記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers Media SA  

Schistosomiasis remains to ha/ve a significant public health impact in the Philippines. The Kato-Katz (K-K) technique is the reference standard and most used technique for definitive diagnosis of intestinal schistosomiasis for control programs in endemic regions. However, this has a very low sensitivity when applied in areas of low endemicity and patients with light infection. Hence, this study determined the diagnostic performance of immunological, molecular, parasitological, and ultrasonographic tests in diagnosing intestinal schistosomiasis in endemic municipalities in the Philippines. We performed a community-based cross-sectional study to determine the positivity of schistosomiasis in Leyte, Philippines. The diagnostic performance of five different detection techniques: (1) three stool K-K with duplicate smears; (2) soluble egg antigen IgG ELISA; (3) urine point-of-care circulating cathodic antigen (POC-CCA) test; (4) detection of Schistosoma japonicum circulating DNA (SjcDNA) in serum and urine samples; (5) focused abdominal ultrasound (US), were also obtained in this study. Multiple stool examinations enhanced the sensitivity of K-K from 26.2% (95% CI [16.4, 38.8]) with single stool to 53.8% (95% CI [41.1, 66.1]) and 69.2% (95% CI [56.4, 80.0]) with two and three stools from consecutive days, respectively. Among the SjcDNA nucleic acid amplification test (NAAT)-based detection assays, loop-mediated isothermal amplification (LAMP) PCR using sera had the highest sensitivity at 92.3% (95% CI [82.2, 97.1]) with LAMP consistently identifying more positive cases in both serum and urine samples. This study showed that single stool K-K, which remains the only diagnostic test available in most endemic areas in the Philippines, had low sensitivity and failed to identify most patients with light infection. SjcDNA detection assay and POC-CCA urine test were more sensitive than stool microscopy in detecting schistosomiasis. On the other hand, US was less sensitive than the widely utilized K-K technique in diagnosing schistosomiasis. This study emphasizes the need to revisit the use of single stool K-K in the surveillance and case detection of schistosomiasis in endemic areas of the Philippines. The availability of advanced and more sensitive diagnostic tests will help better control, prevent, and eliminate schistosomiasis in the country.

DOI： 10.3389/fimmu.2022.899311

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担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

OBJECTIVE: This study examined the knowledge, attitudes, and practices (KAP) of an at-risk population living in Niigata prefecture regarding tick-borne diseases (TBDs) and preventive strategies. METHODS: A cross-sectional questionnaire-based study was conducted to assess the KAP of the community. RESULTS: In total, 186 responses were received. Among the respondents, 130 (69·9%) were men, and the mean age was 51.1 (14·3). Nine (4·8%) respondents reported having experienced tick bites. Of the respondents, 44 (23.7%) knew about both scrub typhus and severe fever with thrombocytopenia syndrome, while 156 (83·9%) and 71 (38·2%) recognized limiting skin exposure and use of insect repellents as preventive measures, respectively. The attitudes towards TBDs: being worried about tick bites (p = 0·018) and interested in preventing TBDs (p = 0·001), were significantly higher among women than men. About 75% of the respondents reported taking preventive measures against tick bites, and limiting skin exposure was the most frequently applied method (69·9%). Insect repellents were used by 58 (31·2%) respondents. Age (p = 0·049), being worried about tick bites (p = 0·046), and knowledge of ticks score (p = 0·024) were the significant independent predictors of practicing countermeasures. CONCLUSION: We identified gaps in knowledge and practices regarding TBDs. Public health interventions should be implemented to improve public awareness of TBDs.

DOI： 10.1371/journal.pone.0270411

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Gastrointestinal parasite infections in livestock and companion animals in Mongolia have not been investigated sufficiently. This study aimed to determine the prevalence of cyclophyllid cestodes (Taeniidae and Mesocestoididae) in Mongolian sheepdogs using copro-DNA analysis. Sheepdog fecal samples (n = 1242) were collected from five ecological zones (mountain taiga, forest-steppe, steppe, desert-steppe, and desert) and four geographical regions (Western, Khangai, Central, and Eastern) within 20 of the country's 21 provinces. Among the 1242 samples, 201 (16.2%) tested positive for mitochondrial cytochrome c oxidase subunit 1 (cox1) and ribosomal 12S rRNA encoding genes of cyclophyllid cestodes. Prevalence in the mountain taiga, forest-steppe, steppe, desert-steppe, and desert zones was 29.2%, 15.4%, 15.1%, 20.1%, and 9.1%, respectively. Prevalence in the Western, Khangai, Central, and Eastern regions was 19.9%, 18.6%, 12.1%, and 12.8%, respectively. Taenia hydatigena, Taenia multiceps, and two Mesocestoides species (Mesocestoides sp.1 and Mesocestoides sp.2) were identified. T. hydatigena was found in the samples from all 20 provinces (all five zones and four regions), while T. multiceps was detected in the samples from 19 provinces (all five zones and four regions). Mesocestoides sp.1 infection was detected in the samples from all zones (except desert) and regions; it was detected in 14 provinces. Mesocestoides sp.2 infection was detected in the samples from all zones (except mountain taiga) and regions and found in seven provinces. Cyclophyllidea infection in sheepdogs is highly prevalent across Mongolia, representing a zoonotic risk. Implementation of a surveillance program for sheepdogs and their owners, and the wild animals in all ecological zones should be considered. In addition, control measures, including public awareness campaigns, especially for sheepdog owners, and periodic deworming of sheepdogs are warranted.

