[Show abstract][Hide abstract] ABSTRACT: The transmission of zoonoses by wildlife, including elephants, is a growing global concern. In this study, we screened for helminth infections among Asian wild elephants (
) of the Salakpra Wildlife Sanctuary, Kanchanaburi, Thailand. Elephant faecal samples (45) were collected from the sanctuary grounds during January through November 2013 and assayed individually using the tetranucleotide microsatellite technique. Microscopic examination indicated a high prevalence of strongylids (93.0%) and low prevalences of trichurids (2.3%) and ascarids (2.3%). To identify the strongylid species, small subunit (SSU) rDNA sequences were amplified from copro-DNA and compared with sequences in GenBank. The generated SSU-rDNA sequences comprised five distinct haplotypes that were closely related to
. A phylogenetic analysis that incorporated related nematodes yielded a tree separated into two main clades, one containing our samples and human and domestic animal hookworms and the other consisting of
. The present results indicate that
in local elephants is a potential source of helminthiasis in human and domestic animals in this wild-elephant irrupted area.
Full-text · Article · Jul 2015 · Journal of Helminthology
[Show abstract][Hide abstract] ABSTRACT: A finding of antibodies to Gnathostoma spinigerum 24-kDa antigen by immunoblot analysis is currently used to confirm a diagnosis of gnathostomiasis. A simple skin test for the diagnosis of gnathostomiasis was developed, and the results were evaluated and compared with the standard Western blot (WB) test.
This cross-sectional study was conducted at the Hospital for Tropical Diseases, Bangkok, Thailand, in 2008-2011. All eligible patients were tested with partially purified proteins of mAb-detected fractions pooled and sterilized by 0.2 μm diameter syringe filter, with a phenol saline solution of 1:10 w/v.
A total of 69 cases, 39 gnathostomiasis cases and 30 controls, were enrolled into the study; the median age (IQR) was 40 (30.5-52.5) years. The most common presenting symptom was edema (56/69, 81%). Gnathostomiasis cases having strong cutaneous reactions to the intradermal test (81%) were also positive by immunoblot. A significant correlation between skin and immunoblot tests was detected (p<0.001). The difference in total IgE levels between cases and controls was not statistically significant (p=0.51). Logistic regression models showed that positive WB and skin-test results were significantly associated with gnathostomiasis (p=0.001 and p=0.007, respectively).
Gnathostoma skin testing, using prepared fractionated antigen solution of Gnathostoma spinigerum, yields good reactivity and significantly correlates with the results of immunoblot testing.
No preview · Article · Feb 2014 · Transactions of the Royal Society of Tropical Medicine and Hygiene
[Show abstract][Hide abstract] ABSTRACT: Helminths use various mechanisms to avoid host immunity and protect themselves from being eliminated. Despite evading host immune responses, immunosuppression and regulation mechanisms elicit functions that diminish the adverse effects of unrelated inflammatory diseases.
We investigated whether helminthic infections can ameliorate inflammatory diseases. Methods: Mice were infected with Trichinella papuae and then subjected to induced colitis through the oral administration of dextran sulfate sodium (DSS). Macroscopic and microscopic examinations measured weight loss, stool consistency, gross bleeding, colon length, and tissue inflammation. In addition, cytokine expression was observed in colon tissue by SYBR real-time RT-PCR to investigate the Th1, Th2, and regulatory cytokines.
The results showed that T. papuae infection decreased the severity of DSS-inducedcolitis, including weight loss, bloody diarrhea, shortening of colon, and colon tissue damage in mice (p <0.05). The expression level of IL-4 was high in the colons of DSS-treated mice without helminthic infection, while infected mice with DSS treatment had lower IL-4 levels (p <0.05). Uninfected DSS-treated mice failed to produce IL-10 mRNA in colon tissue, which may cause more severe colitis. In contrast, prior T. papuae infection DSS-treated mice had IL-10 levels in the colon significant lower than the normal and infected control groups.
Our data provide the evidence that prior T. papuae infection can ameliorate DSS-induced colitis in mice and may be considered for a novel therapeutic strategy against immunological diseases in the future.
No preview · Article · Jun 2013 · Asian Pacific journal of allergy and immunology / launched by the Allergy and Immunology Society of Thailand
[Show abstract][Hide abstract] ABSTRACT: Twelve 924 bp cytochrome c oxidase subunit 1 () mitochondrial DNA sequences from isolates from Thailand were aligned and compared with multiple sequence isolates from Thailand and 6 other countries from the GenBank database. The genetic divergence of was also compared with database sequences from 6 different countries in Asia, including Thailand, and 3 countries from other continents. The results showed that there were minor genetic variations within species, while high intraspecies variation was found in . There were only 2 haplotypes and 1 polymorphic site found in , but 8 haplotypes and 9 polymorphic sites in . Haplotype diversity was very low, 0.067, in and high, 0.700, in . The very low genetic diversity suggested that may be at a risk due to the loss of potential adaptive alleles, resulting in reduced viability and decreased responses to environmental changes, which may endanger the species.
