Throughout this research, 230 Trichoptera specimens were collected in the area of Konavle area, the southernmost region of Croatia. Altogether 21 species, 15 genera and 11 families were identified. DNA barcoding covered 41 specimens represented with 15 species collected in the Konavle region. The highest number of species was recorded at two localities: the spring of the River Ljuta (13) and a spring in the village Vodovađa (8). The highest number of taxa (4) was recorded within the following two families: Hydroptilidae and Hydropsychidae. From a faunistic point of view, the most interesting species in Konavle region are: Agapetus cf. kampos Oláh (the first record for Croatia), Hydroptila martini Marshall (the first record for the Mediterranean part of Croatia), Oxyethira falcata Morton (the first record for Croatia), Tinodes andrasi Oláh (type locality of the species is River Ljuta), Diplectrona cf. afra McLachlan (the first record from the Mediterranean part of Croatia) and Micropterna wageneri Malicky (the second record for Croatia). The first DNA barcoding data for the species Agapetus cf. kampos Oláh and Tinodes andrasi Oláh were entered into the BOLD database. The area of Konavle represents one of the most interesting regions for the fauna of Trichoptera and can be considered as a ”hotspot” for caddisflies in Croatia. Thus, it is necessary to install high standards for their protection.