The Life Cycle of Heterophyes yacyretana n. sp. (Digenea, Heterophyidae), Parasitic in the Endemic Snail Aylacostoma chloroticum (Caenogastropoda, Thiaridae) in Argentina.

Here we describe a new heterophyid species, Heterophyes yacyretana n. sp., and resolve its life cycle experimentally. We found the prosobranch snail Aylacostoma chloroticum in Candelaria, Province of Misiones, Argentina (a sector of the High Paraná River affected by the Yacyretá Dam), naturally infected with opisthorchioid cercariae. These cercariae lacked pigmented eyespots as well as body pigment and possessed 7 pairs of penetration glands arranged in 2 lateral bands, together with 18 pairs of flame cells and a V-shaped excretory vesicle. We exposed specimens of 21 fish species to emerging cercariae and obtained metacercariae from the muscles of the caudal peduncle of 3 species of siluriform fish, and adults from chicks infected with experimentally obtained metacercariae from the albino variety of the bronce corydoras, Corydoras aeneus. The new species differs from other species in the genus by the number of sclerites on the genital sac, the distribution of the vitelline follicles, and the combination of the size relationship of the suckers and the genital sac with respect to the posterior extent of intestinal ceca. Heterophyes yacyretana is the first species of the genus reported from the Americas.

Morphological and Molecular Identification of Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae) in Thailand.

We tried a series of morphological and molecular approaches to identify a new species of Stellantchasmus (Digenea: Heterophyidae) originating from the wrestling half-beaked fish, Dermogenys pusillus of Thailand. Adult worm samples of the new species were recovered from hamsters experimentally infected with the metacercariae from D. pusillus in Thailand. Two isolates (Thai and Korean) of Stellantchasmus falcatus were used as comparative control groups. Worm samples of 3 Stellantchasmus groups were morphologically observed and molecularly analyzed with the mitochondrial cytochrome c oxidase 1 gene. The morphological characteristics of S. dermogenysi n. sp. are similar to S. falcatus originating from brackish water fish, but minor difference was noted including the absence of the prepharynx, position of the ovary near the ceca end, smaller body size, and shorter esophageal length. A phylogenetic tree derived from neighborjoining and maximum-likelihood methods suggests that S. dermogenysi n. sp. is separated from S. falcatus supported by high bootstrap values. The relative divergences persist between these host-specific trematodes, which we suggest should be recognized as 2 distinct species. Comparisons of S. dermogenysi n. sp. with S. falcatus isolated from mullets in Thailand and Korea indicate a genetic divergence of mitochondrial DNA of 19.4% and 21.7%, respectively. By the present study, a new species, Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae), is proposed in Thailand based on molecular evidences, in addition to minor morphological differences between S. falcatus and the new species.

Centrocestus formosanus (Trematoda: Heterophyidae) in Melanoides tuberculata (Gastropoda: Thiaridae) from Vila do Abraão, Ilha Grande, Rio de Janeiro, Brazil / Centrocestus formosanus (Trematoda: Heterophyidae) em Melanoides tuberculata (Gastropoda, Thiaridae) da Vila do Abraão, Ilha Grande, Rio de Janeiro, Brasil

Abstract Pleurolophocercous cercariae found in the invasive gastropod Melanoides tuberculata (Müller, 1774) collected in a stream of the Vila do Abraão, Ilha Grande, Angra dos Reis, Rio de Janeiro, Brazil were used for experimental infection that enabled the identification of the heterophyid trematode Centrocestus formosanus (Nishigori, 1924). The parasite has been found in the locality since 2007, after two years of the introduction of M. tuberculata. Recently, from a sample of 483 specimens collected in June 2013, 101 (21%) were found infected with parasite. The potential environmental impacts caused by the parasite occurrence could be underestimated in the country, and actions to monitor and control both the parasite and the mollusk are necessary.