DOI： 10.1016/j.vprsr.2021.100680

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.meegid.2021.104999

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

We performed tick surveys in all regions (Kaetsu, Chuetsu, Joetsu, and Sado) of the Niigata prefecture, Japan. A total of 105 field surveys were done from 2016 to 2018 in 41 sites, from north to south, in the prefecture. All 4806 ticks collected were identified and classified by species, sex, and developmental stage. Twelve species were recorded: Dermacentor taiwanensis, Haemaphysalis flava, Haemaphysalis hystricis, Haemaphysalis japonica, Haemaphysalis longicornis, Haemaphysalis megaspinosa, Ixodes ovatus, Ixodes nipponensis, Ixodes persulcatus, Ixodes monospinosus, Ixodes columnae, and Ixodes turdus. The major tick species in Niigata prefecture were H. flava, H. longicornis, and I. ovatus and they comprised 93.4% of all samples. These three species have one generation per year. Climatic and anthropogenic factors may be involved in the substantial change of the endemic species composition from a previous tick survey (1959) in the Niigata prefecture. These factors include increasing temperatures, introduction of new hosts such as the wild boar, highway construction, and a rural exodus facilitating animal migration and reproduction. Tick hosts suitable for the transmission of Japanese spotted fever, Lyme borreliosis, and SFTS occur in Niigata prefecture. Heightened awareness of these three tick-borne diseases is needed for preparation and disease prevention.

DOI： 10.1016/j.ttbdis.2021.101683

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

Japanese spotted fever, a tick-borne disease caused by Rickettsia japonica, was firstly described in southwestern Japan. There was a suspicion of Rickettsia japonica infected ticks reaching the non-endemic Niigata Prefecture after a confirmed case of Japanese spotted fever in July 2014. Therefore, from 2015 to 2017, 38 sites were surveyed and rickettsial pathogens were investigated in ticks from north to south of Niigata Prefecture including Sado island. A total of 3336 ticks were collected and identified revealing ticks of three genera and ten species: Dermacentor taiwanensis, Haemaphysalis flava, Haemaphysalis hystricis, Haemaphysalis longicornis, Haemaphysalis megaspinosa, Ixodes columnae, Ixodes monospinosus, Ixodes nipponensis, Ixodes ovatus, and Ixodes persulcatus. Investigation of rickettsial DNA showed no ticks infected by R. japonica. However, three species of spotted fever group rickettsiae (SFGR) were found in ticks, R. asiatica, R. helvetica, and R. monacensis, confirming Niigata Prefecture as a new endemic area to SFGR. These results highlight the need for public awareness of the occurrence of this tick-borne disease, which necessitates the establishment of public health initiatives to mitigate its spread.

DOI： 10.1038/s41598-021-81587-9

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

Toxocariasis is a zoonotic helminthiasis caused by the migrating larvae of Toxocara canis and T. cati, common roundworms of dogs and cats. Our previous study in Savannakhet Province of Lao PDR showed an infection rate of 44.1% of Toxocara spp. in dogs. Thus, we investigate if this previous high prevalence in the definitive hosts influenced the occurrence of human toxocariasis. For that we used a 38 kDa recombinant protein derived from T. canis larvae excretion secretion products in ELISA. Human serum samples were collected in the Lahanam area of Savannakhet province. The population attending the study in Lahanam village were aged from 7 to 59 years old (y/o) 65.9% (54/82) were male and 34.1% (28/82) were female. The total percentage of seropositivity to Toxocara sp. was 30.4% (25/82). Males were more likely to test positive for toxocariasis with a risk ratio of 2.70 (CI95 0.87-4.93). No significant differences between ages were seen. However, it was possible to observe an increase of optical density (OD) values in ELISA according to age. The awareness of the health system on the high prevalence of seropositivity to Toxocara sp. in Savannakhet can prevent irreversible consequences as permanent vision loss and seizures caused by this silent chronic disease revealed in the Lahanam area.

DOI： 10.1016/j.onehlt.2020.100191

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The current study evaluated the anthelmintic effect of different extracts of Leucaena leucocephala, Parkia platycephala, and Senna alata on trichostrongylid eggs and infective larvae and determined the potential active components of each plant. Dried and macerated plant material was concentrated using rotaevaporation to obtain the crude extract (CE), followed by solvent partitioning to obtain hexanic (HexE), acetatic (AcE), and butanolic (BuE) extracts used for phytochemical analysis and anthelmintic efficacy testing in vitro. All the crude and partitioned extracts tested showed inhibition activity in the hatching of trichostrongylid eggs. Larvicidal efficacy was observed at CE concentrations of 2.5, 5.0, and 7.5 mg/mL for P. platycephala and S. alata. However, L. leucocephala CE did not significantly reduce the number of living larvae in the tested concentrations. Chromatographic analysis revealed several active metabolites; gallic acid, ellagic acid, naringin, morin, and kaempferol on AcE of P. platycephala; gallic acid, rutin, and ellagic acid on BuE of P. platycephala; and gallic acid and naringin on BuE of L. leucocephala. The extracts of P. platycephala, L. leucocephala, and S. alata leaves showed egg hatching inhibition and larvicidal activity, probably produced by tannins and flavonoids, which may act alone or by synergism.

DOI： 10.3390/pathogens9120986

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

BACKGROUND: The perpetuation of schistosomiasis japonica in the Philippines depends to a major extent on the persistence of its intermediate host Oncomelania hupensis quadrasi, an amphibious snail. While the malacological survey remains the method of choice in determining the contamination of the environment as evidenced by snails infected with schistosome larval stages, an emerging technology known as environmental DNA (eDNA) detection provides an alternative method. Previous reports showed that O. hupensis quadrasi eDNA could be detected in water, but no reports have been made on its detection in soil. METHODS: This study, thus focused on the detection of O. hupensis quadrasi eDNA from soil samples collected from two selected schistosomiasis-endemic barangays in Gonzaga, Cagayan Valley using conventional and TaqMan-quantitative (qPCR) PCRs. RESULTS: The results show that qPCR could better detect O. hupensis quadrasi eDNA in soil than the conventional method. In determining the possible distribution range of the snail, basic edaphic factors were measured and correlated with the presence of eDNA. The eDNA detection probability increases as the pH, phosphorous, zinc, copper, and potassium content increases, possibly indicating the conditions in the environment that favor the presence of the snails. A map was generated to show the probable extent of the distribution of the snails away from the body of the freshwater. CONCLUSION: The information generated from this study could be used to determine snail habitats that could be possible hotspots of transmission and should, therefore, be targeted for snail control or be fenced off from human and animal contact or from the contamination of feces by being a dumping site for domestic wastes.

