The use of silver nanoparticles (AgNPs) result in an inevitable contact with aquatic environments. Here we study the behavior of AgNPs and the developmental toxicity in zebrafish embryos exposed to these nanoparticles (0–10 mg/L) with and without the presence of HA (20 mg/L), using zebrafish facility water (ZFW) and zebrafish growing media (ZGM). The presence of cations and HA gave rise to a decrease in Ag ion release and ζ-potential, an increase in the hydrodynamic diameter and oxidation of the AgNP surface. The results show that the presence of HA and cations in the media, as well as the silver speciation, i.e., the unusual presence of Ag 3+ , decreases the toxicity of AgNPs (LC50 AgNPs : 1.19 mg/L; LC50 AgNPs + HA : 3.56 mg/L), as well as silver bioavailability and toxicity in zebrafish embryos. Developmental alterations and the LC50 (1.19 mg/L) of AgNPs in ZFW were more relevant ( p ≤ 0.05) than for AgNPs in ZGM (LC50 ˃ 10 mg/L). It was demonstrated that the bioaccumulation and toxicity of AgNPs depends on several factors including AgNPs concentration, nanoparticle aggregation, dissolved silver ions, speciation of silver ions, the amount of salt in the environment, the presence of humic substances and others, and different combinations of all of these factors. [ABSTRACT FROM AUTHOR]