First Report of Molecular Characterization of Oochoristica hemidactyli Johri, 1955 (Cestoda, Linstowiidae) from the Common Wall Lizard, Hemidactylus brooki Gray, 1845 (Reptilia: Gekkonidae) Along with Morphological Redescription.

INTRODUCTION: During an investigation, a rare cestode species belonging to the genus, Oochoristica were found in Meerut, UP, India, from the intestine of the host, Hemidactylus brookii Gray, 1845 (Reptilia: Gekkonidae). The tapeworms were found to be similar in their morphological characters with Oochoristica hemidactyli Johri, 1955 (Cestoda: Linstowiidae). MATERIALS AND METHODS: The host Hemidactylus brookii used in this analysis was caught from May to July 2018 from Department of Zoology, Chaudhary Charan Singh University, Meerut (UP), India. Parasites were examined by microscopy. 18S and ITS1-5.8S-ITS2 region of ribosomal RNA were sequenced and analyzed for the molecular study. RESULTS: This paper provides the first trustworthy confirmation about the occurrence of O. hemidactyli inferred with molecular data using ribosomal 18S and ITS1-5.8S-ITS2 gene cluster DNA sequences. Molecular analysis shows that the studied specimens were clustered separately and did not show a close relationship with any other cestode spp., represented in the GenBank. CONCLUSION: This is the foremost molecular characterization of any species of Oochoristica and of O. hemidactyli after 64 years of its original description that contributing to the systematic analysis of the helminthological biodiversity of reptilian hosts. These results provide a reference for future molecular study of species of Oochoristica.

First molecular evidence of Thelohanellus wallagoi Sarkar, 1985 (Myxozoa) from economically important food fish, freshwater shark Wallago attu (Siluridae) in India.

The freshwater shark Wallago attu (Bl. and Schn.) is a frequent silurid in the River Ganga and one of the most commercially exploit fish in India. In a survey on its infection with myxosporeans, spore type belongs to Thelohanellus species was found in the gills, kidney and intestine respectively. Through morphological and molecular investigations, we identified the spore as Thelohanellus wallagoi Sarkar, 1985. They were pyriform in valvular view and slim in sutural view, and had one pyriform polar capsule with four to five turns. The spores measured 8.0 × 4.0 × 2.3 µm. T. wallagoi developed in small cysts in the gill lamellae, whereas cyst and scattered spores of T. wallagoi were also found in the kidney and intestine respectively. The 18S rDNA sequence of T. wallagoi isolates recovered from gills, kidney and intestine were found similar to each other and differed from any other Thelohanellus species available in GenBank and validated its status after 32 years of original description. Phylogenetic analysis signified that T. wallagoi was placed sister to Myxobolus species in the clade that indicated the polyphyletic nature of the genus Thelohanellus.

Redescription of two species of Thaparocleidus (Monogenea: Dactylogyridae), with the description of T. armillatus sp. n. from Wallago attu and a phylogenetic analysis based on 18S rDNA sequences.

The phylogenetic studies on monogeneans of the genus Thaparocleidus parasitizing W. attu in India was inferred from 18S rDNA gene data. Out of species of Thaparocleidus, one new, T. armillatus sp. n., is described herein which distinguished from its congener species in the shapes and sizes of sclerotized structures and rest of the two species, T. indicus and T. wallagonius are redescribed morphologically as well as validated and confirmed by molecular means also. Besides the above, T. gangus was also included in the molecular analysis as it is recently described on the basis of morphology only. The phylogenetic tree was also reconstructed in order to elucidate the taxonomic status of Thaparocleidus armillatus sp. n., T. indicus, T. wallagonius and T. gangus. This research reports for the first time, the molecular phylogenetic analysis of 18S rDNA gene for four species belonging to the genus Thaparocleidus from W. attu collected off the River Ganga in India. In phylogram, all four species of Thapaocleidus come in a single clade that confirmed their monophyletic status. A very low interspecific variability was observed in T. armillatus sp. n. and T. wallagonius suggesting that these species are highly similar in morphology. Correspondingly, T. indicus and T. gangus also showed low interspecific variability which too signifies their morphological similarities in their sclerotized structures. Our result suggested that all four species taken in the present investigation from India are monophyletic and it is also observed that morphological similarities in the sclerotized structures are well correlated with phylogeny.

Morphological and Molecular Characterization of Metacercaria of Tylodelphys (Digenea: Diplostomidae) from the Piscine Host, Mystus tengara from India.