Resumo Cercárias do tipo pleurolofocerca encontradas no gastrópode invasor Melanoides tuberculata (Müller, 1774) coletados em um riacho da Vila do Abraão em Ilha Grande, Angra dos Reis, Rio de Janeiro, Brasil foram utilizadas em estudos de infecção experimental que possibilitaram a identificação do trematódeo heterofiídeo Centrocestus formosanus (Nishigori, 1924). O parasito vem sendo encontrado na localidade desde 2007, dois anos após a introdução do molusco. Recentemente, de uma amostra de 483 espécimes coletados em junho de 2013, 101 (21%) apresentavam-se infectados pelo parasito. Os potenciais impactos ambientais ocasionados pela ocorrência deste parasito podem estar sendo subestimados no país, sendo necessárias ações visando o monitoramento e controle tanto do parasito quanto do molusco.

Metagonimus suifunensis sp. n. (Trematoda: Heterophyidae) from the Russian Southern Far East: Morphology, life cycle, and molecular data.

The trematode from the Russian Southern Far East, earlier defined as M. yokogawai Katsuradai, 1912, was determined as a representative of the new species Metagonimus suifunensis sp. n. The life cycle of this trematode was experimentally studied and morphometric data for the cercariae, metacercariae and adult worm stages were obtained. The molecular data for samples from six Russian localities were received using the ITS1-5.8S-ITS2 region and 28S gene nuclear ribosomal DNA. The results revealed that M. suifunensis sp. n. does not have significant morphometric differences with M. yokogawai, M. takahashii and M. miyatai. However, the new species validity was confirmed by the molecular data. The phylogenetic relationship analysis of Metagonimus representatives showed that M. suifunensis sp. n. is well-differentiated specie with low variability in the whole area.

Centrocestus formosanus (Trematoda: Heterophyidae) in Melanoides tuberculata (Gastropoda: Thiaridae) from Vila do Abraão, Ilha Grande, Rio de Janeiro, Brazil.

Pleurolophocercous cercariae found in the invasive gastropod Melanoides tuberculata (Müller, 1774) collected in a stream of the Vila do Abraão, Ilha Grande, Angra dos Reis, Rio de Janeiro, Brazil were used for experimental infection that enabled the identification of the heterophyid trematode Centrocestus formosanus (Nishigori, 1924). The parasite has been found in the locality since 2007, after two years of the introduction of M. tuberculata. Recently, from a sample of 483 specimens collected in June 2013, 101 (21%) were found infected with parasite. The potential environmental impacts caused by the parasite occurrence could be underestimated in the country, and actions to monitor and control both the parasite and the mollusk are necessary.

Stellantchasmus falcatus (Digenea: Heterophyidae) in Cambodia: Discovery of Metacercariae in Mullets and Recovery of Adult Flukes in an Experimental Hamster.

Stellantchasmus falcatus (Digenea: Heterophyidae) is first reported from Cambodia through recovery of the metacercariae from mullet fish and adult flukes from an experimentally infected hamster. We purchased 7 mullets, Chelon macrolepis, in a local market of Phnom Penh, Cambodia, and each of them was examined by the artificial digestion method on May 2010. The metacercariae of S. falcatus were detected in all mullets (100%) examined, and their average density was 177 per fish. They were elliptical, 220×168 µm in average size. They were orally infected to an hamster to obtain adult flukes. Adults recovered at day 10 post infection were observed with a light microscope and a scanning electron microscope (SEM). They were small, 450×237 µm in average size, had a small oral sucker (41×50 µm), subglobular pharynx (29×21 µm), slender esophagus (57 µm), long and thick-walled expulsor (119×32 µm), spherical ovary (58×69 µm), and 2 ovoid testes (right: 117×74 µm; left: 114×63 µm). Eggs were small, yellow, and 23×12 µm in average size. In SEM observations, tegumental spines were densely distributed on the whole tegument, and single small type I sensory papillae were distributed around the lip of oral sucker. The small ventral sucker was dextrally located and had 8 type I sensory papillae on the left margin. It has been first confirmed in the present study that the mullet, C. macrolepis, is playing the role of a second intermediate host of S. falcatus in Cambodia.

