Ageing is known to have significantly detrimental effect on mode I fatigue delamination growth (FDG) in unidirectional (UD) composite laminates. However, composite structures are usually designed with multidirectional (MD) layups, which raises the question that is it enough to only conducted fatigue delamination experiments on specimens with a UD layup. The aim of this study is therefore to explore mode I FDG in MD composite laminates with 45//45 interface after different ageing, i.e. at 70 °C 85 % relative humidity (RH) and immersion in 70 °C water bath. Fatigue delamination experiments were conducted at stress ratios R = 0.1 and 0.5. The fatigue data, interpreted via Paris-type fatigue laws, demonstrated that: (1) the change of ageing severity has no influence on mode I FDG in MD composite laminates; (2) FDG remains the same in composite laminates after different ageing, regardless of layups. In all cases, the same master resistance curves can be obtained to determine the intrinsic mode I fatigue delamination resistance of UD and MD composite laminates after different ageing. The physical reasons for these findings were discussed based on the moisture content analysis, Fourier transform infrared (FTIR) analysis, dynamic mechanical thermal analysis (DMTA), and fractographic examinations. It was found that material degradation and delamination mechanisms remain the same for UD and MD layups, as well as for 85 %RH and water bath conditioning.