DOI： 10.3390/pathogens8040160

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Yearly, several reports of unknown boats and corpses brought by the Tsushima Current are found ashore Japanese coast. Niigata prefecture had the highest number of the drifting ashore corpses in Japan with 45.7% (16/35) in 2017. Corpses from North Korea, confirmed by documents and photos were autopsied and in 3/16 was possible to recover worms full of eggs, morphologically identified as ascarids. Further molecular analysis of ITS1, 5.8S rDNA and ITS2 sequences confirmed all specimens were Ascaris lumbricoides. The contamination level by Ascaris lumbricoides eggs in the coast, the health impact and consequences of the epidemiological bridging produced by this forced migration in public health should be investigated. Moreover, control of helminthiases might be a necessary task in North Korea.

DOI： 10.1016/j.parint.2019.01.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI} {AG  

Bovine cysticercosis, caused by Taenia saginata metacestodes, is the cause of significant economic losses to the meat production chain by condemnation and downgrading of infected carcasses. It is also a public health issue causing human taeniasis. This study evaluated the occurrence of bovine cysticercosis at the meat inspection procedures in slaughterhouses of south and north regions of the Tocantins State in Brazil. Specimens identified as cysts of T. saginata were collected and analyzed by molecular (PCR) and histopathological techniques. The cysts were collected from March to December of 2010 in slaughterhouses located in the cities of Alvorada (South) and Araguaína (North). The frequency of cystic lesions during the study was 0.033% (53/164,091) with 69.81% of calcified lesions and 30.9% of live cysts at meat inspection. From 14 samples submitted to molecular analysis, 28.57% (4/14) were positive for T. saginata. The histopathological analysis of the non-T. saginata samples showed lesions suggestive of granuloma and hydatid disease. The results indicated that the identification of the etiological agent is difficult by macroscopic inspection, emphasizing the need to associate specific diagnostic methods at meat inspection in abattoirs. In addition, species-specific PCR would be an effective tool for diagnosis, monitoring, and identifying cysticercosis, assisting the conventional tests.

DOI： 10.3390/pathogens8020066

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Leptospires, which cause the zoonotic disease leptospirosis, persist in soil and aqueous environments. Several factors, including rainfall, the presence of reservoir animals, and various abiotic and biotic components interact to influence leptospiral survival, persistence, and pathogenicity in the environment. However, how these factors modulate the risk of infection is poorly understood. Here we developed an approach using environmental DNA (eDNA) metabarcoding for detecting the microbiome, vertebrates, and pathogenic Leptospira in aquatic samples. Specifically, we combined 4 sets of primers to generate PCR products for high-throughput sequencing of multiple amplicons through next-generation sequencing. Using our method to analyze the eDNA of leptospirosis-endemic areas in northern Okinawa, Japan, we found that the microbiota in each river shifted over time. Operating taxonomic units corresponding to pathogenic L. alstonii, L. kmetyi, and L. interrogans were detected in association with 12 nonpathogenic bacterial species. In addition, the frequencies of 11 of these species correlated with the amount of rainfall. Furthermore, 10 vertebrate species, including Sus scrofa, Pteropus dasymallus, and Cynops ensicauda, showed high correlation with leptospiral eDNA detection. Our eDNA metabarcoding method is a powerful tool for understanding the environmental phase of Leptospira and predicting human infection risk.

DOI： 10.1038/s41598-019-42978-1

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The time is passing, and the worms are still a major struggle for local people in Asian countries, especially the less empowered and in a situation of social vulnerability. We are working in the field in Laos, Thailand, and the Philippines where the usual control programs based only on human treatment are partially effective. Areas with mass drug administration could diminish, but not eliminate STHs of endemic areas. The persistence of helminthic NTDs in the environment and animal hosts makes the eradication a very difficult task. Great changes in the landscapes of endemic areas, such as construction of dams, can change the fauna and the lifestyle of local people. Those changes can improve infrastructure, but it can also lead to social vulnerability. The challenge, then, is to conceive new and directed control programs for helminthiasis based on multi- and transdisciplinary approaches diminishing the health gap in a globalized world. In this short review, we summarize the actual scenario concerning the main helminths in Southeast Asia and how an environmental DNA approach and the use of GIS could contribute to surveillance and control programs.

DOI： 10.1016/j.parepi.2018.e00083

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

In recent years, the prevalence and infection intensity of Schistosoma japonicum in endemic areas of the Philippines have significantly decreased due to yearly population-based treatment strategies, yet transmission rates remain high and uninterrupted. An important indicator of active disease transmission is the presence of Schistosoma japonicum and its snail intermediate host Oncomelania hupensis quadrasi in freshwater habitats. In this study, we sought to apply a species-specific real-time PCR (qPCR) assay for the detection of S. japonicum and O. hupensis quadrasi in freshwater samples using environmental DNA approach that can complement the commonly utilized malacological survey in determining potential transmission foci in order to have a more effective snail surveillance strategy for schistosomiasis japonica in endemic areas. The newly developed assay was specific to S. japonicum and O. hupensis quadrasi with no amplification detected against non-target trematode Fasciola spp. and snails such as Lymnaea spp., Pomacea canaliculata, and Melanoides spp. that typically co-exist in the same environment. The assay effectiveness was determined using 19 environmental water samples collected from Northern Samar (N = 5 sites), Leyte (N = 11 sites) and Compostela Valley (N = 3 sites) and compared to malacological survey for determining O. hupensis quadrasi snail colonies and snail crushing to visualize S. japonicum cercariae. TaqMan qPCR targeting a short fragment of the cytochrome c oxidase subunit 1 (cox1) gene was positive for S. japonicum in 9 sites, for O. hupensis quadrasi in 9 sites, and for both S. japonicum and O. hupensis quadrasi in 5 sampling sites. Moreover, it was able to detect O. hupensis quadrasi in 3 out of 12 sites found negative and 6 out of 7 sites found positive through malacological survey, and in 4 of the 5 snail sites positive for snails with cercariae. Overall, this method can complement malacological surveys for monitoring of schistosomes in endemic areas of the Philippines, especially those with high risk of human infection.

