A host-independent role for Fasciola hepatica transforming growth factor-like molecule in parasite development
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Calves were immunized twice in 4 weeks with a Fasciola/Schistosoma cross-reactive, cross-protective defined immunity antigen (denoted Fh SmIII(M) isolated from F. hepatica adult worm extracts by antibody affinity chromatography and challenged 7 weeks later with F. hepatica metacercariae. F1ukes were recovered at 16 weeks of infection at which time the immunized calves had 55% less F. hepatica than the controls. All of the immunized calves developed high antibody levels of Fh SmIII(M) , detectable in the ELISA, by 4 weeks after a single immunization. By 9 weeks of infection with F. hepatica the immunized calves had lower sorbitol dehydrogenase levels than the unimmunized, F. hepatica -infected control calves, indicating less liver damage in the vaccinated group. These studies demonstrate that subcellular F. hepatica macromolecules cross-reactive and cross-protective against S. mansoni also have the potential to serve as vaccines in cattle exposed to this parasitic disease.
Acquired resistance
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Calves were immunized twice in 4 weeks with a Fasciola/Schistosoma cross-reactive, cross-protective defined immunity antigen (denoted FhSmIII(M)) isolated from F. hepatica adult worm extracts by antibody affinity chromatography and challenged 7 weeks later with F. hepatica metacercariae. Flukes were recovered at 16 weeks of infection at which time the immunized calves had 55% less F. hepatica than the controls. All of the immunized calves developed high antibody levels of FhSmIII(M), detectable in the ELISA, by 4 weeks after a single immunization. By 9 weeks of infection with F. hepatica the immunized calves had lower sorbitol dehydrogenase levels than the unimmunized, F. hepatica-infected control calves, indicating less liver damage in the vaccinated group. These studies demonstrate that subcellular F. hepatica macromolecules cross-reactive and cross-protective against S. mansoni also have the potential to serve as vaccines in cattle exposed to this parasitic disease.
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Fasciola hepatica is a liver parasite of ruminants whose distribution is determined by its intermediate host, the freshwater snail Galba truncatula. In Europe, F. hepatica is mostly associated with lowlands. Infection from sympatric domestic reservoirs is rarely reported in wild mountain ungulates. This study explores F. hepatica in a multi-host system in a European alpine area. Serum samples (n = 1,209) from Pyrenean chamois (Rupicapra p. pyrenaica), European mouflon (Ovis aries musimon), domestic sheep (Ovis aries) and domestic cattle (Bos taurus) were collected in the National Game Reserve of Freser-Setcases (NGRFS) in Catalonia, Northeastern Spain, from 2008 to 2019, and tested for antibodies against F. hepatica. During the same period, the livers of 214 chamois hunted in the NGRFS were inspected for F. hepatica and associated pathological changes. Finally, 907 freshwater snails were collected in summer 2016 between 1559 and 2,224 metres above sea level (asl) in the NGRFS, and F. hepatica DNA sought by PCR. Antibodies against F. hepatica were detected in all four species, with a higher prevalence in cattle and sheep than in chamois. Fasciola hepatica and hepatic lesions were concurrently observed in 13/214 of the chamois livers inspected (6.1%, CI95 2.9%-9.3%). Fasciola hepatica DNA was detected in one out of the 907 snails (0.1%, Cl95 0.1% - 0.3%; Ct value 33.3) and collected at 2054 m asl. Fasciola hepatica was consistently detected in a high mountain multi-host system, suggesting that its life cycle is completed and that it occurs endemically at the highest elevation reported in Europe. Transhumant livestock are the likely source in this alpine ecosystem, which according to rare occurrence of F. hepatica DNA in G. truncatula is still a suboptimal habitat for F. hepatica life cycle. Studying parasites at their highest distribution range can be useful to monitor climate change in seasonal mountain environments.
