Parasites, Hosts and Diseases

"Opisthorchis viverrini"

Brief Communication

Prevalence of Opisthorchis viverrini-Like Fluke Infection in Ducks in Binh Dinh Province, Central Vietnam: Thanh Thi Ha Dao, Emmanuel Nji Abatih, Thanh Thi Giang Nguyen, Ha Thi Lam Tran, Sarah Gabri?l, Suzanne Smit, Phap Ngoc Le, Pierre Dorny; Korean J Parasitol 2016;54(3):357-361.
Published online June 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.3.357

Abstract
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Following the first report of Opisthorchis viverrini infection in a domestic duck in Phu My District of Binh Dinh Province, Central Vietnam, many other cases were observed in the province. We determined the infection rate and intensity of O. viverrini infection in ducks in 4 districts of the province. A total of 178 ducks were randomly selected from 34 farms for examination of flukes in the liver and gall bladder. An infection rate of 34.3% (range 20.7-40.4% among districts) was found; the intensity of infection was 13.8 worms per infected duck (range 1-100). These findings show the role of ducks as a host for O. viverrini, duck genotype, which is sympatric with the human O. viverrini genotype in this province. It also stresses the need for investigations on the zoonotic potential and the life cycle of this parasite.

Citations

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Parasitic Helminth Infections and Intron Sequence Genotyping of Opisthorchis viverrini-like Eggs in Outdoor Domestic Cats and Dogs Across the Chi River Basin, Maha Sarakham Province, Thailand
Kotchaphon Vaisusuk, Wasupon Chatan, Warayutt Pilap, Tongjit Thanchomnang, Chavanut Jaroenchaiwattanachote, Paiboon Sithithaworn, Ross H. Andrews, Chairat Tantrawatpan, Weerachai Saijuntha
International Journal of Molecular Sciences.2025; 26(7): 3005. CrossRef
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Veterinary Quarterly.2024; 44(1): 1. CrossRef
Natural variables separate the endemic areas of Clonorchis sinensis and Opisthorchis viverrini along a continuous, straight zone in Southeast Asia
Jin-Xin Zheng, Hui-Hui Zhu, Shang Xia, Men‐Bao Qian, Hung Manh Nguyen, Banchob Sripa, Somphou Sayasone, Virak Khieu, Robert Bergquist, Xiao-Nong Zhou
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Journal of Parasitology.2022;[Epub] CrossRef
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Weerachai Saijuntha, Paiboon Sithithaworn, Trevor N. Petney, Ross H. Andrews
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Investigation of possible alternate animal reservoir hosts of Opisthorchis viverrini
Sirikachorn Tangkawattana, Piya Sereerak, Songkiad Upontain, Prasarn Tangkawattana, Banchob Sripa
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Animal Health Research Reviews.2021; 22(1): 1. CrossRef
Updated molecular phylogenetic data for Opisthorchis spp. (Trematoda: Opisthorchioidea) from ducks in Vietnam
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Parasites & Vectors.2017;[Epub] CrossRef

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Original Article

Infection Status of Zoonotic Trematode Metacercariae in Fishes from Vientiane Municipality and Champasak Province in Lao PDR: Keeseon S. Eom, Han-Sol Park, Dongmin Lee, Woon-Mok Sohn, Tai-Soon Yong, Jong-Yil Chai, Duk-Young Min, Han-Jong Rim, Bounnaloth Insisiengmay, Bounlay Phommasack; Korean J Parasitol 2015;53(4):447-453.
Published online August 25, 2015; DOI: https://doi.org/10.3347/kjp.2015.53.4.447

Abstract
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The infection status of fishborne zoonotic trematode (FZT) metacercariae was investigated in fishes from 2 localities of Lao PDR. Total 157 freshwater fishes (17 species) were collected in local markets of Vientiane Municipality and Champasak Province in December 2010 and July 2011, and each fish was examined by the artificial digestion method. Total 6 species of FZT metacercariae, i.e., Opisthorchis viverrini, Haplorchis taichui, Haplorchis yokogawai, Haplorchis pumilio, Centrocestus formosanus, and Procerovum varium, were detected in fishes from Vientiane Municipality. The metacercariae of O. viverrini were detected in 50 (49.5%) out of 101 fishes (6 species), and their average number was 154 per fish infected. The remaining 5 species of heterophyid metacercariae were detected in 36.8%, 65.8%, 9.4%, 23.9%, and 5.1% fishes examined, and their average densities were 12, 1,038, 4, 15, and 13 per infected fish, respectively. In fishes from Champasak Province, 3 species of FZT metacercariae, i.e., O. viverrini, H. taichui, and H. yokogawai, were detected. Only 2 O. viverrini metacercariae were found in only 1 Barbonymus schwanefeldi. The metacercariae of H. taichui and H. yokogawai were detected in 60.0% and 50.0% of fishes examined, and their average densities were 47 and 28 per fish infected. By the present study, it has been confirmed that several species of FZT metacercariae are prevalent in fishes from Vientiane Municipality, with P. varium being a new member of FZT in Lao PDR. In comparison, FZT metacercariae are less prevalent in fishes from Champasak Province.

