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Cases of tick-borne diseases have been steadily increasing in the USA, owing in part to tick range expansion, land cover and associated host population changes, and habitat fragmentation. However, the relative importance of these and other potential drivers remain poorly understood within this complex disease system. Ticks are ectotherms with multi-host lifecycles, which makes them sensitive to changes in the physical environment and the ecological community. Here, we describe data collected by the National Ecological Observatory Network on tick abundance, diversity and pathogen infection. Ticks are collected using drag or flag methods multiple times in a growing season at 46 terrestrial sites across the USA. Ticks are identified and enumerated by a professional taxonomist, and a subset of nymphs are PCR-tested for various tick-borne pathogens. These data will enable multiscale analyses to better understand how drivers of tick dynamics and pathogen prevalence may shift with climate or land-use change.