Background & aim: Parasitological methods for diagnosing fascioliasis have limitations such as,Intermittent excretion of parasite eggs is the long stage of presence of parasite eggs in the feces. Also, serological methods based on the detection of antiparasitic antibodies are not able to distinguish past infections from new infections, so the detection of parasitic co-antigens is more reliable than other diagnostic methods. Therefore, the aim of the present study was to determine and detect Fasciola coproantigens using polyclonal antibodies produced against Fasciola hepatica and Fasciola gigantica secretory excretory antigens. Methods: In the present experimental study conducted in 2016-2017, adult Fasciola worms were collected from the livers of infected animals and species were determined by molecular method of enzymatic digestion. Excretory-secretory antigen was prepared from both Fasciola species. In order to produce polyclonal antibodies, 2 rabbits were immunized for each antigen. IgG antibodies produced were purified by affinity chromatography and conjugated with peroxidase enzyme. Coproantigens from 99 animal fecal samples included,30 samples infected with Fasciola, 39 samples with other parasitic diseases and 30 healthy samples were prepared. In the present study, two ELISA sandwich systems were designed and evaluated using polyclonal antibodies produced to identify Fasciola co-antigen. Statistical characteristics of the tests were calculated using Medical Collector 16. 4. 3 software. Kappa coefficient and consistency check between ELISA sandwich tests were calculated using SPSS software version 20. Results: Sandwich ELISA test using polyclonal antibodies produced against Fasciola hepatica and Fasciola gigantica secretory antigens showed 96. 67%, 89. 86% and 96. 67% and 91. 3%, respectively. Moreover, a high agreement (kappa coefficient <0. 9) was observed between polyclonal antibodies produced against both Fasciola species for the diagnosis of fascioliasis. Conclusion: Overall, the findings indicated that polyclonal antibodies produced against excretory-secretory antigens of both Fasciola species have good efficacy for detection of Fasciola co-antigens and diagnosis of fascioliasis.

To determine the electrophoretic patterns of Fasciola hepatita and F gigantica, the former was collected from sheep and buffalo infected liver and the latter from that of cattle and camel and were washed in PBS. Somatic antigens of both species were prepared though homogenization and centrifugation of adult flukes. To show the eclrophoretic patterns of somatic antigens, the prepared antigens were electrophoresed using SDS-PAGE. Our findings revealed that proteins of 18-70 kDa molecular weights were present in both species in different hosts. Although a 57 kDa protein was seen in Fasciola gigantica of cattle and camel only. Meanwhile two bands of 54 and 68 kDa were seen exclusively in Fhepatica of sheep and buffalo. A band of 64 kDa was shown in F.hepatica of buffalo. According to our results in any study on immunological or immunodiagnostic of Fasciola spp, the species trematode as well as their hosts should be taken in consideration.

Background: Fasciolosis is a worldwide disease with major economic and public health consequences. Early detection of the infection is important for the prevention and control of the disease. ELISA allows for early detection of fasciolosis in man and animals. Fasciolosis is caused by Fasciola hepatica and F. gigantica in man and domestic animals respectively. These two species have many similar morphological characteristics. In this study, the crude antigens of these two species are investigated by ELISA test. Methods: The excretory-secretory and somatic antigens of two species were prepared from adult flukes collected from the bile ducts of sheep and stored at -20oc. For the preparation of the antisera, the antigens were injected to laboratory-bred rabbits. Each rabbit received five injections at intervals of seven days, starting with 0.5 ml and ending with 2.5 ml. Ten days after the last injection, the rabbits were bled, and serum samples separated and stored at -20oc. The reaction between homologous and heterologous antigens and antisera was tested by ELISA and optical densities were recorded. Results: Excretory- secretory and somatic antigens of each species showed a strong positive reaction with the antisera of the other species. In a homologous combination of antigens and antisera, a stronger reaction was observed compared to the heterologous combination, therefore many antigenic materials of both species are the same.Conclusion: The differences of these crude antigenic materials of F. hepatica and F. gigantica are insufficient to prevent cross reaction of two species by ELISA. Further investigations are recommended for the identification, detection and purification of antigenic material of each species to improve the specificity of this assay.

Background: Fascioliasis is a parasitic disease caused by Fasciola hepatica, Fasciola gigantica, and intermediate Fasciola, which is present in a wide variety of mammalian species, including humans, throughout the world. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method is one of the best methods for detecting Fasciola species in places where there are two types of Fasciola hepatica and Fasciola gigantica in domestic livestock. Methods: In this study, Fasciola worms were collected from slaughterhouses in five western provinces of Iran. Parasitic species were identified using morphological and molecular methods. A total of 756 livestocks (cattle and sheep) were studied, with 89 livers infected with adult Fasciola worms. 18 worms were morphometrically examined, and DNA from the eggs of these worms were extracted. A fragment of genome containing the ITS1, 5. 8S, ITS2 gene was amplified and utilized by TasI enzyme, and PCR-RFLP was performed on amplified parts. Findings: A total of 178 adult Fasciola worm were isolated from infected cattle and sheep livers. During the spring 2017, the highest and the lowest levels of contamination were reported in Ilam Province (sheep, 28. 53%) and Kermanshah Province (sheep, 7. 9%), Respectively. In Fasciola hepatica, TasI enzyme produced three fragments of 427, 360 and 117 base pair (bp), and for Fasciola gigantica, it produced three fragments of 360, 220, and 117 bp on 1. 5% agarose gel. Conclusion: According to the study, microscopic measurements can be sufficient for microscopic characterization of Fasciola species, but the PCR-RFLP method using TasI enzyme is simpler, faster, and more accurate. The use of PCR-RFLP method and ITS genetic marker is very suitable for identification of Fasciola species.