Epidemiological and molecular data on heterophyid trematode metacercariae found in the muscle of grey mullets (Osteichthyes: Mugilidae) from Sardinia (western Mediterranean Sea).

This study is a contribution to the molecular taxonomy and epidemiology of heterophyid (Trematoda: Heterophyidae) metacercariae found in the muscle of Mugilidae (Osteichthyes) from Sardinia (western Mediterranean Sea). Sixty specimens of mugilids (13 Chelon labrosus, 18 Liza aurata, 6 Liza ramada, 8 Liza saliens, 15 Mugil cephalus) were examined and 17,899 metacercariae isolated in 95 % of the hosts. Four types of metacercariae were identified: Heterophyes sp. (n = 14,113), Heterophyes sp. -small (1225), Stictodora sp. (1606), and Ascocotyle (Phagicola) sp. (955). The experimental infection of a hamster with Heterophyes sp. metacercariae produced six adults identified as Heterophyes heterophyes and two as Heterophyes cf. nocens. The morphology of Heterophyes sp. -small metacercariae matched with that of Heterophyes dispar. The sequence analysis of the ITS2 and 28S portions of rDNA confirmed the morphological identification of metacercariae, showing four clusters. All adults grouped together with the Heterophyes sp. metacercariae, whereas adults of Heterophyes nocens from Korea clustered separately, showing that this species is distinguished from H. heterophyes, and suggesting caution in the exclusive use of the number of rodlets of the genital sucker to separate the two species. The presence of metacercariae was high in all hosts; the highest prevalence is of Heterophyes sp. (prevalence ≥78 %; mean intensity ≥135 metacercariae/100 g muscle), and the most heavily infected host is M. cephalus (prevalence = 100 %; mean intensity = 841 metacercariae/100 g muscle).

The life cycle of Neocladocystis intestinalis (Vaz, 1932) (Digenea: Cryptogonimidae), in Aylacostoma chloroticum (Prosobranchia: Thiaridae), and Salminus brasiliensis (Characiformes: Characidae), in Argentina.

The life cycle of Neocladocystis intestinalis (Vaz, 1932) was resolved experimentally. The prosobranchiate snail Aylacostoma chloroticum Hylton Scott (Thiaridae) collected in the Yacyretá Dam, Province of Misiones, Argentina, was found naturally infected with cercariae that possessed pigmented eye spots, 7 pairs of penetration glands, 12 pairs of flame cells, and a V-shaped, or Y-shaped excretory vesicle with very short stem. The cercariae developed in oval cysts, which were found on fin rays, and under scales of naturally and experimentally exposed tetragonopterid fish species and of experimentally exposed poecilid and prochilodont fish species. Adults were obtained experimentally from juvenile Salminus brasiliensis (Characidae), bred in captivity, and infected with metacercariae from albino Gymnocorymbus ternetzi (Tetragonopteridae), which had been exposed to emerging cercariae.

First molecular identification of Ascocotyle (Phagicola) longa in its first intermediate host the mud snail Heleobia australis.

This is the first study that used species-specific DNA primers to confirm the presence of the heterophyid Ascocotyle (Phagicola) longa Ransom, 1920 in its first intermediate host. The larval stages (rediae and cercariae) of this parasite were morphologically and genetically identified in the gonad of the intertidal mud snail Heleobia australis (d'Orbigny, 1835) (Cochliopidae) in the Bahía Blanca estuary, Argentina. In addition, we asked whether the prevalence in H. australis varied between seasons. Mullets - the second intermediate host of this heterophyid - migrate in estuaries during the warmer seasons and it is expected that piscivorous birds and mammals - the definitive hosts - prey more intensively on this species at those times. Thus, the number of parasite eggs released into the tidal flat within their feces should be higher, thereby increasing the ingestion of the parasite by H. australis.We therefore expected a higher prevalence of A. (P.) longa in H. australis in the Bahía Blanca estuary during spring and summer than autumn and winter. We found that 16 out of 2,744 specimens of H. australis had been infected with A. (P.) longa (total prevalence of 0.58%). Nonetheless, the prevalence showed no significant variation between seasons. Hence, we discuss an alternative scenario where the lack of seasonal changes might be mostly related to the permanent residence of definitive hosts in the estuary and not to the seasonal recruitment of mullets. Finally, we highlight the need for more experimental and comparative approaches in order to understand the diagnosis and geographical distribution of this worldwide heterophyid.

