Parasites, Hosts and Diseases

"Porntip Laummaunwai"

Brief Communication

Effect of Temperature on the Killing of Opisthorchis viverrini Eggs In Vitro: Parichart Boueroy, Kunyarat Duenngai, Chatanun Eamudomkarn, Panupan Sripan, Thidarut Boonmars, Benjamabhorn Pumhirunroj, Atchara Artchayasawat, Jiraporn Songsri, Kanpicha Chomphumee, Panaratana Rattanasuwan, Porntip Laummaunwai, Sukhonthip Khueangchiangkhwang, Sirintip Boonjaraspinyo; Korean J Parasitol 2019;57(1):49-53.
Published online February 26, 2019; DOI: https://doi.org/10.3347/kjp.2019.57.1.49

Abstract
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Contaminated liver fluke egg in the environment has led to the high prevalence of human opisthorchiasis associated with cholangiocarcinoma in Southeast Asia. To find the effective lessening methods of Opisthorchis viverrini eggs in the contaminated environment, we investigated the temperature conditions for killing of these trematode eggs in vitro. Numerous O. viverrini eggs were obtained in the proximal part of uteri of adult worms from experimental hamsters. Mature eggs with miracidium were allocated by experimental groups (2 control: positive and negative and 4 treatment: 50, 60, 70, and 80°C) with 0.85% saline, and treated by the experimental plan. Eggs in each experimental groups were observed under the confocal microscope after stain with Propidium Iodide (PI) to evaluate the effect of temperatures. Eggs in 70 and 80°C groups were all killed after over 10 min heated. Majority of eggs in 60°C (10, 15, and 30 min heated), 70 and 80°C (5 min heated) groups were inactivated. However in 50°C group, below half of eggs were to be killed in all time lapse (10, 15 and 30 min). In order to prevent O. viverrini infection and cholangiocarcinoma, direct treatment of sewage by heating at 70 or 80°C at least 10 min is essential. Therefore, treatment of O. viverrini eggs at a high temperature is a potential method for controlling egg contamination in sewage.

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Development of a Dielectric Heating System for Selective Thermal Targeting of Liver Fluke Regions in Cirrhinus microlepis
Supatinee Kornsing, Sirigiet Phunklang, Chanchai Thongsopa, Piyaporn Krachodnok, Nuchanart Santalunai, Samran Santalunai
Applied Sciences.2025; 15(10): 5466. CrossRef
Viability of Trichinella spiralis in traditional sour pork fermentation and its inactivation by microwave heating: Implications for zoonotic risk and food safety
Atchara Artchayasawat, Benjamabhorn Pumhirunroj, Sukhonthip Khueangchiangkhwang, Thidarut Boonmars, Parichart Boueroy, Porntip Laummaunwai, Panaratana Rattanasuwan
Veterinary World.2025; : 1660. CrossRef

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Original Article

An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis: Krongkaew Seesui, Kanokwan Imtawil, Phimphakon Chanetmahun, Porntip Laummaunwai, Thidarut Boonmars; Korean J Parasitol 2018;56(1):25-32.
Published online February 28, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.1.25

Abstract
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Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris?EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was 40 parasites/μl for P. falciparum and 35.2 parasites/μl for P. vivax, whereas for Sn-PCR the limit of detection was 1.6 parasites/μl for P. falciparum and 1.4 parasites/μl for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

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Roland Dieki, Elsa-Rush Eyang-Assengone, Patrice Makouloutou-Nzassi, Félicien Bangueboussa, Edouard Nsi Emvo, Jean Paul Akue, Ricardo Santos
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Brief Communication

Status and Risk Factors of Strongyloides stercoralis Infection in Rural Communities of Xayaburi Province, Lao PDR: Phasouk Senephansiri, Porntip Laummaunwai, Sakorn Laymanivong, Thidarut Boonmar; Korean J Parasitol 2017;55(5):569-573.
Published online October 31, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.5.569

