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The use of adenosine triphosphate (ATP) as a universal biomass indicator is built on the premise that ATP concentration tracks biomass rather than the physiological condition of cells. However, reportedly high variability in ATP in response to environmental conditions is the main reason the method has not found widespread application. To test possible sources of this variability, we used the diatom Thalassiosira weissflogii as a model and manipulated its growth rate through nutrient limitation and through exposure to three different temperatures (15°C, 20°C, and 25°C). We simplified the ATP protocol with hot‐water or chemical extraction methods, modified a commercially available luciferin‐luciferase assay, and employed single‐photon counting in a scintillation counter, all of which increased sensitivity and throughput. Per‐cell ATP levels remained relatively constant despite changes in growth rates by approximately 10‐fold in the batch culture (i.e., nutrient limitation) experiments, and approximately 2‐fold in response to temperature. The re‐examination of related literature values revealed that average cellular ATP levels differed little among taxonomic groups of aquatic microbes, even at the domain level, and correlated well with bulk properties such as elemental carbon or nitrogen. Fulfilling multiple cellular functions in addition to being the universal energy currency requires ATP to be maintained in a millimolar concentration range. Consequently, ATP relates directly to live cytoplasm volume, while elemental carbon and nitrogen are constrained by an indeterminate pool of detrital material and intracellular storage compounds. The ATP‐biomass indicator is sensitive, economical, and can be readily standardized among laboratories and across environments. [ABSTRACT FROM AUTHOR]

Two frequently used universal eukaryote probes, EUK1209 and EUK516, are not consistent with one branch of the eukaryotic phylogenetic tree, the Kinetoplastida, which has undergone rapid evolution of their small subunit rRNA gene. Kinetoplastids include medically important parasitic organisms (e.g. Trypanosoma, Leishmania) and free-living flagellates that occur in all aquatic environments and in soils (e.g. Bodo, Neobodo, Rhynchomonas). A modified probe presented here as KIN516, now based on the kinetoplastid sequence, provides a strong signal with Neobodo designis, Leishmania donovani, and Trypanosoma cruzi using the catalyzed reporter deposition protocol. EUK516 and KIN516 function as competitor probes, thereby greatly increasing discriminatory power when used in combination. The probe pair was tested in field samples collected in a freshwater pond in Norfolk, the mesohaline Elizabeth River, Norfolk, Virginia, and a tropical lagoon in Belize. The combined probes bound to 58–84% of organisms identified as eukaryotic based on having large DAPI-stained nuclei. The contribution of kinetoplastids to total eukaryotes (positive signal of EUK516+KIN516) was much higher in marine samples (ca. 17%) than in either the freshwater or brackish water sites (<0.2%). [ABSTRACT FROM AUTHOR]

The combination of gut contents and evacuation rate is an important tool with which to determine the in situ feeding rates of many organisms. Traditionally used equations of gut evacuation models, however, have made comparisons among models difficult. For consistency, we changed the notation of the linear gut evacuation model to provide a rate-constant parameter ( r) similar to the one commonly used in the exponential model. In a previous analysis using examples from the literature, we demonstrated that prolonged retention of food after a true linear pattern of gut evacuation may be mistakenly identified as an exponential pattern. We explored the linear gut evacuation rate model empirically and in more detail using the radiated shanny as the model species. Rate constants based on field data were similar to those determined in the laboratory. The basic assumption of nonfeeding during darkness was met, indicating that foraging was primarily based on visual cues. As predicted by our model, both laboratory and field data confirmed that gut evacuation was linear and characterized by a rate constant (i.e., instantaneous rate of gut evacuation) that was independent of initial gut fullness. Rate constants were, however, influenced by fish size and in some cases by the size of food particles. Analysis of triplicate time courses of gut evacuation in the field showed that larger larvae evacuated food at a relatively higher rate than did smaller larvae. [ABSTRACT FROM AUTHOR]

We investigated the interaction of the copepod Acartia hudsonica in relation to thin layers of the diatom Skeletonema costatum. Thin layers have recently received much attention, since they are common and persistent features in the water column, often overlooked by traditional sampling methods. Their frequent abundance in coastal oceans and the high biomass associated with them has led to the assumption that they are important grazing sites of calanoid copepods. We employed 2-m tall tower tanks that allowed us to simulate thin layers. Three variables representative of three time scales were considered: the distribution of copepods in the tanks (time scale of minutes), fecal pellet production as a proxy for ingestion rate (time scale of hours), and egg production rate (time scale of >12 h). A. hudsonica responded significantly but very little to the thin layers in terms of their distribution. Given a choice, there was a slightly higher tendency to swim through a patch of diatoms than to swim around it. Fecal pellet production was slightly lower in the thin-layer treatments than in the homogeneous controls. Egg production was not influenced by differential distribution of diatoms in the tanks, which indicated that the copepods dealt equally well with patchy food as when the same numbers of cells were available in a homogeneous distribution. Time series experiments showed that ingested carbon is integrated over time scales of >12 h. Therefore, small-scale fluctuations of food in space and time do not necessarily translate into small-scale fluctuations in reproductive output. [ABSTRACT FROM AUTHOR]