Molecular characterization of Stictodora tridactyla (Trematoda: Heterophyidae) from Kuwait Bay using rDNA ITS and mtCO1.

Stictodora tridactyla is an intestinal fluke in the family Heterophyidae that parasitizes shorebirds and mammals, including humans. Its metacercarial cyst stage was reported in the Arabian killifish, Aphanius dispar, at Kuwait Bay. In the present study, Cerithidea cingulata was found to serve as the first intermediate host of S. tridactyla. In order to establish the snail-fish link in the life cycle of S. tridactyla, complete sequences of ribosomal DNA internal transcribed spacer region 1 and 2 (rDNA ITS1 and ITS2) and partial sequence of cytochrome oxidase subunit 1 were obtained for metacercarial cysts isolated from the fish A. dispar and rediae isolated from the snail C. cingulata. Sequence alignment demonstrated that these larval stages belong to the same heterophyid species, S. tridactyla. Phylogenetic analysis based on rDNA ITS1, ITS2, and mtCO1 confirmed the position of S. tridactyla within the Heterophyidae and found it to cluster with Haplorchis spp. The present study represents the first molecular study correlating the larval stages of S. tridactyla using rDNA ITS1, ITS2, and mtCO1 and examining the phylogenetic relationships of S. tridactyla with different heterophyid species.

Host preference of an introduced 'generalist' parasite for a non-native host.

Parasites can invade new ecosystems if they are introduced with their native hosts or if they successfully infect and colonise new hosts upon arrival. Here, we ask to what extent an introduced parasite demonstrates specialisation among novel host species. Infection surveys across three field sites in Gatun Lake, Panama, revealed that the invasive peacock bass, Cichla monoculus, was more commonly infected by the introduced trematode parasite Centrocestus formosanus than were three other common cichlid fishes. Laboratory infection experiments were conducted to determine whether parasitism might be driven by differential encounter/exposure to parasites or by differential infection susceptibility/preference across different host species. These experiments were performed by controlling for parasite exposure in single host (compatibility) experiments and in mixed host (preference) experiments. In all cases, the peacock bass exhibited higher infection rates with viable metacercariae relative to the other potential fish hosts. Our experiments thus support that an introduced generalist parasite shows apparent specialisation on a specific novel host. Further studies are needed to determine whether these patterns of specialisation are the result of local adaptation following invasion by the parasite.

Developmental and Phylogenetic Characteristics of Stellantchasmus falcatus (Trematoda: Heterophyidae) from Thailand.

This study aimed to investigate the infection status, worm development, and phylogenetic characteristics of the intestinal trematode, Stellantchasmus falcatus. The metacercariae of S. falcatus were detected only in the half-beak (Dermogenus pusillus) out of the 4 fish species examined. Their prevalence was 90.0%, and the intensity of infection was 919 metacercariae on average. Worms were recovered from 33 (97.1%) of 34 chicks that were experimentally infected with 200 S. falcatus metacercariae each, and the average recovery rate was 43.0%. The body size and inner organs of S. falcatus quickly increased in the experimental chicks over days 1-2 post-infection (PI). In addition, ITS2 sequence data of this parasite were analyzed to examine the phylogenetic relationships with other trematodes using the UPGMA method. The results indicated that the ITS2 sequence data recorded from trematodes in the family Heterophyidae appeared to be monophyletic. This study concluded that D. pusillus serves as a compatible second intermediate host of S. falcatus in Thailand and that S. falcatus can develop rapidly in the experimental chicks. Data collected from this study can help to close the gap in knowledge regarding the epidemiology, biology, and phylogenetic characteristics of S. falcatus in Thailand.

