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<i>Cherax destructor</i> is a crayfish endemic to south-eastern Australia and one of the last alien crayfish to be introduced in Italy. In the Laghi di Ninfa Natural Reserve (Latium region, Central Italy), the species was probably introduced in 1999, but only reported for the first time in 2008. Nearby this area, the most widespread alien crayfish is the red swamp crayfish <i>Procambarus clarkii</i>. In the Natural Reserve, between 2008 and 2013 and during spring and summer, crayfish sampling was carried out with baited traps to assess the distribution of <i>C. destructor</i> and its possible relationship with <i>P. clarkii</i>. <i>Cherax destructor</i> was first recorded in 2008; few <i>P. clarkii</i> were detected in the cultivation ponds where <i>C. destructor</i> was present in 2012 and 2013. Moreover, crayfish plague analyses evidenced a positive result in two out of the 12 sampled <i>P. clarkii</i>. <i>Cherax destructor</i> is now completely absent from the Natural Reserve, while <i>P. clarkii</i> has spread in the area and was probably responsible for this eradication since <i>C. destructor</i> is vulnerable to crayfish plague which was also detected in the area. An ecosystem restoration project in the area favoured the spread of. <i>P. clarkii</i>; the implications of this intervention are discussed.