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Acta Tropica.2022; 225: 106216. CrossRef
Similarities and differences among the Opisthorchiidae liver flukes: insights from Opisthorchis felineus
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Ei Ei Phyo Myint, Amornpun Sereemaspun, Joacim Rocklöv, Choosak Nithikathkul
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Jong-Yil Chai, Bong-Kwang Jung, Keon Hoon Lee, Jin-Youp Ryu, Hyeon-Seung Kim, Sung-Jong Hong, Thi Thi Htoon, Htay Htay Tin, Byoung-Kuk Na, Woon-Mok Sohn
The Korean Journal of Parasitology.2020; 58(6): 701. CrossRef
Infection status with zoonotic trematode metacercariae in fish from Lao PDR
Jong-Yil Chai, Soon-Hyung Lee, Han-Jong Rim, Woon-Mok Sohn, Bounlay Phommasack
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Ju Yeong Kim, Tai-Soon Yong, Han-Jong Rim, Jong-Yil Chai, Duk-Young Min, Keeseon S. Eom, Woon-Mok Sohn, Jae Hoon Lim, Dongil Choi, Sithat Insisiengmay, Bounlay Phommasack, Bounnaloth Insisiengmay
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Ha Thanh Thi Dao, Veronique Dermauw, Sarah Gabriël, Apiporn Suwannatrai, Smarn Tesana, Giang Thanh Thi Nguyen, Pierre Dorny
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Jian-Hua Qiu, Ying Zhang, Xiao-Xuan Zhang, Yuan Gao, Qi Li, Qiao-Cheng Chang, Chun-Ren Wang
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Jong-Yil Chai, Bong-Kwang Jung
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Atcharaphan Wanlop, Chalobol Wongsawad, Pongphol Prattapong, Pheravut Wongsawad, Thapana Chontananarth, Jong-Yil Chai
The Korean Journal of Parasitology.2017; 55(4): 445. CrossRef
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Jong-Yil Chai, Woon-Mok Sohn, Byoung-Kuk Na, Jong-Bok Park, Hoo-Gn Jeoung, Eui-Hyug Hoang, Thi Thi Htoon, Htay Htay Tin
The Korean Journal of Parasitology.2017; 55(6): 631. CrossRef

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Brief Communication

Chronic Opisthorchis viverrini Infection and Associated Hepatobiliary Disease Is Associated with Iron Loaded M2-like Macrophages: Moses T. Bility, Banchob Sripa; Korean J Parasitol 2014;52(6):695-699.
Published online December 23, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.6.695

Abstract
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Chronic Opisthorchis viverrini-induced hepatobiliary disease is associated with significant leukocyte infiltration, including activated macrophages; however, the polarization of infiltrating macrophages remains to be fully characterized. In this study, we characterized macrophage polarization and phenotype in chronic O. viverrini-induced hepatobiliary disease in humans and hamsters using gene expression and histochemical analysis. Chronic O. viverrini infection and associated hepatobiliary diseases were associated with iron loaded M2-like macrophages in both humans and hamsters. This study provides suggestive evidence that iron loaded M2-like macrophages promote hepatobiliary disease in chronic O. viverrini infection.

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Anna Kovner, Yaroslav Kapushchak, Oxana Zaparina, Dmitry Ponomarev, Maria Pakharukova
International Journal of Molecular Sciences.2024; 25(22): 12002. CrossRef
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Susel Loli Quinteros, Bronwyn O'Brien, Sheila Donnelly
Parasitology.2022; 149(10): 1364. CrossRef
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Anna V. Kovner, Alena A. Tarasenko, Oxana Zaparina, Olga V. Tikhonova, Maria Y. Pakharukova, Viatcheslav A. Mordvinov
Scientific Reports.2022;[Epub] CrossRef
The pathogenic potential of the combined action of chronic Opisthorchis felineus infection and repeated social defeat stress in C57BL/6 mice
Damira Avgustinovich, Anna Kovner, Elena Kashina, Natalia Shatskaya, Galina Vishnivetskaya, Natalia Bondar, Maria Lvova
International Journal for Parasitology.2021; 51(5): 353. CrossRef
High macrophage activities are associated with advanced periductal fibrosis in chronic Opisthorchis viverrini infection
Kanin Salao, Krongkarn Watakulsin, Eimorn Mairiang, Sutas Suttiprapa, Sirikachorn Tangkawattana, Steven W. Edwards, Banchob Sripa
Parasite Immunology.2019;[Epub] CrossRef
Chronic hepatitis C infection–induced liver fibrogenesis is associated with M2 macrophage activation
Moses T. Bility, Kouki Nio, Feng Li, David R. McGivern, Stanley M. Lemon, Eoin R. Feeney, Raymond T. Chung, Lishan Su
Scientific Reports.2016;[Epub] CrossRef

