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Pramod Kumar Prasad - One of the best experts on this subject based on the ideXlab platform.
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© 2009 Biomedical Informatics Publishing Group
2016Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena T, Lalit Mohan GoswamiAbstract:Use of sequence motifs as barcodes and secondary structures of Internal Transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae
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Use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae).
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (
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use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 its2 rdna for identification of the indian liver fluke fasciola trematoda Fasciolidae
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (trematode species identification. Using the novel approach of molecular morphometrics that is based on ITS2 secondary structure homologies, phylogenetic relationships of the various isolates of fasciolid species have been discussed.
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PCR-based determination of internal transcribed spacer (ITS) regions of ribosomal DNA of giant intestinal fluke, Fasciolopsis buski (Lankester, 1857) Looss, 1899.
Parasitology research, 2007Co-Authors: Pramod Kumar Prasad, Veena Tandon, Anupam Chatterjee, S. BandyopadhyayAbstract:Fasciolopsis buski, the zoonotic intestinal flukes of pigs in South and Southeast Asia, is commonly prevalent in regions across Northeast India. Populations of the fluke collected from different parts of the region exhibit variations in morphology. The main objective of our study was to provide molecular characterization of the parasite so as to supplement morphological criteria, using ribosomal DNA cluster (rDNA), which is flanked by more conserved internal transcribed spacer (ITS) regions. We describe herein the ITS sequences of the parasite collected from swine hosts of Assam region. The ITS sequences of both egg and adult origins were found to be identical in length and composition. Phylogenetically, F. buski resembles closely the other members of family Fasciolidae, showing significant expectation value in the alignment. The results corroborate that the ITS sequences are not stage specific and are conserved through different stages of development of the fluke, and thus could be used as species markers.
Scott P Lawton - One of the best experts on this subject based on the ideXlab platform.
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Comparative mitogenomics of the zoonotic parasite Echinostoma revolutum resolves taxonomic relationships within the 'E. revolutum' species group and the Echinostomata (Platyhelminthes: Digenea).
Parasitology, 2020Co-Authors: Linh Thi Khanh Pham, Huong Thi Thanh Doan, Weerachai Saijuntha, R.p.v. Jayanthe Rajapakse, Scott P LawtonAbstract:The complete mitochondrial sequence of 17,030 bp was obtained from Echinostoma revolutum and characterized with those of previously reported members of the superfamily Echinostomatoidea, i.e. six echinostomatids, one echinochasmid, five fasciolids, one himasthlid, and two cyclocoelids. Relationship within suborders and between superfamilies, such as Echinostomata, Pronocephalata, Troglotremata, Opisthorchiata, and Xiphiditata, are also considered. It contained 12 protein-coding, two ribosomal RNA, 22 transfer RNA genes and a tandem repetitive consisting non-coding region (NCR). The gene order, one way-positive transcription, the absence of atp8 and the overlapped region by 40 bp between nad4L and nad4 genes were similar as in common trematodes. The NCR located between tRNAGlu (trnE) and cox3 contained 11 long (LRUs) and short repeat units (SRUs) (seven LRUs of 317 bp, four SRUs of 207 bp each), and an internal spacer sequence between LRU7 and SRU4 specifying high-level polymorphism. Special DHU-arm missing tRNAs for Serine were found for both tRNAS1(AGN) and tRNAS2(UCN). Echinostoma revolutum indicated the lowest divergence rate to E. miyagawai and the highest to Tracheophilus cymbius and Echinochasmus japonicus. The usage of ATG/GTG start and TAG/TAA stop codons, the AT composition bias, the negative AT-skewness, and the most for Phe/Leu/Val and the least for Arg/Asn/Asp codons were noted. Topology indicated the monophyletic position of E. revolutum to E. miyagawai. Monophyly of Echinostomatidae and Fasciolidae was clearly solved with respect to Echinochasmidae, Himasthlidae, and Cyclocoelidae which were rendered paraphyletic in the suborder Echinostomata.
