Over the period from 1994 to 2007, air and water temperatures in the Australian state of New South Wales (NSW) increased while rainfall and river flows declined. Data on the occurrence of stream macroinvertebrate families in bioassessment samples collected in NSW during this period were examined to see whether a biological response to these climatic and hydrological trends could be discerned. Multiple logistic regression was used to test for long-term trends in the probability of detection of individual macroinvertebrate families within the samples, taking account of the latitude, longitude, altitude, hydraulic habitat, time of year and subsampling method associated with each sample. Of the 124 families and family groups tested, 33 had statistically significant increasing trends, 37 had significantly declining trends and 54 had no significant trend; however, many of the last group were seldom collected and their trend estimates had wide confidence limits. Significant relationships were found between the thermophily and rheophily of the families and the estimated strength and direction of their long-term trends, with families that favour colder waters and faster-flowing habitats more likely to have declined. Although many families showed trends of increasing detection within samples, such trends do not necessarily equate to increasing prevalence in the environment because the extent of wetted habitat has probably declined, especially flowing habitat. In addition, because of likely intrafamilial trait diversity, increasing or apparently increasing families may include species in decline. Many freshwater macroinvertebrate species in NSW may be threatened by anthropogenic climate change, exacerbated by water withdrawals. The thermal tolerances, rheophily and other pertinent traits of individual species need to be determined to identify those most at risk.