DOI： 10.1371/journal.pone.0224617

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

OBJECTIVES: Schistosomiasis is an important disease in Madagascar, and several studies on the disease have focused on the occurrence of the parasite in humans. However, the range of the pathogen in the environment and its impact on human infection is difficult to predict. An environmental DNA (eDNA) detection system for Schistosoma mansoni was developed to improve schistosomiasis eco-epidemiology studies. METHODS: Primers and probes were designed and tested in experimental biotopes. The field study was conducted in Maevatanana District of Madagascar. Seven water sources with human use were sampled, with a total of 21 water samples collected. Snails were collected, and patients were examined by ultrasound to determine the occurrence of schistosomiasis in the study area. RESULTS: One water source with active transmission was identified through the detection of S. mansoni eDNA in the water and the intermediate host Biomphalaria pfeifferi collected from the same water source. People with clinical schistosomiasis were found in the area, reinforcing the findings. CONCLUSIONS: The application of eDNA in eco-epidemiology enables the determination of hot spots and safe spots in endemic areas, constituting an alternative ecological tool for follow-up and monitoring of control programs for schistosomiasis, and contributing information on water safety for improving the standard of living of the people in endemic areas.

DOI： 10.1016/j.ijid.2018.08.018

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記述言語：日本語   出版者・発行元：新潟大学医学部保健学科  

壮年期にある２型糖尿病女性患者の自己管理における負担感情と更年期症状との関連を明らかにすることを目的とした。対象者107名，平均年齢58.0歳に厚生労働省栄養摂取状況調査票に基づく質問のほか，糖尿病問題領域質問表（PAID），クッパーマン更年期障害指数安部変法（KKSI）による自記式アンケート調査を実施した。その結果，糖尿病に対する負担感情は更年期症状と弱い正の相関関係を示し，更年期症状の「ゆううつ」はやや強い正の相関関係を示した。更年期前・中・後の３群間で糖尿病に対する負担感情と更年期症状に有意差が認められ，更年期中群は更年期後群と比較し糖尿病に関する負担感情と更年期症状が有意に高かった。糖尿病に対する負担感情を高める影響要因として更年期症状とHbA1c値が推測された。

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その他リンク： http://hdl.handle.net/10191/49813

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Public Library of Science  

Most part of Southeast Asia is considered endemic for human-infecting Taenia tapeworms
Taenia solium, T. saginata, and T. asiatica. However, until now there was no report of the occurrence of human cases of T. asiatica in Lao PDR. This study, conducted in Savannakhet Province, Lao PDR, microscopically examined a total of 470 fecal samples by Kato Katz method and found 86% of people harboring at least one helminth. Hookworms were detected in 56% of the samples besides Opisthorchis like eggs (42%), Trichuris trichiura (27%), Ascaris spp. (14%), and Taenia spp. (4%) eggs. Serology for cysticercosis showed 6.8% positives with results varying from 3% to 14.3% in Ethnic School students and Kalouk Kao village respectively. Species-specific PCR targeting mitochondrial DNA (mtDNA) of 28 tapeworms, recovered from 16 patients, revealed T. solium (n = 2), T. saginata (n = 21), and T. asiatica (n = 5). Two patients were confirmed to be coinfected with T. saginata and T. asiatica, indicating the endemicity of the 3 human Taenia in Lao PDR. However, nucleotide sequencing of a nuclear DNA gene, DNA polymerase delta (pold) revealed that all the tapeworms identified as T. asiatica using mtDNA had T. saginata type allele at pold locus, demonstrating that they are not “pure T. asiatica” but the hybrid descendants between the two species, confirming the wide distribution of hybrids of T. saginata/ T. asiatica in Southeast Asia. The high prevalence of several helminthic NTDs in east Savannakhet area even with conventional control measures indicates the importance to establish wide and multifaceted health programs to sustainably improve the quality of life of the populations living in these communities.

DOI： 10.1371/journal.pntd.0006260

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Public Library of Science  

Background: Amphimeriasis is a fish-borne disease caused by the liver fluke Amphimerus spp. that has recently been reported as endemic in the tropical Pacific side of Ecuador with a high prevalence in humans and domestic animals. The diagnosis is based on the stool examination to identify parasite eggs, but it lacks sensitivity. Additionally, the morphology of the eggs may be confounded with other liver and intestinal flukes. No immunological or molecular methods have been developed to date. New diagnostic techniques for specific and sensitive detection of Amphimerus spp. DNA in clinical samples are needed. Methodology/Principal findings: A LAMP targeting a sequence of the Amphimerus sp. internal transcribed spacer 2 region was designed. Amphimerus sp. DNA was obtained from adult worms recovered from animals and used to optimize the molecular assays. Conventional PCR was performed using outer primers F3-B3 to verify the proper amplification of the Amphimerus sp. DNA target sequence. LAMP was optimized using different reaction mixtures and temperatures, and it was finally set up as LAMPhimerus. The specificity and sensitivity of both PCR and LAMP were evaluated. The detection limit was 1 pg of genomic DNA. Field testing was done using 44 human stool samples collected from localities where fluke is endemic. Twenty-five samples were microscopy positive for Amphimerus sp. eggs detection. In molecular testing, PCR F3-B3 was ineffective when DNA from fecal samples was used. When testing all human stool samples included in our study, the diagnostic parameters for the sensitivity and specificity were calculated for our LAMPhimerus assay, which were 76.67% and 80.77%, respectively. Conclusions/Significance: We have developed and evaluated, for the first time, a specific and sensitive LAMP assay for detecting Amphimerus sp. in human stool samples. The procedure has been named LAMPhimerus method and has the potential to be adapted for field diagnosis and disease surveillance in amphimeriasis-endemic areas. Future large-scale studies will assess the applicability of this novel LAMP assay.

