Identification of circulating antibodies in fasciolosis and localization of 66 kDa antigenic target using monoclonal antibodies.
We identified three specific circulating antibodies in serum of cattle naturally infected with Fasciola gigantica. Two of the antibodies were found to react specifically to 97 and 66 kDa antigenic molecules of adult worm tegumental membrane extract. The third antibody was identified by the reaction with 26-28 kDa molecule of the excretory/secretory antigens. Monoclonal antibody against 66 kDa protein was developed and used for localization of its antigenic target in adult worm frozen sections. The experiment demonstrated that 66 kDa protein is a component on the outer surface membrane and on the membrane lining of the caecal epithelial of adult worm. The 66 kDa antigen was considered as a promising candidate for immunodiagnosis and vaccine. (+info)
Fasciola gigantica-specific antigens: purification by a continuous-elution method and its evaluation for the diagnosis of human fascioliasis.
Immunodominant antigens of an approximate molecular mass of 27 kD were obtained from an excretory-secretory product of adult Fasciola gigantica by a continuous-elution method. An indirect ELISA using the antigens obtained by this relatively simple procedure was developed for detecting specific antibodies from patients infected with F. gigantica. Sera from patients with other parasitic infections, healthy volunteers, and cholangiocarcinoma were also analyzed. The sensitivity, specificity, and positive and negative predictive values for this ELISA using the fractionated antigens were 100%. The data indicated a possible correlation of antibodies to F. gigantica with cholangiocarcinoma. (+info)
Detection of circulating E/S antigens in the sera of patients with fascioliasis by IELISA: a tool of serodiagnosis and assessment of cure.
An IELISA was developed to evaluate the performance of Fasciola E/S antigens in diagnosis and cure assessment of human Fasciola infection. Twenty patients with acute (prepatent) fascioliasis and another 20 with patent infection were enrolled in the study. Patients were treated with TCZ and followed at 1, 3 and 6 months after therapy. At inspection, the sensitivity of the test to diagnose prepatent cases was 100% compared to 70% for patent infections. There was a gradual decrease of antigenaemia over the follow-up period in acute cases. In chronic cases antigen disappeared from 13 cases (65%) at 1 month; this proportion did not change at 3 or 6 months. (+info)
Parasite-specific antibody profile in human fascioliasis: application for immunodiagnosis of infection.
The antibody isotype response to an adult Fasciola worm antigen preparation (FWAP) was examined in sera from 60 Egyptians with parasitologically confirmed fascioliasis by an ELISA. The FWAP-specific IgG1 and IgG4 antibodies were found in 97-100% of the patients. The ratio of the mean absorbance values between infected patients and healthy controls was 9.7 and 29.7 for IgG1 and IgG4 antibodies, respectively. The IgM, IgA, IgG2, and IgG3 antibodies were less dominant. In contrast to IgG1 antibodies, which were often detected in sera from patients infected with Schistosoma, Echinococcus granulosus, Ascaris lumbricoides, Ancylostoma duodenale, or Hymenolepis nana, FWAP-specific IgG4 antibodies were detected exclusively in the sera of patients with fascioliasis. The data thus support the conclusion that an IgG4/ELISA with crude FWAP as antigen may be used for sensitive and accurate immunodiagnosis of human fascioliasis. (+info)
Comparative study of the reproductive organs of Fasciola groups by optical microscope.
Reproductive organs of stained and mounted whole specimens of different types of Fasciola (F. hepatica, F. gigantica, and parthenogenetic diploid and triploid flukes) were observed to clarify the structure of their reproductive organs. The results are as follows; 1. Basic structure differences could not be identified. 2. The flukes without sperm, or those with an extremely small quantity in the seminal vesicle. are parthenogenetic Fasciola sp. 3. It was newly discovered that the surface of the cirrus is surrounded by many shallow gutters, and that spines form a line in the gutters. 4. The structure of the reproductive organ on the genus Fasciola are shown in detail in the figures. (+info)
Preliminary study of therapeutic efficacy of a new fasciolicidal drug derived from Commiphora molmol (myrrh).
Myrrh (from the stem of the Commiphora molmol tree) is an oleo gum resin that may prove efficacious for the treatment of fascioliasis. We studied 7 patients who were passing Fasciola eggs in their stools and treated them with myrrh. The drug (a formulation consisting of 8 parts of resin and 3.5 parts of volatile oils, all extracted from myrrh) was given in a dose of 12 mg/kg per day for 6 consecutive days in the morning on an empty stomach. Patients were followed for 3 months. The therapy proved to be effective, with pronounced improvement of the general condition and amelioration of all symptoms and signs. A dramatic drop in the egg count was detected at the end of treatment. Eggs were no longer detectable in the feces 3 weeks after treatment and after a follow-up period of 3 months. High eosinophilic counts, elevated liver enzymes, and Fasciola antibody titers returned to nearly normal. No signs of toxicity or adverse effects were observed. We conclude that the formulation of myrrh is safe, well tolerated, and effective for treating fascioliasis. (+info)
The influence of different food sources on cercarial production in Lymnaea truncatula experimentally infected with Digenea.
Experimental infections of Lymnaea truncatula with a digenean species (Fasciola gigantica, F. hepatica, or Paramphistomum daubneyi) were performed under laboratory conditions to study the effect of four sources of food (microalgae, romaine lettuce, wheat germs, or modified Boray diet) on cercarial production. The mean number of F. hepatica, F. gigantica, or P. daubneyi cercariae was significantly greater when snail food was microalgae or Boray diet. The lowest cost prices for 100 metacercariae of F. hepatica (9.7-10.1 euros) were noted when algae of Boray diet were used as food for snails. In contrast, the highest prices (18.0-18.5 euros) were found in lettuce-reared snails. The choice of a source of food for breeding the intermediate hosts of F. hepatica and collecting metacercariae must take into account the importance of the commercial demand for metacercariae and will depend on the strategy the producer wishes to develop. (+info)
Production and characterization of a monoclonal antibody against recombinant glutathione S-transferase (GST) of Fasciola gigantica.
A monoclonal antibody (MoAb) against a recombinant glutathione S-transferase (rGST) of F. gigantica was produced in BALB/c mice. Reactivity and specificity of this monoclonal antibody was assessed by ELISA and immunoblotting. Six stable clones, namely 3A3, 3B2, 3C6, 4A6, 4B1 and 4D6 were obtained, All these MoAb reacted with rGST and native GST at a molecular weight of 28 kDa and found to be IgG1, kappa-light chain isotypes. These MoAb cross-reacted with Schistosoma mansoni and Schistosoma japonicum antigens at molecular weights of 28 and 26 kDa, respectively, but no cross-reactions were detected with antigens of Eurytrema and Paramphistomum spp. The localization of GST in metacercaria, 7-week-old juvenile and adult F. gigantica was performed by immunofluorescence technique, using MoAb as well as polyclonal antibody (PoAb) to the native protein as probes. In general, all clones of MoAb gave similar results and the pattern was quite similar to staining by PoAb. The fluorescence was intense, which implied the presence of a high concentration of GST in the parenchymal tissue in all stages of the parasite. However, the parenchymal cells were not evenly stained which implied the existence of subpopulations of this cell type with regard to GST production and storage. In addition, in adult and juvenile stages a moderate fluorescence was present in the basal layer of the tegument, while light fluorescence was observed in the caecal epithelium, cells in the ovary, testis and vitelline gland of the adult. In the metacercaria stage, in addition to parenchymal tissue, the tegument and tegumental cells were stained relatively more intense with MoAb and PoAb than in other stages. (+info)