| Summary: | This study aimed to assess the presence of thermotolerant <i>Campylobacter</i> spp., as one of the most important foodborne zoonotic pathogens, in three shellfish species: mussels (<i>Mytilus galloprovincialis</i>), oysters (<i>Ostrea edulis</i>) and queen scallops (<i>Aequipecten opercularis</i>). The samples were collected from nine locations in the Istrian aquatory, Croatia. Isolation of <i>Campylobacter</i> was done according to standard ISO method, and species were identified using multiplex PCR. Isolates identified as <i>C. jejuni</i> and <i>C. lari</i> were genotyped using multilocus sequence typing (MLST) to determine the potential source of contamination. Among 108 examined samples of bivalve molluscs, mussels dominated and were the only ones found positive for the presence of <i>Campylobacter</i> (25.6%). In total, 19 <i>C. lari</i> and 1 <i>C. jejuni</i> strains were isolated. <i>C. lari</i> isolates found in this study belong to 13 sequence types (STs), and 9 of them are newly described in this paper. Two out of the four previously described <i>C. lari</i> STs that were found in this study were previously found in human stool. The only <i>C. jejuni</i> isolate was found to be sequence type 1268, which belongs to ST-1275 clonal complex that is almost exclusively found in seabirds and can sporadically cause infection in humans. Regarding the obtained results, introducing surveillance of thermotolerant <i>Campylobacter</i> in shellfish in the Republic of Croatia is advised as an improvement for public health safety.
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