King Tutankhamun was most likely the child of the "heretic" pharaoh Akhenaten, fathered two stillborn girls and was afflicted by several diseases, including malaria, according to a major genetic investigation into the boy king's family.
Published in the Journal of the American Medical Association, the international study used the modern tools of molecular genetics and X-ray analysis to provide details on the ancient pharaoh's life and early death at the age of 19. The research was led by Dr. Zahi Hawass, the head of Egypt's Supreme Council of Antiquities.
"We have found so many curious conditions and pathologies in King Tut that it is really a problem to define what killed him," author Carsten Pusch at the Institute of Human Genetics of Tubingen University, Germany, told Discovery News.
Even though King Tut's mummy had been X-rayed several times, the researchers during this examination found a series of malformations in the pharaoh's feet. The second toe lacked the middle bone, making it shorter, and one of his feet was clubbed.
In addition, they found that a degeneration of the second and third long bones in the left foot could have been consistent with Köhler disease II, a foot bone disorder.
The researchers found the DNA of Plasmodium falciparum, the malaria parasite, in four royal mummies, including Tutankhamun's. That finding represents the oldest genetic proof of malaria in precisely dated mummies.
"We should not forget the leg fracture, which we know from the 2005 CT scans. It did not heal, so it occurred close to King Tut's death," Pusch said.
"It is very likely that the bone necrosis and the clubfoot required King Tut to use canes. Maybe he just fell and broke his leg," the researcher said.
Indeed, about 130 walking sticks found in King Tut's treasure-packed tomb would support the diagnosis.
Since King Tut had multiple disorders, but none alone would have caused death, there is the possibility that some of them might have turned into an inflammatory, immuno-suppressive, weakening syndrome, the researchers concluded.
"He might be envisioned as a young but frail king who needed canes to walk because of the bone-necrotic and sometime painful Köhler disease II," Hawass and colleagues wrote.
The leg fracture, possibly caused by a fall, might have resulted in a life-threatening condition when a malaria infection occurred.
"You have a bone necrosis, malaria, a fracture: all of this together is not in agreement with a long life," Pusch said.
The genetic analysis, conducted in a DNA lab funded by the Discovery Channel, took place between Sept. 2007 and Oct. 2009, when the researchers selected 10 mummies (1410-1324 B.C.) closely related in some way to Tutankhamun.
Of these, the identities were certain for only three. In addition to these 11 mummies, five other royal individuals dating to the early New Kingdom (1550-1479 B.C.) were chosen as a control group.
Using partial Y chromosomal information, the researchers produced the most reliable five-generation pedigree of Tutankhamun's immediate lineage to date.
Yuya and Thuya were recognized as King Tut's great-grandparents. Pharaoh Amenhotep III and the mummy known as the Elder Lady (KV35EL) were found to be his grandparents, while the mummy known as KV55 -- most likely Akhenaten -- and KV35YL, the Younger Lady, were identified as siblings, as well as King Tut's parents.
"We know that the Younger Lady (KV35) is King Tut's mother. But we can't tell whether she is Nefertiti or Kiya or another queen or princess. We have just opened the door, and more work must be done. Moreover, the high degree of inbreeding in this family makes the DNA even more homogeneous, and analysis is of course more difficult," Pusch said.
Yuya and Thuya were also affected by malaria, but surprisingly both reached the advanced age of 50 and older. According to the researchers, this means that either the infection took place quite late in their lifetime, or they acquired a partial immunity to the pathogen.
The researchers also established that the two female fetuses buried in the tomb of Tutankhamun were most likely his offspring.
"The mother is not yet genetically identified, but the data obtained from KV21A points to this mummy as the mother of the fetuses. Unfortunately we are not yet able to identify her as Ankhesenamun, Nefertiti's daughter," Pusch said.
Morfological and radiological examination did not reveal signs of a feminine or androgynous physique.
"It is unlikely that either Tutankhamun or Akhenaten actually displayed a significantly bizarre or feminine physique. It is important to note that ancient Egyptian kings typically had themselves and their families represented in an idealized fashion," Hawass and colleague wrote.