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Original Articles

Prevalence of Helminthic Infections among Inhabitants of Lao PDR: Keeseon S. Eom, Tai-Soon Yong, Woon-Mok Sohn, Jong-Yil Chai, Duk-Young Min, Han-Jong Rim, Hyeong-Kyu Jeon, Virasack Banouvong, Bounnaloth Insisiengmay, Bounlay Phommasack; Korean J Parasitol 2014;52(1):51-56.
Published online February 19, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.1.51

Abstract
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The prevalence of helminthic infections was surveyed on a total of 6,178 residents (males 2,549 and females 3,629) in 102 villages of 9 provinces in Lao PDR between 2007 and 2011 under the project of Korea-Laos Collaborative Project for Control of Foodborne Trematode Infections in Lao PDR. Fecal specimens were collected and examined by the Kato-Katz thick smear and Stoll's egg counting techniques. The overall liver/intestinal helminth egg positive rate was 71.9% with a single or mixed infections with Opisthorchis viverrini and minute intestinal flukes (Ov/MIF), Ascaris lumbricoides, hookworms, Trichuris trichiura, Trichostrongylus sp., echinostomes, Taenia spp., and others. Ov/MIF revealed the highest prevalence (55.6%) followed by hookworms (27.8%) and T. trichiura (6.5%). The endemic regions with the highest prevalence of Ov/MIF were Savannakhet, Khammouane, Vientiane (Nam Ngum), Champasak (Khong Island), and Saravane Province. High prevalences of A. lumbricoides (33.8%), hookworms (47.8%), and T. trichiura (32.6%) were observed in Phongsaly, Luang Prabang, and Vientiane (Nam Ngum) areas, respectively. The results of this study highlight helminth parasites of current public health significance in different areas of Lao PDR.

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Zoonotic Trematode Metacercariae in Fish from Phnom Penh and Pursat, Cambodia: Jong-Yil Chai, Woon-Mok Sohn, Byoung-Kuk Na, Tai-Soon Yong, Keeseon S. Eom, Cheong-Ha Yoon, Eui-Hyug Hoang, Hoo-Gn Jeoung, Duong Socheat; Korean J Parasitol 2014;52(1):35-40.
Published online February 19, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.1.35

Abstract
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A survey was performed to investigate the infection status of freshwater fish with zoonotic trematode metacercariae in Phnom Penh and Pursat Province, Cambodia. All collected fish with ice were transferred to our laboratory and examined using the artificial digestion method. In fish from Phnom Penh, 2 kinds of metacercariae (Opisthorchis viverrini and Haplorchis yokogawai) were detected. O. viverrini metacercariae were positive in 37 (50.0%) of 74 fish in 11 species (average no. metacercariae/fish, 18.6). H. yokogawai metacercariae were detected in 23 (57.5%) of 40 fish in 5 species (average no. metacercariae/fish, 21.0). In fish from Pursat Province, 5 kinds of metacercariae (O. viverrini, H. yokogawai, Haplorchis pumilio, Centrocestus formosanus, and Procerovum sp.) were detected; O. viverrini metacercariae (n=3) in 2 fish species (Henicorhynchus lineatus and Puntioplites falcifer), H. yokogawai metacercariae (n=51) in 1 species (P. falcifer), H. pumilio metacercariae (n=476) in 2 species (H. lineatus and Pristolepis fasciata), C. formosanus metacercariae (n=1) in 1 species (H. lineatus), and Procerovum sp. metacercariae (n=63) in 1 species (Anabas testudineus). From the above results, it has been confirmed that various freshwater fish play the role of a second intermediate host for zoonotic trematodes (O. viverrini, H. yokogawai, H. pumilio, C. formosanus, and Procerovum sp.) in Cambodia.

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A Cross-Sectional Study on Intestinal Parasitic Infections in Rural Communities, Northeast Thailand: Sirintip Boonjaraspinyo, Thidarut Boonmars, Butsara Kaewsamut, Nuttapon Ekobol, Porntip Laummaunwai, Ratchadawan Aukkanimart, Nadchanan Wonkchalee, Amornrat Juasook, Pranee Sriraj; Korean J Parasitol 2013;51(6):727-734.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.727

