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272 M. J. JENNINGS, L. S. FORE AND J. R. KARR <br />How well does the current RFAI meet these three tests? The first challenge is grounded in the observation <br />that human actions, from the construction of dams to the introduction of toxic chemicals or exotic species, <br />cause changes in the local biota. That is, biological systems are responsive-humans do something and the <br />system changes in return. The success of the general multimetric framework in a variety of types of environ- <br />ments, with various taxa and in diverse geographical areas give us confidence in the RFAI concept and in the <br />metrics selected for this project. Expectations for biomonitoring applications must be tempered with the <br />reality that the development of ecologically meaningful, statistically sound indexes is an iterative process. <br />Further, TVA biologists are currently testing these hypotheses and tuning the index based on their knowledge <br />of biological patterns and new data. The challenge is to formalize and refine that knowledge in the context of <br />the unique aspects of reservoir environments. <br />The second challenge is more troublesome for the RFAI as it stands today. The observation that RFAI <br />scores were not consistent over time within a year, and that no consistent seasonal trend accounted for the <br />differences, suggests that the index as presently constructed is subject to considerable sampling error. Neither <br />natural nor anthropogenic variation can be invoked to explain the pattern. Further, we do not accept the <br />premise that fish assemblage health changed in any meaningful way during a single year, as did index scores <br />from the 1992 supplemental data set. Therefore, we doubt that the annual variation in reservoir ranks <br />observed over the four years of the study reflects real changes among reservoirs in fish assemblage health. <br />This concern is reinforced by the pattern of RFAI variation derived from bootstrap analysis. <br />Sampling error occurs when the entire population (e.g. all fish in the reservoir) is not sampled. For electro- <br />fishing in reservoirs, the sampling error is caused by differences in sampling locations-that is, placement of <br />electrofishing runs within the zone. A comparison of sampling error and measurement error for sites eval- <br />uated as part of the 1992 supplemental studies reveals that variability among run locations (sampling error) <br />contributes more to the overall variance of RFAI scores than measurement error (Figure 4) for 13 of 16 <br />reservoir months. We conclude that sampling variability is the cause of the high variability of RFAI scores. <br />Three lines of evidence support this conclusion: (1) the high sampling variability revealed by resampling <br />analysis; (2) the failure of species accumulation curves to level off, and (3) the inconsistent ranking of reser- <br />voirs using TAXA and ABUN. These results indicate that sampling effort at a site needs to be improved to <br />ensure a representative sample. <br />An ancillary problem stems from the need for more comprehensive tests of the individual metrics. Some <br />metrics included in our RFAI are probably not appropriate. The FHAI may fail because it was originally <br />developed to reflect the effects of bacterial and fungal infections and other stresses associated with hatchery <br />fish, populations that are widely known for carrying such problems (Goede, 1986). The method may be less <br />appropriate in largely healthy wild fish, especially in view of the sample bias associated with small sample sizes. <br />Much of the decision-making with respect to biological goals in reservoirs may be directed towards <br />improvement in fisheries. In that situation, we suggest the direct but selective inclusion of metrics such as <br />the age structure of sport fish species in the next round of RFAI development. Such efforts should avoid <br />the temptation to only use metrics based on harvestable fishery concepts, and should emphasize the value <br />of naturally reproducing, self-sustaining fisheries-that is, RFAI may include fishery concepts, but should <br />always be ecologically broader. <br />The third challenge has not yet been adequately explored. Our use of best values across the reservoirs to <br />establish scoring criteria is defensible, but not very satisfying. The multimetric approach relies on an ability <br />to sample systematically sites that span a broad range of conditions, especially a range of human influences. <br />The linear connection among the TVA reservoirs presents an important problem in extrapolating our <br />analysis. Further, we consider our efforts to document the associations between human actions and reser- <br />voir biology to be preliminary. A more systematic effort at defining these patterns must come in concert <br />with a refinement of the sampling protocols (e.g. the appropriate number of electrofishing runs). <br />In addition to defining sampling protocols, more careful study design is necessary to document the influ- <br />ence of habitat type (e.g. sand, rip-rap, emergent vegetation) and variation among sites (within a reservoir <br />zone), zones and reservoirs. These sources of biological pattern need to be integrated with the effect of land <br />use and other human influences, such as the management of hydrological conditions (e.g. water-level <br />fluctuations within a reservoir and the level of water release from upstream reservoirs).