In vitro development of Haplorchis taichui (Trematoda: Heterophyidae)

Newly excysted metacercariae of Haplorchis taichui were cultured in a candle jar set at 37 degrees C. Both monophasic culture media [0.85% NaCl, RPMI 1640, RPMI 1640+10% fetal calf serum (FCS)] and diphasic culture media [RPMI 1640 + egg yolk agar, RPMI 1640 + 5%, 10% or 15% blood in blood agar (BA), RMPI 1640 + 5%, 10% and 15% FCS with 5% blood in BA] were used in vitro. Parasites survived for only 1 day in 0.85% NaCl without any development. In RPMI 1640 with egg yolk agar and RMPI 1640 + 5%, 10% FCS, the parasite survived for 3-5 days. In contrast, worms survived for 12-14 days in RPMI 1640 with blood agar without any change in result in a different concentration of blood in BA. The ovary and testes were observed after 3 days incubation in this media. Nevertheless, only 1 parasite in RPMI 1640 with 15% blood in BA had vitellaria and eggs at day 6. RPMI 1640 with blood agar can be used as short-term maintenance for the in vitro culture of H. taichui. However, further studies are needed.     

In vitro excystation of Haplorchis taichui (Trematoda: Heterophyidae)

The effects of trypsin, bile extract, temperature and acid-based condition for the in vitro excystation of Haplorchis taichui metacercariae were studied. At 37 degrees C, approximately half the number of metacercariae excysted when exposed to 1% trypsin for 15 minutes with no more excystation found beyond this time. Increasing trypsin concentration seemed to reduce the excystation rate while bile extract was, however, unlikely to be an absolute requirement. A temperature of 37 degrees-41 degrees C yielded a similar excystation result in combination with 1% trypsin; however, less excystation occurred at a lower temperature of 35 degrees C. The acid-based environment of pH 8 gave the best excystation result in association with 1% trypsin at a temperature of 39 degrees C. Higher and lower basicity produced a smaller excystation rate. An environmental condition of 1% trypsin at pH 8 and a temperature of between 37 degrees-41 degrees C was recommended for the in vitro excystation of H. taichui metacercariae. The relatively broad temperature and pH range condition for the excystation of H. taichui corresponded with various definitive hosts that were infected naturally by this fluke.     

Recovery and growth of Haplorchis taichui (Trematoda: Heterophyidae) in chicks

An experimental study was performed to observe the recovery and growth of a minute intestinal fluke, Haplorchis taichui in chicks (Gallus gallus domesticus). Metacercariae of H. taichui were isolated from Jullien's mud carp, Henicorhynchus siamensis, which were collected in the Chiang Mai Province, Thailand. Two hundred metacercariae were orally force-fed to each chick. The intestine of the chicks were examined from day 1 to day 54 post-infection (PI). The incidence of infection was 84.9% (28/33) and the mean intensity was 19.9 (656/33), with the range 0-59. The worm recovery rate was the highest at day 11 PI (29.5%). On day 3 PI, mature adult worms were recovered and 1-200 eggs were observed in the uterus of the worms. The worms grew rapidly in the chicks and the genital organs were fully developed in 14 days. This parasite can survive in chicks up to day 48 PI. It is concluded that they are a suitable definitive host for infection with H. taichui.     

Development of Haplorchis taichui (Trematoda: Heterophyidae) in Mus musculus mice

The development of Haplorchis taichui was studied in sixteen mice, Mus musculus. Metacercarial cysts of H. taichui were obtained from the freshwater fish, Thynnichthys thynnoides, collected in Chiang Mai Province, north Thailand. Approximately 200 active metacercariae were orally introduced into each mouse. Two mice were randomly sacrificed and necropsied daily from day 2-9 postinfection (pi). Two peaks of increment in the length and width of worms were found at day 3 and 7 pi. H. taichui was rapid in maturation, similar to other minute intestinal flukes. Rudimentary sex organs were found at day 2 pi. Spermatozoa in a seminal receptacle, vitellaria and eggs were seen as early as 3 days pi. The number of eggs increased daily to approximately 50-60 eggs thereafter. Mice can serve as a suitable experimental definitive host for harvesting adult H. taichui, especially in 1-week pi.     