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Molecular characterization of 'Fasciola jacksoni' from wild elephants ('Elephas maximus maximus') of Sri Lanka : a taxonomic evaluation
Parasitology, 2019Co-Authors: R.p.v.j. Rajapakse, Scott P Lawton, K.j.k. Karunathilake, B V P Perera, N T B NguyenAbstract:Fasciola jacksoni is a significant contributor to the health and mortality of Asian elephants, particularly those in Sri Lanka. Despite the impact of fascioliasis on elephant populations, it is a neglected veterinary disease with limited taxonomic understanding. Molecular characterization and phylogenetic analysis of F. jacksoni were carried out to evaluate its suggested basal position in the Fasciolidae. Adult worms were collected during post-mortem of elephants, and eggs were collected from living elephants in National parks across Sri Lanka. Using the mitochondrial genes nicotinamide dehydrogenase subunit 1 (nad1) and cytochrome oxidase subunit 1 (cox1), and a partial 28S ribosomal DNA (28S rDNA), DNA sequences were generated from the F. jacksoni adult and egg material. Maximum likelihood (ML) phylogenetic analyses did not resolve F. jacksoni to be basal to the Fasciolidae. Furthermore, the ML analyses showed that the genus Fasciola was not monophyletic and that F. jacksoni was a sister species to the deer liver fluke Fascioloides magna. A clear framework is required to determine the taxonomic status of F. jacksoni and this current study provides the first detailed application of molecular techniques from multiple hosts across Sri Lanka with the production of reference DNA sequences for this important parasite.
Anupam Chatterjee - One of the best experts on this subject based on the ideXlab platform.
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Use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae).
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (
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use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 its2 rdna for identification of the indian liver fluke fasciola trematoda Fasciolidae
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (trematode species identification. Using the novel approach of molecular morphometrics that is based on ITS2 secondary structure homologies, phylogenetic relationships of the various isolates of fasciolid species have been discussed.
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PCR-based determination of internal transcribed spacer (ITS) regions of ribosomal DNA of giant intestinal fluke, Fasciolopsis buski (Lankester, 1857) Looss, 1899.
Parasitology research, 2007Co-Authors: Pramod Kumar Prasad, Veena Tandon, Anupam Chatterjee, S. BandyopadhyayAbstract:Fasciolopsis buski, the zoonotic intestinal flukes of pigs in South and Southeast Asia, is commonly prevalent in regions across Northeast India. Populations of the fluke collected from different parts of the region exhibit variations in morphology. The main objective of our study was to provide molecular characterization of the parasite so as to supplement morphological criteria, using ribosomal DNA cluster (rDNA), which is flanked by more conserved internal transcribed spacer (ITS) regions. We describe herein the ITS sequences of the parasite collected from swine hosts of Assam region. The ITS sequences of both egg and adult origins were found to be identical in length and composition. Phylogenetically, F. buski resembles closely the other members of family Fasciolidae, showing significant expectation value in the alignment. The results corroborate that the ITS sequences are not stage specific and are conserved through different stages of development of the fluke, and thus could be used as species markers.
David Blair - One of the best experts on this subject based on the ideXlab platform.
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adaptive radiation of the flukes of the family Fasciolidae inferred from genome wide comparisons of key species
Molecular Biology and Evolution, 2020Co-Authors: Young Jun Choi, Santiago Fontenla, David Blair, Peter U Fischer, Alicia Costabile, Paul J Brindley, Jose F Tort, Miguel M Cabada, Makedonka MitrevaAbstract:Liver and intestinal flukes of the family Fasciolidae cause zoonotic food-borne infections that impact both agriculture and human health throughout the world. Their evolutionary history and the genetic basis underlying their phenotypic and ecological diversity are not well understood. To close that knowledge gap, we compared the whole genomes of Fasciola hepatica, Fasciola gigantica, and Fasciolopsis buski and determined that the split between Fasciolopsis and Fasciola took place similar to 90 Ma in the late Cretaceous period, and that between 65 and 50 Ma an intermediate host switch and a shift from intestinal to hepatic habitats occurred in the Fasciola lineage. The rapid climatic and ecological changes occurring during this period may have contributed to the adaptive radiation of these flukes. Expansion of cathepsins, fatty-acid-binding proteins, protein disulfide-isomerases, and molecular chaperones in the genus Fasciola highlights the significance of excretory-secretory proteins in these liver-dwelling flukes. Fasciola hepatica and Fasciola gigantica diverged similar to 5 Ma near the Miocene-Pliocene boundary that coincides with reduced faunal exchange between Africa and Eurasia. Severe decrease in the effective population size similar to 10ka in Fasciola is consistent with a founder effect associated with its recent global spread through ruminant domestication. G-protein-coupled receptors may have key roles in adaptation of physiology and behavior to new ecological niches. This study has provided novel insights about the genome evolution of these important pathogens, has generated genomic resources to enable development of improved interventions and diagnosis, and has laid a solid foundation for genomic epidemiology to trace drug resistance and to aid surveillance.
