In shrimp hatcheries, giving enriched feed to larval stages is considered an important part, and it has been fulfilled by live organism like brine shrimp Artemia salina is in practice. The objective of this study was to investigate the pathogenicity of Enterocytozoon hepatopenaei (EHP) to Artemia and to identify whether it acts as reservoir for EHP infection in penaeid shrimp. Transmission of EHP in Artemia and penaeid shrimp was carried out by immersing with EHP infected tissue homogenate. Polymerase chain reaction was carried out to confirm the EHP infection in Artemia, revealing positive in all developmental stages without any incidence of mortality. Histopathological analysis of challenged adult Artemia showed spores within late plasmodium stages in tissue sections. EHP exposed live-Artemia was fed to healthy post larvae (EHP negative post larvae) and was demonstrated positive towards EHP as confirmed by polymerase chain reaction. Therefore, Artemia can be accepted as a possible vector for transmission of Enterocytozoon hepatopenaei in penaeid shrimp. © 2020 Elsevier Ltd