Abstract
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Despite the existence of effective anthelmintics, parasitic infections remain a major public health problem in Southeast Asia, including Thailand. In rural communities, continuing infection is often reinforced by dietary habits that have a strong cultural basis and by poor personal hygiene and sanitation. This study presents a survey of the prevalence of intestinal parasitic infections among the people in rural Thailand. The community-based cross-sectional study was conducted in villages in Khon Kaen Province, northeastern Thailand, from March to August 2013. A total of 253 stool samples from 102 males and 140 females, aged 2-80 years, were prepared using formalin-ethyl acetate concentration methods and examined using light microscopy. Ninety-four individuals (37.2%) were infected with 1 or more parasite species. Presence of parasitic infection was significantly correlated with gender (P=0.001); nearly half of males in this survey (49.0%) were infected. Older people had a higher prevalence than younger members of the population. The most common parasite found was Opisthorchis viverrini (26.9%), followed by Strongyloides stercoralis (9.5%), Taenia spp. (1.6%), echinostomes (0.4%), and hookworms (0.4%). The prevalence of intestinal protozoa was Blastocystis hominis 1.6%, Entamoeba histolytica 0.8%, Entamoeba coli 0.8%, Balantidium coli 0.4%, Iodamoeba b?tschlii 0.4%, and Sarcocystis hominis 0.4%. Co-infections of various helminths and protozoa were present in 15.9% of the people. The present results show that the prevalence of parasitic infections in this region is still high. Proactive education about dietary habits, personal hygiene, and sanitation should be provided to the people in this community to reduce the prevalence of intestinal parasite infections. Moreover, development of policies and programs to control parasites is needed.

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R. Rucksaken, O. Haonon, P. Pinlaor, C. Pairojkul, S. Roytrakul, P. Yongvanit, C. Selmi, S. Pinlaor
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Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-Cancer Drugs: Nattapong Puthdee, Kulthida Vaeteewoottacharn, Wunchana Seubwai, Orasa Wonkchalee, Worasak Kaewkong, Amornrat Juasook, Somchai Pinlaor, Chawalit Pairojkul, Chaisiri Wongkham, Seiji Okada, Thidarut Boonmars, Sopit Wongkham; Korean J Parasitol 2013;51(6):711-717.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.711

Abstract
PDF

Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo.

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A tumorigenic cell line derived from a hamster cholangiocarcinoma associated with Opisthorchis felineus liver fluke infection
Viatcheslav A. Mordvinov, Galina A. Minkova, Anna V. Kovner, Dmitriy V. Ponomarev, Maria N. Lvova, Oxana Zaparina, Svetlana A. Romanenko, Alexander G. Shilov, Maria Y. Pakharukova
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Mucin-producing hamster cholangiocarcinoma cell line, Ham-2, possesses the aggressive cancer phenotypes with liver and lung metastases
Piyanard Boonnate, Kulthida Vaeteewoottacharn, Ryusho Kariya, Sawako Fujikawa, Thidarut Boonmars, Somchai Pinlaor, Chawalit Pairojkul, Seiji Okada
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Anticancer effect of berberine based on experimental animal models of various cancers: a systematic review and meta-analysis
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Nanoencapsulated curcumin and praziquantel treatment reduces periductal fibrosis and attenuates bile canalicular abnormalities in Opisthorchis viverrini-infected hamsters
Lakhanawan Charoensuk, Porntip Pinlaor, Supason Wanichwecharungruang, Kitti Intuyod, Kulthida Vaeteewoottacharn, Apisit Chaidee, Puangrat Yongvanit, Chawalit Pairojkul, Natthakitta Suwannateep, Somchai Pinlaor
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Parichart Boueroy, Chariya Hahnvajanawong, Thidarut Boonmars, Sunitta Saensa-Ard, Natthinee Anantachoke, Kulthida Vaeteewoottacharn, Vichai Reutrakul
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Distribution and Abundance of Opisthorchis viverrini Metacercariae in Cyprinid Fish in Northeastern Thailand: Somchai Pinlaor, Sudarat Onsurathum, Thidarut Boonmars, Porntip Pinlaor, Nuttanan Hongsrichan, Apisit Chaidee, Ornuma Haonon, Wutipong Limviroj, Smarn Tesana, Sasithorn Kaewkes, Paiboon Sithithaworn; Korean J Parasitol 2013;51(6):703-710.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.703

Abstract
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To increase public health awareness for prevention of opisthorchiasis caused by eating raw freshwater fish, the distribution and abundance of Opisthorchis viverrini metacercariae (OV MC) was investigated in freshwater fish obtained from 20 provinces in northeastern Thailand between April 2011 and February 2012. A cross-sectional survey was conducted on 12,890 fish consisting of 13 species randomly caught from 26 rivers, 10 dams, and 38 ponds/lakes. Fish, were collected in each of the rainy and winter seasons from each province. Fish were identified, counted, weighed, and digested using pepsin-HCl. Samples were examined for OV MC by a sedimentation method, and metacercariae were identified under a stereomicroscope. OV MC were found in 6 species of fish; i.e., Cyclocheilichthys armatus, Puntius orphoides, Hampala dispar, Henicorhynchus siamensis, Osteochilus hasselti, and Puntioplites proctozysron from localities in 13 provinces. Among the sites where OV MC-infected fish were found, 70.0% were dams, 23.7% were ponds/lakes, and 7.7% were rivers. The mean intensity of OV MC ranged from 0.01 to 6.5 cysts per fish (or 1.3-287.5 cysts per kg of fish). A high mean intensity of OV MC per fish (>3 cysts) was found in 5 provinces: Amnat Charoen (6.5 cysts), Nakhon Phanom (4.3), Mukdahan (4.1), Khon Kaen, (3.5) and Si Sa Ket (3.4). In conclusion, OV MC are prevalent in natural cyprinid fish, with the infection rate varying according to fish species and habitats.