Full-text · Article · Feb 2013 · The Korean Journal of Parasitology
[Show abstract][Hide abstract] ABSTRACT: 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 faeces 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 faecal 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 faeces, 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 faecal 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 faeces, 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.
[Show abstract][Hide abstract] ABSTRACT: Angiostrongylus cantonensis infection is the major cause of eosinophilic meningitis. Successful migration and evasion of the immune system by infective-stage larvae (L3) rely heavily on secreted proteases, which activate human pro-matrix metalloprotease (MMP-9) into active MMP-9. This study showed that the proteases in excretory-secretory (ES) products of A. cantonensis third stage larvae degraded recombinant and native human proMMP-9 in a dose- and time-dependent manner. Protease inhibitory assays showed that metalloproteases were the key enzymes involved in the degradation of human proMMP-9. To assess the effects of ES products on inflammation, ES products were incubated with THP-1 human monocytic cells, which showed induction of MMP-2 and not MMP-9 production. These results indicated that degradation of human MMP-9 was due to metalloproteases present in ES of A. cantonensis L3, which may be involved in suppressing the host's immune response to allow parasite migration to the host central nervous system.
No preview · Article · Sep 2012 · The Southeast Asian journal of tropical medicine and public health
[Show abstract][Hide abstract] ABSTRACT: Serine protease inhibitors, known as serpins, are mainly expressed in newborn and early-stage Trichinella spiralis larvae, suggesting that T. spiralis serpin (TsSERP) could be used as antigen for the immunodiagnosis of swine trichinosis. We produced His-tagged recombinant TsSERP (rTsSERP) in Escherichia coli and purified it using a Co(2+)-affinity column. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were performed to determine T. spiralis-infected swine sera samples (n = 5), negative controls (n = 26), and other parasite-infected samples (n = 83). WB showed that T. spiralis-infected sera initially reacted with rTsSERP at day 6 post-infection (dpi), and more strongly in late infection (62 and 84 dpi). However, other parasite-infected sera also elicited cross-reactivity to rTsSERP. On the other hand, indirect ELISA showed that TsSERP was an appropriate antigen for detecting late (> 60 dpi) but not early infection. No cross-reaction was observed with other parasite-infected sera. Sensitivity and specificity of TsSERP-ELISA at 62 dpi was 80% and 100%, respectively, and at 84 dpi 100% and 100%, respectively. These preliminary results show that TsSERP-ELISA method is suitable for the diagnosis of swine trichinosis, and could become the standard test for diagnosis of trichinosis in several hosts, including humans.
No preview · Article · Sep 2012 · The Southeast Asian journal of tropical medicine and public health
[Show abstract][Hide abstract] ABSTRACT: Angiostrongylus cantonensis is the causative agent of angiostrongyliasis, which is widely distributed throughout the world. It can specifically infect many species of intermediate and definitive hosts. This study examined the genetic differentiation and population structure using the RAPD-PCR method of parasites obtained from 8 different geographical areas of Thailand. Based on 8 primers, high levels of genetic diversity and low levels of gene flow among populations were found. Using genetic distance and neighbor-joining dendrogram methods, A. cantonensis in Thailand could be divided into two groups with statistically significant genetic differentiation of the two populations. However, genotypic variations and haplotype relationships need to be further elucidated using other markers.
Full-text · Article · May 2012 · The Southeast Asian journal of tropical medicine and public health
[Show abstract][Hide abstract] ABSTRACT: Metacercariae, morphologically similar to those of small liver flukes, were found to parasitize red-tailed snakehead fish, Channa limbata, collected from the city of Vientiane, Lao People's Democratic Republic. Adult worms that were recovered from experimentally-infected hamsters showed characteristics distinctly different from Opisthorchis viverrini, but closely similar to Opisthorchis lobatus, which was first reported in poultry (Anas sp.) from Pakistan. The present study aimed to redescribe O. lobatus based on the adult worms recovered from experimentally-infected hamsters. Additionally, it aimed to document the genetic relationships among O. lobatus and other opisthorchiid liver flukes using the mitochondrial cytochrome c oxidase subunit 1 (COI) gene and the internal transcribed spacer 2 (ITS2) region. DNA alignment of the O. lobatus and O. viverrini COI partial sequences (330 bp) showed 3.03% fixed differences (2.72% of amino acids changed) while the ITS2 region (350 bp) indicated a 0.86% difference for nucleotides. Species boundaries between the 2 parasites were determined by neighbor-joining analysis using the molecular sequence data. The phenogram confirmed that O. lobatus was distinctly different from O. viverrini, representing the first reported instance of O. lobatus in the Greater Mekong Sub-region (GMS) and the first record of C. limbata as the second intermediate host of a small liver fluke. Questions regarding human infection and the extent of the geographic distribution of these species should be investigated further.
