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Table 1.

Biological response variables measured in the field surveys and corresponding response variables tested in laboratory and instream nutrient limitation assays.

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Table 1 Expand

Table 2.

Summary of seasonal nutrient characteristics of streams and stream waters tested.

Streams are listed in order of increasing average total phosphorus (P) concentrations.

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Fig 1.

Matched views of a nutrient-diffusing substrate (NDS) rack at deployment and after 21-days of colonization in a nitrogen and phosphorus (N+P) co-limited stream, the Big Wood River.

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Fig 1 Expand

Fig 2.

Green algae growth results from six streams with Nitrogen (N), Phosphorus (P), or N+P additions.

Averages ± 95th percentile confidence intervals (CI) of the mean (n = 3). Primary nutrient limitation is concluded when the CI of the single nutrient additions does not overlap the mean of the controls. Co-limitation of N+P is determined by significant (p<0.05) interaction term in 2-factor ANOVA test.

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Fig 3.

Green algae growth in ambient waters (no additions).

The asymptotic relation with total P suggests saturation with a half-saturation value (50% of maximum) of about 65 μg/L TP and above about 120 μg/L TP, no further growth increases with P were noted. Error bars show standard deviation.

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Table 3.

Percent phosphorus (P) and nitrogen (N) removed from the water column during the 11-day P and N growth series.

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Table 3 Expand

Fig 4.

Duckweed and periphyton responses to Phosphorus (P) enrichment (left) and Nitrogen (N) enrichment (right).

Error bars show standard deviations. EC50 (± SE): the concentration causing an increase in growth to 50% of maximum growth; saturation concentrations–further increases in P or N result in little further growth, calculated as 95% of the absolute asymptote.

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Table 4.

Nutrient limitation conclusions from the nutrient-diffusing substrate (NDS), average (ranges) of ambient nutrient concentrations in the streams at the time of the tests, and ancillary chemistry and channel measurements.

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Table 4 Expand

Fig 5.

Chlorophyll (a) accrual on nutrient-diffusing substrates (NDS) with Nitrogen (N), Phosphorus (P), or N+P additions.

Averages ± 95th percentile confidence intervals (CI) of the mean (n = 6). Nutrient limitation is interpreted as with Fig 2.

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Table 5.

Nutrient limitation conclusions from streams tested under similar conditions with both the green algae bottle test and nutrient-diffusing substrates.

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Fig 6.

Nitrogen to phosphorus ratios corresponding with empirically determined nutrient limitation from the different stream water experiments.

(N-nitrogen limited; NP-nitrogen and phosphorus co-limited; P-phosphorus limited).

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Fig 7.

Benthic chlorophyll a density over time in a nitrogen-limited stream.

A. Benthic chlorophyll a relative to both nitrogen (N) and phosphorus (P); B. Overlain with the three nutrient limitation experiments conducted on the stream that demonstrated consistent N limitation; C. A simplified comparison without P shows nearly perfect inverse correspondence between chlorophyll a and N.

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Fig 8.

Benthic chlorophyll a on artificial substrates at nitrogen (N) limited sites versus field samples.

Curve fit to chlorophyll a from NDS controls (no nutrient additions), excluding P-limited sites, following 21-day accrual (top), and the same line fit overlaying 62 benthic chlorophyll a samples collected from natural substrates in associated field collections (bottom). Field data from [28, 39].

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Fig 9.

Benthic chlorophyll a on artificial substrates at phosphorus (P) limited sites versus field samples.

Curve fit to chlorophyll a from NDS controls (no nutrient additions), excluding N-limited sites, following 21-day accruals overlain with 62 benthic chlorophyll a samples collected from natural substrates.

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