From 2011-2013, a large-scale ecotoxicological study was conducted in several Chesapeake Bay tributaries (Susquehanna River and flats, the Back, Baltimore Harbor/Patapsco River, Anacostia/ middle Potomac, Elizabeth and James Rivers), and Poplar Island as a mid-Bay reference site. Osprey (Pandion haliaetus) diet and the transfer of contaminants from fish to osprey eggs were evaluated. The most bioaccumulative compounds (biomagnification factor >5) included p,p'-DDE, total PCBs, total PBDEs, and BDE congeners 47, 99, 100 and 154. This analysis suggested that alternative-brominated flame-retardants and other compounds (methoxytriclosan) are not appreciably biomagnifying. A multivariate analysis of similarity indicated major differences in patterns among study sites were driven by PCB congeners 105, 128, 156, 170/190 and 189, and PBDE congeners 99 and 209. An integrative redundancy analysis showed that osprey eggs from Baltimore Harbor/Patapsco River and the Elizabeth Rivers had high residues of PCBs and p,p'-DDE, with PBDEs having a substantial contribution to overall halogenated contamination on the Susquehanna and Anacostia/middle Potomac Rivers. The redundancy analysis also suggested a potential relation between PBDE residues in osprey eggs and oxidative DNA damage in nestling blood samples. Results of this study also indicate that there is no longer a discernible relation between halogenated contaminants in osprey eggs and their reproductive success in Chesapeake Bay. Osprey populations are thriving in much of the Chesapeake, with productivity rates exceeding those required to sustain a stable population. This article is protected by copyright. All rights reserved.