.9–56.4% dry wt). No significant change in lipid levels occurred with growth stage, but tended to increase in N. oleoabundans. Mean lipid content differed significantly in the order: N. oleoabundans > P. tricornutum > B. grandis. Total fatty acids (TFA) were higher at LS stage compared to other stages in the two green algae, and stationary stage in the diatom. Mean unsaturated fatty acids (UFA) as %TFA was significantly higher in N. oleoabundans than the other species. The green algae contained high percentages of C-18 polyunsaturated fatty acids (PUFAs), while the diatom was abundant in C-16 saturated and mono-unsaturated fatty acids and C-20 PUFA fatty acids. Growth stage had no effect on sterol concentration of any species. B. grandis showed significantly higher sterol levels than the other species except P. tricornutum at S stage. B. grandis was characterized by predominantly Δ5, C-29 sterols, while N. oleoabundans synthesized Δ5,7, Δ5,7,22 , and Δ7, C-28 sterols. P. tricornutum produced primarily a Δ5,22, C-28 sterol, and a small amount of a Δ7,22, C-28 sterol.

Seasonal utilization of different seston carbon sources by the ribbed mussel, Geukensia demissa (Dillwyn) in a mid-Atlantic salt marsh

Seston in salt marshes contains a temporally and spatially complex mixture of natural microparticulate organic material, including phytoplankton, vascular plant detritus, bacteria, heterotrophic nanoflagellates and benthic diatoms. Quantitative information is available concerning how suspension-feeding consumers, such as the ribbed mussel, Geukensia demissa (Dillwyn), utilize some of these components to satisfy their carbon demands. Despite this information there is still a limited understanding of how the relative nutritive contribution of these different dietary items may shift during the year associated with variations in both seston composition and the mussels physiological condition. To investigate if the mussels ability to use specific constituents of natural seston varies seasonally, we ran a series of pulse-chase 14C feeding experiments under ambient conditions in March, May, August and November 1996. Phytoplankton, cellulosic detritus, bacteria, heterotrophic nanoflagellates and benthic diatoms were radiolabeled and supplemented in small amounts to natural marsh water for feeding to mussels. The fate of 14C in mussel tissues, feces, respiration and excretion was quantified and contrasted among the different diet types and seasons. Microcapsules containing radiolabeled carbohydrate and protein were used as standards to differentiate possible between-experiment variations in seston composition from seasonal changes in the mussels feeding and digestive physiology. Mussel clearance rates for all diets were highest in summer and autumn and lowest in winter and spring. In contrast, seasonal shifts in digestive physiology were only found for certain diets. The seasonal range of assimilation efficiencies for microcapsule standards (18-29%) and field-collected microheterotrophs (bacteria 76-93% and heterotrophic nanoflagellates 87-94%) did not differ significantly during the year, whereas summer and autumn assimilation efficiencies for cellulosic detritus (22-24%), phytoplankton (71-79%) and benthic diatoms (89-93%) were up to twofold greater than those in winter and spring (13%, 40-59% and 45-81%, respectively). We conclude that the digestive physiology (e.g., digestive enzyme production) of mussels responds to shifts in dietary components during the year.

Trophic Complexity Between Producers and Invertebrate Consumers in Salt Marshes

Salt marshes on the Atlantic coast of North America are characterized by having a high biomass of smooth cordgrass, Spartina alterniflora. Because of the refractory nature of the lignocellulosic structure of this angiosperm, invertebrates utilize C from these plants with very low efficiency, if at all. This is true for both living cordgrass and post-senescent plant detritus. To balance their C demands, invertebrate consumers living in salt marshes must utilize a wide variety of other resources, including microheterotrophs (bacteria and bacterivorous flagellates) either associated with detritus or free in the water column, fungi colonizing decaying vascular plants, surface-associated algae (e.g., microphytobenthic diatoms and cyanobacteria, epiphytes, surface film algae) and phytoplankton. This high degree of trophic complexity is likely to be an important source of community stability. As an example, we estimate that ribbed mussels, Geukensia demissa, in a Delaware marsh must rely on a variety of different food resources since no single food type can meet their nutritional demands for either C or N. To balance their C demands, mussels appear to rely mainly on microheterotrophs, followed by phytoplankton > microphytobenthos > cellulosic detritus. Non-detrital foods are even more important for maintaining positive N balance in G. demissa. Previous and emerging evidence from other studies suggests that other important marsh consumers have a similar general diet. Although cordgrass may dominate overall rates of primary production and detritus from cordgrass contributes significantly to secondary production, we challenge the paradigm that salt marshes have a “detritus-based food web.” Further research is needed to deduce the importance of microphytobenthos and microheterotrophs as sources of C and N for dominant animal consumers in these marsh systems.