Macrobenthos abundance is a major factor to consider when assessing environmental quality of a river. To analyze macrobenthos abundance and its influencing factors, both field and laboratory experiments were done. A total of 6 macrobenthos species were collected, of which, 36.66% were chironomids, 60.69% were tubificids, 0.72% were snails, 0.73% were shrimps, 0.50% were caddisflies, 0.65% were shellfish，and 0.05% were unidentified species. Macrobenthos were collected, and in situ measurements of stream flow, water temperature and vertical hydraulic conductivity of riverbed sediments were done simultaneously at 5 research sites and 14 sampling sites along the Weihe River in Shaanxi Province. Water quality was determined in the laboratory. The type and abundance of macrobenthos were also analyzed. Data analysis was mainly done by de-trended correspondence analysis (DCA), principal component analysis (PCA), canonical correspondence analysis (CCA), and Student's t-test. It was found that abundance of chironomids was the highest in Huaxian and Lintong; abundance of snails and shrimps were the highest in Lintong; abundance of tubificids and caddisflies were the highest in Xianyang and Meixian; and abundance of shellfish was the highest in Caotan. In addition to variation in the number of macrobenthos across different research sites, their proportion and frequency of total individuals also varied, due to differences across research sites. In order to obtain more objective and reliable data, environmental factors considered in this work included both physical and chemical parameters. Quality of water in Weihe River was not a critical factor affecting macrobenthos abundance, because macrobenthos sensitivity to environmental factors were different. The main environmental factors influencing macrobenthos distribution are: total nitrogen, alkalinity, chemical oxygen demand, hexavalent chromium, total dissolved solids, stream-flow, temperature and vertical hydraulic conductivity of the riverbed sediments.