DOI： 10.1371/journal.pntd.0005672

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WALTER DE GRUYTER GMBH  

Dogs have been bred since ancient times for companionship, hunting, protection, shepherding and other human activities. Some canine helminth parasites can cause significant clinical diseases in humans as Opisthorchis viverrini causing cholangiocarcinoma in Southeast Asian Countries. In this study, socio-cultural questionnaire, canine parasitological analysis, necropsy, parasite molecular confirmation and dog roaming data were evaluated in Savannakhet, Lao-PDR, a typical Mekong Basin area. Dog owners comprised 48.8% of the studied population, with 61.2% owning one dog, 25.1% 2 dogs, 8.5% 3 dogs and 1.8% owning more than 4 dogs. Data from GPS logger attached to dogs showed they walked from 1.4 to 13.3 km per day, covering an area of 3356.38m2 average, with a routine of accessing water sources. Thirteen zoonotic helminth species were observed. Causative agents of visceral and cutaneous larva migrans occurred in 44.1% and 70% of the samples respectively. Spirometra erinaceieuropaei was detected in 44.1% of samples. Importantly, O. viverrini was found in 8.8% of samples. Besides the known importance of dogs in the transmission of Ancylostoma spp., Toxocara spp. and S. erinaceieuropaei, the observed roaming pattern of dogs confirmed it as an important host perpetuating O. viverrini in endemic areas; their routine access to waterbodies may spread O. viverrini eggs in a favorable environment for the fluke development, facilitating the infection of fishes, and consequently infecting humans living in the same ecosystem. Therefore, parasitic NTDs control programs in humans should be done in parallel with parasite control in animals, especially dogs, in the Mekong River basin area.

DOI： 10.1515/ap-2017-0047

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Opisthorchiasis, which can lead to cholangiocarcinoma in cases of chronic infection, is a major public health problem in Southeast Asian countries. The trematode, Opisthorchis viverrini, is the causative agent of the disease. Accurate and rapid monitoring of O. viverrini is crucial for disease prevention and containment. Therefore, in this study we sought to develop a novel species -specific real-time PCR assay for detecting O. viverrini using environmental DNA (eDNA). The diagnostic sensitivity of the newly developed real-time PCR assay was similar to that of the traditional PCR assay for 50 fecal samples collected in Lao PDR (21 and 19 samples were positive by real-time PCR and traditional PCR, respectively). The efficacy of eDNA analysis and its applicability in the field were tested using a total of 94 environmental water samples collected from 44 sites in Savannakhet, Lao PDR during May and October 2015 and February 2016. O. viverrini eDNA was detected in five samples by real-time PCR, indicating the presence of the fluke in the area and the risk of infection for individuals consuming fish from these water sources. The application of eDNA analysis would facilitate the identification of O. viverrini endemic hotspots and contribute to the ecological control of opisthorchiasis, and this strategy can be applied to other eukaryotic water pathogens. (C)2017 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.actatropica.2017.01.008

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：KOREAN SOC PARASITOLOGY, SEOUL NATL UNIV COLL MEDI  

In the present study, we report on the occurrence of paramphistomes, Fischoederius cobboldi and Paramphistomum epiclitum, in Lao PDR with the basis of molecular data. Parasite materials were collected from bovines bred in Ban Lahanam area, Savannakhet Province, Lao PDR at Lahanam public market. Morphological observations indicated 2 different species of paramphistomes. The mitochondrial gene cox1 of the specimens was successfully amplified by PCR and DNA sequencing was carried out for diagnosis of 11 specimens. Pairwise alignment of cox1 sequences were performed and confirmed F. cobboldi and P. epiclitum infecting bovines in Laos. Although there were many limiting points, as the small number of worm samples, and the restricted access of the animal host materials, we confirmed for the first time that 2 species of paramphistomes, F. cobboldi and P. epiclitum, are distributed in Lao PDR. More studies are needed to confirm the paramphistome species present in Savannakhet and its hosts to clear the natural history of these parasites of ruminants in the region and measure the impact of this parasite infection in the life and health of the local people.

DOI： 10.3347/kjp.2016.54.4.543

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BioMed Central Ltd.  

Background: Infections with the food-borne trematode Opisthorchis viverinni are common in Southeast Asia. In Lao PDR alone, two million people are supposed to be infected. Opisthorchiasis may cause severe liver disease, eventually leading to cholangiocarcinoma. The objective of this study is to assess the eating habits, complaints, symptoms, signs and ultrasonographical findings in three different areas of Savannakhet Province. Methods: Study participants were recruited in Lahanam village in the flood-prone lowland of, Sonkhone district, Savannakhet Province (group A)
in Non Somboon village, a community located on a hilly plateau in the same district (group B)
and in staff of Savannakhet Province Hospital, Savannakhet town (group C). Eating habits, complaints and symptoms were recorded by standardized structured questionnaires. Participants were thoroughly examined clinically, and ultrasonography was performed. O. viverrini eggs were looked for in stool and in duodenal fluid. An array of biochemical and haematological parameters potentially related to liver disease was determined. Group A consisted of 45, group B of 31 and group C of 18 individuals. Results: Eating habits were similar in the three groups, except that participants from group C tended to consume less high-risk types of fish dishes and more frequently ate beef and pork. Average intensity of infection (eggs per gram of stool) was low, but significantly higher in group A than in group B and C (p &lt
0.001). Medical history and complaints were similar in the three groups. Ultrasonography did not reveal any bile duct pathology. The only pathological finding was a slight elevation of ASAT and gamma-GT in a few participants in groups A and B. Conclusions: The study shows that eating habits favouring the infection with O. viverrini are common in south Laos. Although the average intensity of infection was low, there was a significant difference between the groups, paralleling slightly different eating habits. Clinically, this corresponded to a paucity of liver disease-associated complaints and signs. The low intensity of infection probably explains why no alterations of bile ducts were detectable by ultrasonography.

