Lysterfield Park consists of approximately one third remnant vegetation and two thirds plantation. Within these areas about 300 plant species have been recorded. Some of the over-storey species native to the area include: Yellow Box, Narrow-leaved Peppermint, Stringybark, Manna Gum, River Red Gum, Swamp Gum and Long-leaved Box.
There are eight plantation types in the park:River Red Gum, Southern Mahogany, Spotted Gum, Swamp Gum, Blue Gum, Forest Red Gum, Long-leaved Box and Sugar Gum.
Lysterfield Park provides a diverse range of habitats and is a valuable wildlife refuge. The list of bird species recorded is about 140. Of these, 37 are waterbirds and include common species such as Eurasian Coots and Eastern Swamphens to the uncommon visitors such as Japanese Snipe and Red-necked Avocet.
Eastern Grey Kangaroos and Black Wallabies are readily seen. Koalas, antechinus and echidnas are much harder to find. At night the park comes alive with Brushtail and Ringtail Possums, Sugar Gliders and various species of bats and owls.