Susceptibility, metacercarial burden, and mortality of juvenile silver barb, common carp, mrigal, and tilapia following exposure to Haplorchis taichui.

Fish-borne zoonotic trematodes (FZT) including heterophyids and opisthorchiids are prevalent in native and aquaculture fish in Southeast Asia. FZT are reported highly prevalent in juvenile aquaculture fish that belonged to the family Heterophyidae, particularly Haplorchis taichui. Aquaculture fish are reported to have varying levels of natural infection with H. taichui, but data for their susceptibility as well as resistance to infection are not available. The aim of this study was to determine the susceptibility of five aquaculture fish to H. taichui. Experimental infections were performed by exposing fish individually to 0 (control), 50, 100, or 200 H. taichui cercariae for 12 h. Metacercarial burden was measured at 45 days postinfection. Three out of five fish species, silver barb (Barbonymus gonionotus), common carp (Cyprinus carpio), and mrigal (Cirrhinus mrigala), were highly susceptible to H. taichui infection with percentage ranging from 93.33 to 100%. The Nile and red tilapia were not susceptible to infection. Among the susceptible fish species, silver barb had higher intensity of H. taichui metacercariae than common carp and mrigal (P < 0.001). Metacercarial burden significantly correlated with the dose of cercarial infection (P < 0.001). Our findings that common aquaculture fish species have varying degrees of susceptibility to H. taichui infection provide important information to reduce parasite transmission in aquaculture fish.

[A woman with long-lasting diarrhoea and fatigue]. / En ung kvinne med langvarig diaré og slapphet.

BACKGROUND: Intestinal trematodes are common parasites in man and many mammals. Infection is often asymptomatic and unrecognised. CASE PRESENTATION: A woman in her twenties presented with loose stools of variable intensity over six months. Additionally, she had experienced considerable fatigue during this period. There was no weight loss and initial blood tests were normal. Further testing at the second visit included stool microscopy, and small trematode eggs consistent with H. heterophyes infection were found. A more thorough anamnesis revealed the onset of symptoms on the day she returned from a week's holiday, and the probable exposure occurred from eating sushi twice during this holiday. After one day of treatment with praziquantel 40 mg/kg administered in three doses, the patient recovered completely within two to four weeks. Her asymptomatic partner had consumed the same food and had the same eggs in his stool sample. He was successfully treated with the same treatment dose. INTERPRETATION: A detailed travel history may provide important information relating to the diagnosis of diarrhoea and fatigue. Symptoms of H. heterophyes infection are variable. A single day's dose of 40 mg/kg of praziquantel was sufficient to eradicate infection in the two cases presented.

Survival of heterophyid metacercaria in common carp (Cyprinus carpio).

Heterophyidae are small intestinal trematodes that infect vertebrates worldwide. Common carp (Cyprinus carpio) is one of the most preferred freshwater fish species by consumers in Asia, the region where fish-borne trematodes like Heterophyidae are most prevalent. How long Heterophyidae survive in common carp is unknown. The objective of this study was to quantify survival of Heterophyidae in common carp after experimental exposure. Fish of 0.18 g were either used as controls or exposed to 250 heterophyid cercaria for 24 h. Control fish did not become infected. Percentage infection of exposed fish at 0-2 (n = 53), >2-10 (n = 15), >10-20 (n = 11), and >20-27 (n = 33) weeks post exposure was 98, 80, 100, and 100 % respectively. The number of metacercaria per fish did not significantly decrease (P = 0.19) during 27 weeks after exposure: exp [3.6200-0.0193 × weeks post exposure]. All developed metacercaria were identified as Haplorchis spp. It was concluded that Heterophyidae may persist in carp for a long time, implying that harvestable carp are a risk to human health.

