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Angiostrongylus cantonensis is an emerging infectious agent causing eosinophilic meningitis or meningoencephalitis in humans with clinical manifestation of severe headache. Molecular genetic studies on classification and phylogeny of A. cantonensis in Thailand are limited. This study surveyed A. cantonensis larvae prevalence in natural intermediate hosts across Thailand and analyzed their phylogenetic relationships. A total of 14,032 freshwater and land snails were collected from 19 provinces of Thailand. None of Filopaludina sp, Pomacea sp, and Cyclophorus sp were infected with Angiostrongylus larvae, whereas Achatina fulica, Cryptozona siamensis, and Megaustenia siamensis collected from Kalasin, Kamphaeng Phet, Phetchabun, Phitsanulok, and Tak Provinces were infected, with C. siamensis being the common intermediate host. Based on morphology, larvae isolated from 11 samples of these naturally infected snails preliminarily were identified as A. cantonensis. Comparison of partial nucleotide sequences of cytochrome c oxidase subunit I gene revealed that four sequences are identical to A. cantonensis haplotype ac4 from Bangkok and the other seven to that of A. cantonensis isolate AC Thai, indicating two independent lineages of A. cantonensis in Thailand.

Ko Ae Sub-district of Khueang Nai, Ubon Ratchathani Province, Thailand is located in an endemic area for Opisthorchis viverrini and other fish-borne zoonotic trematodes (FZT) infection. This study shows the status in Ko Ae Sub-district of FZT infection based on availability of intermediate hosts and necessary requirements for the transmission of FZT. A cross-sectional survey of intermediate hosts of FZT, including Bithynia siamensis goniomphalos and cyprinoid fish, was conducted from April 2013 to December 2014. Examination of 1,000 snails revealed 3.4% were infected with trematode cercariae, with a density of infection greater than 100 cercariae per infected snail. Six groups of morphologically-distinguishable trematode cercariae were identified, namely, cystophorous, echinostome, furcocercous, mutabile, parapleurolophocercous, and xiphidio, the latter being the most predominant type. Among 250 cyprinoid fish samples with metacercariae present at their caudal fins and examined for FZT by pepsin digestion, metacer- cariae of Haplorchis taichui, H. pumilio, and Centrocestus formosanus were found. Unidentified metacercariae collected from fish caudal fins were subsequently shown using a PCR-based assay to be C. formosanus. No infection by O. viverrini in the intermediate hosts, Bithynia siamensis goniomphalos and cyprinoid fish was evident. The study provides new information regarding trematode larvae infection in the primary and secondary intermediate hosts of FZT in this area of Thailand.

Trematode cercariae are commonly found in many freshwater gastropods. These cercariae can serve to identify the occurrence of such trematodes as Centrocestus formosanus, Haplorchis taichui, Haplorchoides sp, and Stellantchasmus falcatus, which are important parasites in Chiang Mai Province, Thailand. As the species of these cercariae cannot be identified accurately based on morphology, this study employed sequencing of a fragment of 28S ribosomal DNA and phylogenetic analysis to identify the trematode cercariae found in freshwater gastropods in Chiang Mai Province. Eight types of trematode cercariae were identified, namely, distome cercaria (grouped with Philophthalmus spp clade), echinostome cercaria (grouped with Echinostoma spp clade), furcocercous cercaria (grouped with Posthodiplostomum sp/Alaria taxideae/Hysteromorpha triloba clade), monostome cercaria (grouped with Catatropis indicus clade), parapleurolophocercous cercaria (grouped with Haplorchoides sp clade), pleurolophocercous cercaria (grouped with Centrocestusformosanus clade), transversotrema cercaria (grouped with Transversotrema spp clade), and xiphidiocercaria (grouped with Prosthodendrium spp clade). These results provide important information that can be used for identifying these parasites in epidemiological surveys.

We conducted a malacological investigation in four districts of the Nam Theun 2 (NT2) hydroelectric dam project area, Khammouane Province, central Lao PDR (Nakai, Gnommalath, Mahaxai and Xe Bang Fai), after the first and second years of full operation in March 2010 and November 2011 to determine health risks for humans. A total 10,863 snail specimens (10 families/23 species) from 57 sampling stations and 12,902 snail specimens (eight families/21 species) from 66 sampling stations were collected in 2010 and 2011, respectively. Neotricula aperta (gamma race), the intermediate host for Schistosoma mekongi, was found in large numbers (5,853 specimens) in 2010 in Nam Gnom (downstream) at Station 25 (Mueang Gnommalath: Gnommalath District) and in fewer numbers (170 specimens) at Station 26 (Ban Thathod: Gnommalath District). In 2011, significantly fewer numbers (434 specimens) of N. aperta were found at Station 25. No snails were found to be infected with S. mekongi; however, 3.6% and 0.45% of Bithynia (D.). s. goniomphalos specimens collected were found to be infected with Opisthorchis viverrini (human liver fluke) during 2010 and 2011, respectively. Pomacea canaliculata, the rice crop pest, the intermediate host of Angiostrongylus (Parastrongylus) cantonensis, was found in the greatest numbers during 2010 and 2011; the prevalence increased significantly from 1.3% in 2010 to 53.3% in 2011. We also found seasonal variation in snail populations in terms of abundance and diversity. The snail fauna and risk for transmission of parasitic diseases need to be monitored continuously to evaluate the long-term impact of the dam project.

Opisthorchis viverrini is restricted to and requires for its aquatic life cycle only Bithynia snail as first intermediate host but many species of cyprinid fish as second intermediate hosts. A survey in Thailand of trematode infection in freshwater snails of the family Bithyniidae carried out during October 2008 - July 2009 found a total of 5,492 snails, classified into ten species distributed in various geographic areas. Bithyniafuniculata and Gabbia pygmaea were localized to the north, B. s. goniomphalos, Wattebledia siamensis and W. crosseana to northeast and B. s. siamensis, Hydrobioides nassa and G. wykoffi to central region. W. baschi and G. erawanensis was found only in the south and Erawan waterfall, Kanchanaburi Province, respectively. Trematode infection rate was 3.15%. Cercariae were identified as belonging to six types, namely, amartae , monostome, mutabile, O. viverrini, virgulate, and unknown. The prevalence of cercarial infection in B. s. goniomphalos of amartae, mutabile, O. viverrini, virgulate, and unknown type cercaria was 0.55%, 0.74%, 1.07%, 2.87%, and 0.37%, respectively, and in B. s. siamensis monostome (1.10%) and virgulate (0.55%). Only virgulate cercariae were shed from W. crosseana (3.85%) and W. siamensis (5.19%). Cercariae of the unknown type were found in G. wykoffi (1.69%). No infection of O. viverrini cercariae was detected in the other species.