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Curcumin Prevents Bile Canalicular Alterations in the Liver of Hamsters Infected with Opisthorchis viverrini: Prapaporn Jattujan, Somchai Pinlaor, Lakhanawan Charoensuk, Channarong Arunyanart, Jariya Umka Welbat, Wunnee Chaijaroonkhanarak; Korean J Parasitol 2013;51(6):695-701.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.695

Abstract
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Opisthorchis viverrini infection causes inflammation and liver injury leading to periductal fibrosis. Little is known about the pathological alterations in bile canaliculi in opisthorchiasis. This study aimed to investigate bile canalicular alterations in O. viverrini-infected hamsters and to examine the chemopreventive effects of curcumin on such changes. Hamsters were infected with O. viverrini and one group of animals was fed with 1% dietary curcumin supplement. Animals were examined during the acute infection phase, days 21 and 30 post-infection (PI) and chronic infection phase (day 90 PI). Scanning electron microscopy revealed that in the infected group fed with a normal diet, bile canaliculi became slightly tortuous by 30 day PI and more tortuous at day 90 PI. Transmission electron microscopy showed a reduction in microvilli density of canaliculi starting at day 30 PI, with a marked loss of microvilli at day 90 PI. These ultrastructral changes were slightly seen at day 21 PI, which was similar to that found in infected animals fed with 1% curcumin-supplemented diet. Notably, curcumin treatment prevented the reduction of microvilli density, reduced the dilation of bile canaliculi, and decreased the tortuosity of the bile canaliculi relative to non-infected animals on a normal diet at days 30 and 90 PI. These results suggest that curcumin reduces alteration of bile canaliculi and may be a promising agent to prevent the onset of bile duct abnormalities induced by O. viverrini infection.

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Experimental Parasitology.2020; 209: 107810. CrossRef
Nanoencapsulated curcumin and praziquantel treatment reduces periductal fibrosis and attenuates bile canalicular abnormalities in Opisthorchis viverrini-infected hamsters
Lakhanawan Charoensuk, Porntip Pinlaor, Supason Wanichwecharungruang, Kitti Intuyod, Kulthida Vaeteewoottacharn, Apisit Chaidee, Puangrat Yongvanit, Chawalit Pairojkul, Natthakitta Suwannateep, Somchai Pinlaor
Nanomedicine: Nanotechnology, Biology and Medicine.2016; 12(1): 21. CrossRef
Bile canalicular changes and defective bile secretion in Opisthorchis viverrini-infected hamsters
Lakhanawan Charoensuk, Porntip Pinlaor, Umawadee Laothong, Puangrat Yongvanit, Chawalit Pairojkul, Yukifumi Nawa, Somchai Pinlaor
Folia Parasitologica.2014; 61(6): 512. CrossRef

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Molecular Differentiation of Opisthorchis viverrini and Clonorchis sinensis Eggs by Multiplex Real-Time PCR with High Resolution Melting Analysis: Worasak Kaewkong, Pewpan M. Intapan, Oranuch Sanpool, Penchom Janwan, Tongjit Thanchomnang, Porntip Laummaunwai, Viraphong Lulitanond, Pham Ngoc Doanh, Wanchai Maleewong; Korean J Parasitol 2013;51(6):689-694.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.689

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Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at 82.4±0.09℃ and 85.9±0.08℃ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

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Multiparasitism in Spain in a Korean Visiting Friends and Relatives: Case Report and Review of Imported Cases of Clonorchis sinensis in Europe
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Study of the population genetic structure of Opisthorchis-like eggs in northern Thailand using mitochondrial genes
Picha Suwannahitatorn, Mathirut Mungthin, Ittisak Subrungruang, Lakhanawan Charoensuk, Nithikoon Aksorn, Saiwasan Buathong, Krystyna Cwiklinski
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Histopathological Changes in Tissues of Bithynia siamensis goniomphalos Incubated in Crude Extracts of Camellia Seed and Mangosteen Pericarp: Ratchadawan Aukkanimart, Thidarut Boonmars, Somchai Pinlaor, Smarn Tesana, Surasit Aunpromma, Chantana Booyarat, Pranee Sriraj, Porntip Laummaunwai, Wiyada Punjaruk; Korean J Parasitol 2013;51(5):537-544.
Published online October 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.5.537

