Avian schistosomes belonging to the genus Austrobilharzia (Digenea: Schistosomatidae) are among the causative agents of cercarial dermatitis in humans. In this paper, ribosomal and mitochondrial DNA sequences were used to study schistosome cercariae from Kuwait Bay that have been identified morphologically as Austrobilharzia sp. Sequence comparison of the ribosomal DNA (rDNA) 28S and 18S regions of the collected schistosome cercariae with corresponding sequences of other schistosomes in GenBank revealed high sequence similarity. This confirmed the morphological identification of schistosome cercariae from Kuwait Bay as belonging to the genus Austrobilharzia. The finding was further supported by the phylogenetic tree that was constructed based on the combined data set 18S-28S-mitochondrial cytochrome oxidase I (mtCO1) sequences in which Austrobilharzia sp. clustered with A. terrigalensis and A. variglandis. Sequence comparison of the Austrobilharzia sp. from Kuwait Bay with A. variglandis and A. terrigalensis based on mtCO1 showed a variation of 10% and 11%, respectively. Since the sequence variation in the mtCO1 was within the interspecific range among trematodes, it seems that the Austrobilharzia species from Kuwait Bay is different from the two species reported in GenBank, A. terrigalensis and A. variglandis.

Stictodora tridactyla is an intestinal fluke in the family Heterophyidae that parasitizes shorebirds and mammals, including humans. Its metacercarial cyst stage was reported in the Arabian killifish, Aphanius dispar, at Kuwait Bay. In the present study, Cerithidea cingulata was found to serve as the first intermediate host of S. tridactyla. In order to establish the snail-fish link in the life cycle of S. tridactyla, complete sequences of ribosomal DNA internal transcribed spacer region 1 and 2 (rDNA ITS1 and ITS2) and partial sequence of cytochrome oxidase subunit 1 were obtained for metacercarial cysts isolated from the fish A. dispar and rediae isolated from the snail C. cingulata. Sequence alignment demonstrated that these larval stages belong to the same heterophyid species, S. tridactyla. Phylogenetic analysis based on rDNA ITS1, ITS2, and mtCO1 confirmed the position of S. tridactyla within the Heterophyidae and found it to cluster with Haplorchis spp. The present study represents the first molecular study correlating the larval stages of S. tridactyla using rDNA ITS1, ITS2, and mtCO1 and examining the phylogenetic relationships of S. tridactyla with different heterophyid species.

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Microphallid trematodes are common parasites in marine snails and crustacean hosts at Kuwait Bay. The larval stages of two microphallids, Maritrema eroliae and Probolocoryphe uca, are difficult to differentiate morphologically. In this study, two PCR-based techniques were established for quick and accurate discrimination between the larval stages of the two microphallid species, employing restriction fragment length polymorphism (PCR-RFLP) and species-specific primers. Both techniques utilized nucleotide differences in the second internal transcribed region (ITS2) of the ribosomal DNA (rDNA) in the two species. For the PCR-RFLP technique, restriction enzyme AvaII was selected and it generated different restriction profiles among the two microphallids. In addition, species-specific primers were prepared for each microphallid species that amplified distinctive fragments. Both techniques showed that the larval stages of the two microphallid species can be identified accurately. However, direct PCR amplification using species-specific primers was more advantageous than the PCR-RFLP technique since it allowed rapid and specific discrimination between the two species. This technique provides a useful tool that can be used in future studies for the study of the distribution of microphallid species and their definitive hosts at different localities of Kuwait Bay.