The diversity and taxonomy of diplostomids infecting freshwater fishes of the Indian region are predominantly poorly known. Yet compared to other trematodes, identification and discrimination of diplostomid metacercaria are difficult using morphology and site of infection. The present study provides the first molecular study of a Tylodelphys sp. from India. Specimens of Tylodelphys were collected from the vitreous humor of the eyes and from the brains of siluriform fish, Mystus tengara (Siluriformes: Bagridae) from Hastinapur, Meerut (U.P.), India. Taxonomic studies were performed on the basis of morphological, morphometrical, and molecular analysis. The dissimilarities in morphological and sites of infection between 2 forms of Tylodelphys, designated as Tylodelphys sp. 1 and Tylodelphys sp. 2, suggested that the forms were different species but, after performing molecular analysis, we conclude that despite morphological differences both morphotypes are conspecific.

First report on the molecular characterization of Diaphorocleidus armillatus Jogunoori et al. 2004 (Monogenea: Dactylogyridae) infecting the gills of introduced fish, Gymnocorymbus ternetzi in India.

During an investigation of the diversity of monogenean parasites in Meerut, U.P., India, the exotic freshwater fish Gymnocorymbus ternetzi was found infected with the gill dactylogyrid species of Diaphorocleidus Jogunoori et al. 2004. Using morphological study, the present monogeneans were found similar in morphology of anchors, bars and male copulatory organ with D. armillatus Jogunoori et al. 2004. This is the first genetic characterization of D. armillatus based on the analysis of 18S ribosomal RNA sequence (1800 bp) and associated with morphological redescription. Our study suggests that more molecular investigation is required on Diaphorocleidus species for deducing correct phylogenetic position of D. armillatus.

First Molecular Characterization of Aspiculuris tetraptera (Nematoda: Heteroxynematidae) from Mus musculus (Rodentia: Muridae) in India.

Mus musculus (Rodentia: Muridae) were infected by a nematode belonging to the order Oxyurida. According to external morphological characters it was found to be Aspiculuris tetraptera Schulz, 1924. Present communication deals with molecular characterization of A. tetraptera infecting intestine of Mus musculus in Meerut (U.P.), India. A partial sequence of 18S ribosomal and cytochrome c oxidase subunit 1 (Cox 1) mitochondrial DNA analyses were performed. Sequence corresponding to 18S rRNA and mt Cox 1 gene was identical to sequences reported for A. tetraptera on GenBank. These results confirm the taxonomic validation of A. tetraptera parasitizing Mus musculus. This is the first report of molecular study of A. tetraptera in India.

Molecular characterization of Gastrothylax crumenifer (Platyhelminthes: Gastrothylacidae) from goats in the western part of India by LSU of nuclear ribosomal DNA.

The rumen parasite, Gastrothylax crumenifer (Platyhelminthes: Gastrothylacidae), is a highly pathogenic trematode parasite of goat (Capra hircus). It sucks blood that causes acute disease like anemia, and severe economic losses occur due to morbidity and mortality of the ruminant infected by these worms. The study of these rumen paramphistomes, their infection, and public health importance remains unclear in India especially in the western part of state Uttar Pradesh (U.P.), Meerut, India, where the goat meat consumption is very high. This paper provides the molecular characterization of G. crumenifer recovered from the rumen of Capra hircus from Meerut, U.P., India by the partial sequence of 28S rDNA. Nucleotide sequence similarity searching on BLAST of 28S rDNA from parasites showed the highest identity with those of G. crumenifer from the same host Capra hircus. This is the first report of molecular identification of G. crumenifer from this part of India.

PCR-based molecular characterization, phylogenetic analysis and secondary structure of the 28S rDNA of Thaparocleidus wallagonius (Monogenea: Dactylogyridae) - the most primitive species of this genus from India.

Species of the monogenean genus Thaparocleidus are specific to freshwater siluriform fish. The infection caused by these gill parasites are a major health problem to fish. But, to focus the control strategies of these parasites, first it is important to establish an accurate discrimination by molecular methods. In the present study, phylogenetic and structural analysis of 28S region of ribosomal DNA of T. wallagonius species collected from fish Wallago attu from Meerut (U.P.), India, was carried out. In the first step, we amplified, sequenced 28S region of ribosomal DNA of T. wallagonius to establish the phylogenetic relationship with other species of this genus. T. wallagonius found on gill filaments of fish W. attu, is the most primitive parasite of this genus from India, was unequivocally discriminate from other species of the same genus in this study. A secondary-structure model of the large subunit rDNA was also predicted using a combined comparative and thermodynamic approach. Molecular morphometric and phylogenetic relationship of T. wallagonius are discussed in detailed that based on molecular analysis using bioinformatic tools.