Abstract
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The present study was performed to observe histopathological changes in tissues of Bithynia siamensis goniomphalos (Gastropoda, Bithyniidae) incubated in crude extract solutions of camellia (Camellia oleifera) seed and mangosteen (Garcinia mangostana) pericarp, and furthermore to estimate the molluscicidal effects of 2 plant substances. Substantial numbers of bithyniid snails were incubated in various concentrations of 2 plant solution for 24 hr. As the positive control, snails incubated in various concentrations of niclosamide, a chemical molluscicide, were used. The histopathological findings were observed in sectioned snail specimens of each experimental and control groups. The results showed that both camellia and mangosteen extracts had molluscicidal effects at 24 hr with 50% lethal concentration (LC₅₀) at concentrations of 0.003 and 0.002 g/ml, respectively, while niclosamide had LC₅₀ at concentrations 0.599 ppm. B. siamensis goniomphalos snail tissues (foot, gill, and digestive system) showed disruption of columnar muscle fibers of the foot, reduction of the length and number of gill cilia, numerous mucous vacuoles, and irregularly shaped of epithelial cells. Irregular apical and calciferous cells, dilatation of the digestive gland tubule, and large hemolymphatic spaces, and irregular apical surfaces, detachment of cilia, and enlargement of lysosomal vacuoles of epidermis were also shown in all groups. By the present study, it is confirmed that 2 plants, camellia and mangosteen, are keeping some substance having molluscicidal effects, and histopathological findings obtained in this study will provide some clues in further studies on their action mechanisms to use them as natural molluscicides.

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Hyperendemicity of Haplorchis taichui Infection among Riparian People in Saravane and Champasak Province, Lao PDR: Jong-Yil Chai, Tai-Soon Yong, Keeseon S. Eom, Duk-Young Min, Hyeong-Kyu Jeon, Tae-Yun Kim, Bong-Kwang Jung, Lay Sisabath, Bounnaloth Insisiengmay, Bounlay Phommasack, Han-Jong Rim; Korean J Parasitol 2013;51(3):305-311.
Published online June 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.3.305

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In this study, we found that Haplorchis taichui, a heterophyid intestinal fluke, is highly prevalent, with heavy worm loads, among riparian people in Saravane and Champasak province, Lao PDR. Fecal specimens were collected from 1,460 people (717 men and 743 women) in 12 riparian (Mekong river) districts and were examined by the Kato-Katz fecal smear technique. The overall helminth egg positive rate was 78.8% and 66.4% in Saravane and Champasak province, respectively. The positive rate for small trematode eggs (STE), which included H. taichui and other heterophyids, Opisthorchis viverrini, and lecithodendriids, was 69.9% and 46.3% in Saravane and Champasak province, respectively. To obtain adult flukes, 30 STE-positive people were treated with 40 mg/kg praziquantel and then purged. Whole diarrheic stools were collected 4-5 times for each person and searched for fluke specimens using a stereomicroscope. Mixed infections with various species of trematodes (H. taichui, Haplorchis pumilio, O. viverrini, Prosthodendrium molenkampi, Centrocestus formosanus, and Echinochasmus japonicus) and a species of cestode (Taenia saginata) were found. However, the worm load was exceptionally high for H. taichui compared with other trematode species, with an average of 21,565 and 12,079 specimens per infected person in Saravane and Champasak province, respectively, followed by H. pumilio (41.9 and 22.5, respectively) and O. viverrini (9.4 and 1.5, respectively). These results show that diverse species of intestinal and liver flukes are prevalent among riparian people in Saravane and Champasak province, Lao PDR, with H. taichui being the exceptionally dominant species.

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Fishborne Trematode Metacercariae in Luang Prabang, Khammouane, and Saravane Province, Lao PDR: Han-Jong Rim, Woon-Mok Sohn, Tai-Soon Yong, Keeseon S. Eom, Jong-Yil Chai, Duk-Young Min, Soon-Hyung Lee, Eui-Hyug Hoang, Bounlay Phommasack, Sithat Insisiengmay; Korean J Parasitol 2013;51(1):107-114.
Published online February 18, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.1.107

Abstract
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Fishborne trematode (FBT) metacercariae were investigated in fish from 3 Provinces of Lao PDR. Total 242 freshwater fish of 40 species were collected in local markets of Luang Prabang (59 fish of 16 species), Khammouane (81 fish of 19 species), and Saravane (97 fish of 14 species), and each of them was examined by artificial digestion method. Four species of metacercariae (Opisthorchis viverrini, Haplorchis taichui, Haplorchis yokogawai, and Centrocestus formosanus) were detected. O. viverrini was detected in 35 fish (14.5%), and their density was 252 per infected fish (Luang Prabang, 88 metacercariae in 5 fish; Khammouane, 187 in 6 fish; Saravane, 303 in 24 fish). H. taichui was found in 102 fish (42.1%), and their density was 485 per infected fish (Luang Prabang, 260 metacercariae in 38 fish; Khammouane, 1,084 in 23 fish; Saravane, 359 in 41 fish). H. yokogawai was detected in 92 fish (38.0%), and their density was 222 per infected fish (Luang Prabang, 362 metacercariae in 17 fish; Khammouane, 126 in 20 fish; Saravane, 214 in 55 fish). Metacercariae of C. formosanus were found in 8 fish (3.3%), and their density was 3 per infected fish. In the present study, it has been confirmed that FBT metacercariae, in particular, H. taichui, H. yokogawai, and O. viverrini, are highly prevalent in fish from Luang Prabang, Khammouane, and Saravane Province, Lao PDR.