Opisthorchiasis in northeastern Thailand is an important etiology of cholangiocarcinoma. To form the infectious stage, free swimming cercariae penetrate cyprinid fish, shed their tails, and then secret a cystic substance to cover their larval stage to form metacercariae in the fish body. We determined the location of the cystogenous glands in Opisthorchis viverrini cercariae. The cercariae and metacercariae were obtained from the naturally infected snail host, Bithynia siamensis goniomphalos and from cyprinid fish, respectively. The cyst walls of the metacercariae were separated and used to immunize inbred male BALB/c mice to obtain cyst wall antibodies. The general characteristics of the O. viverrini cercariae and metacercariae were studied by hematoxylin and eosin (H&E) staining of sections. The location and ultrastructure of the cystogenous glands of cercariae were studied by immunoperoxidase, immunofluorescence and transmission electron microscopy. The structures and organelles of cercariae and metacercariae could be identified, but the cystogenous glands could not be detected in H&E sections. The immunoperoxidase and immunofluorescence sections revealed positive reactions for cystogenous glands predominanted in the lateral part of the cercariae and were clearly seen in the cyst wall of the metacercariae. The ultrastructure of the cystogenous glands contained semitranslucent electron dense oval shaped granules. If interference occurs during the formation of the cysts by fish immune response, the metacercariae may not develop to maturity. It may be easily digested or degraded by human stomach acid and pepsin. This may be an efficient method for control of O. viverrini infection which requires further detailed study.

Angiostrongyliasis has been frequently reported from the People's Republic of China during the last decade. An outbreak of angiostrongyliasis among migrant laborers in Guangning, Guangdong Province is described here. A questionnaire was developed to collect epidemiological and clinical information about 17 migrant laborers from the Bai ethnic group in Dali, Yunnan Province. Serum samples were collected and tested by enzyme-linked immunosorbent assay. Rats and mollusks from the same area where patients had collected Pomacea canaliculata were examined for presence of Angiostrongylus cantonensis. All 17 Bai migrant laborers consumed P. canaliculata and six had meningitis 3-19 days after consumption of P. canaliculata. Headache, myalgia and fatigue were the most common symptoms. Blood samples from 5 patients were positive for antibodies to A. cantonensis. The places where the migrant laborers collected P. canaliculata were identified as endemic areas for A. cantonensis. This outbreak highlights the vulnerability of migrants to angiostrongyliasis.

The trematode, Stellantchasmus falcatus, is one of the members in the Family Heterophyidae, which is reported to be an endemic fluke in northern region of Thailand. A sensitive and specific detection of this parasite is required to determine the epidemiology at larval stages. Specific primers to determine the presence of S. falcatus were investigated using high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR, with 10 arbitrary primers to generate different polymorphic DNA profiles. Eleven parasite species were used for comparison. A 380 bp HAT-RAPD S. falcatus-specific marker was found, and was cloned and sequenced, allowing a pair of primers (St-F 5'-GGCCAACG CAATCGTCATCC-3' and St-R 5'-GCGTCGGGTTTCAGACATGG- 3') to be designed to produce a 320 bp amplicon specific for S. falcatus. It revealed no cross-reaction with any of the other tested parasite species. The S. falcatus-specific primers can be used for epidemiological monitoring and for detection in snail intermediate hosts, which serve as usefulness tools in management and epidemiological control programs.

The tsunami and non-tsunami affected areas of Takua Pa District, Phang-Nga Province were investigated for fresh- and brackish-water snails that transmit human parasitic diseases during 2006 and 2007. Among 46 snail species found, 17 species of 8 families were freshwater snails, 28 species of another 7 families were brackish-water snails, and 1 species was a land snail. Of these species, 11 freshwater snails, 4 brackish-water snails and 1 land snail were of medical importance. The fresh-water snails were Pomacea canaliculata, Pila angelica, P. gracilis, P. polita, Filopaludina (S.) martensi, F. (F.) s. polygramma, Melanoides tuberculata, Indoplanorbis exuxtus, Radix rubiginosa, Helicorbis umbilicalis, Gyraulus convexiusculus. Four brackish-water snails were Cerithidea cingulata, C. djadjarensis, C. alata, Sermyla riqueti and Achatina fulica was the land snail. I. exutus, M. tuberculata and F. (F.) s. polygramma harbored Xiphidio, Microcercus, Furocercus, Echinostome cercariae, and cercaria without eyespots or tail with hair. Three species of brackish-water snails, Cerithidia cingulata, C. djadjariensis, and C. alata presented with 6 types of trematode cercariae and rediae. Knowledge of medically important snails and their parasitic diseases, and prevention were given to Takua Pa people by poster, pamphlets and broadcasting through community radio.

Snail and fish intermediate hosts were collected from rice fields in 3 districts; Mueang, Mae Taeng and Mae Rim of Chiang Mai Province during April-July 2008. For identification of larval trematode infection, standard (cracked for snail and enzymatically digested for fish) and molecular methods were performed. The results showed that three types of cercariae were found, pleurolophocercus, cotylocercous, and echinostome among 4 species of snail with a prevalence of 29, 23 and 3% respectively. Melanoides tuberculata snail was the most susceptible host for cercariae infection. Four species of metacercariae, Haplorchis taichui, Stellantchasmus falcatus, Haplorchoides sp and Centrocestus caninus, were found with a prevalence of 67, 25, 60 and 20%, respectively. The Siamese mud carp (Henicorhynchus siamensis) was the most susceptible fish host for H. taichui, and half- beaked fish (Dermogenys pusillus) for S. falcatus metacercariae infection, whereas Haplorchoides sp and C. caninus were concomitantly found in Puntius brevis. HAT-RAPD profile confirmed that pleurolophocercus cercariae found in Melanoides tuberculata from Mae Taeng District belonged to H. taichui and in Tarebia granifera from Mueang District were S. falcatus.