Abstract
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The present study was performed to reveal the current status and risk factors of Strongyloides stercoralis infections in the villages of Kenethao district, Xayaburi Province, Lao PDR. Fecal specimens were collected and examined for S. stercoralis using Koga-agar plate culture technique. Among 516 individuals, the prevalence of S. stercoralis and hookworm infection was 44.2% and 17.1%, respectively. Co-infection was detected in 13.2% of the cases. The prevalence did not significantly differ between males and females (P=0.193). However, the prevalence of S. stercoralis infection increased significantly with age (P=0.041). Of the risk factors examined, both performing farming activities (P=0.001) and walking barefoot when going outside of the house (P=0.003) showed significant correlations with S. stercoralis infections. Our results suggest that S. stercoralis is highly endemic in this area. The National Helminth Control Program of Lao PDR should take actions to control S. stercoralis infection. In addition, provision of health education about the benefits of wearing shoes would be important for reducing infection in the study area. Moreover, the application of high-sensitivity diagnostic approaches is needed to obtain the true impact of S. stercoralis infections in all rural communities in order to provide surveillance activities in Lao PDR.

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Mary Y Yeh, Sanjana Aggarwal, Margaret Carrig, Ahad Azeem, Anny Nguyen, Shannon Devries, Chris Destache, Toan Nguyen, Manasa Velagapudi
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Cesar G. Berto, Christina M. Coyle, Lianna Friedman, Patricia F. Walker
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Toni Wandra, Dewi Masyithah Darlan, Hemma Yulfi, Ivan Elisabeth Purba, Marcello Otake Sato, Christine M Budke, Akira Ito
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The American Journal of Tropical Medicine and Hygiene.2020; 102(2): 339. CrossRef
Current status of Opisthorchis viverrini, minute intestinal fluke and Taenia spp. infections among inhabitants in the Kenethao district of northern Lao PDR
Phasouk Senephansiri, Thaksaporn Thongseesuksai, Phaviny Sithay, Sakhone Laymanivong, Thidarut Boonmar, Porntip Laummaunwai
Transactions of The Royal Society of Tropical Medicine and Hygiene.2020; 114(5): 397. CrossRef
Status of Strongyloidiasis in Japan, 2000–2017
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Somaphone Chankongsin, Rahel Wampfler, Marie-Therese Ruf, Peter Odermatt, Hanspeter Marti, Beatrice Nickel, Valy Keoluangkhot, Andreas Neumayr
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Tropical Medicine and Health.2018;[Epub] CrossRef

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Original Article

Animal Models for Echinostoma malayanum Infection: Worm Recovery and Some Pathology: Jiraporn Songsri, Ratchadawan Aukkanimart, Thidarut Boonmars, Panaratana Ratanasuwan, Porntip Laummaunwai, Pranee Sriraj, Panupan Sripan; Korean J Parasitol 2016;54(1):47-53.
Published online February 26, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.1.47

Abstract
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Echinostomes are intestinal trematodes that infect a wide range of vertebrate hosts, including humans, in their adult stage and also parasitize numerous invertebrate and cold-blooded vertebrate hosts in their larval stages. The purpose of this study was to compare Echinostoma malayanum parasite growth, including worm recovery, body size of adult worms, eggs per worm, eggs per gram of feces, and pathological changes in the small intestine of experimental animals. In this study, 6-8-week-old male hamsters, rats, mice, and gerbils were infected with echinostome metacercariae and then sacrificed at day 60 post-infection. The small intestine and feces of each infected animal were collected and then processed for analysis. The results showed that worm recovery, eggs per worm, and eggs per gram of feces from all infected hamsters were higher compared with infected rats and mice. However, in infected gerbils, no parasites were observed in the small intestine, and there were no parasite eggs in the feces. The volume of eggs per gram of feces and eggs per worm were related to parasite size. The results of histopathological changes in the small intestine of infected groups showed abnormal villi and goblet cells, as evidenced by short villi and an increase in the number and size of goblet cells compared with the normal control group.

