The shipping industry is considered the main vector of introduction of marine non-indigenous species (NIS). NIS distributions are often a consequence of frequent trade activities that are affected by economic trends. A dominant trend in the shipping industry is the operation of Ultra Large Container Vessels (ULCV), which are over 395 m long and sail mostly on the East-Asia - northern-Europe route. Understanding the risk of NIS introduction by this emerging shipping category is needed for devising strategies for sustainable shipping. Here, we conducted a controlled simulation of key abiotic factors that determine marine bioinvasion success: temperature, salinity, and food availability along selected routes, under two treatments: ULCV and intermediate-size vessels. We tested the effect of each treatment and the varying environmental conditions on the survival of two invasive ascidians (Chordata, Ascidiacea). We used survival analysis methods to locate predictors of ascidian mortality; Environmental conditions at ports with high mortality were used to identify similar major ports on a global scale as potential abiotic barriers. The key factors in ascidian mortality varied between the two species, but for both species, the treatment and salinity were dominant predictors for survival. We identified Port Klang, Rotterdam, and Dammam as ports with high mortality and located several globally distributed major ports that present similar environmental conditions. Our results highlight the potential role of selected major ports as abiotic barriers to fouling organisms during ocean voyages. The tolerance of the tropical-origin Microcosmus exasperatus to the northern-Europe conditions, and of the temperate/sub-tropical origin Styela plicata, to high temperature conditions, point out the urgent need to modify international fouling regulations in view of global change. Further studies on the survival of fouling organisms during a cascade of changing environmental conditions will contribute to the advancement of science-based regulations to reduce the adverse effects of NIS.