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.

The freshwater snail family Thiaridae was studied at five different locations: water sources for the Khek River, Thailand. Snail samples were collected by hand using counts per unit of time sampling method between December 2004 and October 2005. The physico-chemical quality of the water changed with the seasons and affected the sampling areas during both the dry season and the flood season. A total of 9,568 snail samples comprised of 14 species were found. These were 284 Tarebia granifera, 24 Melanoides tuberculata, 86 Thiara scabra, 3,295 Paracrostoma pseudosulcospira pseudosulcospira, 736 P. paludiformis paludiformis, 3,266 P. paludiformis dubiosa, 117 P. morrisoni, 304 Brotia (Brotia) binodosa binodosa, 1,250 B. (Brotia) microsculpta, 146 B. (Senckenbergia) wykoffi, 1 B. (Brotia) pagodula, 5 B. (Brotia) binodosa spiralis, 5 B. (Brotia) insolita and 49 B. (Brotia) manningi. The cercariae were investigated using shedding and crushing methods where they were categorized into two types and five species. The first type, Parapleurolophocercous cercariae, were comprised of Haplorchis pumilio Looss, 1899 and Centrocestus formosanus Nishigori, 1924. The second type, Xiphidiocercariae were comprised of Acanthatrium hitaense Koga, 1953, Loxogenoides bicolor Kaw, 1945 and Haematoloechus similis Looss, 1899. The cercarial infection rates in the above 5 species were 0.1% (5:9,568), 0.2% (15:9,568), 0.3% (24:9,568), 0.4% (37:9,568) and 0.1% (5:9,568), respectively. Five species of snails were susceptible to trematode infections. They were T. granifera, M. tuberculata, T. scabra, P. paludiformis paludiformis and B. (Senckenbergia) wykoffi; infections were found in 26.1% (74:284), 33.3% (8:24), 1.2% (1:86), 0.3% (2:736) and 0.7% (1:146), respectively.

Intertidal snail-trematode communities in southern Thailand were examined before and after the South Asia tsunami. Infection rates and species diversity of cercaria in the host snail Cerithidea in tidal zones did not change significantly from one year before to one month after the tsunami. However, the host snails C. quadrata, C. alata and C. obtusa disappeared from greatly damaged sites. It is important to follow up on the intertidal snail-trematode community recovery process after destruction of the intertidal ecosystem.

The study was conducted at 75 collecting loci in 15 districts of 11 provinces in Thailand during 1999-2004. A total of 12,079 live mollusks were collected, 11,874 were snails and 205 were clams. The snails were comprised of 39 species and classified into 9 families: Ampullariidae, Bithyniidae, Buccinidae, Potamiopsidae, Stenothyridae, Thiaridae, Viviparidae, Planorbidae and Lymnaeidae. The clams were comprised of 14 species classified into 2 families: Amblemidae and Corbiculidae. Fifteen species were medically important snails: Pomacea canaliculata, Pila ampullacea, P. pesmei, P. polita, Bithynia (Digoniostoma) funiculata, B. (D.) siamensis goniomphalos, B. (D.) s. siamensis, Filopaludina (Siamopaludina) martensi martensi, F. (Filopaludina) sumatrensis polygramma, Melanoides tuberculata, Tarebia granifera, Helicorbis umbilicalis, Gyraulus convexiusculus, Indoplanorbis exustus and Radix rubiginosa. Of these 3 snail species harbored trematode cercariae. I. exustus harbored Echinostoma malayanum, Xiphidio and Schistosoma spindale, and R. rubiginosa and B. (D.) siamensis goniomphalos harbored Xiphidio and intestinal flukes, respectively.

The morphology of cercariae of freshwater snails from Erawan Waterfall, Erawan National Park, Kanchanaburi Province was studied between December 2002 and August 2003. The snail samples were collected by handpicking using a counts per unit of time sampling method. The cercariae, larva stage of a trematode, were investigated using the shedding method where they were categorized into three groups and six species. The first group, Pleurolophocercous cercariae, consists of Haplorchis pumillo (C1) and Stictodora tridactyla (C3). The second group, Furcocercous cercariae, consisted of Mesostephanus appendicalatus (C2), Transversotrema laruei (C6) and Cardicola alseae(C4). The third group, Xiphidio cercariae, has only one species which is Loxogenoides bicolor (C5). Out of 1163 snails, only 62 were found to be infected by cercariae, equivalent to a 5.33% infection rate. The infections grouped by species of the cercariae are as follows: C, 22 (1.9%), C, 29 (2.5%), C2 1 (0.1%), C6 1 (0.1%), C4 6 (0.5%) and C5 3 (0.3%). The freshwater snail samples consist of four species. From a total of 1163 samples, there are 687 Melanoides jugicostis, 91 Tarebia granifera, 296 Thiara scabra and 89 Melanoides tuberculata. Infections were found in 45 (6.5%), 6 (6.6%), 1 (0.3%) and 10 (11.2%), respectively.

A survey of freshwater snail, Bithynia funiculata, was conducted in four locations, Doi Saket, Mueang, Saraphi and Mae Rim Districts of Chiang Mai Province, northern Thailand, between June and October, during the rainy season of 2004. A total of 2,240 snails was collected and classified into 7 families and 15 genera; of which 352 B. funiculata were obtained. B. funiculata was found most abundant in July and September. The infection rate of trematode cercariae in B. funiculata was 9.6% (19/352), while that of pleurolophocercous was 0.3% (1/352). Virgulate cercaria was the most common type, followed by lophocercous, monostome and pleurolophocercous. B. funiculata prefers habitats with clear water, which was less than 30 cm depth, temperatures between 24.48 and 31.78 degrees C, dissolved oxygen 2.03-7.66 mg/l, saturated dissolved oxygen 26.70-95.00%, conductivity 0.000-0.2642 mS/cm, turbidity 16.00-288.00 NTU and pH 6.58-7.56. Geographic Information System (GIS) analysis using loose soil mixed with clay revealed that the breeding grounds of this snail species was in the paddy fields and village environs of the Ping, Kuang and Fang river basins.