Full-text · Article · Jun 2011 · Journal of Parasitology
[Show abstract][Hide abstract] ABSTRACT: In Lahanam Village, Savannakhet Province, Laos, 125 of 253 villagers (49.4%) 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.
Full-text · Article · Jan 2011 · The American journal of tropical medicine and hygiene
[Show abstract][Hide abstract] ABSTRACT: While human gnathostomiasis cases have been reported sporadically in Lao PDR since 1975, little is known about the disease in this country. We aimed to investigate sero-prevalence of gnathostomiasis and Gnathostoma species in Lao PDR. One village each in the north, central and south regions of Lao PDR was selected as the study sites. Overall, 125 (29.8%) of 420 sera from the randomly selected participants were sero-positive by immunoblot technique, with anti-Gnathostoma IgG antibody against the 24 kDa fraction. The sero-prevalence was high in the central (47.1%) and south (38.6%), but low (3.6%) in the north. Risk factor analyses revealed that the consumption of raw/undercooked fish was significantly associated with Gnathostoma sero-positivity (95% CI 1.05-17.05, P=0.042). The sero-positivity significantly increased with the age of the participants. Several fish, swamp eels, and frogs collected from central and southern Lao were infected with G. spinigerum advanced 3rd-stage larvae. Channa limbata (red-tailed snakehead fish) was identified as a natural second intermediate host of G. spinigerum. Eggs of G. spinigerum were found in dog feces collected in the south. Gnathostomiasis is endemic in central and southern Laos, so that preventive measures should be introduced for people living in these regions.
No preview · Article · Dec 2010 · Parasitology International
[Show abstract][Hide abstract] ABSTRACT: The aim of this study is to clarify the clinical features of Haplorchis taichui infection in humans in Nan Province, Thailand, and to correlate the clinical features with irritable bowel syndrome (IBS)-like symptoms. In this study area, only H. taichui, but neither other minute intestinal flukes nor small liver flukes were endemic. The degree of infection was determined by fecal egg counts and also by collecting adult worms after deworming. The signs and symptoms of individual patients together with their hematological and biochemical laboratory data were gathered to evaluate the relationship between the clinical features and the severity of infection. Special emphasis was made to elucidate the possible similarities of the clinical features of H. taichui infection and IBS-like symptoms. The results showed useful clinical information and the significant (> 50%) proportion of haplorchiasis patients complained of abdominal pain, lassitude, and flatulence, which were the important diagnostic symptoms of IBS. This study has reported a possible link between H. taichui and IBS, and H. taichui might probably play a role in the etiology of these IBS-like symptoms.
Full-text · Article · Sep 2010 · The Korean Journal of Parasitology
[Show abstract][Hide abstract] ABSTRACT: Ivermectin 2 mg/kg single dose reduced worm load by 83.3 % (75.0 – 91.7 %) for rats experimentally infected with 20 Gnathostoma spinigerum advanced third-stage larvae administered at week 1, with sacrifice at week 15 post-infection. Some larvae recovered from
the muscles of treated rats were found to be grossly deformed in appearance. Morphological changes in the Gnathostoma larvae were clearly observed. The internal structures of the deformed larvae showed significant degeneration in the esophageal
glandular tissues. The esophageal lumen branched irregularly and occupied a wider area in the center of the esophagus. The
epithelial cells thickened along these branches. The esophageal gland ducts were completely destroyed. The excretory ability
of the esophageal cell glands may be depressed, causing loss of function. It is suggested that ivermectin is effective against
Gnathostoma larvae due to worm recovery reductions. The drug efficacy appears to generate by esophageal destruction of the worms.
Key wordsEsophagus tissues-
[Show abstract][Hide abstract] ABSTRACT: A human trichinellosis outbreak caused by Trichinella papuae occurred in the Uthai Thani Province of Thailand in September 2007. A total of 34 villagers suffering at least one of the symptoms suggestive of trichinellosis, or those who were asymptomatic but had a history of ingesting raw wild pig meat, were enrolled in the study. Twenty-two villagers had ingested undercooked pork from a hunted wild pig (Sus scrofa). One patient with a severe clinical picture was hospitalised and more than 80 non-encapsulated larvae were detected in the muscle biopsy. The larvae were identified as T. papuae by molecular analyses of the mitochondrial cytochrome c oxidase subunit I (COI) gene and the expansion segment 5 (ES5) of the large subunit rRNA. Of the 34 suspected cases, 27 agreed to be subjected to haematological and serological tests. Immunoblot analysis using crude antigens from T. spiralis muscle larvae revealed anti-Trichinella IgG in 20 of the 26 serum samples (1 serum sample could not be analysed). All infected people were successfully treated with mebendazole; the one patient with severe symptomatology was treated successfully with prednisolone.
