Molecular identification of the strongyloid nematode Oesophagostomum aculeatum in the Asian wild elephant Elephas maximus.

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 (Elephas maximus) 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 Oesophagostomum aculeatum. 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 Strongyloides. The present results indicate that O. aculeatum in local elephants is a potential source of helminthiasis in human and domestic animals in this wild-elephant irrupted area.

School-based health education for the control of soil-transmitted helminthiases in Kanchanaburi province, Thailand.

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.

Strongyloides stercoralis infection and chronological changes of other soil-transmitted helminthiases in an endemic area of southern Thailand.

A field survey was conducted in 4 primary schools in Nakhon Si Thammarat Province, southern Thailand. By Sasa modified Harada-Mori cultivation method, 1.8% of the schoolchildren were found to be infected with Strongyloides stercoralis, and 25.1% had hookworm infection. By Kato's thick smear method, the overall prevalence of soil-transmitted helminths was 46.8%, being Trichuris trichiura 28.5%, hookworm 18.0%, and Ascaris lumbricoides 5.7%. Fecal examination, performed by Kato's thick smear and culture method, indicated that the prevalence of hookworm infection was 26.9%. The prevalence in the present study was very much lower than many previous reports in the past decade. This may indicate the partial success of the parasite control project in Thailand by mass treatment, improving the sanitation and personal hygiene of the people in the endemic area. In light infection with Trichuris, albendazole administered at a dosage of 200 mg daily for 3 days showed a 48.7% cure rate. When mebendazole was given at 100 mg twice daily for 3 days, its effectiveness was 88.5%. A lower cure rate was obtained (70.0%) in moderate to heavy infection.

Morphological variation and abnormality of cephalic hooklets of Gnathostoma spinigerum hepatic stage larvae from laboratory infected mice.

One thousand advanced third-stage larvae of Gnathostoma spinigerum from laboratory infected mice, two to five weeks after being fed with infected cyclops, were examined specifically for the morphology of their cephalic hooklets. Among these, only the 15-day old (early hepatic-stage) larvae and the 30-day old (late hepatic-stage) larvae were measured for the size of their body and hooklets. The average body size of the 15-day old and 30-day old larvae were 3.4 +/- 0.4 x 0.4 +/- 0.04 mm and 4.9 +/- 0.4 x 0.5 +/- 0.04 mm, respectively. The average size of the hooklets from rows one to four of the 15-day old larvae was 14.6 +/- 1.7 x 6.8 +/- 0.6 microm, 15.6 +/- 2.0 x 7.2 +/- 0.5 microm, 16.0 +/- 1.8 x 7.4 +/- 0.6 microm and 15.9 +/- 1.9 x 7.3 +/- 0.6 microm, respectively. Those of the 30-day old larvae were 15.1 +/- 1.7 x 7.1 +/- 0.6 microm, 16.3 +/- 1.6 x 7.5 +/- 0.7 microm, 16.5 +/- 1.7 x 7.8 +/- 0.6 microm and 16.3 +/- 1.7 x 7.6 +/- 0.8 microm, respectively. The average number of cephalic hooklets from rows one to four of the two- to five-week old larvae were 42.8 +/- 2.6, 45.3 +/- 2.8, 46.9 +/- 2.8 and 50.2 +/- 2.9, respectively. Several types of morphological variation and abnormality of the cephalic hooklets were observed. The most common ones were extra rudimentary hooklets below row four or within the four rows of hooklets (10.8%), the present of a fifth row of hooklets (1.9%), abnormal hooklets in only row four (1.2%), lobed or branched hooklets (0.5%), spiral arrangement of the four rows of hooklets (0.4%), and fragmented hooklets (0.4%).

Seasonal variation in the intensity of Gnathostoma larvae in swamp eels (Fluta alba) sold in a local market in Bangkok.

The viscera of swamp eels were obtained from a local market in Bangkok twice a month from June 1996 to May 1997. The livers were separated, weighed and counted. Gnathostome larvae were recovered from the livers by the digestion technic, examined, identified, and counted. A total of 12,278 Gnathostoma larvae were obtained from 18,561.1 g (15,264 pieces) of eel livers. The overall average number of larvae/g liver and the overall average number of larvae/liver are 0.91 and 0.94, respectively. The greatest number of larvae/g liver (on average) was in December (high levels of infection during the months of October to December) whereas the lowest was in April (lowest levels of infection during the months of March to April). Thus there was a marked decrease in the average number of larvae/g liver during January to April, which then started to rise in May. This finding suggests that the level of infection abruptly decreases soon after the completion of the rainy season, starts to rise when the rain has come, and reaches its peak when the amount of rainfall is highest. More than 99% of the total gnathostome larvae recovered were identified to be G. spinigerum, and 25.4% of the entire larvae recovered bore variant or abnormal cephalic hooklets. The most common unusual feature was that there were extra rudimentary hooklets above row one, below row four and in between the four rows of hooklets which comprised 21.4%. In addition, the body size and the number of cephalic hooklets of G. spinigerum are also discussed.