Three malacological surveys were conducted in the Takua Pa District of Phang-Nga Province, southern Thailand, before and after the Indian Ocean Tsunami disaster. Twenty-nine species of fresh- and brackish-water snails were found, in which 10 species of freshwater snails were present, including live Pila polita; 8 species were of medical importance. Two brackish-water snails, Nerita articulata and Littorinopsis scabra, were absent after the tsunami disaster, while brackish-water Cerithidea cingulata and C. djadjariensis harbored 9 types of trematode cercariae.

The objective of this study was to observe the molluscicidal activities of Euphorbia milli, known as "poysean" in Thailand, against Indoplanorbis exustus. Latex from 12 different E. milii hybrids was screened for its molluscicidal activities. Indoplanorbis exustus were exposed for 24 and 48 hours to the latex at various concentrations ranging from 6 to 25 ppm and mortality rates were recorded. Eight hybrids of latex were effective. The six most effective hybrids were E. milii Dang-udom, E. milii Arunroong, E. milii Raweechotchuong, E. milii Srisompote, E. milii Sri-umporn and E. milii Tongnopakun, which killed all snails after 24 hours of exposure. Under the same conditions, latex of E. milii Dowpraket and E. milii Promsatid killed 50% of the snails. Such results indicate that these 6 hybrids seem promising as natural molluscicidal agents.

Brackish-water mollusks inhabiting the mangrove areas along the Gulf of Thailand of Surat Thani Province were investigated for distribution, abundance and natural infections. Nine families and 32 species of brackish-water snails were recovered from 14 sampling stations. Species belonging to the genus Ceritidea of the family Potamididae were mainly examined and Cerithidea (Cerithideopsilla) Cingulata, C. (C.) djadjariensis, and C. (Cerithidea) charbonnieri were naturally infected with 2 types of trematode cercariae, and one which was undetermined. C. (C.) cingulata had the highest infection rate (38.5%). Viewing two snail communities, the first community on the mainland and the second on Samui Island in Surat Thani Province, 28 brackish-water mollusk species were present on the mainland, 15 species were evident on Samui Island, and 11 snail species were common to both the mainland and Samui Island. Measurement of community similarity based on species presence revealed an index of similarity of 0.51. Concerning land use by the local people in the station areas investigated, brackish-water snails in Surat Thani Province are facing habitat degradation by human use.

Under natural conditions, the emergence of Opisthorchis viverrini cercariae from naturally infected Bithynia (Digoniostoma) siamensis goniomphalos showed diurnal periodicity, peaking between 8:00-10:00 AM. The cercariae did not emerge during darkness, but low-intensity light could induce a release. Cercariae shedded from each field infected B.(D.) s. goniomphalos was recorded daily. The maximum output from one snail was 1,728 cercariae in a day. The total cercarial output from all five infected snails was 56,555 and the maximum of total cercariae shed from one snail was 27,692. The field-infected B. (D.) s. goniomphalos could survive for 70 days after the snails were collected.

We have shown previously that anti-fecundity immunity can be induced experimentally against recombinant 26 kDa glutathione S-transferase (reSjc26GST) in Chinese water buffaloes (Bos buffelus), important reservoir hosts for Schistosoma japonicum in China. In the field study described here, we immunized buffaloes with reSjc26GST to induce protective immunity against S. japonicum and to evaluate its effectiveness in controlling schistosomiasis japonica. We selected two villages as test and control groups in inside-embankment areas endemic for schistosomiasis japonica. The buffaloes in the test village were vaccinated with reSjc26GST, whereas those in the control village were not. The indicators of the effect of the vaccine included the generation of specific IgG antibodies in the vaccinated buffaloes, changes in the prevalence and infection intensity in buffaloes and village children, changes in the density of infected snails, and changes in the infectivity of water bodies (assessed by sentinel mice) in transmission areas adjacent to both villages. Twenty months after vaccination, the infection rate of buffaloes in the test village was decreased by 60.4% (from an initial prevalence of 13.5% to 5.4%), and 67.9% when compared with that in the control village (initial prevalence of 16.7%). However, the infection rate in village children remained unchanged. The density of infected snails decreased by 71.4%, from 0.0049/0.11 m2 to 0.0014/0.11m2 in the high transmission area outside the embankment in the test village. There was no change in the infectivity of the water body transmission areas between the test and control villages. The levels of specific antibodies to reSjc26GST showed a continuous increase after vaccination. These results indicate that protective immunity was induced and maintained in buffaloes after vaccination with reSjc26GST. The vaccine could thus play a significant role in reducing S. japonicum transmission caused by water buffaloes in the Lake region of China.

Paludomus petrosus, the freshwater snails found in Pa La-U Waterfall, were examined for cercarial infection of trematodes. The snails were collected every other month from April, 2001 to February, 2002. Collections were taken from two sampling stations. The counts per unit of time' method was used for collection of the snails. The density of snails was highest in June 2001, and the highest of parasite infection rate was in February 2002. Four types of cercariae were found in the snails: Xiphidiocercariae, Amphistome, Furcocercous cercariae type I, and Furcocercous cercariae type II. Xiphidiocercariae were found in April 2001 to February 2002. Amphistome, Furcocercous cercariae type I and Furcocercous cercariae type II were found in February 2002.

To determine the genetic basis of the resistance of Schistosoma mansoni to praziquantel (PZQ) and to understand whether the resistance is dominant or recessive trait, a schistosome cross was undertaken between a PZQ-susceptible and a PZQ-resistant isolate using infections of the single-sex cercariae which were identified by a direct W1-specific PCR technique. The resistances of F1 and F2 generation to PZQ were evaluated using in vitro egg, miracidial and cercarial tests. The F1 hybrid progeny from crosses between the susceptible and resistant isolates were resistant to PZQ. The resistant phenotype reappeared in the F2 progeny. It can thus be considered that the PZQ resistance behaves like a dominant trait.

