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PubMed1996

Detection of circulating Fasciola gigantica antigen in experimental and natural infections of sheep with fasciolosis.

Detection of circulating Fasciola gigantica antigen was performed in sera of sheep with experimental and natural F. gigantica infections using the direct enzyme-linked immunosorbent assay method. Sera from sheep with monoinfections of Schistosoma bovis, Dicrocoelium hospes and Paramphistomum microbothrium were included in the assay to ascertain specificity. Circulating F. gigantica antigen (CFA) was detected as early as 1 week after infection in the experimentally infected sheep. No detectable CFA was observed 2 weeks after chemotherapy. Positivity rates of 82.5%, 12.5%, 10% and 10% were found in sera with monospecific infections of F. gigantica, P. microbrothrium, D. hospes and S. bovis, respectively. Acid treatment of the sera did not enhance the sensitivity of the assay.

Animals↗
PubMedDate not supplied

Promutagen activation in parasite-infected organisms: preliminary observations with Fasciola hepatica-infected mice and aflatoxin B1.

Mutagenic activity of aflatoxin B1 (AFB1) was followed in the Ames Salmonella/microsome test using liver S-9 from uninfected and Fasciola hepatica infected mice. The S-9 preparations from parasite-infected animals were capable of inducing significantly greater AFB1 mutagenic activity to strain TA100 than S-9 preparations from either control, uninfected mice or Aroclor 1254-induced mice. The differences in activity between infected and uninfected mice were consistent between sex and strains of mice studied. The observed increase in biological activity was apparently due to an induction phenomenon in the host tissues rather than due to metabolism of AFB1 by F. hepatica enzymes.

Aflatoxin B1↗
PubMed1996

Molluscicidal activity of neem (Azadirachta indica A. Juss).

Molluscicidal property of Azadirachta indica A. Juss (neem) against the snails Lymnaea acuminata and Indoplanorbis exustus was studied. It was observed that the molluscicidal activity of the leaf, bark, cake, neem oil and the neem-based pesticides, achook and nimbecidine, was both time- and dose-dependent. The toxic effect of pure azadirachtin against both the snails was greater than the synthetic molluscicides.

Animals↗
PubMed1991

Quantification and partial characterisation of regulatory peptides in the liver fluke, Fasciola hepatica, from different mammalian hosts.

1. Extracts of the liver fluke, Fasciola hepatica from three different hosts (cow, sheep, rat) have been subjected to radioimmunoassay using antisera to 6 mammalian regulatory peptides. 2. Immunoreactivity was measured to pancreatic polypeptide, substance P, peptide histidine isoleucine and gastrin-releasing peptide. Levels of each peptide varied considerably in flukes from different hosts. 3. Reverse-phase HPLC of rat and sheep fluke extracts revealed three molecular forms of tachykinin immunoreactivity and single peaks of pancreatic polypeptide and peptide histidine isoleucine immunoreactivity. No GRP-immunoreactivity was detected by RIA of HPLC fractions.

Animals↗
PubMed2004

Human fasciolosis in Van province, Turkey.

This investigation was carried out to determine the frequency of the fasciolosis in Erciş, a town in the Van province located in the Eastern Turkey. The study includes 500 asymptomatic subjects (322 males and 178 females) whose ages ranged from 5 to 75 years. In all subjects, stool samples were examined at initiation of the study and one week later to rule out pseudoparasitosis. The methods of flotation (in saturated saline solution) and sedimentation (in formalin-ether solution) were used in examination of the stool samples. Fasciola hepatica eggs were detected in nine (1.8%) out of 500 subjects. In conclusion, our data showed that the prevalence of human fasciolosis was high in our region because watercress consumption was common. We also think that human fasciolosis is more frequent than expected in Turkey. To determine the exact prevalence of the disease, extensive serological investigations should be performed in different regions of Turkey.

Adolescent↗
PubMed2004

Detection and genetic distance of resistant populations of Pseudosuccinea columella (Mollusca: Lymnaeidae) to Fasciola hepatica (Trematoda: Digenea) using RAPD markers.

