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AbstractThe U.S. Geological Survey, in cooperation with the Massachusetts Department of Conservation and Recreation, Massachusetts Department of Environmental Protection, and the Massachusetts Department of Fish and Game, conducted an investigation of fish assemblages in small-to medium-sized Massachusetts streams. The objective of this study was to determine relations between fish-assemblage characteristics and anthropogenic factors, including impervious cover and estimated flow alteration, relative to the effects of environmental factors, including physical-basin characteristics and land use. The results of this investigation supersede those of a preliminary analysis published in 2010. Fish data were obtained for 669 fish-sampling sites from the Massachusetts Division of Fisheries and Wildlife fish-community database. A review of the literature was used to select fish metrics-species richness, abundance of individual species, and abundances of species grouped on life history traits-responsive to flow alteration. The contributing areas to the fish-sampling sites were delineated and used with a geographic information system to determine a set of environmental and anthropogenic factors that were tested for use as explanatory variables in regression models. Reported and estimated withdrawals and return flows were used together with simulated unaltered streamflows to estimate altered streamflows and indicators of flow alteration for each fish-sampling site. Altered streamflows and indicators of flow alteration were calculated on the basis of methods developed in a previous U.S. Geological Survey study in which unaltered daily streamflows were simulated for a 44-year period (water years 2 1961-2004), and streamflow alterations were estimated by use of water-withdrawal and wastewater-return data previously reported to the State for the 2000-04 period and estimated domestic-well withdrawals and septic-system discharges.A variable selection process, conducted using principal components analysis and Spearman rank correlation, was used to select a set of 15 non-redundant environmental and anthropogenic factors to test for use as explanatory variables in the regression analyses. Twenty-one fish species were used in a multivariate analysis of fish-assemblage patterns. Results of nonmetric multidimensional scaling and hiera...