Climate change is gradual and long-term, consistently collected data are required to detect resulting biological responses and to separate such responses from local effects of human activities that monitoring programs usually are designed to assess. The reference-condition approach is commonly used in freshwater assessments that use predictive modeling, but a consistent reference condition is required to maintain the relevance and integrity of results over the long term. We investigated whether external influences, such as climate change, inhibited clear interpretation of bioassessment results in a study design using reference vs test sites. Macroinvertebrates were collected from 16 sites (11 sites affected by ski resorts and 5 reference sites) on 5 streams in 4 seasons each year from 1994 to 2008 within Kosciuszko National Park, Australia. We analyzed trends over 15 y to address questions regarding climate-change and macroinvertebrate bioindicators of stream condition (observed/expected [O/E] taxa; Stream Invertebrate Grade Number Average Level [SIGNAL] 2 scores; Simpson's Diversity; Ephemeroptera, Plecoptera, Trichoptera [EPT] richness ratio; and Oligochaeta abundance). Climate became slightly warmer and less humid (p < 0.0001), but no significant relationships between climate variables and bioindicators were evident. All bioindicators consistently distinguished between test and reference sites in all seasons. All bioindicators except for O/E taxa scores differed among streams (regardless of site type). O/E taxa are inherently adjusted for specific stream characteristics, and, thus, were robust to differences in stream type while remaining sensitive to reference and test site variation. Generally, reference and test sites did not respond differently to any gradual climate changes. Furthermore, the reference sites sampled through time remained in a condition equivalent to the previously defined reference condition and provided a valid comparison for current test sites of unknown condition. The bioindicators used here were insensitive to the small but significant changes in climate detected over the 15-y study. However, extreme climate-related events (such as severe drought and extensive bushfire) were detected by the chosen bioindicators at both reference and test sites. Ecological outcomes of climate change can be accounted for only by an appropriate study design that includes standardized sampling of fixed sites (both test and reference) over long periods.
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