DOI： 10.1186/s41182-016-0001-1

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記述言語：日本語   出版者・発行元：新潟県保健環境科学研究所  

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その他リンク： http://search.jamas.or.jp/link/ui/2018088550

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BIOMED CENTRAL LTD  

Background: Diagnostic techniques based on PCR for the detection of Plasmodium DNA can be highly sensitive and specific. The vast majority of these techniques rely, however, on the invasive sampling of blood from infected hosts. There is, currently, considerable interest in the possibility of using body fluids other than blood as sources of parasite DNA for PCR diagnosis.
Methods: Urine and faeces were obtained from a Plasmodium knowlesi infected-Japanese macaque (Macaca fuscata) over the course of an experimentally induced infection. P. knowlesi DNA (PkDNA) extracted from urine and faeces were monitored by nested PCR targeting the P. knowlesi specific cytochrome b (cytb) gene.
Results: Urinary PkDNA was detected on day 2, but was not amplified using DNA templates extracted from the samples on day 4, day 5 and day 6. Subsequently, urinary PkDNA was detected from day 7 until day 11, and from day 20 until day 30. PkDNA in faeces was detected from day 7 until day 11, and from day 20 until day 37. Moreover, real-time quantitative PCR showed a remarkable increase in the amount of urinary PkDNA following anti-malarial treatment. This might have been due to the release of a large amount of PkDNA from the degraded parasites as a result of the anti-malarial treatment, leading to excretion of PkDNA in the urine.
Conclusions: The cytb-PCR system using urine and faecal samples is of potential use in molecular epidemiological surveys of malaria. In particular, monkey faecal samples could be useful for the detection of zoonotic primate malaria in its natural hosts.

DOI： 10.1186/1475-2875-13-373

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記述言語：日本語   出版者・発行元：医歯薬出版  

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その他リンク： http://search.jamas.or.jp/link/ui/2014214572

掲載種別：研究論文（学術雑誌）   出版者・発行元：Cambridge University Press (CUP)  

Abstract

The liver fluke, Opisthorchis viverrini, and the minute intestinal fluke, Haplorchis taichui, are prevalent in many Asian countries. This study analysed the patterns of infections of O. viverrini and H. taichui in Lahanam and Thakhamlien villages (Savannakhet Province, Lao PDR), in two cross-sectional investigations. Out of a total of 207 human participants, post-anthelmintic treatment positivity rates for expelled worms were 170 (82.1%) for H. taichui and 65 (31.4%) for O. viverrini. Both these species co-exist in the study villages. When each parasite was analysed separately, H. taichui infections reached a plateau among people aged >20 years. Opisthorchis viverrini infection rates were highest in the age group 21–30 years, with decreasing infection rates after the age of 30. Our findings indicated that fish-borne trematode infections were more prevalent among adults. Fish, common intermediate hosts, were acquired in the study area for analysis. The examination of 35 species of fish as intermediate hosts found O. viverrini metacercariae in only six species, and these were found mostly during the month of November. Many farmers who live on the rice fields obtain their food from their immediate environment, including these intermediate-host fish, potentially putting them at greater risk of O. viverrini infection. By contrast, H. taichui metacercariae were found in three species of fish obtained from the market, meaning that anyone could consume them and become infected. If people who work in rice fields limit the species of fish they consume, or avoid consuming raw fish during the month of November, they may reduce their risk of O. viverrini infection.

DOI： 10.1017/s0022149x14000261

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BRAZILIAN COLL VETERINARY PARASITOLOGY  

Trichostrongylids infection has gained significant public health importance since Trichostrongylus spp. infections have been reported in humans in Lao PDR. In this study, gastrointestinal nematodes were identified and the intensity of infections was determined in goats and cattle, which are animals greatly used for meat production in Lahanam Village, Lao PDR. The total number of goats and bovines was 23 and 29, respectively, pertaining to 32 households surveyed in the area. Feacal samples were randomly collected from 14 goats and 11 bovines. Ninety three percent (13/14) of goats and 36% (3/11) of cattle were infected, with an average of 1,728 and 86 eggs per gram of faeces (EPG), respectively. Coproculture showed Trichostrongylus spp. (goats 16%; bovines 48%), Haemonchus spp. (goats 69%; bovines 37%), Cooperia spp. (bovines 8%) and Oesophagostomum spp. (goats 15%; bovines 6%). After performing the necropsy on an adult goat, Trichuris spp. was also found. We confirmed the presence of Oesophagostomum spp., H. contortus and T. colubriformis by morphology and DNA sequencing analysis of the ITS region of rDNA. Due to interactions between humans and goats in Lahanam Village and high EPG results, the diagnosis of species and the intensity of gastrointestinal nematode infection in these animals are important public-health issues. Other ruminant parasites, such as Oesophagostomum and Haemonchus, found in caprines and bovines, are reported to be causes of zoonosis and their presence in humans should be investigated in future field surveys in this area.

