A systematic checklist of fish and shellfishes of the Poonthura estuary, Kerala, India is provided including notes on their conservation status. This checklist includes 66 finfish and five shellfish, belonging to 17 orders, 35 families, and 60 genera. Carangiformes is the richest order (11 species, eight genera, and three families), representing 15.4% of the total fish diversity. Carangidae, is the most diverse family with nine representatives, contributing to 12.6% of the total fish diversity. Following the IUCN Red List Categories, of the total 69 species (excluding both exotic and transplanted fish species), 59 belong to the ‘Least Concern’, while one species Pampus argenteus is listed as ‘Vulnerable’, four are ‘Data Deficient’ (Megalops cyprinoides, Arius maculatus, Cynoglossus semifasciatus, and Epinephelus tauvina) and five are ‘Not Evaluated’ (Nuchequula blochii, Channa pseudomarulius, Penaeus indicus, P. monodon, and Scylla serrata). Around 94% of the recorded fish fauna have commercial value and contribute to subsistence fisheries throughout the year. Taxonomy and diversity of fish fauna of least studied or isolated estuarine ecosystems should be updated with proper documentation of their conservation status, in order to design and implement pragmatic management and conservation programs.