Epidemiology of food-borne trematodes and other parasite infections in a fishing community on the Nam Ngum reservoir, Lao PDR

Food-borne trematodes (FBT) are important causes of parasitic infections in many Asian countries. Parasitological surveys in Xai Udom, a small fishing community on the Nam Ngum reservoir, Lao People's Democratic Republic, revealed an overall parasitic infection rate in May 1999 of 68.8% (n = 173) and in December 1999 of 65.9% (n = 261). The liver fluke, Opisthorchis viverrini accounted for most of the infections (prevalences of 53.8% and 42.1%, during the first and second surveys, respectively). The prevalence and intensity showed increasing trends with age. Minute intestinal flukes were also present but with relatively low infection rates (3.8-10.9%). The second common group of parasites comprised soil-transmitted nematodes, Trichuris trichiura, Ascaris lumbricoides, hookworm and Strongyloides stercoralis, with prevalences of 22.4 and 17.6%, 20.8 and 8.0%, 16.8 and 13.4%, and 4.0 and 15.3% (first and second surveys, respectively). Most people had no or only light infections, with a few people having heavy infections. Coexisting intestinal protozoa were Giardia lamblia (5.2 and 4.9%) and Entamoeba coli (6.9 and 6.5%). Concurrent tapeworm infections were Taenia (1.7 and 1.1%) and Hymenolepis nana (0.7 and 0.6%) (first and second surveys, respectively). Despite the availability of effective anthelmintic drugs, the results of our study reaf-firm that FBT are still a major health threat in this fishing area of Lao PDR, similar to a decade ago.     

Melanoides tuberculata (Mollusca: Thiaridae) as an intermediate host of Centrocestus formosanus (Trematoda: Heterophyidae) in Brazil

Pleurolophocercous cercariae emerged from naturally infected Melanoides tuberculata from Minas Gerais State, Brazil, were used to perform experimental infection of laboratory-reared Poecilia reticulata. Mature metacercariae were obtained from the gills of fishes and force-fed to Mus musculus. The adult parasites which recovered from small intestines of mice were identified as Centrocestus formosanus. This is the first report of M. tuberculata as intermediate host of this heterophyid in Brazil.     

Melanoides tuberculatus (Gastropoda: Thiaridae) as intermediate host of Heterophyidae (Trematoda: Digenea) in Rio de Janeiro metropolitan area, Brazil

In the late 1960s, Melanoides tuberculatus snails were introduced in Brazil from North/East Africa and Southeast Asia. The first records of specimens infected with cercariae were registered in Rio de Janeiro State in 2001. The present study reports the occurrence of M. tuberculatus infected with larval trematodes in Rio de Janeiro City. Bottom sediment was collected with dip nets and sieved through 0.25 inch-mesh screening. Snails were transported to the laboratory in vials with stream water, then measured and individually isolated in glass vials with distilled water. They were exposed to artificial light and temperature to induce cercarial emergence. The most actively emerging cercariae were processed by differential staining and silver nitrate impregnation methods. Negative snails were subsequently dissected. Approximately 700 snails were collected. Snail total lengths ranged from 1.2 to 3.3 cm. The prevalence rate was 15.76% although 53.76% of the snails were found infected in one of the sites. Infected snails were infected with rediae and pleurolophocercous cercariae. Cercarial morphology and chaetotaxy were consistent with those of the family Heterophyidae mostly due to the presence of median dorsal and ventral fins on the tail and the absence of CI dorsal sensory receptors.     

Study of the body surface of Haplorchis yokogawai (Katsuta, 1932) and H. taichui (Nishigori, 1924) (Trematoda:Heterophyidae).

The morphology of the surface of two heterophyid flukes, Haplorchis yokogawai and H. taichui, that can infect humans eating raw fish flesh, has been studied using scanning and transmission electron microscopy. The body surface of both species is covered with scale-like, serrated spines with the exception of areas around the oral sucker, ventrogenital opening and excretory pore. The species slightly differ by the dentation pattern of tegumental spines: H. yokogawai has finer and more numerous spine teeth (maximum tooth number 14-16) while H. taichui has more serrated spines, mostly with a lower number (up to 10-12) of somewhat more robust teeth. However, because of the variability, the shape and dentation of spines do not seem to be reliable diagnostic features enabling mutual differentiation of both species.     

Human parasitism by Phagicola sp (Trematoda, Heterophyidae) in Cananéia, S?o Paulo state, Brazil

We report one case of parasitism by Phagicola sp. (Trematoda, Heterophyidae) in a 31 year-old woman who, in 1987, travelled and stayed several months in the municipality of Cananéia (SP), where she ingested, in various occasions, raw mullet (Mugil sp.). The patient referred mild intestinal pain and laboratory examinations showed eggs of Phagicola sp. in the stools and a slight increase in eosinophil blood levels (8%). After treatment with praziquantel (75 mg/kg per day for three days) all the symptoms and signs disappeared. This is, certainly, the first record of human infection by Phagicola sp. in Brazil and, perhaps, in countries other than the U.S.A. where unclear references to a few human cases were reported in the South-eastern region.     

