Marine fungi communities play a crucial role in the recycling of nutrients, restoration of biological systems, and the overall functioning of ecosystems. While aquatic fungal communities do react to pollution, there is a significant lack of information regarding the changes in the fungal community's structure, caused by marine pollution. In this study, we aim to address this gap in knowledge by investigating the range and makeup of fungal species present in marine environments in a polluted bay in Tunisia, spanning a biodiversity hotspot (Monastir Bay). Sequence analysis of the internal transcribed spacer region from culturable mycobiome and physicochemical parameters were investigated at seven sites in the bay. A total of 32 fungal taxa were identified at the genus and/or species levels and were assigned to four major groups (Aspergillacae 37.5%, Dothiomyceta 21.87%, Sordariamyceta 28.12%, and Yeasts 12.5%) with a remarkable predominance of Aspergillus genus. Assessment of the Shannon-Wiener diversity index and the Simpson dominance index revealed that the highest species diversity index (0.84) was recorded at the Kheniss site. Our results suggest the existence of diverse fungal communities, can be considered a useful community model for further ecological and evolutionary study of fungi in the bay.