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The Mugger Crocodile (Crocodylus palustris) is a threatened reptile inhabiting the Indian Sub-continent and Western Asia. Despite its "Vulnerable" conservation status, data about population genetic structure and connectivity are unavailable. This study makes a preliminary assessment of the genetic diversity, population structure and habitat connectivity of C. palustris in Iran. Ten tissue samples collected along the Sarbaz-Bahukalat basins were analysed and a set of 12 microsatellites was genotyped. Genetic diversity indices were estimated and population substructuring was assessed through Bayesian clustering analysis. Potential connectivity was verified through Remote Sensing water indexes, further implemented in a circuit analysis. Low genetic diversity was observed (mean observed heterozygosity = 0.35; mean expected heterozygosity = 0.43) and no population structure was found (K = 1). Water index and circuit analysis suggested possible connection among sites. This study highlights the potential vulnerability of crocodile populations and the importance of habitat connectivity for their persistence in the arid regions of Iran.