Metagonimus yokogawai: metacercariae survey in fishes and its development to adult worms in various rodents.

A parasitological survey for Metagonimus yokogawai metacercariae was carried out by examining a total of 321 freshwater fish comprising of 7 species. Of the 321 fish samples examined, 182 (56.7%) were found to be infected with M. yokogawai metacercariae. The prevalence of M. yokogawai metacercariae in Opsariichthys pachycephalus was 93.4% (86/92), Zacco platypus 75.0% (30/40), Distoechodon turmirostris 61.3% (38/62), Varicorhinus barbatulus 56.5% (13/23), Hemibarbus labeo 33.3% (1/3), Acrossocheilus formosanus 15.9% (14/88), and 0% in Sinibrama macrops (0/13), respectively. This is the first record of M. yokogawai infection in Z. platypus, D. turmirostris, V. barbatulus, and H. labeo in Taiwan. The major site of predilection of the metacercariae in the fishes was in the scale, but some metacercariae were also observed in the flesh and fins. The M. yokogawai metacercariae were orally inoculated into mice, rat, gerbil, and golden hamster to study their infectivity and also to obtain the adult worms for taxonomic study. Worm recovery in hamsters was 75.3%, in mice was 70.0%, in rats was 23.3%, and in gerbils was 6.0%, respectively. Moreover, larger worms were recovered from the golden hamster. Golden hamster was thus found to be the most susceptible experimental rodent host for the infectivity study of Metagonimus. Besides M. yokogawai, metacercariae of Centrocestus formosanus was also observed in the fishes examined.

Metagonimoides oregonensis (Heterophyidae: Digenea) infection in Pleurocerid snails and Desmognathus quadramaculatus salamander larvae in Southern Appalachian streams.

Metagonimoides oregonensis (Heterophyidae) is a little-known digenetic trematode that uses raccoons and possibly mink as definitive hosts, and stream snails and amphibians as intermediate hosts. Some variation in the life cycle and adult morphology in western and eastern populations has been previously noted. In the southern Appalachians, Pleurocera snails and stream salamanders, e.g., Desmognathus spp., are used as intermediate hosts in the life cycle. We completed a series of studies in this system examining some aspects of larval trematode morphology and first and second intermediate host use. Molecular sequencing of the 28S rDNA of cercariae in our survey placed them clearly within the heterophyid family. However, light and scanning electron microscopy revealed both lateral and dorso-ventral finfolds on the cercariae in our region, whereas original descriptions of M. oregonensis cercariae from the west coast indicate only a dorso-ventral finfold, so further work on the systematics of this group may be warranted. A survey of first intermediate host, Pleurocera proxima, from 7 streams in the region identified only M. oregonensis, virgulate-type cercariae, and cotylomicrocercous-type cercariae in the streams, with M. oregonensis having the highest prevalence, and the only type present that use amphibians as second intermediate hosts. Based on clearing and staining of 6 Desmognathus quadramaculatus salamander larvae, we found that individual salamanders could have over 600 metacercariae, which form between muscle fibers throughout the body. Histological observations suggest that the metacercariae do not cause excessive tissue damage or inflammation, and likely persist through metamorphosis, thereby transmitting potentially large numbers of worms to definitive host raccoons foraging along streams.

Fluctuations in densities of the invasive gill parasite Centrocestus formosanus (Trematoda: Heterophyidae) in the Comal River, Comal County, Texas, U.S.A.