A survey of trematode larvae in the fresh water snail, Bithynia goniomphalos in the main water reservoir of Huai Thalaeng district, Nakhon Ratchasima Province, Thailand, was conducted. The study site is in an area with a high prevalence of Opisthorchis viverrini infection. Malacological samplings were made during the year 2000 to study the prevalence and intensity of infection. Parasitological examinations of 527 live snails were performed at the Veterinary Parasitology Laboratory, Chulalongkorn University. Of the 527 snails, only 16 (3.04%) were found to be infected with cercariae. Of these 16 snails, 75% were infected with Amphistome cercariae, 6.25% with Armatae cercariae and 18.75% with Virgulate cercariae. Most infected snails (75%) had a density of infection of less than 100 cercariae/snail. No snail was infected with cercariae of Opisthorchis spp. This snail was, therefore, unlikely to be the intermediate host of Opisthorchis viverrini in the main water reservior of Huai Thalaeng district.

A serosurvey of various indigenous interior tribes (Orang Ulu) in upper Rejang River Basin Sarawak Malaysia, the site of a multibillion Ringgit hydroelectric power project, found 6.8% of the individual surveyed were seropositive for schistosomiasis, as determined by ELISA method using the soluble egg antigen of Schistosoma malayensis Baling strain. In all age group, the seroprevalence rate is higher (9.5%) in males than in females (4.5%) except for the 31-40 age group. Seroprevalence of schistosomiasis was found to increase with age with the above 60 age group having the highest rate followed by the 31-40 age group. Seroprevalence rate among the tribes ranges from 4.1% among the Penan to 11.6% among the Kajang. There was no seroevidence of schistosomiasis among the Ukits. A snail survey found four snail species including Brotia species, the intermediate host of the lung fluke Paragonimus westermani, however no schistosome snail host was identified. Although schistosomiasis malayensis-like infection may be endemic in the area, its public health significance remains undetermined.

A region-wide sampling survey was conducted in 1995 in order to evaluate the current epidemiological status of schistosomiasis japonica in Hunan Province, China. A total of 45,590 humans and 3,726 domestic animals, from 52 villages, were examined parasitologically and/or serologically for current Schistosoma japonicum infections. In uncontrolled endemic areas (43 villages) the overall human prevalence of S. japonicum was 7.81% across the different geographical subtypes. The geometric mean intensity of infection was 17.71 eggs per gram (epg) among infected individuals and only 1.25 epg in the general population. The bovine prevalence, as determined by the hatching test, was 9.63% in the uncontrolled endemic villages. Only one sero-positive (by indirect hemagglutination assay) child was found among 1,072 children tested aged 10-14 years in the 9 endemic villages under effective control. No infection was confirmed by the Kato-Katz thick smear stool examination. When the results of this survey were compared to those seen at baseline (1989) an overall reduction of 45.65% was seen in the human prevalence but no significant change was apparent in the lake-beach ecotype. Additionally, there was more than a 60% reduction in the prevalence among bovines over the same sampling period. The results demonstrate that the World Bank Loan Schistosomiasis Program was successful in achieving its most basic objectives for this province - to reduce human and bovine infections by 40%.

With the construction of the Three Gorges high dam on the Yangtze River in China in mind, a serious of ecological environmental factors that might affect the transmission of Schistosoma japonicum in Jian Han Plain were investigated by means of data collection, field surveys and observation in Hubei Province. Several ecological factors including water level of the Yangtze River; riparian water table, annual rainfall and yearly evaporation were investigated in relation to the prevalence of schistosomiasis. The results suggest that after the dam construction, middle water level flows (ie between flood flows and dry-weather flows) will persist in the flood season due to a rise in the water table. The investigation indicated that snail distribution and human schistosomiasis prevalence differed significantly between years which had typically high, middle and low typical water levels in the Yangzte. Moreover, the prevalence of the disease showed a significant linear regression relationship with density of snail intermediate hosts, water table, annual rainfall, yearly evaporation and ground altitude in the survey area. Systematic and careful monitoring and surveillance is necessary to investigate the impact of the environmental changes brought about by the dam construction on schistosomiasis transmission.

This study examined the prevalence of Schistosoma japonicum infections in field rats, Rattus rattus mindanensis, according to different trapping locations. Between October 1995 and January 1996, traps were set in the municipality of Palo, Leyte, Philippines to determine the correlation of rats infected with schistosomiasis to the proximity of the intermediate snail host, Oncomelania hupensis quadrasi, colonies. Of the 22 rats that were caught within a snail colony, 21 (95.5%) were positive for schistosomiasis. Of the 23 rats that were caught 100 meters from a snail colony, 13 (56.5%) were positive for schistosomiasis. Of the 17 rats that were caught approximately 1 km from a snail colony, zero (0%) were positive for schistosomiasis. Infection rates were highest within the habitat of the intermediate host and lowest in rats captured far from snail colonies. Captured rats were also examined for the presence of other endoparasites. Infections of the following were found: Angiostrongylus cantonensis, Gonylonema neoplasticum, Hymenolepis diminuta, Nippostrongylus muris, Strongyloides ratti, Syphacia obvelata, Taenia crassicollis and Trichuris muris, but there was no correlation between trapping location and prevalence. None of the rats were infected with Moniliformis moniliformis, Trichinella spiralis, Trypanosoma lewisi or Vampirolepis nana.

