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Trematode Infection of Freshwater Snail, Family Bithyniidae in Thailand

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Trematode Infection of Freshwater Snail, Family Bithyniidae in Thailand SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH TREMATODE INFECTION OF FRESHWATER SNAIL, FAMILY BITHYNIIDAE IN THAILAND Jutharat Kulsantiwong1,2, Sattrachai Prasopdee1,3, Supawadee Piratae1, Panita Khampoosa1, Chalida Thammasiri1, Apiporn Suwannatrai1, Thidarut Boonmars1, Vithoon Viyanant4, Jiraporn Ruangsitichai5, Pairat Tarbsripair6 and Smarn Tesana1 1Food-Borne Parasite Research Group, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen; 2Department of Biology, Faculty of Science, Udon Thani Rajabhat University, Udon Thani; 3Chulabhorn International College of Medicine, Thammasat University, Klong Luang, Pathum Thani; 4Center of Excellence for Research in Biomedical Sciences, Thammasat University, Klong Luang, Pathum Thani; 5Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University, Bangkok; 6Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand Abstract. 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. Bithynia funiculata 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, Kanchana- buri 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. sia- mensis 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. Keywords: Opisthorchis viverrini, cercarial type, family Bithyniidae, snail, Thailand INTRODUCTION Correspondence: Dr Smarn Tesana, Food- Borne Parasite Research Group, Department of Freshwater snails of the family Bithy- Parasitology, Faculty of Medicine, Khon Kaen niidae serve as intermediate hosts for the University, Khon Kaen 40002, Thailand. multiplication of intramolluscan stages Tel: +66 (0) 43 363 434; Fax: +66 (0) 43 202 475 of many types of trematodes. Snails in E-mail: [email protected], tessmarn@yahoo. this family were first described for native com snails in Europe and Asia in the early 396 Vol 46 No. 3 May 2015 TREMATODE INFECTION IN SNAILS OF THE FAMILY BITHYNIIDAE 1870s (Brandt, 1974; Mills et al, 1994). A Thailand and Vietnam) (IARC, 1994; Keiser number of species have been reported as and Utzinger, 2005; IARC, 2011). The intermediate hosts for trematode parasites aquatic life cycle of O. viverrini requires (Wykoff et al, 1965; Chitramvong and Bithynia snail as the first intermediate Upatham, 1989; Ellen et al, 2008). host and many species of cyprinid fish as In Thailand, three genera of Bithy- second intermediate hosts (Wykoff et al, niidae were classified by Brandt (1974), 1965; Harinasuta et al, 1984; WHO, 1995; viz. Bithynia (2 subgenera of Digoniostoma Waikagul, 1998). In the endemic regions and Gabbia), Hydrobioides and Wattebledia. of northeast Thailand, the prevalence of O. Chitramvong and Upatham (1989) clas- viverrini infection in humans and cyprinid sified Gabbia as a valid genus. According ranges from 2.1% to 97% (Brockelman et al, to Chitramvong (1991, 1992), 10 species/ 1986; Haswell-Elkins et al, 1992; Sriamp- subspecies, namely, Bithynia funiculata, orn et al, 2004), but is lower in snail hosts B. siamensis goniomphalos, B. s. siamensis, (0.083%-1.6%) (Wykoffet al, 1965; Upatham Gabbia wykoffi, G. pygmaea, G. erawanen- and Sukhapanth, 1980; Upatham et al, sis, Wattebledia crosseana, W. siamensis, W. 1983). Moreover, epidemiological surveys baschi and Hydrobioides nassa make up during 1981-1991 indicated that there is an the family Bithyniidae as identified by increase in incidence of human O. viverrini morphological characteristics of shells, infection in the north and central regions internal organs and radula cusp patterns. of the country, whilst in the northeastern Their ecological habitats have been re- region the incidence is decreasing due ported in various types of water bodies in part to the implementation of a drug and a number of species share the same control strategy (Jongsuksuntigul and ecological niches. Within Bithyniidae, Imsomboon, 1998). only the genus Bithynia has been identified Experimental infection studies show- as being of medical importance (Wykoff ed that B. funiculata and B. s. siamensis have et al, 1965; Harinasuta et al, 1984; WHO, higher susceptibility to O. viverrini than 1995; Waikagul, 1998; Sri-aroon et al, 2005). B. s. goniomphalos, and within the same In Thailand, two species of snails, B. taxon immature laboratory bred snails funiculata and B. siamensis (2 subspecies are more susceptible to O. viverrini infec- of B. s. goniomphalos and B. s. siamensis, tion than mature field snails (Chanawong respectively) have been reported to serve and Waikagul, 1991). Trematode infection as the first intermediate hosts of human in B. s. goniomphalos showed that it could liver fluke, Opisthorchis viverrini (Wykoff serve as an intermediate host for 7 types of et al, 1965; Brockelman et al, 1986; Sri-aroon cercariae (amartae, amphistome, furcocer- et al, 2005). Opisthorchiasis is believed to cariae, monostome, O. viverrini, virgulate predispose the onset of cholangiocarci- and xiphidiocercaria) when shedding is noma , a cancer of the bile duct (Thamavit induced by illumination (Adam et al, 1993; et al, 1978; Haswell-Elkins et al, 1992; IARC, Nithiuthai et al, 2002; Sri-aroon et al, 2005). 