1.Molecular Markers for Identification of Stellantchasmus falcatus and a Phylogenic Study using the HAT-RAPD Method.
Chalobol WONGSAWAD ; Pheravut WONGSAWAD
The Korean Journal of Parasitology 2010;48(4):303-307
Stellantchasmus falcatus is a minute intestinal fluke in the family Heterophyidae. Metacercariae, the infective stage, were reported in a marine fish, mullet Liza subviridis, and a fresh water fish, Dermogenus pusillus, in Thailand. Adults were found in chicks, rats, cats, and humans. Morphological studies were done for comparing Stellantchasmus sp. worms found in 2 different fish hosts; their shapes and organ arrangements were very similar except for the prepharynx length. Therefore, the present study aimed to compare their DNA fingerprints using the HAT-RAPD method for both types of Stellantchasmus and several other related species. Ten arbitrarily selected primers (OPA-04, OPA-09, OPN-02, OPN-03, OPN-09, OPN-12, OPP-11, OPR-15, OPX-13, and OPAD-01) were used. It was found that OPA-09, OPN-03, and OPAD-01 were able to generate S. falcatus specific fragments in mullets which consisted of 200, 760, and 280 bp, respectively. In addition, the results of morphologic, DNA fingerprinting, and phylogenetic analyses strongly suggest that the fresh water and marine specimens of Stellantchamus may be different species.
Animals
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DNA Fingerprinting/*methods
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DNA Primers/genetics
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DNA, Helminth/genetics
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Fish Diseases/*parasitology
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Heterophyidae/anatomy & histology/*classification/*genetics/isolation & purification
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Phylogeny
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Thailand
2.Prevalence of Haplorchis taichui in Field-Collected Snails: A Molecular Approach.
Thapana CHONTANANARTH ; Chalobol WONGSAWAD
The Korean Journal of Parasitology 2010;48(4):343-346
The prevalence of the cercarial stage of an intestinal trematode, Haplorchis taichui, in thiarid snails (Gastropoda: Thiaridae) was investigated using light microscope and species-specific PCR procedures. A total of 988 snails were collected from Mae Taeng district, Chiang Mai province, northern Thailand, which comprised of 3 species; Melanoides tuberculata, Tarebia granifera, and Thiara scabra. The overall prevalence of pleurolophocercous cercariae was 21.7% as determined by the morphology. For genetic detection of H. taichui infection in snails, 2 primers Hapt_F (5'-GGCCAACGCAATCGTCATCC-3') and Hapt_R (5'-GCGTCGGGTTTCAGACATGG-3'), were used. The genomic DNA of H. taichui, which was used as a positive control, gave an amplification of the 256 bp fragment. The overall prevalence of H. taichui from specific PCR was 9.7%. The proportion of H. taichui among the pleurolophocercous cercariae in this study was 44.9%.
Animals
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DNA Primers/genetics
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DNA, Helminth/genetics
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Heterophyidae/*isolation & purification
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Microscopy/methods
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Parasitology/methods
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Polymerase Chain Reaction/methods
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Prevalence
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Snails/*parasitology
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Thailand
3.Complete Mitochondrial Genome of Haplorchis taichui and Comparative Analysis with Other Trematodes.
Dongmin LEE ; Seongjun CHOE ; Hansol PARK ; Hyeong Kyu JEON ; Jong Yil CHAI ; Woon Mok SOHN ; Tai Soon YONG ; Duk Young MIN ; Han Jong RIM ; Keeseon S. EOM
The Korean Journal of Parasitology 2013;51(6):719-726
Mitochondrial genomes have been extensively studied for phylogenetic purposes and to investigate intra- and interspecific genetic variations. In recent years, numerous groups have undertaken sequencing of platyhelminth mitochondrial genomes. Haplorchis taichui (family Heterophyidae) is a trematode that infects humans and animals mainly in Asia, including the Mekong River basin. We sequenced and determined the organization of the complete mitochondrial genome of H. taichui. The mitochondrial genome is 15,130 bp long, containing 12 protein-coding genes, 2 ribosomal RNAs (rRNAs, a small and a large subunit), and 22 transfer RNAs (tRNAs). Like other trematodes, it does not encode the atp8 gene. All genes are transcribed from the same strand. The ATG initiation codon is used for 9 protein-coding genes, and GTG for the remaining 3 (nad1, nad4, and nad5). The mitochondrial genome of H. taichui has a single long non-coding region between trnE and trnG. H. taichui has evolved as being more closely related to Opisthorchiidae than other trematode groups with maximal support in the phylogenetic analysis. Our results could provide a resource for the comparative mitochondrial genome analysis of trematodes, and may yield genetic markers for molecular epidemiological investigations into intestinal flukes.