Oncomelania snails are the intermediate hoste of Schistosoma japonicum in Asian countries. In order to understand the genetic and morphological variation of Oncomelania snails in mainland China, field snails from 31 localities were collected and investigated by means of allele enzyme electrophoresis and numerical taxonomical technics. Results demonstrated that out of 17 loci examined, seven polymorphic loci were presented. Genetic distance (Nei, 1978) among the populations varied from 0.03 to 0.27. The phenogenetic tree based on UPGMA cluster analysis showed that genetic diversity corresponded to geographic distribution along the Yangtze River, which provided supplementary genetic data about the evolution of Oncomelania spp. A morphological study showed that Mahalanobis' morphological distance ranged from 1.53 to 346.7. Both genetic and morphological data indicated that the diversity among populations of smooth shelled snails was higher than that among populations of ribbed shelled snails. A positive correlation (r = 0.80) between Mahalanobis' morphological distance and genetic distance supports the hypothesis that the different shell phenotypes represent different species or subspecies.

Clinical and stool examinations for clonorchiasis were carried out in an endemic area, Kim Son District, Ninh Binh Province, Vietnam. Stool examination with the Kato-Katz technic revealed that in 306 residents selected randomly, 42 people (13.7%) were infected with Clonorchis sinensis. The rate was biased towards men (23.4%) as opposed to women (1.5%) and increased with age. No children younger than 10 years old were infected, reflecting difference in a chance for acquisition of infection through a habit of eating raw fish. Few clinical abnormalities were found by blood and urine examinations of the patients. Treatment with praziquantel decreased the infection rate to 5.3% at 6 weeks later. Snails, Melanoides tuberculatus, collected from ponds around the settlements were infected with cercariae at a rate of 13.3%. Farmed fish (Hypophthalmichtys molitrix) in the ponds were infected with metacercariae at rates of 56.4% in small individuals and 100% in large ones. The life cycle of C. sinensis is exclusively completed in the ponds and the traditional habit of eating raw fish in summer was thought to be a major route of infection.

Four batches of crude somatic antigens from: (1) Opisthorchis viverrini adult worms, (2) Bithynia funiculata-whole body, (3) B. funiculata-head-foot, and (4) B. funiculata-visceral mass were assayed against sera from 81 opisthorchiasis patients, 30 parasite-free healthy individuals, and 50 individuals infected with other helminthic infections, and their antibody levels determined. By IgG-ELISA, the antigenic reactive proteins were found in both the head-foot and the visceral mass of B. funiculata snails, but the whole snail antigens gave the best results. Furthermore, it was as good as when O. viverini antigens were used. Antibody levels of sera from patients with opisthorchiasis assayed against antigens from whole B. funiculata snails were significantly higher than those of the other two groups. The cut-off value for positivity at 0.228 gave 80.2% sensitivity and 81.2% specificity. Cross reactions were observed with sera from patients with paragonimiasis and strongyloidiasis. No cross reactions were found to occur with sera from healthy individuals.

The population dynamics and production of cercariae of Schistosoma japonicum in Oncomelania hupensis are reported. The experiments covered the whole life span of positive snails and different intervals of cercariae shedding. The results indicated that two patterns of the dynamics of cercariae shedding had been found in the life span of positive snails. The first was a long-time interval (4-7 days) and progressive decline pattern. The cercariae shedding of positive snails lasted 18-19 weeks in males and for 32-33 weeks (once a week). The second was a short-time interval (1-3 days) and continued release pattern. The cercariae shedding of positive snails lasted for 20-36 days (every day shedding). Shedding cercariae stimulate cercariae development.

A survey of Angiostrongylus infections in rats and snails was carried out in the provinces of Ubon Ratchathani, Udon Thani. Kalasin, Chaiyaphum and Khon Kaen in northeast Thailand. Only two Rattus norvegicus (3.8%) and one Bandicota indica (1.4%) out of 151 R. rattus, 52 R. norvegicus and 69 B. indica examined were infected with adult lung worms. All worms recovered were A. cantonensis. Prevalence of infection in 423 Pila polita was 0.9% while all of 77 P. ampullacea were negative for larvae. In contrast to this 36.4% of 500 Achatina fulica harbored L3 of Angiostrongylus (with variations of between 29% and 46% in the five provinces). The average infection intensity in A. fulica was 13.6 L3 (1 to 441). Experimental infection of Wistar rats with L3 (isolated from A. fulica resulted in a recovery rate of 48.3% of adult worms of which 91.7% and 8.3% were identified as A. cantonensis and A. malaysiensis, respectively. This is the first proven finding of A. malaysiensis in northeast Thailand.

Opisthorchiasis due to Opisthorchis viverrini and transmitted through infected freshwater cyprinoid fish (carps) affects more than 8 million people in Thailand, People's Democratic Republic of Lao, and Vietnam. The Hazard Analysis Critical Control Point (HACCP)-concept has been recommended by FAO and WHO to be included in programs to control foodborne trematode infections (FBT). HACCP is a multifactorial approach to control food hazards through surveillance of diseases, foods, and operations and education. This study describes the first attempt to apply HACCP to the prevention and control of Opisthorchis viverrini in pond culture carp (Puntius gonionotus). The experiment was designed and carried out by a multidisciplinary "HACCP team" including experts in the field of public health, parasitology, epidemiology, aquaculture, fisheries extension and fish inspection. The investigation was performed in two fish ponds in the District of Sun Pa Tong, Chiang Mai, Thailand. In the experimental pond, fish was cultured according to HACCP principles and compared with the control pond, which followed conventional aquaculture practices. Water supply to the pond, fish fry, fish feed and pond conditions during the growing period were identified as critical control points (CCPs). Hazards were identified and analyzed, as well as control measures, critical limits, monitoring procedures, corrective actions, and record keeping developed for each one of the above CCPs. Complete pond preparation, particularly aiming to eliminate contamination of pond water with O. viverrini eggs, fish infected with parasite meacercariae and the first intermediate host (Bithynia spp), was conducted. After the pond was filled with water, O. viverrini metacercaria-free fry were released into the pond. The preliminary results obtained indicate that HACCP-based principles applied to carp pond culture could be used as a strategy to prevent and control O. viverrini. Further studies should be undertaken aiming to confirm these preliminary results.