1994; Sithithaworn et al, 1994; Vatanasapt Four types of cercariae (lophocercous, et al, 2000; Watanapa and Watanapa, 2002; monostome, pleurolophocercous and Honjo et al, 2005; IARC, 2011). O. viverrini is virgulate) were found in B. funiculata in a medically important parasite of humans the north but no O. viverrini infection was in the Greater Mekong subregion (Cam- detected (Ngern-klun et al, 2006). bodia, Lao People’s Democratic Republic, There is no report of human liver Vol 46 No. 3 May 2015 397 SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH fluke, O. viverrini, or other cercarial in- time shedding. Cercariae were preserved fections in the other species of family in 10% formalin and then observed under Bithyniidae apart from the genus Bithynia a stereo microscope (40x magnification). even though they are in the same family Snail specimens were observed for emerg- and share the same ecological niche. The ing cercariae at least 2 times within one present study examined the prevalence of month. Morphology study of cercariae trematode infections particularly O. viver- was undertaken using both unstained rini, in snail species of the family Bithy- and stained [with 0.1% (w/v) Fast Red B niidae in various localities in Thailand. or 0.1% (w/v) Malachite Green dye] under a light microscope and photographed. MATERIALS AND METHODS Cercaria types were identified based on available morphological description Snail collection (Wykoffet al, 1965; Schell, 1970; Frandsen The study localities for snail collec- and Christensen, 1984). For confirmation tion were in the north, south, west, north- of O. viverrini cercariae, species specific east and central areas of Thailand. The PCR was used (Wongratanacheewin et al, locations were selected based on informa- 2001). tion from previous reports (Brandt, 1974; Chitramvong, 1989, 1992; Nithiuthai et al, RESULTS 2002; Ngern-klun et al, 2006). The snail A total of 5,492 snails were collected survey was conducted during October from 17 locations of Thailand: Nong 2008 to July 2009. The locations of snail Khai, Nong Bua Lam Phu, Khon Kaen, collection were recorded using the Global Maha Sarakham, Yasothon, and Roi Et in Positioning System (Garmin model Nuvi the northeast; Chiang Mai, Lamphun in 203, Taiwan) and plotted on a map (Fig the north; Phitsanulok, Phichit, Nakhon 1). All snails were collected by wire-mesh Sawan, Sing Buri, Saraburi, Suphan Buri, scoop or by hand, and each specimen Bangkok in central; Kanchanaburi in the was cleaned, air-dried, labeled and kept west; and Surat Thani in the south (Fig 1). in porous plastic bags. The snail samples Snails were classified into ten species/sub- were brought back to the laboratory to be species in family Bithyniidae (Fig 2). The examined for trematode infection. Snails distribution of the snails were as follows: were identified based on shell morphol- B. funiculata (n = 120) and G. pygmaea (120) ogy for species following available keys from the north; B. s. goniomphalos (2,159), and descriptions (Brandt, 1974; Upatham W. siamensis (154) and W. crosseana (104) et al, 1983; Chitramvong, 1992). from the northeast; B. s. siamensis (1,813), Examination for trematode infection H. nassa (160) and G. wykoffi (592) from Trematode infection of snails were central; W. baschi (150) from the south conducted using a cercarial shedding and G. erawanensis (120) from Erawan method (day time and night time shed- waterfall, Kanchanaburi Province in the ding) as follows. Snails were placed in west (Table 1). plastic containers with 50 ml of dechlori- No night time cercarial shedding was nated tap-water, exposed to electric light observed. B. s. goniomphalos, B. s. siamensis, for 2 hours for day time shedding, or G. wykoffi, W. crosseana and W. siamensis were covered with a black plastic sheet shed cercariae in day time, with 6 types of overnight at room temperature for night cercariae, namely, amartae, monostome, 398 Vol 46 No. 3 May 2015 TREMATODE INFECTION IN SNAILS OF THE FAMILY BITHYNIIDAE no fin fold.
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