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Brief Communication

High Prevalence of Opisthorchis viverrini Infection in a Riparian Population in Takeo Province, Cambodia: Tai-Soon Yong, Eun-Hee Shin, Jong-Yil Chai, Woon-Mok Sohn, Keeseon S. Eom, Dong-Min Lee, Keunhee Park, Hoo-Gn Jeoung, Eui-Hyug Hoang, Yoon-Hee Lee, Hyun-Ju Woo, Ji-Hwa Lee, Sin-Il Kang, Jae-Ku Cha, Keon-Hoon Lee, Cheong-Ha Yoon, Muth Sinuon, Duong Socheat; Korean J Parasitol 2012;50(2):173-176.
Published online May 24, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.2.173

Abstract
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Opisthorchis viverrini infection was found to be highly prevalent in 3 riverside villages (Ang Svay Chek A, B, and C) of the Prey Kabas District, Takeo Province. This area is located in the southern part of Cambodia, where the recovery of adult O. viverrini worms was recently reported. From May 2006 until May 2010, fecal examinations were performed on a total of 1,799 villagers using the Kato-Katz thick smear technique. In the 3 villages, the overall positive rate for helminth eggs ranged from 51.7 to 59.0% (av. 57.4%), and the percentage positive for O. viverrini was 46.4-50.6% (47.5%). Other helminths detected included hookworms (13.2%), echinostomes (2.9%), Trichuris trichiura (1.3%), Ascaris lumbricoides (0.6%), and Taenia spp. (0.06%). The prevalence of O. viverrini eggs appeared to reflect a lower infection in younger individuals (<20 years) than in the adult population (>20 years). Men (50.4%) revealed a significantly higher (P=0.02) prevalence than women (44.3%). The Ang Svay Chek villages of the Prey Kabas District, Takeo Province, Cambodia have been confirmed to be a highly endemic area for human O. viverrini infection.

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Original Articles

High Prevalence of Haplorchis taichui, Phaneropsolus molenkampi, and Other Helminth Infections among People in Khammouane Province, Lao PDR: Jong-Yil Chai, Eun-Taek Han, Eun-Hee Shin, Woon-Mok Sohn, Tai-Soon Yong, Keeseon S. Eom, Duk-Young Min, Jin-Young Um, Min-Sung Park, Eui-Hyug Hoang, Bounlay Phommasack, Bounnaloth Insisiengmay, Soon-Hyung Lee, Han-Jong Rim; Korean J Parasitol 2009;47(3):243-247.
Published online August 28, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.3.243

Abstract
PDF

The prevalence of liver and intestinal helminth infections, including Opisthorchis, Haplorchis, Phaneropsolus, hookworms, Enterobius, and Taenia, was surveyed in Khammouane province, Lao PDR. Fecal specimens were collected from 1,242 people (590 men and 652 women) in 3 Mekong riverside villages and were examined by the Kato-Katz thick smear technique. The overall helminth egg positive rate was 81.1%. The positive rate for small trematode eggs, including Opisthorchis viverrini, heterophyids, and lecithodendriids, was 81.1% and the positive rate for hookworms was 6.7%. To obtain adult worms, 35 people who were positive for small trematode eggs were treated with 20-30 mg/kg praziquantel and 10-15 mg/kg pyrantel pamoate, and then purged. Diarrheic stools were collected from 33 of these people and searched for helminth parasites using a stereomicroscope. Mixed infections with various helminths (Haplorchis taichui, Haplorchis yokogawai, Prosthodendrium molenkampi, Phaneropsolus bonnei, echinostomes, hookworms, Trichostrongylus spp., Trichuris trichiura, Enterobius vermicularis, and/or Taenia saginata) were found. The total number of helminth specimens collected was 20,907 (approximately 634 per person). The most common species was H. taichui, followed by P. molenkampi, O. viverrini, P. bonnei, E. vermicularis, hookworms, and Trichostrongylus spp. These results show that diverse species of intestinal nematodes, trematodes, and cestodes are infecting humans in Khammouane province, Lao PDR.