Twelve natural populations of Pseudosuccinea columella snails, sampled in the western and central regions of Cuba, were analyzed using the RAPD-PCR technique to screen for resistance to Fasciola hepatica. Ten OPA primers previously shown to produce marker bands for resistance and susceptibility were tested. A new population of P. columella (El Azufre, Pinar del Río) exhibited the amplification patterns of resistant snails, and its resistant status was confirmed after experimental exposure to miracidia. No genetic variability was detected across or within the susceptible isolates. Similarly, the novel resistant isolate displayed an RAPD profile identical to the profile of two other isolates previously identified as resistant to F. hepatica. However, clear differences in RAPD banding patterns and genetic distance were observed between resistant and susceptible isolates.

Animals↗
PubMed2006

Immuno-diagnosis of bubaline fasciolosis with Fasciola gigantica cathepsin-L and recombinant cathepsin L 1-D proteases.

Fasciola gigantica cathepsin-L cysteine proteinase and recombinant cathepsin L 1-D were assessed for their potential in the immuno-diagnosis of F. gigantica infection in buffaloes. A diagnostic ELISA, based on these two antigens, was developed to detect antibodies against F. gigantica in water buffaloes. Sensitivity of the ELISA was assessed using sera from buffaloes experimentally or naturally infected with F. gigantica from F. gigantica endemic areas and its specificity by probing the sera of the host from F. gigantica non-endemic area. Our earlier studies under experimental setting showed 100% sensitivity of cathepsin-L ELISA in the diagnosis of fasciolosis in buffaloes, with the earliest detection of infection at 4 weeks post-infection. However, under field situation of natural F. gigantica infection, this sensitivity declined to 97.1% but specificity of the test remained 100%. Cross-reactivity of the antigen was checked with Schistosoma indicum, S. spindale, Paramphistomum epiclitum, Gastrothylax spp., Gigantocotyle explanatum, hydatid and Strongyloides papilossus in the bubaline host, naturally infected with these helminths. F. gigantica cathepsin-L and the recombinant cathepsin L-1D does not cross-react with these helminth parasites in natural mono or mixed infection of the host. The present ELISA contributes a relatively sensitive and reliable tool for the early serodiagnosis of bubaline fasciolosis.

Amino Acid Sequence↗
PubMed2006

Fasciola hepatica in capybara.

The purpose of this report was to register the presence of Fasciola hepatica in an adult female capybara (Hydrocherus hydrocheris). The macro and microscopical characteristics of hepatic lesions, together with the finding of F. hepatica in mature forms indicate the possible infection of capybara by the trematode.

Animals↗
PubMed2006

High risk of bacterobilia in advanced experimental chronic fasciolosis.

Fasciolosis is recognized as an important human disease. Wistar rats experimentally infected with Fasciola hepatica were examined using data obtained in the advanced chronic state of the disease (200, 300 and 400 days post-infection, dpi). Pigment stones (PS) and bile specimens were collected. The same procedure was applied in control rats. Liver tests were determined using stored serum samples. Bacteriological bile culture revealed viable bacteria (Escherichia coli, 45% of cases, Enterococcus faecalis, 45% and Klebsiella pneumoniae, 10%). The presence of bacterobilia was associated with liver serum enzymes, including aspartate aminotransferase (AST or SGOT), alanine aminotransferase (ALT or SGPT), alkaline phosphatase (AP) and total bilirubin levels. Multivariate analysis suggested an association between bacterobilia and the following factors: duration of parasitic infection and intensity of parasitic infection supported the impression that the obstruction caused by advanced chronic fasciolosis in the rat may be related to biliary sepsis. Extrapolation to human infection in fasciolosis hyperendemic areas is discussed. In conclusion, the results of the rodent model should lead to a reconsideration of treatment features in human disease, i.e. therapeutic strategies should not only include a parasitic treatment but also consider the possibility of bacterial co-infection.

Animals↗
PubMed2005

Fasciolosis (liver fluke) of the breast in a male patient: a case report.

Fasciolosis is an uncommon disease in this region, as are parasitic infections of the breast. This report describes a 56-year-old man with a previous history of liver abscess presenting with a painful breast mass. He underwent a mastectomy and is well. The pathological findings revealed chronic granulomatous mastitis with Fasciola spp. ova. The literature on this very rare condition is reviewed.

Fascioliasis↗
PubMed2005

Promoter hypermethylation is a major event of hMLH1 gene inactivation in liver fluke related cholangiocarcinoma.

