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Genetic testing was performed on the pups as part of research related to a breeding program for the species and the researchers were stunned to discover Bubbles was the sole parent of the baby sharks, having reproduced via parthenogenesis, or "virgin birth," where a female will fertilize eggs with her own genetic material.
The aquarium said zebra sharks have been known to reproduce asexually on rare occasions, but it was believed to be a response to a lack of available male mates.
"This discovery throws a wrench in what we thought we knew about how and why parthenogenesis happens, and it illustrates a key aspect of science: we're continually learning," Kevin Feldhim, a researcher at Chicago's Field Museum, said in a Shedd Aquarium news release.
Feldhim co-authored a study on the case, published in the Journal of Fish Biology.