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Fishborne Trematode Metacercariae Detected in Freshwater Fish from Vientiane Municipality and Savannakhet Province, Lao PDR: Han-Jong Rim, Woon-Mok Sohn, Tai-Soon Yong, Keeseon S. Eom, Jong-Yil Chai, Duk-Young Min, Soon-Hyung Lee, Eui-Hyug Hoang, Bounlay Phommasack, Sithat Insisengmay; Korean J Parasitol 2008;46(4):253-260.
Published online December 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.4.253

Abstract
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Freshwater fish from Vientiane Municipality and Savannakhet Province, Lao PDR were examined by the muscle compression and artificial digestion methods to know the infection status with trematode metacercariae. In the fish from Savannakhet, 2 species of metacercariae, Opisthorchis viverrini and Haplorchis taichui, were detected. O. viverrini metacercariae were found in 6 species of fish, Puntius brevis, Hampala dispar, Esomus metallicus, Mystacoleucus marginatus, Puntioplites falcifer, and Cyclocheilichthys armatus. H. taichui metacercariae were detected in 3 species of fish, P. brevis, P. falcifer, and M. marginatus. In the fish from Vientiane, 4 species of metacercariae, O. viverrini, H. taichui, Haplorchis yokogawai, and Centrocestus formosanus, were detected. Among them, O. viverrini metacercariae were found in 7 species of fish, Onychostoma elongatum, C. armatus, H. dispar, P. brevis, Cyclocheilichthys repasson, Osteochilus hasseltii, and Hypsibarbus lagleri. The metacercariae of H. taichui were detected in 6 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, and Cirrhinus molitorella. The metacercariae of H. yokogawai were found in 9 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, O. hasseltii. C. molitorella, Hypsibarbus wetmorei, and H. lagleri. The metacercariae of C. formosanus were detected in 4 species of fish, C. repasson, P. brevis, O. hasseltii, and C. molitorella. From these results, it is confirmed that fishborne trematode metacercariae, i.e. O. viverrini, H. taichui, H. yokogawai and C. formosanus, are prevalent in various species of freshwater fish from Savannakhet Province and Vientiane Municipality, Lao PDR.

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High prevalence of liver and intestinal fluke infections among residents of Savannakhet Province in Laos: Jong-Yil Chai, Eun-Taek Han, Sang-Mee Guk, Eun-Hee Shin, Woon-Mok Sohn, Tai-Soon Yong, Keeseon S. Eom, Keon-Hoon Lee, Hoo-Gn Jeong, Yong-Sang Ryang, Eui-Hyug Hoang, Bounlay Phommasack, Bounnaloth Insisiengmay, Soon-Hyung Lee, Han-Jong Rim; Korean J Parasitol 2007;45(3):213-218.
Published online September 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.3.213

Abstract
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The prevalence of liver and intestinal fluke infections was surveyed on residents of Savannakhet Province, Laos. Fecal specimens were collected from a total of 981 residents in 4 Mekong riverside villages and examined by the Kato-Katz thick smear technique. The results revealed that the overall helminth egg positive rate was 84.2%, and the positive rate for small trematode eggs, including Opisthorchis viverrini, heterophyids, or lecithodendriids, was 67.1%. To obtain adult flukes, 38 small trematode egg positive cases were treated with a 20-30 mg/kg single dose of praziquantel and purged. Diarrheic stools were then collected from 29 people and searched for helminth parasites using stereomicroscopes. Mixed infections with O. viverrini and 6 kinds of intestinal flukes were found, namely, Haplorchis taichui, Haplorchis pumilio, Haplorchis yokogawai, Prosthodendrium molenkampi, Phaneropsolus bonnei, and echinostomes. The total number of flukes collected was 7,693 specimens (av. no. per treated person; 265.3). The most common species was O. viverrini, followed by H. taichui, P. molenkampi, echinostomes, H. pumilio, P. bonnei, and H. yokogawai. The results indicate that foodborne liver and intestinal fluke infections are prevalent among residents of Savannakhet Province, Laos.

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Specific and common antigens of Clonorchis sinensis and Opisthorchis viverrini (Opisthorchidae, Trematoda): Min-Ho Choi, Jin-Sook Ryu, Mejeong Lee, Shunyu Li, Byung-Suk Chung, Jong-Yil Chai, Paiboon Sithithaworn, Smarn Tesana, Sung-Tae Hong; Korean J Parasitol 2003;41(3):155-163.
Published online September 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.3.155

Abstract
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The antigenic characterizations and serological reactions of human liver flukes, Clonorchis sinensis and Opisthorchis viverrini, were analyzed by immunoblot. The antigenic profiles of the crude extract of Clonorchis contained major proteins of 8, 26-28, 34-37, 43, and 70 kDa, and those of Opisthorchis 34-37, 43, 70, and 100 kDa. Of these, the 8, 26-28 and 34-37 kDa bands of Clonorchis and the 100 kDa of Opisthorchis were major components of each excretory-secretory antigen. The 8 and 26-28 kDa bands were specific to Clonorchis but the 100 kDa of Opisthorchis cross-reacted with the sera of clonorchiasis, and the 34-37, 70 and 100 kDa bands cross-reacted with sera of other helminthiases. The frequency and intensity of the immunoblot reactions were positively correlated with the intensity of the liver fluke infection.

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