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Immune pathogenesis in pigeons during experimental Prohemistomum vivax infection
Asmaa M. I. Abuzeid, Mahmoud M. Hefni, Yue Huang, Long He, Tingting Zhuang, Guoqing Li
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Molecular and phylogenetic analysis and risk assessment of a trematode parasite, Artyfechinostomum sufrartyfex Lane, 1915 with a new host record from India
Dimple Mandla, Neena Singla, Sukhmanpreet Kaur Brar, Lachhman Das Singla
Biologia.2022; 78(1): 119. CrossRef
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Woon-Mok Sohn, Tai-Soon Yong, Keeseon S. Eom, Muth Sinuon, Hoo-Gn Jeoung, Jong-Yil Chai
The Korean Journal of Parasitology.2017; 55(3): 341. CrossRef

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Brief Communications

Evaluation of IgG4 Subclass Antibody Detection by Peptide-Based ELISA for the Diagnosis of Human Paragonimiasis Heterotrema: Pewpan M. Intapan, Oranuch Sanpool, Penchom Janwan, Porntip Laummaunwai, Nimit Morakote, Yoon Kong, Wanchai Maleewong; Korean J Parasitol 2013;51(6):763-766.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.763

Abstract
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A synthetic peptide was prepared based on the antigenic region of Paragonimus westermani pre-procathepsin L, and its applicability for immunodiagnosis for human paragonimiasis (due to Paragonimus heterotremus) was tested using an ELISA to detect IgG4 antibodies in the sera of patients. Sera from other helminthiases, tuberculosis, and healthy volunteers were used as the references. This peptide-based assay system gave sensitivity, specificity, accuracy, and positive and negative predictive values of 100%, 94.6%, 96.2%, 100%, and 88.9%, respectively. Cross reactivity was frequently seen against the sera of fascioliasis (75%) and hookworm infections (50%). Since differential diagnosis between paragonimiasis and fascioliasis can be easily done by clinical presentation and fascioliasis serology, this cross reaction is not a serious problem. Sera from patients with other parasitoses (0-25%) rarely responded to this synthetic antigen. This synthetic peptide antigen seems to be useful for development of a standardized diagnostic system for paragonimiasis.

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An immunochromatographic test using whole blood for rapid diagnosis of human paragonimiasis and its diagnostic usefulness
Patcharaporn Boonroumkaew, Lakkhana Sadaow, Penchom Janwan, Rutchanee Rodpai, Oranuch Sanpool, Tongjit Thanchomnang, Hiroshi Yamasaki, Pewpan M. Intapan, Wanchai Maleewong
Food and Waterborne Parasitology.2024; 37: e00246. CrossRef
Development of point-of-care testing tool using immunochromatography for rapid diagnosis of human paragonimiasis
Lakkhana Sadaow, Oranuch Sanpool, Hiroshi Yamasaki, Wanchai Maleewong, Pewpan M. Intapan
Acta Tropica.2020; 203: 105325. CrossRef
Paragonimus and paragonimiasis in Asia: An update
Ayako Yoshida, Pham Ngoc Doanh, Haruhiko Maruyama
Acta Tropica.2019; 199: 105074. CrossRef
A Case of Pulmonary Paragonimiasis with Chronic Abdominal Pain and Erythematous Rash in a 6-year-old Girl
Ju Young Kim, Min Kyu Park, Yong Ju Lee, Sun Huh, Ky Young Cho
Pediatric Infection and Vaccine.2018; 25(1): 54. CrossRef
A tool for mass-screening of paragonimiasis: an enzyme-linked immunosorbent assay with urine samples
Xu Guang Qiu, Fukumi Nakamura-Uchiyama, Yukifumi Nawa, Makoto Itoh
Tropical Medicine and Health.2016;[Epub] CrossRef
Expression characteristics and specific antibody reactivity of diverse cathepsin F members of Paragonimus westermani
Chun-Seob Ahn, Byoung-Kuk Na, Dong-ll Chung, Jeong-Geun Kim, Jin-Taek Kim, Yoon Kong
Parasitology International.2015; 64(1): 37. CrossRef
North American paragonimiasis: epidemiology and diagnostic strategies
Peter U Fischer, Gary J Weil
Expert Review of Anti-infective Therapy.2015; 13(6): 779. CrossRef

