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Along with other varanid lizards, such as the Komodo dragon and the lace monitor, megalania belongs to the proposed clade Toxicofera, which contains all known reptile clades possessing toxin-secreting oral glands, as well as their close, non-venomous relatives, including Iguania, Anguimorpha, and Serpentes.

Closely related varanids use a potent venom found in glands inside the jaw.

Judging from its size, it would have fed mostly upon medium to large sized animals, including any of the giant marsupials like Diprotodon along with other reptiles and small mammals, as well as birds and their eggs and chicks They note that the "marsupial lion" (Thylacoleo carnifex) has been implicated with the butchery of very large Pleistocene mammals, while Megalania has not.

In addition, they note that Megalania fossils are extremely uncommon, in contrast to Thylacoleo carnifex with its wide distribution across Australian Pleistocene deposits.

Several studies have attempted to establish the phylogenetic position of megalania within the Varanidae.

An affinity with the perentie, Australia's largest living lizard, has been suggested based on skull-roof morphology.

Unless other Australian monitor species were each also classified their own exclusive genera, Megalania would not be a valid genus name.

If it had a long thin tail like the lace monitor (Varanus varius), then it would have reached a length of 7.9 metres (26 ft), while if its tail-to-body proportions were more similar to that of the Komodo dragon (Varanus komodoensis), then a length of around 7 m (23 ft) is more likely.

Recent paleontological analysis using accelerator mass spectrometry (AMS) 14C dating of known fossils shows megalania to have been alive circa Pleistocene epoch 50,000 years ago.

An affiliate hypothesis to these datings is that anthropogenic extirpation was the cause of the downfall of megalania and other Australian megafauna in the similar vein as to how a large factor of the extinction of the northern hemisphere’s megafauna at the end of the Pleistocene epoch was caused by early humans.

This would rapidly decrease the prey's blood pressure and lead to systemic shock.

Being a member of Anguimorpha, megalania may have been venomous and if so, would be the largest venomous vertebrate known.

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