Toxocara and Gnathostoma among stray canines in Bangkok.

Stomachs and intestines of 88 adult and 112 young stray dogs were obtained from the Rabies Control Subdivision, Bangkok, and examined especially for the presence of Gnathostoma spinigerum and Toxocara canis. Forty-five dogs were found positive for T. canis (overall prevalence 22.5%) but none were found infected with G. spinigerum. The prevalence of T. canis in young dogs was 37.5% (42 of 112) whereas in adult dogs was only 3.4% (3 of 88). The total number of T. canis recovered from the 45 positive dogs was 272 (averaging 6.0 worms/dog). This includes 268 worms from 42 young dogs (averaging 6.4 worms/dog) and four worms from three adult dogs (averaging 1.3 worms/dog). The average number of worms, according to sex and stage, per young dog were as follows: male worms 2.4 +/- 3.5 (range 0-15), female worms 2.8 +/- 3.5 (0-16), immature worms 1.2 +/- 2.5 (0-9), and all worms 6.4 +/- 8.2 (1-34). The maximum number of worms per young dog was 34 while the minimum was one, and 35.7% (15/42) of these young dogs harbored only one worm. The body length of the recovered T. canis were as follows: males measuring 3.0-12.0 cm (averaging 7.1 +/- 2.1 cm), females 4.1-18.2 cm (11.0 +/- 4.1 cm), and immature worms 0.7-3.7 cm (2.1 +/- 0.8 cm).

Effect of albendazole and mebendazole on soil-transmitted helminth eggs.

Primary school children from Nakhon Si Thammarat Province, Thailand, on endemic area of soil-transmitted helminths, were selected for study. The infected children were divided into two groups and pair-matched according to intensity of infections: group I were given albendazole (400mg) single dose and group II were given mebendazole (100mg) twice daily for 3 days. On the day following treatment, the number of Trichuris eggs in the stool markedly increased and the egg shape was also altered. These phenomena did not occur in Ascaris infections since 100% cure rate were obtained using both drugs. Incomplete ovicidal effect of the drugs to Trichuris and Ascaris eggs were demonstrated, embryos were observed to develop within the treated eggs and they hatched after feeding them to experimental animals. In hookworm infection, albendazole stimulated the females to release more eggs after medication, but both drugs showed complete ovicidal effect upon examining the eggs from the second bowel movement.

The impact of primary health care intervention on reinfection of soil-transmitted helminths in the community.

A study to evaluate the impact of primary healthy (PCH)care practices on the prevention of reinfection of soil-transmitted helminths (STH) was carried out in Nakhon Si Thammarat Province, southern Thailand. Fecal samples from the inhabitants of Ban Paruhas, the implementing village, and Ban Pungsing, the control village, were examined following mass treatment with a three-day regimen of albendazole (400 mg). Training of health volunteers, implementation of a health education program through village broadcasting system and improvements in sanitation including increasing the number of latrines were carried out at Ban Paruhas village. After one year of implementation, prevalence and intensity of STH in both villages were determined using Kato-Katz's method. Pre- and post intervention stool examination results were compared. The overall prevalence of STH at post-intervention decreased significantly in both villages, with a greater degree of reduction appearing in ascariosis and hookworm infections in the village where PHC was implemented.

Circumoval and larval microprecipitation reactions in experimental and human gnathostomiasis.

Mice, rats and cats were infected either orally or percutaneously with a number of early or advanced third-stage larvae (EL3 or AL3, respectively) of G. spinigerum. Sera obtained from these infected animals and 10 human gnathostomiasis cases were tested against various developmental stages of the parasite which were prepared and used while being alive (fresh) or dead (air-dried) for the circumoval and larval microprecipitation (COP and LMP) reactions. No precipitin reactions were observed in all sera tested against unembryonated eggs, embryonated eggs and first stage larvae neither air-dried nor fresh preparations. Sera were merely reactive giving various degrees of membranous or filamentous precipitates against the air-dried preparation of AL3.

Detection of humoral immune response to Gnathostoma spinigerum in mice.

The humoral immune response to early third stage larvae (EL3) and advanced third stage larvae (AL3) of Gnathostoma spinigerum infection was studied in mice by Ouchterlony gel diffusion technique. The antibodies was detected at week 3 in mice infected with EL3 and remained up to week 10 after infection. Highest positive sample of sera were demonstrated at week 4 to week 7. Similar results were obtained from AL3 infected sera except the antibodies was found and disappeared earlier (week 2 to week 6). G. spinigerum larvae recovery from mice in both groups showed that the number of advanced third stage larvae located in muscle correlated to the peak of positive sera. No cross reaction was observed on positive sera of G. spinigerum and antigens of A. cantonensis, P. siamensis, T. spiralis, O. viverrini and A. ceylanicum. Cross reaction was shown on the G. spinigerum antigen against rat sera with angiostrongyliasis and bandicoot sera with paragonimiasis.