Human fascioliasis due to unknown species and animal fascioliasis caused by both or one of Fasciola spp. are commonly seen in Iran. To compare electrophoretic patterns of somatic and excretory-secretory antigens of F. hepatica and F. gigantica by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), the adult flukes were collected from infected slaughtered bovine livers. E/S and somatic antigens were prepared by incubation and homogenizing of adult flukes, respectively. The antigens were electrophoresed using SDS-PAGE. Following SDS-PAGE, E/S proteins of F. hepatica and F. gigantica were characterized by the presence of 6 common major peptide bands with molecular weights of 15, 16, 20, 24, 33 and 42 kDa. Differences between F. hepatica and F. gigantica somatic proteins were noticed. F. gigantic had 11 major protein bands with molecular weights of 18, 22, 24, 33, 36, 42, 46, 57, 60, 62 and 68 kDa, whereas F. hepatica had proteins characterized by 8 distinct bands with molecular weights of 18, 22, 24, 33, 36, 42, 46 and 62 kDa.

Introduction: Successful development of free-living stages of parasitic helminths depends on larva ability to survive, develop, and hatch. In this study, we aimed to study the host role in the hatching process of Fasciola species. Methods: Fasciola hepatica and Fasciola gigantica eggs were collected from adult worms that originated from naturally infected sheep and cattle livers and were incubated at 26± 1° C for 15 days. The percentage of hatched and developed eggs were obtained for each isolate under a light microscope. A polymerase chain reaction followed by restriction fragment length polymorphism (PCRRFLP) was applied to identify the F. hepatica and F. gigantica species. Results: Our findings showed no significant differences in the development rates of F. gigantica and F. hepatica eggs in sheep (69. 32% and 72. 71%) and cattle (73. 56% and74. 69%). However, the rates of hatched eggs of F. gigantica and F. hepatica originated from cattle (69. 19% and 62. 36%) were almost twice the rates in sheep (31. 69% and 32. 59%), indicating a significant difference. Conclusion: This study demonstrated that host species significantly affect the hatching of Fasciola eggs as the hatching rates of F. gigantica and F. hepatica originated from cattle were higher than those taken from sheep did not affect their larval development. Thus, in addition to environmental factors, the hatching phenomenon is influenced by host species.

Background and purpose: Fasciola hepatica and Fasciola gigantica are common liver flukes which affect both human and livestock worldwide. In this study we evaluated the loop-mediated isothermal amplification (LAMP) assay for detection and discrimination of Fasciola species.Materials and methods: Fifty adults of Fasciola worms were isolated from sheep and cattle liver form abattoirs in Mazandaran province. A total of 8 primer sets for LAMP was designed to amplify the 28S ribosomal RNA gene of Fasciola sp. Conventional LAMP was carried out in a 20mI reaction mixture under isothermal condition at 60oC for 90 minutes. Amplification result was observed by monitoring the turbidity by naked-eye, using fluorescent dye and gel electrophoresis. The specificity of LAMP method for detecting Fasciola sp. was tested by amplification of Dicrocoelium dendriticum, Trichostrongylus colubriformis, and Echinococcus granulosus DNA templates. To evaluate the detection limit of LAMP assay in detecting Fasciola genus, serial dilution of the extracted DNA was used.Results: A positive LAMP reaction by the specific primers of two species produced many bands of different sizes in 600C after 90 min. The optimal assay conditions were established with no reaction with other parasites’ DNA. The detection limit of this LAMP assay was 1 pg DNA/tube. The result of turbidity and fluorescent dye detection were consistent with agarose gel electrophoresis. Conclusion: Our results demonstrated that LAMP is a rapid, cost-effective, highly specific, easy, and reliable method for differentiation of Fasciola sp. in epidemiological and clinical researches on human and domestic animals in endemic regions of fasciolosis.

The patient was a 5-year-old boy with malaise, abdominal discomfort, intermittent fever and vomiting since two years ago. Complete blood count was relatively normal except for mild eosinophilia Open Access What is your diagnosis? See the next page for your diagnosis. (white blood cells = 8. 05 ×106/L, hemoglobin = 116 g/L, platelet = 464×106/L, neutrophil = 42. 8%, lymphocyte = 43%, monocyte = 8. 1%, eosinophil = 5. 7%, basophil = 0. 4%)....