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A Recombinant Matrix Metalloproteinase Protein from Gnathostoma spinigerum for Serodiagnosis of Neurognathostomiasis: Penchom Janwan, Pewpan M. Intapan, Hiroshi Yamasaki, Porntip Laummaunwai, Kittisak Sawanyawisuth, Chaisiri Wongkham, Chatchai Tayapiwatana, Amnat Kitkhuandee, Viraphong Lulitanond, Yukifumi Nawa, Wanchai Maleewong; Korean J Parasitol 2013;51(6):751-754.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.751

Abstract
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Neurognathostomiasis is a severe form of human gnathostomiasis which can lead to disease and death. Diagnosis of neurognathostomiasis is made presumptively by using clinical manifestations. Immunoblotting, which recognizes antigenic components of molecular mass 21 kDa and 24 kDa in larval extracts of Gnathostoma spinigerum (Gs 21/24), has high sensitivity and specificity for diagnosis of neurognathostomiasis. However, only very small amounts of the Gs 21/24 antigens can be prepared from parasites harvested from natural or experimental animals. To overcome this problem, we recently produced a recombinant matrix metalloproteinase (rMMP) protein from G. spinigerum. In this study, we evaluated this rMMP alongside the Gs 21/24 antigens for serodiagnosis of human neurognathostomiasis. We studied sera from 40 patients from Srinagarind Hospital, Khon Kaen University, Thailand, with clinical criteria consistent with those of neurognathostomiasis, and sera from 30 healthy control adults from Thailand. All sera were tested for specific IgG antibodies against both G. spinigerum crude larval extract and rMMP protein using immunoblot analysis. The sensitivity and specificity for both antigenic preparations were all 100%. These results show that G. spinigerum rMMP protein can be used as an alternative diagnostic antigen, in place of larval extract, for serodiagnosis of neurognathostomiasis.

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Development of Immunochromatographic Test Kit for Rapid Detection of Specific IgG4 Antibody in Whole-Blood Samples for Diagnosis of Human Gnathostomiasis
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Molecular Detection of Ancylostoma duodenale, Ancylostoma ceylanicum, and Necator americanus in Humans in Northeastern and Southern Thailand: Issarapong Phosuk, Pewpan M. Intapan, Tongjit Thanchomnang, Oranuch Sanpool, Penchom Janwan, Porntip Laummaunwai, Witthaya Aamnart, Nimit Morakote, Wanchai Maleewong; Korean J Parasitol 2013;51(6):747-749.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.747

Abstract
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The 2 principal species of hookworms infecting humans are Necator americanus and Ancylostoma duodenale. Case studies on zoonotic hookworm infections with Ancylostoma ceylanicum and/or Ancylostoma caninum are known mainly from Asian countries. Of these 2 zoonotic species, only A. ceylanicum can develop to adulthood in humans. In the present study, we report a molecular-based survey of human hookworm infections present in southern and northeastern Thailand. Thirty larval hookworm samples were obtained from fecal agar plate cultures of 10 patients in northeastren Thailand and 20 in southern Thailand. Partial ITS1, 5.8S, and ITS2 regions of the ribosomal DNA genes were amplified using PCR. The amplicons were sequenced, aligned, and compared with other hookworm sequences in GenBank database. The results showed that, in Thailand, N. americanus is more prevalent than Ancylostoma spp. and is found in both study areas. Sporadic cases of A. ceylanicum and A. duodenale infection were seen in northeastern Thailand.

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Original Articles

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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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

Abstract
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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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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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