DOI： 10.1590/S1984-29612014011

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

There have been few studies on human trichostrongyliasis in Southeast Asia, information on its clinical manifestations is also sparse. Trichostrongyliasis occurs predominantly in areas where poor hygiene is common especially where human/animal feces are used as a fertilizer, thereby contaminating vegetables and stream water. The intimate coexistence of domestic animals and humans explains the prevalence of Trichostrongylus infection in such areas. The goal of the current study was to determine the prevalence of trichostrongyliasis among villagers in Thakamrien village, Sonkon district, Savannakhet province, Laos, and to investigate potential relationships between clinical features, laboratory data, and severity of infection. Of 272 villagers examined, 160 (58.8%) were determined positive for helminthic infections by fecal examination, and 59 (36.9%) of these were infected with Trichostrongylus. Only 58 cases were in the inclusion criteria of the study and then underwent further assessment, including a questionnaire on personal behaviors, physical examination, and laboratory tests. Villagers in the trichostrongyliasis group were more likely than the control group to have consumed fresh vegetables, not washed their hands before meals or after using the toilet, and to have had close contact with herbivorous animals (goats and cows). Similarly, villagers in the trichostrongyliasis group were more likely than the control group to have a history of loose feces, rash, or abdominal pain
however, no obvious clinical symptoms were observed during physical examination of the trichostrongyliasis patients. The degree of infection was determined by both fecal egg counts and quantification of adult worms after deworming. Laboratory data were evaluated for any relationship with severity of infection. No significant differences were found in laboratory values between the trichostrongyliasis and control groups, with most values being within normal limits
however, both groups had high eosinophil counts. This study demonstrated that the useful clinical characteristics of trichostrongyliasis patients include history of loose feces, rashes, and abdominal pain, as well as in personal behaviors, such as the regular consumption of fresh vegetables, lack of hand washing, and close contact with cattle. © 2013 Elsevier B.V.

DOI： 10.1016/j.actatropica.2013.01.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOUTHEAST ASIAN MINISTERS EDUC ORGANIZATION  

Sixty-eight residents of Ban Luang and Ban Pang Kae villages, in Nan Province, northern Thailand, visited our mobile field station in September 2006 and March 2007, seeking treatment for taeniasis. After treatment, 22 cases discharged tapeworm strobila in their fecal samples and 17 scolices were recovered. Among these, 3 were morphologically abnormal, with six suckers on the scolex. To confirm the species of these tapeworms, the mitochondrial cytochrome c oxidase subunit I (COI) gene was used as a molecular marker. The partial COI sequences (800 bp) of the abnormal tapeworms were identical to the sequences of Taenia saginata deposited in Genbank.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC TROP MED & HYGIENE  

In Lahanam Village, Savannakhet Province, Laos, 125 of 253 villagers (49 w4%) were found by fecal examination to harbor hookworm eggs The eggs were heterogeneous in morphology and size, suggesting infections of mixed nematode species To confirm the hookworm egg species, on a voluntary basis, 46 hookworm egg-positive participants were treated with albendazole, and post-treatment adult worms were collected from purged fecal samples The common human hookworm was found in only 3 participants, 1 case of Necator americanus, and 2 cases of Ancylostoma duodenale In contrast, adult Trichostrongylus worms were expelled from most participants (43 of 46, 93 5%) The Trichostrongylus species were confirmed by morphology and internal transcribed spacer 2 sequences, all worms were of the same species (T colubriformis) In addition, some Trichostrongylus worms were obtained from a goat in the same village and identified as T colubriformis The results suggested that T colubriformis was the main zoonotic species causing hookworm infections in the village

DOI： 10.4269/ajtmh.2011.10-0385

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER IRELAND LTD  

A community-based field survey on taeniasis and cysticercosis was performed in two villages in Thong Pha Phum District, Kanchanaburi Province, central Thailand, where 3 Taenia species, T. solium, T. saginata and T. asiatica, are sympatrically occurring. Four (0.6%) out of 667 stool samples were egg-positive for Taenia sp. by Kato-Katz technique. Three out of those four persons and other three persons who were Taenia egg-negative but having a recent (<1 year) history of discharging worms in stool were treated with niclosamide. One Taenia egg-positive woman was not treated because of severe ascites. After treatment, three persons expelled long strobilae with scolices and two persons expelled strobilae without scolex. One Taenia egg-positive person did not expel any worms post-treatment. Among 5 persons, four expelled a single worm, whereas one expelled multiple worms, may be 6 worms but not confirmed by detection of scolices. One scolex was armed with hooklets, whereas 2 others did not. Multiplex PCR of 10 expelled proglottids (including 6 estimated worms from one patient) revealed that one sample was T. solium, one T. saginata, and 8 T. asiatica. A total of 159 residents agreed to receive a serological test for cysticercosis. By ELISA using partially purified glycoprotein antigen, 9 cases, 5 and 4 from villages A and B respectively, were found to be sero-positive. The five and an additional sample on the border line from village A were evaluated using confirmative immunoblot using recombinant chimeric antigen. Among the six samples, four including the border line sample were confirmed to be cysticercosis by immunoblotting. One of the 4 persons had neurological symptoms with nodular lesions in the brain by computed tomography. These 4 confirmed or suspected cysticercosis cases were free of T. solium worms, but two of them including confirmed NCC case had a past (>1 year) history of expelling proglottids in the stool. (c) 2010 Published by Elsevier Ireland Ltd.

