The data presented include concentrations of 122 organic, 23 inorganic constituents, and estrogenicity in water collected from 13 off-stream non-point source sites (natural and created wetlands) and eight on-stream point source sites (streams and impoundments) in the New Jersey Pinelands, 2017-2020. Native and non-native fish health data collected from the on-stream sites as well as native frog data collected from the off-stream sites are also included. For the on-stream point-source component, we analyzed water chemistry, hormone activity, and collected native and non-native fish from two locations upstream and one downstream from two municipal wastewater treatment plants (six total sites) and from two minimally impacted locations to serve as reference conditions. For the off-stream non-point source component, we analyzed water chemistry, hormone activity, and collected Green Frogs (Lithobates clamitans) from eight stormwater basins and ponds embedded in altered landscapes and five ponds surrounded by forest land to serve as reference sites. Animals were assessed histologically for measures of endocrine disruption and surface water was analyzed for hormone activity and known or suspected endocrine disrupting compounds (EDCs) including hormones, phytoestrogens, waste indicator compounds and some metals.