No preview · Article · Jun 2010 · Transactions of the Royal Society of Tropical Medicine and Hygiene
[Show abstract][Hide abstract] ABSTRACT: 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 (thetaw) and -pi (thetapi) 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.
Full-text · Article · Feb 2010 · Parasitology International
[Show abstract][Hide abstract] ABSTRACT: Aquatic snails, Pila ampullacea and Pomacea canaliculata were experimentally found to be suitable paratenic hosts for advanced third-stage larvae (L3) of the nematode Gnathostoma spinigerum, the causative parasite of gnathostomiasis in humans. G. spinigerum (L3) were found to be encapsulated in the tissue of the snail's foot and its internal organs. The infection, intensity and survival of third-stage larvae of G. spinigerum in both species of aquatic snails are described. This is the first evidence to reveal that not only vertebrates but also invertebrates (snails) can serve as paratenic hosts to this parasite. Aquatic snails are one of several sources of human gnathostomiasis in Thailand.
Full-text · Article · Apr 2009 · The Southeast Asian journal of tropical medicine and public health
[Show abstract][Hide abstract] ABSTRACT: A 54-year-old male Thai patient from Prachin Buri Province presented with a history of chronic watery diarrhea for many years. He passed stool five to ten times per day with occasionally colicky pain, abdominal distension, nausea and vomiting. He had visited hospitals and private clinics and received treatment but with no improvement. He presented to the Hospital for Tropical Diseases, Bangkok, Thailand, where on physical examination, he had moderate dehydration, weakness, abdominal distension and a gurgling abdomen. The eggs, larvae and adult worms of Capillaria philippinensis were found on stool examination. The patient was admitted and treated with Mebendazole for 20 days, whereupon his symptoms resolved. Two months previously, he had ingested a raw small fresh-water fish dish called "Phra-Pla Siw/Soi". Small fresh-water fish near the patient's home were collected and examined for Capillaria philippinensis larva. The results were negative for parasitic organisms.
Preview · Article · Dec 2008 · The Southeast Asian journal of tropical medicine and public health
[Show abstract][Hide abstract] ABSTRACT: Soil-transmitted helminthiases are major parasitic diseases that cause health problems world-wide. School-based health education is one of several basic interventions currently recommended by the World Health Organization for the control of these infections. A 3-year programme of health education for the control of soil-transmitted helminths (STH) has recently been completed in four primary schools in the Hauykayeng subdistrict of Thong Pha Phum district, in the Kanchanaburi province of Thailand. Overall, the percentage of the schoolchildren found infected with STH increased between the start of year 1 of the intervention (16.6%) and the end of year 2 (23.8%) but showed signs of falling by the end of year 3 (19.4%). Although none of these year-on-year changes in overall prevalence was statistically significant, some significant trends were detected when the six school grades (i.e. age-groups) were considered separately. The grade showing the highest prevalence of STH infection changed, from grade 6 (representing the oldest children investigated) at the start of year 1 (when grade-1 children were excluded from the survey) to grade 1 (representing the youngest children) at the ends of year 2 and year 3. By the end of year 3, the children in grades 5 and 6 had significantly lower prevalences of infection than the grade-1 subjects. The prevalence of STH infection in the grade-1 children was significantly higher than that in any of the older grades at the end of year 2 and significantly higher than that in grades 3-6 at the end of year 3. These results indicate that the health education had a greater impact on the children in the higher grades (who, presumably had better levels of understanding and practised better, personal, infection prevention) than on the younger children. Although school-based interventions can serve as a useful entry point for parasite control, more effort, including anthelminthic treatment, may be required among the youngest children. The activities need to be sustainable and supported by appropriate school-health policies.
No preview · Article · Oct 2008 · Pathogens and Global Health
[Show abstract][Hide abstract] ABSTRACT: We confirmed sympatric occurrence of Taenia solium, T. saginata, and T. asiatica in western Thailand. DNA analysis of morphologically identified T. saginata, in a dual infection with T. solium, indicated it was T. asiatica. To our knowledge, this report is the first of T. asiatica and a dual Taenia infection from Thailand.
Full-text · Article · Oct 2007 · Emerging infectious diseases