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To determine the intermediate host of Fasciola hepatica in Dali of Yunnan Province, and investigate its development and characteristics.F. hepatica eggs from cattle were collected from July 2012 to July 2013, and placed in 28 degrees C water bath for incubation. Galba pervia, Radix swinhoei, and Physa acuta were collected from Dali, and used to be infected with F. hepatica in the laboratory. Trematode infections were excluded from the snails before experiment. All the snails were infected with F. hepatica miracidia, reared in mud pots. Dead snails were dissected for observing the development of F. hepatica. The metacercariae were collected and identified by PCR amplification of partial sequence of mitochondrial cytochrome oxidase subunit I (COX1) gene.A total of 1 146 R. swinhoei, 996 P. acuta, and 3 307 G. pervia snails were infected with F. hepatica, respectively. Mother rediae were found in two R. swinhoei snails, but no child rediae were observed in the snails. No larval forms were found in P. acuta. G. pervia was infected by F. hepatica with an infection rate of 27.2% (900/3307). The miracidium escaped from the egg and penetrated into G. pervia at temperature 22 degrees C, developed into a sporocyst after 7-15 days, which transformed into mother redia at the 11 st-20th day post-infection. The mother redia developed into daughter redia at the 30th-37th day, and produced cercaria with longtail, and became metacercaria at the 42nd-55th day. PCR confirmed that the metacercariae were that of F. hepatica, with an obvious band (approximately 500 bp).Among the three potential intermediate hosts in Dali, G. pervia is experimentally infected with F. hepatica.
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Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea. Key words: Fasciola hepatica, snail, prevalence, Korea, ITS-1, ITS-2, PCR
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When F. hepatica adult worm antigen is fractionated with Sephadex G-200, four major peaks are obtained. The first peak, when used as antigen in counterelectrophoresis, reacts with the serum of normal rats as well as from rats infected with F. hepatica for 5 weeks or longer. The crest and descending portion of peak II and the third peak are nonreactive with normal rat serum, but highly reactive with the serum of rats with F. hepatica. In addition, F. hepatica antigens cross-reacting with S. mansoni adult worm antisera are absent in these fractions. It is suggested, then, that these fractions of F. hepatica adult worm antigen be utilized for the immunodiagnosis of fascioliasis by CEP. Optimum antigen concentration for this test has been determined to be approximately 1.80 mg/ml protein.
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Fasciola hepatica: Infection Status of Freshwater Snails Collected from Gangwon-do (Province), Korea
Fasciola hepatica is a trematode that causes zoonosis, mainly in cattle and sheep, and occasionally in humans. Few recent studies have determined the infection status of this fluke in Korea. In August 2015, we collected 402 samples of freshwater snails at Hoenggye-ri (upper stream) and Suha-ri (lower stream) of Song-cheon (stream) in Daegwalnyeong-myeon, Pyeongchang-gun in Gangwon-do (Province) near many large cattle or sheep farms. F. hepatica infection was determined using PCR on the nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 402 samples, F. hepatica 1TS-2 marker was detected in 6 freshwater snails; thus, the overall prevalence in freshwater snails was 1.5%. The prevalence varied between collection areas, ranging from 0.0% at Hoenggye-ri to 2.9% at Suha-ri. However, F. gigantica ITS-2 was not detected in the 6 F. hepatica-positive samples by PCR. The nucleotide sequences of the 6 F. hepatica ITS-2 PCR-positive samples were 99.4% identical to the F. hepatica ITS-2 sequences in GenBank, whereas they were 98.4% similar to F. gigantica ITS-2 sequences. These results indicated that the prevalence of F. hepatica in snail intermediate hosts was 1.5% in Gangwon-do, Korea; however the prevalence varied between collection areas. These results may help us to understand F. hepatica infection status in natural environments.
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Fasciola gigantica
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The aim of this study was to determine the effect of an extremely low-frequency magnetic field (ELFMF) on the production of liver fluke larvae in a parasite-host system: Fasciola hepatica--Galba truncatula. Both F. hepatica eggs and F. hepatica-infected snails were exposed to an ELFMF (50 Hz, 2.0 mT) for 14 days and 36 days, respectively. F. hepatica-infected snails were divided into 4 groups, 10 specimens each. The snails of groups I and II were infected with F. hepatica larvae--miracidia obtained from control cultures, while the snails of groups III and IV were infected with miracidia reared from eggs that had been incubated in an ELFMF. After infection, the snails of groups II and IV were placed in an ELFMF, while those of groups I (control) and III were housed outside the ELFMF. At 36 days post-infection (dpi) there were no statistically significant differences between the number of F. hepatica larvae--cercariae and metacercariae, obtained from G. truncatula snails in the control group (group I) and the snail groups exposed to ELFMF (groups II, III and IV). However, a statistically significant difference between the average number of F. hepatica larvae in snail groups III and IV may indicate that the duration of exposure to ELFMF, i.e. embryogenesis period vs. the entire larval development, played a role in the production of F. hepatica larvae, and resulted in a reduction of their number.
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Fasciola
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