DOI： 10.1016/j.parint.2010.03.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER IRELAND LTD  

Phylogenetic relationships of 6 species in the trematode subfamily Haplorchiinae were analyzed using small and large subunit of ribosomal DNA genes (185 rDNA and 28S rDNA) and internal transcribed spacer subunit II (ITS2) region as molecular markers. Maximum Likelihood and Bayesian inference analyses of combined rDNAs and ITS2 indicated a close relationship between the genera Haplorchis and Procerovum, while these two genera were distinct from Stellantchasmus falcatus. These phylogenetic relationships were consistent with the number of testes but not with the characters of the modification of the seminal vesicle or of the ventral sucker. Although three Haplorchis spp. were, together with Procerovum, in the same cluster, their mutual topology was incongruent between rDNA and ITS2 trees. Phylogenetic analyses using other molecular markers with more species are necessary to work out solid phylogenetic relationships among the species in this subfamily. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.parint.2010.06.009

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

The prevalence of hookworm eggs in fecal samples, by modified cellophane thick-smear technique, was found to be 30.0% (61/203 samples) in Lahanam Village, Savannakhet Province, Lao PDR. The hookworm eggs were morphologically heterogeneous, so that identification was confirmed by copro-PCR with specific primers for hookworms and Trichostrongylus spp.; 12 samples were positive for Necator americanus, 19 for Ancylostoma spp., with one mixed infection of both. Sequencing of the Ancylostoma spp. copro-PCR products found A. duodenale, and also the animal hookworms, A. caninum and A. ceylanicum. Moreover, Trichostrongylus spp.-specific copro-PCR revealed a significant rate of infection (43/203; 21.2%). Sequencing confirmed the zoonotic species, T. colubriformis. PCR/sequencing is useful for differentiating parasite species. The positives by the Kato-Katz method, and by the combined hookworm/Trichostrongylus specific-PCR, were comparable, although the agreement between the two methodologies was only 50%. (c) 2010 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.trstmh.2010.06.006

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER IRELAND LTD  

The liver fluke. Opisthorchis viverrini, causes serious public-health problems in the Lower Mekong Basin. This study aimed to clarify whether O. viverrini populations may be genetically divided into sub-specific taxa. We collected 6 populations of O. viverrini from different places in Cambodia, Lao PDR, and Thailand, along both sides of the Mekong River, and analyzed the population structure of these using the mitochondrial nad1 gene as a marker. The results of the DNA polymorphism measurements, by theta-w (theta w) and -pi (theta pi) values, neutrality tests, and mismatch distribution, suggested that the population of O. viverrini has expanded under the influence of purifying selection and selective sweep. The analysis of molecular variance (AMOVA) test revealed no significant genetic differences among the O. viverrini populations on opposite sides of the Mekong River. O. viverrini haplotypes occurred in multiple populations, and no distinct geographical clade. The star-like haplotype network confirmed a demographic expansion of the O. viverrini population. Overall, the genetic data from these populations suggested that the postulated existence of an O. viverrini species complex should be rejected. The bio-geographical diversity of O. viverrini populations should be explored further, using other appropriate markers and a wider range of samples from geographically different areas. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.parint.2010.02.009

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOUTHEAST ASIAN MINISTERS EDUC ORGANIZATION  

The utility of differential copro-DNA diagnosis using modified sample preparation steps of small liver and minute intestinal fluke infections was tested. Fecal samples containing parasite eggs were washed extensively with diluted detergent Solution. Parasite eggs were concentrated by sedimentation and broken by microwaving before DNA extraction. PCR targeting ITS1 and ITS2 regions were performed using primer specific for Opisthorchis viverrini, Haplorchis taichui and other related species. Of 125 fecal samples, 94 were positive for small trematode eggs by a modified cellophane thick smear method. By ITS1-PCR, 52 samples were positive for O. viverrini, 12 H. taichui and 7 mixed infection. By ITS2-PCR, 63 were positive for O. viverrini, 17 H. taichui, and 19 mixed infection. The ITS-PCR assay identified a higher number of opisthorchiasis cases than those with O. viverrini expelled after treatment, but for H. taichui, ITS-PCR identified less than half of the worm expelled cases. These results showed that copro-DNA diagnosis was useful for the differential diagnosis of O. viverrini and H. taichui infection, which could not be discriminated by microscopy.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：KOREAN SOC PARASITOLOGY, SEOUL NATL UNIV COLL MEDI  

Quantitative fecal egg counts represented as the number of eggs per gram of feces (EPG) are generally a reliable parameter to estimate the worm burden of intestinal and hepatic parasitoses. Although Haplorchis taichui ( Digenea: Heterophyidae) is one of the most common minute human intestinal flukes, little is known about the relationship between EPG and the actual worm burden in patients or the severity of the disease. In the present study, fecal samples were collected from 25 villagers in northern Thailand before and after praziquantel treatment. The EPG values of each participant were determined by the modified cellophane thick smear method, and adult worms were collected from the whole stool after the treatment. Eggs per day per worm (EPDPW) of H. taichui were estimated 82 from egg counts and expelled worms. The EPG was not well correlated with the worm burden, and a reverse correlation was observed between the EPDPW and the worm burden.

DOI： 10.3347/kjp.2009.47.3.315

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Small liver and minute intestinal flukes are highly prevalent in Southeast Asia, and in mixed infections, their eggs are difficult to differentiate morphologically in fecal samples. PCR assays targeting the ITS regions in ribosomal DNA were designed to identify and differentiate species. The PCR amplicons of Opisthorchis viverrini, Clonorchis sinensis, Haplorchis pumilio, and Haplorchis taichui were 800, 820, 1250, and 930 bp for the ITS1 region, and 380,390,380, and 530 bp for ITS2, respectively. The ITS1-region amplicon sizes successfully differentiated 4 species, while only H. taichui were significantly different from the other 3 species in the ITS2 region. PCR assays were employed for preliminary analysis using fecal samples diagnosed as having "small trematode eggs" by modified thick smear, showing 76.2% sensitivity for ITS1 and 95.2% for ITS2. (C) 2008 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.actatropica.2008.09.015

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掲載種別：研究論文（学術雑誌）  

In this study, adult patients were treated with praziquantel to expel intestinal flukes. Unexpectedly, dozens of adult Enterobius vermicularis worms with disfigured morphology, which had not been detected on fecal examination using Kat's modified thick-smear technique, were expelled from 6 of 33 patients.

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