Animals
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Asia
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Codon, Initiator
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DNA, Mitochondrial/chemistry/genetics
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Gene Order
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Genes, Helminth
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*Genome, Mitochondrial
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Heterophyidae/*genetics/isolation & purification
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Humans
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Molecular Sequence Data
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Sequence Analysis, DNA
4.Developmental and Phylogenetic Characteristics of Stellantchasmus falcatus (Trematoda: Heterophyidae) from Thailand.
Pralongyut SRIPALWIT ; Chalobol WONGSAWAD ; Thapana CHONTANANARTH ; Somboon ANUNTALABHOCHAI ; Pheravut WONGSAWAD ; Jong Yil CHAI
The Korean Journal of Parasitology 2015;53(2):201-207
This study aimed to investigate the infection status, worm development, and phylogenetic characteristics of the intestinal trematode, Stellantchasmus falcatus. The metacercariae of S. falcatus were detected only in the half-beak (Dermogenus pusillus) out of the 4 fish species examined. Their prevalence was 90.0%, and the intensity of infection was 919 metacercariae on average. Worms were recovered from 33 (97.1%) of 34 chicks that were experimentally infected with 200 S. falcatus metacercariae each, and the average recovery rate was 43.0%. The body size and inner organs of S. falcatus quickly increased in the experimental chicks over days 1-2 post-infection (PI). In addition, ITS2 sequence data of this parasite were analyzed to examine the phylogenetic relationships with other trematodes using the UPGMA method. The results indicated that the ITS2 sequence data recorded from trematodes in the family Heterophyidae appeared to be monophyletic. This study concluded that D. pusillus serves as a compatible second intermediate host of S. falcatus in Thailand and that S. falcatus can develop rapidly in the experimental chicks. Data collected from this study can help to close the gap in knowledge regarding the epidemiology, biology, and phylogenetic characteristics of S. falcatus in Thailand.
Animals
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Chickens
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Fish Diseases/*parasitology
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Fishes
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Heterophyidae/*classification/genetics/growth & development/*isolation & purification
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Metacercariae/classification/genetics/*growth & development/isolation & purification
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*Phylogeny
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Poultry Diseases/*parasitology
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Thailand
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Trematode Infections/parasitology/*veterinary
5.Human Infections with Liver and Minute Intestinal Flukes in Guangxi, China: Analysis by DNA Sequencing, Ultrasonography, and Immunoaffinity Chromatography.
Hyeong Kyu JEON ; Dongmin LEE ; Hansol PARK ; Duk Young MIN ; Han Jong RIM ; Hongman ZHANG ; Yichao YANG ; Xueming LI ; Keeseon S EOM
The Korean Journal of Parasitology 2012;50(4):391-394
The prevalence of liver and intestinal fluke infections was determined by surveying inhabitants of Hengxuan, Fusui, and Shanglin villages which were known to be endemic for liver flukes in Guangxi, China in May 2010. A total of 718 people were examined for helminth eggs by the Kato-Katz thick smear technique, ultrasonography, immunoaffinity chromatography, and DNA sequencing. The overall egg positive rate was found to be 59.6% (28.0-70.6%) that included mixed infections with liver and intestinal flukes. Cases showing higher than 20,000 eggs per gram of feces (EPG) were detected between 1.3% and 16.2%. Ultrasonographic findings exhibited overall 28.2% (72 of 255 cases) dilatation rate of the intrahepatic bile duct. Clonorchis sinensis infection was detected serologically in 88.3% (38 of 43 cases) among C. sinensis egg positive subjects by the immunoaffinity chromatography using a specific antigen for C. sinensis. For differential diagnosis of the liver and intestinal flukes, more precise PCR and nucleotide sequencing for copro-DNA were performed for 46 egg positive cases. Mixed infections with C. sinensis and Metagonimus yokogawai were detected in 8 of 46 egg positive cases, whereas 29 specimens were positive for Haplorchis taichui. Ultrasonographic findings and immunoaffinity chromatography results showed usefulness, even in a limited way, in figuring out of the liver fluke endemicity.
Animals
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China/epidemiology
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Chromatography, Affinity
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Clonorchiasis/*epidemiology/parasitology/ultrasonography
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Clonorchis sinensis/genetics/immunology/*isolation & purification
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Coinfection
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DNA, Helminth/chemistry/genetics
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Feces/parasitology
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Female
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Heterophyidae/genetics/*isolation & purification
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Humans
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Intestines/*parasitology
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Liver/*parasitology
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Parasite Egg Count
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Polymerase Chain Reaction
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Prevalence
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Sequence Analysis, DNA
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Trematode Infections/*epidemiology/parasitology/ultrasonography