Three species of the pulmonate snails of the family Planorbidae have been reported from Korea; Gyraulus convexiusculus, Hippeutis (Helicorbis) cantori and Segmentina (Polypylis) hemisphaerula. Of these 3 species, only H. cantori is reported as the molluscan intermediate host of Neodiplostomum seoulensis, one of the important snail-borne human intestinal trematodes in Korea. However, S. hemisphaerula was also found to be an intermediate host for N. seoulensis. In field-collected snails, H. cantori and S. hemisphaerula were found shedding bifurcated cercariae of N. seoulensis, whereas no G. convexiusculus was found shedding cercariae. In experiments with laboratory-bred snails, only S. hemisphaerula was susceptible to miracidia of N. seoulensis. Tadpoles of Rana nigromaculata and R. rugosa were exposed to cercariae shed from field-collected and laboratory-bred S. hemisphaerula. All tadpoles of R. nigromaculata were found to be massively infected, but none of the tadpoles of R. rugosa were infected with larvae of N. seoulensis. Metacercariae from tadpoles of R. nigromaculata and the snake Rhabdophis tigrinus tigrinus were fed to rats, and eggs of N. seoulensis were detected in the rat feces one-week later. These rats were killed and adult N. seoulensis recovered from the small intestines. This is the first report of S. hemisphaerula as a molluscan intermediate host for N. seoulensis in Korea.

In Korea, soybean-sauce soaked freshwater crabs (kejang) have been a favorite delicacy when eating a bowl of rice. This traditional food has been a main source of human paragonimiasis. Until the late 1960s, human paragonimiasis had been prevalent; at least two million people had contracted the infection as determined by intradermal tests. About 40% of these were egg positive. In the turmoil of the green revolution and industrialization in the 1970s/1980s, ecological damage occurred widely. In many streams, populations of snail and crustacean hosts were reduced to levels almost of extinction. Population reduction of the intermediate hosts was followed by lowered endemicity. Attitudes of people, changed during the period, also reduced chances of paragonimiasis. Survey data in the 1990s indicated that prevalence of human paragonimiasis has lowered to about one 100th of that in the early 1970s. In a referral system, however, about one hundred clinical cases have been diagnosed annually by antibody test (ELISA), undertaken for clinical differentiation from tuberculosis. At least 10% of freshwater crabs sold in local markets are infected with the metacercariae. Paragonimiasis control has benefited in Korea mainly by the untoward effects of water pollution. To place P. westermani infections as enzootic, health education and surveillance systems should have a priority.

Thirteen enzymes encoded by 16 loci of six population of Oncomelania hupensis in Zhejiang, China, were investigated by means of starch gel electrophoresis. Ten loci (AO, 6PGD, ME, AKP, OCT-1, HBDH-1, HBDH-2, XDH, MDH and MPI) were monomorphic and 6 loci (OCT-2, PGI, AAT, PGM-1, PGM-2 and ACP) were polymorphic. Three enzymes (OCT, HBDH and PGM) were encoded by 2 loci. The results indicated that there were allozyme variations in two subspecies, O.h. hupensis and O.h. fausti in Zhejiang, China. Nei's multilocus genetic distances (D) between subspecies ranged from 0.167 to 0.265. Minor genetic distances were detected between populations of the same subspecies. The results indicated that the enzyme acid phosphatase (ACP) is a possible marker to measure the degree of susceptibility of O. hupensis to S. Japonicum.

Beta and gamma races of Neotricula aperta were exposed to naturally raw waters of the Sirindhorn reservoir, Mun River and Mekong River in laboratory conditions and non supplement on food. Snails of two races could withstand well in variable qualities of water within 48 hours. The survival rates of N. aperta in three sources of raw water were 100% for the beta race and 97 to 100% for the gamma race.

Five subspecies of Oncomelania snails, Oncomelania hupensis nosophora, O.h. hupensis, O.h.chiui, O.h.formosana and O.h.quadrasi, were experimentally exposed to the first stage larvae of Angiostrongylus cantonesis. The presence of third stage larvae was observed in all of the five subspecies of Oncomelania snails 20 days after infection. Infection rates of the third stage larvae of the parasite in Oncomelania snails were 38.0-40.0%. There were no differences in preferences among Oncomelania snails. The third stage larvae in Oncomelania snails almost distributed in kidney and intestine region, and most of the larvae were active and free in tissues. The distribution pattern of the larvae in Oncomelania snails was quite different from that in Achatina fulica and Ampullarium sp. These third stage larvae were ingested by rats, and developed to adults. These data suggest that Oncomelania snails may play important role when A. cantonensis will spread, and indicate the possibility of human infection with A. cantonensis.

Twenty-two yellow tree monitors (Varanus bengalensis) were trapped and transported from 5 provinces, namely Lampang, Phitsanulok, Kamphaeng Phet, Tak and Prachin Buri to look for the infective larvae stage of Angiostrongylus cantonensis. In 4 provinces all of the yellow tree monitors were infected with the infective stage larvae of A. cantonensis altogether. Twenty-one of the 22 yellow tree monitors (95.5%) in the five provinces were infected. Our results of this study extend our knowledge of natural prevalence of A. cantonensis in yellow tree monitors. The snail eating habit of the yellow tree monitor might possibly be the source of the larvae.

The first intermediate host of six-known the Paragonimus species in Thailand had not been found until the Filopaludina (Siamopaludina) martensi martensi snail was discovered to maintain the cercariae of a Paragonimus species. An extensive study examined cercarial development through to adult worms by infecting 3 genera of 7 crab species with penetration of cercariae and feeding of snails containing such cercariae. These crabs provided many metacercariae which were fed to cats and bandicoots. The animals gave many Paragonimus adult worms which were characterized as Paragonimus siamensis by the following criteria: 6-lobed ovary and cuticular spines in groups. It is concluded that the Filopaludina martensi martensi snail is a susceptable natural first intermediate host of P. siamensis. Second intermediate hosts Somanniathelphusa brandti, S. sexpunctatum and S. bangkokensis were experimentally infected; prior to this study only S. germaini and S. dugasti had ever been naturally infected with metacercariae of this species.