Centrocestus formosanus (Trematoda: Heterophyidae) is an invasive fish parasite in the Comal River, Texas, and is considered a threat to the federally endangered fountain darter, Etheostoma fonticola . Monitoring densities of C. formosanus cercariae is crucial to determining levels of infection pressure. We sampled 3 sites in the Comal River during 2 sampling periods, the first during 2006-2007, and again during 2009-2010. Two of the sites were located in the upstream reach of Landa Lake, sites HS and LA, and the third site was located downstream of Landa Lake in the old channel of the river. Cercariae densities were highest at the downstream most site (EA), followed by sites LA and HS, during both sampling periods, but a significant decline in cercariae density was observed between the first and second sampling periods. Several abiotic factors were monitored, including total stream discharge, wading discharge, temperature, and dissolved oxygen, but no river-wide trends were observed. Therefore, we speculate that these factors do not adequately explain the observed long-term decline in cercariae density. We propose that the decline is simply a reflection of a typical pattern followed by most invasive species as they gradually become integrated into the local community following an initial explosive growth in population size. Although cercariae densities may be abating, fountain darters in the Comal River are still threatened by the parasite, and conservation efforts must focus on reducing levels of infection pressure from the parasite whenever possible.

Co-infection with Opisthorchis viverrini and Haplorchis taichui detected by human fecal examination in Chomtong district, Chiang Mai Province, Thailand.

Diseases caused by the liver fluke, Opisthorchis viverrini and the minute intestinal fluke, Haplorchis taichui, are clinically important, especially in the Northeast and North regions of Thailand. It is often difficult to distinguish between these trematode species using morphological methods due to the similarity of their eggs and larval stages both in mixed and co-infections. A sensitive, accurate, and specific detection method of these flukes is required for an effective epidemiological control program. This study aimed to determine the prevalence of O. viverrini and H. taichui infections in human feces by using formalin-ether sedimentation and high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR methods. Fecal specimens of people living along the Mae Ping River, Chomtong district were examined seasonally for trematode eggs using a compound microscope. Positive cases were analyzed in HAT-RAPD, DNA profiles were compared with adult stages to determine the actual species infected, and specific DNA markers of each fluke were also screened. Our results showed that out of 316 specimens, 62 were positive for fluke eggs which were pre-identified as O. viverrini and H. taichui. In addition, co-infection among these two fluke species was observed from only two specimens. The prevalence of H. taichui infections peaked in the hot-dry (19.62%), gradually decreased in the rainy (18.18%), and cool-dry seasons (14.54%), respectively. O. viverrini was found only in the hot-dry season (6.54%). For molecular studies, 5 arbitrary primers (Operon Technologies, USA) were individually performed in HAT-RAPD-PCR for the generation of polymorphic DNA profiles. The DNA profiles in all 62 positives cases were the same as those of the adult stage which confirmed our identifications. This study demonstrates the mixed infection of O. viverrini and H. taichui and confirms the extended distribution of O. viverrini in Northern Thailand.

The life cycle of Ascocotyle (Phagicola) longa (Digenea: Heterophyidae), a causative agent of fish-borne trematodosis.

The complete life cycle of the trematode Ascocotyle (Phagicola) longa (Digenea: Heterophyidae) is elucidated by natural observation validated by experimental infections. The natural first intermediate host of A. (P.) longa, an agent of human heterophyiasis in Brazil, is the cochliopid snail Heleobia australis (new first intermediate host). Metacercariae were found encysted in the body musculature, heart, stomach, liver, kidney, spleen, gonads and mesentery of mullets Mugil liza. Hamsters Mesocricetus auratus were experimentally infected with metacercariae of A. (P.) longa obtained from the mullets, and the adults recovered were used to infect the snails H. australis. Rediae and cercariae of A. (P.) longa are described for the first time. The ultrastructure of the tegument of A. (P.) longa shows a change in spination pattern from the cercaria with single-pointed spines to the metacercaria and adult with multipointed, brush-shaped spines. The life cycle of A. (P.) longa is related to estuaries and coastal lagoons where the recruitment of mugilid juveniles occurs. The high prevalence (100%) of A. (P.) longa encysted in the mullets examined within the urban area of Rio de Janeiro indicates the potentially great public health impact of the consumption of raw mullets.