Cholangiocarcinoma is a crucial health problem in Northeast Thailand where liver fluke infection is endemic. However, molecular genetic and epigenetic mechanisms involved in carcinogenesis of this cancer remain unclear. We attempted to study genetic and epigenetic alterations of hMLH1 gene in 65 intrahepatic cholangiocarcinoma using polymerase chain reaction (PCR) based microsatellite marker D3S1611 and methylation-specific PCR, respectively. Of 65 cases, 29 (44.6%) showed hypermethylation of hMLH1 promoter. Loss of heterozygosity (LOH) of hMLH1 was detected in 12 of 51 informative cases (23.5%). Five out of 29 (17.2%) methylated cases demonstrated LOH. Aberrant methylation of hMLH1 promoter was significantly associated with poorly differentiated type (P=0.013). Our study suggests that both genetic and epigenetic mechanisms cause the inactivation of hMLH1 where epigenetic is a major event resulting in mismatch repair deficiency and contributing to carcinogenesis of liver fluke related cholangiocarcinoma. Since, gene silencing by methylation is an early event in carcinogenesis, promoter hypermethylation of hMLH1 may be a molecular targeted therapy and prevention of liver fluke related cholangiocarcinoma.

Adaptor Proteins, Signal Transducing↗
PubMed2005

Alterations in carbohydrates and the protein metabolism of the harmful freshwater vector snail Lymnaea acuminata induced by the Euphorbia tirucalli latex extract.

To know the short- as well as long-term effect of aqueous latex extracts of Euphorbia tirucalli on carbohydrate and protein metabolism, the snail Lymnaea acuminata was exposed to sublethal doses of 0.37 and 0.55 mg/L for a 24-h and 0.20 and 0.31 mg/L for a 96-h exposure period. Significant (P<0.05) alterations in the glycogen, pyruvate, lactate, total protein, and free amino acid level, as well as in the activity of enzyme lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase, and alanine aminotransaminase were observed in the nervous, hepatopancreatic, and ovotestis tissues of the freshwater vector snail L. acuminata exposed to sublethal doses of E. tirucalli latex extract. The alterations in all biochemical parameters were significantly (P<0.05) time and dose dependent. After the 7th day of the withdrawal of treatment, there was significant (P<0.05) recovery in glycogen, pyruvate, lactate, total protein, and the free amino acid level and in the activity of the lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase and alanine aminotransaminase enzymes in all three of the studied tissues of the snail, which supports the view that the plant product is safe for use as a molluscicide for the control of harmful freshwater vector snails in the aquatic environment.

Animals↗
PubMed2005

Fasciola hepatica: humoral and cytokine responses to a member of the saposin-like protein family following delivery as a DNA vaccine in mice.

The humoral and cellular responses to DNA vaccination of BALB/c mice with a novel antigen from the Fasciola hepatica saposin-like protein family (FhSAP-2) have been investigated. Two constructs were produced containing the FhSAP-2 DNA sequence, one intended for extracellular secretion of FhSAP-2 protein, and one expressing FhSAP-2 in the cytoplasm of a transfected cell. The constructs were tested in HEK 293T cells, with the secretory construct producing less detectable FhSAP-2 relative to cytoplasmic construct when observed by fluorescence. The size of expressed protein was confirmed by Western blot of cell lysate, but FhSAP-2 was undetectable in cell supernatants. Both, secretory and cytoplasmic constructs as well as FhSAP-2 recombinant protein were tested in mice. The antibody response elicited in mice vaccinated with the rFhSAP-2 induced high levels of IgG(1), IgG(2), and IgE as well as high levels of IL-10 and IFNgamma indicating a mixed Th1/Th2 response. Vaccination of mice intramuscularly with the cytoplasmic FhSAP-2 construct resulted in a dominant IgG(2a) isotype antibody as well as a dominant IFNgamma cytokine, with significant IgE, IgG(1), and IL-10 responses also present, suggesting a mixed Th1/Th2 profile. Isotype and cytokine profiles elicited by the FhSAP-2 secretory construct were similar to those obtained with the cytoplasmic construct but at levels that were significantly lower. The results demonstrate that FhSAP-2 can be delivered as a DNA vaccine construct and induces a stronger Th1 response than the recombinant protein alone. This could result in an improvement in the immunoprophylactic potential of this candidate vaccine against F. hepatica.

Animals↗