Matthew R. First

Stream nutrient enrichment has a greater effect on coarse than on fine benthic organic matter

Abstract. Nutrient enrichment affects bacteria and fungi associated with detritus, but little is known about how biota associated with different size fractions of organic matter respond to nutrients. Bacteria dominate on fine (<1 mm) and fungi dominate on coarse (>1 mm) fractions, which are used by different groups of detritivores. We measured the effect of experimental nutrient enrichment on fungal and bacterial biomass, microbial respiration, and detrital nutrient content on benthic fine particulate organic matter (FPOM) and coarse particulate organic matter (CPOM). We collected FPOM and CPOM from 1 reference and 1 enriched stream. CPOM substrates consisted of 2 litter types with differing initial C:nutrient ratios (Acer rubrum L. and Rhododendron maximum L.). Fungal and bacterial biomass, respiration, and detrital nutrient content changed with nutrient enrichment, and effects were greater on CPOM than on FPOM. Fungal biomass dominated on CPOM (∼99% total microbial biomass), whereas bacterial biomass dominated on FPOM (∼95% total microbial biomass). These contributions were unchanged by nutrient enrichment. Bacterial and fungal biomass increased more on CPOM than FPOM. Respiration increased more on CPOM (up to 300% increase) than FPOM (∼50% increase), indicating important C-loss pathways from these resources. Microbial biomass and detrital nutrient content were positively related. Greater changes in nutrient content were observed on CPOM than on FPOM, and changes in detrital C:P were greater than changes in detrital C:N. Threshold elemental ratios analyses indicated that enrichment may reduce P limitation for shredders and exacerbate C limitation for collector-gatherers. Changes in CPOM-dominated pathways are critical in predicting shifts in detrital resource quality and C flow that may result from nutrient enrichment of detritus-based systems.

Ciliate Ingestion and Digestion: Flow Cytometric Measurements and Regrowth of a Digestion-Resistant Campylobacter jejuni

We measured ingestion and digestion rates of the pathogenic bacterium Campylobacter jejuni by a freshwater ciliate Colpoda sp. to determine whether Campylobacter is able to resist protist digestion. Campylobacter and the nonpathogenic bacterium Pseudomonas putida LH1 were labeled with a 5‐chloromethylfluorescein diacetate, which fluoresces in intact and active cells but fades when exposed to low pH environments, such as protistan food vacuoles. Ingestion and digestion rates were measured via flow cytometry as the change in ciliate fluorescence over time, which corresponded to the quantity of intracellular bacteria. The rate of Campylobacter ingestion exceeded the digestion rate. Ciliates retained labeled Campylobacter 5 h after ingestion was stopped. In contrast, ciliates grazing upon P. putida returned to baseline fluorescence within 5 h, indicating that P. putida were completely digested. The ability of intracellular Campylobacter to remain viable after ingestion was tested by sorting individual ciliates and bacterial cells into Campylobacter‐selective media. Campylobacter growth occurred in 15% (± 5 SE) of wells seeded with highly fluorescent ciliates, whereas only 4% (± 1) of wells seeded with free‐living Campylobacter exhibited growth. A key advantage of this approach is that it is rapid and should be applicable to other phagocytotis studies.

Nutrient enrichment alters the magnitude and timing of fungal, bacterial, and detritivore contributions to litter breakdown

Microbial decomposers and metazoan detritivores rely on and affect processing rates of detrital C in streams. Nutrient enrichment can affect the biomass of both groups, but we lack a quantitative understanding of how these groups affect breakdown rates of C under elevated nutrient conditions. We quantified the relative contribution of decomposers (fungi, bacteria) and detritivores (shredders) to breakdown rates in a reference vs a nutrient-enriched stream. We measured breakdown rates and litter-associated shredder, fungal, and bacterial biomass on Rhododendron maximum L. and Acer rubrum L. and modeled the contribution of these groups over the decay sequence for R. maximum and for a single time period for A. rubrum. We used published metabolic parameters for fungi and bacteria under high- and low-nutrient conditions and shredder feeding rates based on organisms and leaf litter from our study area. Breakdown rates of both litter types were higher (∼4–5×) in the treatment than reference stream and were associated with large changes in biomass of fungi, bacteria, and shredders. Changes in microbial metabolic parameters (growth rate, growth efficiency) affected estimates of mass loss. Fungal and shredder contributions were roughly equivalent and greater than bacterial contributions under reference and treatment conditions, but nutrient enrichment accelerated the colonization sequence of fungi and shredders, and shredders had relatively greater effects on mass loss in later stages of breakdown. Our results suggest facilitation of detritivores by decomposers and large combined effects on breakdown with nutrient enrichment. Decomposer contributions could exceed those of detritivores with changes in microbial metabolic rates. Large increases in breakdown rates could be attributed to biotic response to nutrient enrichment, are sensitive to microbial metabolic rates, and have the potential to alter residence times of litter in streams.