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Family Paragonimidae Dollfus, 1939.
2008Co-Authors: David BlairAbstract:[Extract] Adult paragonimids are large, fleshy trematodes occurring in pairs in capsules within the lungs (usually) of mammals. They differ from other members of the Gorgoderoidea Looss, 1899 by a suite of characters, including the absence of a cirrus-sac, extensive dendritic vitellarium and the habit of encysting in pairs in the lungs of mammals. Kerbert (1878) described as Distoma westermaniil Kerbert, 1878 worms from the lungs of a tiger that had died in Amsterdam Zoological Gardens. Braun (1899) made D. westermanii the type of his new genus Paragonimus Braun, 1899. Shortly afterwards, Looss (1899) placed it in his new genus Polysarcus Looss, 1899, which must fall as a synonym (and was in any case preoccupied). Parasitologists at that time (e.g. Stiles & Hassall, 1900) assigned Paragonimus to the Fasciolidae Railliet, 1895, whereas Odhner (1914) placed it in the Troglotrematidae Odhner, 1914. Dollfus (1939) erected the family Paragonimidae Dollfus, 1939 for it. However, some later authors (e.g. Dawes, 1946; Baer & Joyeux, 1961) continued to place it in the Troglotrematidae.
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Family Pachypsolidae Yamaguti, 1958
2008Co-Authors: David BlairAbstract:This publication does not have an abstract. The first paragraph of the Introduction is displayed as the abstract. Pachypsolidae Yamaguti, 1958 contains only the type-genus, within which are a few species found in the digestive tract of marine chelonians and freshwater crocodilians. It is characterized by the presence of four to six anteriorly or laterally directed diverticula arising from the anterior portions of the caeca. Rudolphi (1819) described Distoma irroratum Rudolphi, 1819 from a sea turtle,identified as Testudo midas, from the Adriatic Sea (see Blair & Limpus, 1982). Looss (1901b) placed the species within his new genus Pachypsolus Looss, 1901. Pratt (1902b) placed Pachypsolus as one of several 'related genera' close to members of his 'new' subfamily Plagiorchiinae Looss, 1901 in the family Fasciolidae Railliet, 1895. Many subsequent authors have accepted that Pachypsolus has plagiorchiid affinities, although there was not always agreement on which family and subfamily were most appropriate for the genus.
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Complete DNA sequence and gene organization of the mitochondrial genome of the liverfluke, Fasciola hepatica L. (Platyhelminthes; Trematoda).
Parasitology, 2001Co-Authors: David Blair, Donald P. McmanusAbstract:The complete nucleotide sequence of the mitochondrial (mt) DNA molecule of the liverfluke, Fasciola hepatica (phylum Platyhelminthes, class Trematoda, family Fasciolidae), was determined, It comprises 14462 bp, contains 12 protein-encoding, 2 ribosomal and 22 transfer RNA genes, and is the second complete flatworm (and the first trematode) mitochondrial sequence to be described in detail. All of the genes are transcribed from the same strand. Of the genes typically found in mitochondrial genomes of eumetazoans, only atp8 is absent. The nad4L and nad4 genes overlap by 40 nt. Most intergenic sequences are very short. Two larger non-coding regions are present. The longer one (817 nt) is located between trnG and cox3 and consists of 8 identical tandem repeats of 85 nt, rich in G and C, followed by 1 imperfect repeat. The shorter non-coding region (187 nt) exhibits no special features and is separated from the longer region by trnG. The gene arrangement resembles that of some other trematodes including the eastern Asian Schistosoma species (and cyclophyllidean cestode species) but it is strikingly different from that of the African schistosomes, represented by Schistosoma mansoni. The genetic code is as inferred previously for flatworms. Transfer RNA genes range in length from 58 to 70 nt, their products producing characteristic 'clover leaf' structures, except for tRNA(S-VON) and tRNA(S-AGN) lacking the DHU arm.