Phylogenetic analysis reveals cryptic species diversity within minute intestinal fluke,Stellantchasmus falcatus Onji and Nishio,1916(Trematoda,Heterophyidae)

Objective:To examine the taxonomic boundaries in the Stellantchasmus populations in different hosts and reveal the cryptic speciation of the Stellantchasmus in Thailand based on both morphological and molecular approaches.Methods:Stellantchasmus falcatus(S.falcatus) s.l.was collected from different hosts throughout Thailand.The worms were examined and measured.The nuclear ITS2 gene and the mitochondrial COI gene were used to investigate the S.falcatus s.l.from Dermogenus pusillus and Liza subviridis.Results:Stellantchasmus was one of food-borne trematode that widely distribute throughout Thailand.The comparison of S.falcatus s.l.from Dermogenus pusillus and Liza subviridis indicated a genetic divergence of nuD NA with 3.6% and mtD NA with 19.3%,respectively.Morphological characteristics were fairly different in the ratio of body size,length of prepharynx,and ratio of organ size.Conclusions:Our results provide initial evidence that S.falcatus s.l.from different hosts tend to be a different species based on both molecular and morphological characters.Cryptic species complexes are generally found among parasites that tend to have large populations and/or rapid evolution.The degree of genetic diversity existing would suggest the practice of targeted regimes to design and minimize these lections of anthelmintic resistance in the future.     

Human infection by Phagicola sp. (Trematoda, Heterophyidae) in the municipality of Registro, S?o Paulo State, Brazil.

In 1988, nine cases of human parasitism by Phagicola sp. were diagnosed in the municipality of Registro (S?o Paulo State, Brazil) by stool examinations, in patients who ate raw mullet (Mugil sp.). Six (66%) of the nine patients suffered from flatulence and four (44%) had diarrhoeal episodes; six (66%) showed slight eosinophilia. On the same occasion, 61 dogs and 11 cats from Registro were also submitted to stool examination. Only one dog (1.6%) showed Phagicola eggs in the stool. All patients were treated with a single dose of praziquantel (50 mg kg-1 body weight) and control stool tests performed on the 15th, 30th and 60th days post-treatment showed no trematode eggs.     

Fauna and abundance of medically important flies of Muscidae and Fanniidae (Diptera) in Tehran, Iran

ObjectiveTo determine the faunal diversity of Muscidae and Fanniidae flies in Tehran, Iran.MethodsA net-capturing by direct observing method and fly trap were used to capture adult flies. To determine the fauna of flies in different habitats, 4 biotopes including corpse (human, birds, livestock), garbage and decaying organic matters, animal carcasses and human indoor habitat were selected. Big hashing nets (95 cm in diameter) have been used for adult flies capture in these biotopes.ResultsIn this study, totally 2 418 adult flies from 8 families including Muscidae, Fanniidae, Calliphoridae, Sarcophagidae, Tachinidae, Syrphidae, Anthomyiidae and Conopidae were captured. Among these, 1 279 belong to Muscidae and Fanniidae families. Four genera and 5 medically important species captured from two above mentioned families.ConclusionsBased on scientific documentation, 2 species of Muscina stabulans (M. stabulans) and Fannia scalaris (F. scalaris) are reported for the first time in Iran. However M. stabulans is a cosmopolitan species and its presence in Iran was probable.     

Evolutionary origins, diversification, and biogeography of liver flukes (Digenea, Fasciolidae)

Fasciolid flukes are among the largest and best known digenetic trematodes and have considerable historical and veterinary significance. Fasciola hepatica is commonly implicated in causing disease in humans. The origins, patterns of diversification, and biogeography of fasciolids are all poorly known. We have undertaken a molecular phylogenetic study using 28S, internal transcribed spacer 1 and 2 (ITS-1 and ITS-2) of nuclear ribosomal DNA, and mitochondrial nicotinamide dehydrogenase subunit 1 (nad1) that included seven of the nine recognized species in the family. The fasciolids examined comprise a monophyletic group with the most basal species recovered from African elephants. We hypothesize fasciolids migrated from Africa to Eurasia, with secondary colonization of Africa. Fasciolids have been conservative in maintaining relatively large adult body size, but anatomical features of their digestive and reproductive systems are available. These flukes have been opportunistic, with respect to switching to new snail (planorbid to lymnaeid) and mammalian hosts and from intestinal to hepatic habitats within mammals.     

Mekong schistosomiasis. III: a parasitological survey of domestic water buffalo (Bubalus bubalis) on Khong Island, Laos.

Of 103 water buffalo (Bubalus bubalis) examined on Khong Island by means of the M.I.F.C. and hatching techniques, none were passing eggs resimbling those of the Mekong schistosome. One buffalo calf was infected with Orientobilharzia harinasutai and another with Schistosoma spindale; this is the first time these parasites have been reported from Laos. Since the buffalo that were examined had constant and year-round access to a part of the Mekong River that has been shown to be a site of heavy transmission of schistosomiasis to humans and dogs, it was considered that the buffalo would have acquired the infection with the human Mekong schistosome if this were possible. In the absence of buffalo necropsies, and since no eggs of the Mekong schistosome were detected in the stools of these animals, we assumed that they had either not become infected with this parasite or, if they had, that the infections did not produce eggs in the faeces which were detectable by the methods employed. On the basis of our examinations, it would not seem that domestic water buffalo are involved as reservoirs in the epidemiology of human schistosomiasis on Khong Island.