Chronic, 21-28-day toxicity tests of Macondo source (Massachusetts, or MASS) and weathered Slick A (CTC) and Slick B (Juniper) oils field collected during the 2010 Deepwater Horizon (DWH) Incident in the Gulf of Mexico (GOM) were conducted using standardized procedures. Standard species, Americamysis bahia and Menidia beryllina, were evaluated for changes in survival and growth during daily static-renewal tests. Both species demonstrated an increased sensitivity to low-energy water accommodated fractions (WAFs) of un-weathered MASS oil, with growth and survival decreasing as oil loading rate increased from 0.01 to 1.0 g/L. Survival and growth of mysid shrimp exposed to weathered oil (Slick A and Slick B) did not differ from that of test controls. In contrast, survival and growth of inland silversides declined relative to that of test controls at loading rates of 1 g/L for both weathered oils. Based on the concentration of total polycyclic aromatic hydrocarbons (TPAH42), no observed effect concentrations were lower for inland silverside survival (5.00-7.61 µg/L) and growth (<2.02 to <7.61 µg/L) in chronic exposures to Slick B and Slick A weathered oils compared with mysids (4.75-17.9 µg/L). Average TPAH concentrations in full strength WAFs followed the weathering trend, with 165 ± 17.2, 17.9 ± 0.480, and 4.75 ± 0.521 µg/L for MASS, Slick A, and Slick B oils, respectively. The effect (LOEC, IC25) and no-effect exposure concentrations (in TPAHs) from the standardized laboratory toxicity studies with un-weathered and weathered oils are discussed relative to the actual exposure concentrations in the GOM in 2010. The exposures evaluated in the laboratory toxicity tests represent the highest concentrations of total PAHs that were rarely observed in water column samples collected in the GOM during the release and post release periods of the DWH incident.