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comparison of the second internal transcribed spacer ribosomal dna from populations and species of Fasciolidae digenea
International Journal for Parasitology, 1993Co-Authors: R D Adlard, Stephen C Barker, David Blair, Thomas H CribbAbstract:The nucleotide sequence of the 3' end of the second internal transcribed spacer region (ITS2) was determined for four isolates of Fasciola hepatica, two isolates of Fasciola gigantica, one isolate of Fascioloides magna and one isolate of Fasciola sp. from Japan. Intraspecific sequence divergence was negligible. Sequence divergence between F. hepatica and F. gigantica was 2.8% and between Fasciola hepatica and Fascioloides magna was 13.2%. The sequence of Fasciola sp. from Japan matched closely that of F. gigantica. This study demonstrates variability in nucleotide sequence within the ITS2 region which allows discrimination between species of Fasciolidae.
Veena Tandon - One of the best experts on this subject based on the ideXlab platform.
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Venn diagram showing the number of homologs found in the Fasciola buski transcriptome compared to transcriptome/EST datasets of major trematode families of Opisthorchiidae (Opisthorchis viverrini and Clonorchis sinensis), Fasciolidae (F. hepatica and
2016Co-Authors: Devendra Kumar Biswal, Manish Debnath, Graciously Kharumnuid, Welfrank Thongnibah, Veena TandonAbstract:Venn diagram showing the number of homologs found in the Fasciola buski transcriptome compared to transcriptome/EST datasets of major trematode families of Opisthorchiidae (Opisthorchis viverrini and Clonorchis sinensis), Fasciolidae (F. hepatica and Fasciola gigantica) and Schistosomatidae (Schistosoma mansoni and Schistosoma japonicum).
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Use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae).
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (
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use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 its2 rdna for identification of the indian liver fluke fasciola trematoda Fasciolidae
Bioinformation, 2009Co-Authors: Pramod Kumar Prasad, Devendra Kumar Biswal, Veena Tandon, Lalit Mohan Goswami, Anupam ChatterjeeAbstract:Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (trematode species identification. Using the novel approach of molecular morphometrics that is based on ITS2 secondary structure homologies, phylogenetic relationships of the various isolates of fasciolid species have been discussed.
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PCR-based determination of internal transcribed spacer (ITS) regions of ribosomal DNA of giant intestinal fluke, Fasciolopsis buski (Lankester, 1857) Looss, 1899.
Parasitology research, 2007Co-Authors: Pramod Kumar Prasad, Veena Tandon, Anupam Chatterjee, S. BandyopadhyayAbstract:Fasciolopsis buski, the zoonotic intestinal flukes of pigs in South and Southeast Asia, is commonly prevalent in regions across Northeast India. Populations of the fluke collected from different parts of the region exhibit variations in morphology. The main objective of our study was to provide molecular characterization of the parasite so as to supplement morphological criteria, using ribosomal DNA cluster (rDNA), which is flanked by more conserved internal transcribed spacer (ITS) regions. We describe herein the ITS sequences of the parasite collected from swine hosts of Assam region. The ITS sequences of both egg and adult origins were found to be identical in length and composition. Phylogenetically, F. buski resembles closely the other members of family Fasciolidae, showing significant expectation value in the alignment. The results corroborate that the ITS sequences are not stage specific and are conserved through different stages of development of the fluke, and thus could be used as species markers.