Golden apple snails, Pomacea canaliculata, were collected at two localities having different ecological environments. In both canal and pond, P. canaliculata males were found more than females during the dry season (summer and winter). In the canal, the male snails were highest in number (86.67%) in May. When rain started, they began decreasing and were lowest at 33.33% in August. Of 575 snails collected, 30.6% were infected by one or more of the three groups of amphistome, distome and echinostome metacercariae. There were two high peaks of infection in April and October, as 60.7% and 68.4%, respectively, during which there were more males than females. The average number of parasites per snail which was highest at 54 was found in the medium-sized males (25 out of 35 males) in October. The number of parasites per snail was significantly correlated with the collected males (p < 0.01), but such relationship was not occurred with the females. Of the females, only the large-sized individuals were infected. In the pond, the female snails were present in much greater numbers than the males during the reproductive time (June-September). The females were highest (94.23%) in August. Only 24 (4.0%) of 605 snails were infected; most of the infected snails were large.

Cercariae from Bithynia siamensis goniomphalus were studied in an area endemic for opisthorchiasis. Snails emitted different types of cercariae when shedding was induced by illumination. In addition to two lophocercous cercariae, a monostome cercaria (probably Notocotylus sp), two different furcocercariae and a xiphidiocercaria were found. Two similar types of lophocercous cercariae were distinguishable and one could be identified as Opisthorchis viverrini. To date the cercaria of O. viverrini has been regarded as the only lophocercous cercaria in the endemic area of opisthorchiasis. Therefore, differences between these cercariae were further characterized by scanning electron microscopy to enable us to avoid misinterpretations of O. viverrini in Bithynia.

Freshwater snails, Stenomelania denisoniensis (Brot) from Tinaroo Dam, North Queensland, Australia were found to be infected with a heterophyid cercaria identified as Procerovum sp. The tail of the cercaria has finfolds which are bilateral anteriorly and dorso-ventral posteriorly, features which separate it from other genera in the Haplorchis-group. This group is differentiated from the cercariae of all the other heterophyid genera by the presence of the penetration glands that extend to the posterior end of the body lateral to the excretory bladder. This paper presents a full description of the cercaria, together with comparisons with other known species of Procerovum.

Golden apple snails, Pomacea canaliculata, were collected once a month during a year to search for their natural parasites. The collections were made at two localities having different ecological environments. Of 576 collected snails from a canal, 176 individuals (30.6%) were infected by three groups of metacercariae. These parasites were amphistome, distome and echinostome metacercariae, which had prevalences of 23.5, 19.5 and 0.5%, respectively. The incidence of infection was highest (68.4% in October) when the snail population was composed of the old, juvenile and young Pomacea. Amphistome metacercariae were found most frequently and echinostome metacercariae the least frequently; both parasites were localized in the foot muscle of the snails and had a Shannon index of zero. The range of amphistomes was 1 to 115 with the mean +/- SD of 1 +/- 2 and 95% CL of 1, 2. Distome metacercariae were found primarily in the heart (range: 1-13), and also in the foot muscle (range: 1-5) and kidney (range: 1-14), with a Shannon index of 0.4. The means +/- SD (with 95% CL) were 3 +/- 4 (95% CL = 1, 5), 3 +/- 4 (95% CL = 2, 4) and 2 +/- 1 (95% CL = 1, 2) for the foot muscle, heart and kidney, respectively. The snails from a pond, another locality, had a low proportion of infected individuals. Of 605 snails, only 24 individuals (4.0%) were infected, with the prevalence of amphistomes, distomes and echinostomes being 0.8, 1.8 and 2.1%, respectively. The incidence of infection for each month was zero or less than 10%, except in May when it was 30.2%.

Stool samples from a total of 1,008 persons were examined for intestinal parasites during a comprehensive study on the epidemiology of small fluke infections in Laos. The prevalence of small fluke eggs in the stool was seen to peak at age 20 years, particularly in men from villages (90.4%). Three quarters of infections belong to the category of light infections and only 0.6% to the category of very heavy according to eggs per gram of feces (EPG). The highest EPG was in the 11-15 year age group. In addition, the following parasites were diagnosed: Sarcocystis hominis (prevalence more than 10% in the group over 20 years of age), Taenia sp. (12.4% for the village people over 20 years), Fasciolopsts buski (3.8% for the same group). The habit of Laos people to eat raw fish, beef and pork flesh, is reflected in significant epidemiological consequences. Cercariae of Opisthorchis viverrini occurred in 0.5% of Bithynia siamensis goniomphalus examined, Haplorchis sp. cercariae were found in 0.9% of Tarebia granifera snails. Metacercariae of O. viverrini were found in flesh of 7 species of cyprinid fish. Haplorchis taichui in 4 species of these fish, and H. pumilio of two cyprinid species. Hampala macrolepidota harbored larvae of all above mentioned species. Stellantchasmus falcatus was recovered in fins of belonid fish Xenentodon cancila. Adults of O. viverrini were found in 36% of domestic and stray cats, Heterophyid flukes were found in 24% cats. The most frequent species were H. taichui and H. yokogawai. Eight Laotian students were treated in Czechoslovakia with praziquantel to determine fluke infection. Three were infected only by O. viverrini, four only by H. taichui and one by both O. viverrini and H. taichui. These results demonstrate the problem of correct differential diagnosis of food-borne small fluke infections and the need to assess the clinical course and public health aspects of infections.

Observations on transmission of the rat lung worm, Angiostrongylus cantonensis, from rats to the snail intermediate host. Achatina fulica, in a vacant lot in Bangkok are described. The prevalence of A. cantonensis increased with snail age until 200 days of age when it attained a plateau of 50-60%. The overall prevalence was 53%. The worm burden slowly rose with age until 200 days of age beyond which it remained relatively constant. The highest mean worm burden of 5,478 was observed in the oldest age group. The parasite distribution in the snail population was highly aggregated both within each age class and in the overall population. Experiments on susceptibility of snails to laboratory infection revealed that worm recovery was dependent on dose of first stage larval infection but was independent of snail size in the range of 4-8 cm. The percent worm recovery of third stage larvae was negatively correlated with dose of infection, and no density-dependent effects of worm burden on worm size were observed.