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Antigenicity of a proteolytic enzyme of Fasciola hepatica.

The possibility was examined of using a haemoglobinase released during in vitro incubation of adult Fasciola hepatica for immunodiagnosis of fascioliasis. By SDS gel electrophoresis the enzyme appeared as two closely migrating bands with a molecular weight of approximately 27,000 daltons. After Western blotting the bands reacted with serum from rats infected with F. hepatica and mice infected with Schistosoma mansoni. The enzyme was therefore not a good antigen for immunodiagnosis.

Animals↗

The relationship between the size of Lymnaea truncatula naturally infected with Fasciola hepatica and the intensity and maturity of the redial infection.

Of the 2,652 Lymnaea truncatula collected from sites in Cumbria and Gwynedd during 1973-75, when the prevalence of fascioliasis in the primary host was low and declining, only 123 were infected with Fasciola hepatica. Dissection of these snails revealed that the proportion infected, the mean redial burden and the proportion of mature rediae in each snail increased with shell length. The results are compared with a similar data set acquired when the prevalence of infection in both the primary and intermediate hosts was uncharacteristically high. Although the results are qualitatively similar, there are important quantitative differences. The mean redial burden of infected snails at times of high disease prevalence was generally twice as high as that reported in the present study. It is suggested that differences in habitat microclimate could not account entirely for the observed differences in redial burden and the role of multiple miracidial infection is discussed.

Animals↗

Evaluation of a monoclonal antibody-based competition ELISA for the diagnosis of human hydatidosis.

An antibody competition enzyme-linked immunosorbent assay (ELISA) using 4 different monoclonal antibodies (MAb) raised against major antigens (Antigen 5 and Antigen B) of Echinococcus granulosus was evaluated for the diagnosis of human hydatidosis. The competition assay, using anti-Ag5 MAb 24.14, detected specific antibodies in 70% (131/188) of sera from patients with surgically confirmed E. granulosus infection and 38.5% (10/26) of sera from patients with E. multilocularis infection. None of the sera from patients with Taenia solium cysticercosis (10), T. saginata (2), filariasis (22), strongyloidiasis (19), fascioliasis (4), bilharziasis (4) and amoebiasis (2) tested were positive using a cut-off point established through reaction between MAb 24.14 and normal human sera. The combined use of the MAb 24.14-based competition ELISA with the conventional antibody-binding assay provides a highly sensitive (92.8%) and specific screening system for human hydatid disease diagnosis.

Antibodies, Helminth↗

[The worm in the liver].

Patients with parasitic involvement of the liver are a small group of patients in European countries. In the differential diagnosis of "hepatic mass" "pain in the right upper abdomen" "biliary obstruction" or "abnormal liver function tests" parasites are, if at all, considered last. In most health care centers diagnostic and management experience is limited with respect to these diseases. They should therefore been taken care of by specialized centers. In most countries these are centers for tropical diseases. Important components of the work up of patients with parasitic diseases are: risk assessment (place of origin, travel and exposure), geographic distribution of the parasites, incubation period, prepatent period, laboratory investigations (eosinophilia, IgE, direct parasitological diagnosis) and imaging studies. Important parasites with liver involvement will be presented: schistosomiasis, echinococcosis, fascioliasis, toxocariasis.

Anthelmintics↗

Insights into the relationships of Palearctic and Nearctic lymnaeids (Mollusca: Gastropoda) by rDNA ITS-2 sequencing and phylogeny of stagnicoline intermediate host species of Fasciola hepatica.

Fascioliasis by Fasciola hepatica is the vector-borne disease presenting the widest latitudinal, longitudinal and altitudinal distribution known. F. hepatica shows a great adaptation power to new environmental conditions which is the consequence of its own capacities together with the adaptation and colonization abilities of its specific vector hosts, freshwater snails of the family Lymnaeidae. Several lymnaeid species only considered as secondary contributors to the liver fluke transmission have, however, played a very important role in the geographic expansion of this disease. Many of them belong to the so-called "stagnicoline" type group. Stagnicolines have, therefore, a very important applied interest in the Holarctic region, to which they are geographically restricted. The present knowledge on the genetics of stagnicolines and on their parasite-host interrelationships is, however, far from being sufficient. The present paper analyses the relationships between Palaearctic and Nearctic stagnicoline species on the base of the new light furnished by the results obtained in nuclear rDNA ITS-2 sequencing and corresponding phylogenetic studies of the lymnaeid taxa Lymnaea (Stagnicola) occulta, L. (S.) palustris palustris (topotype specimens) and L. (S.) p. turricula from Europe. Natural infections with F. hepatica have been reported in all of them. Surprisingly, ITS-2 length and GC content of L. occulta were similar and perfectly fitted within the respective ranges known in North American stagnicolines. Nucleotide differences and genetic distances were higher between L. occulta and the other European stagnicolines than between L. occulta and the North American ones. The ITS-2 sequence of L. p. turricula from Poland differed from the other genotypes known from turricula in Europe. The phylogenetic trees using the maximum-parsimony, distance and maximum-likelihood methods confirmed (i) the inclusion of L. occulta in the branch of North American stagnicolines, (ii) the link between the North American stagnicolines-L. occulta group with Galba truncatula, and (iii) the location of the L. p. turricula genotype from Poland closer to L. p. palustris than to other European L. p. turricula genotypes. The Palaearctic species occulta is included in the genus Catascopia, together with the Nearctic species catascopium, emarginata and elodes. The results suggest a potential of transmission capacity for C. occulta higher than that of other European stagnicolines or Omphiscola glabra. The relatively low genetic distances between C. occulta and G. truncatula and the clustering of both species in the same clade suggest that C. occulta may be potentially considered as the second lymnaeid intermediate host species of F. hepatica in importance in eastern and northern Europe, and probably also western and central Asia, after G. truncatula. L. p. turricula may be considered as a potential secondary vector of F. hepatica, at a level similar to that of L. p. palustris.

Animals↗

Laboratory assessment of the molluscicidal and cercaricidal activities of the Egyptian weed, Solanum nigrum L.

The molluscicidal properties of Solanum nigrum L. were tested against three Egyptian snail species (Biomphalaria alexandrina, Bulinus truncatus and Lymnaea natalensis), each an intermediate host of parasites causing human schistosomiasis or fascioliasis. The plant was collected in two regions within Egypt: Fayium and Giza. Snails were exposed for 24 and 48 h, to the dry powdered fruits and leaves or to crude water extracts of the powders, and mortality was recorded. The water extract of the leaves collected in Fayium (FLWE) had the highest molluscicidal activity, with median lethal concentrations (LC50) of 18.6 mg/litre for Bi. alexandrina, 14.5 mg/litre for Bu. truncatus and 17.7 mg/litre for L. natalensis. When Bi. alexandrina infected with Schistosoma mansoni were exposed to FLWE (20 or 25 mg/litre), they shed significantly fewer cercariae than unexposed snails (P < 0.02). The cercaricidal properties of FLWE were directly tested against S. haematobium, S. mansoni and Fasciola gigantica cercariae and a time-concentration relationship was observed; the concentrations needed to kill all cercariae (LC100) within 30 min of exposure were 30 mg/litre for both S. haematobium and S. mansoni and 40 mg/litre for F. gigantica.

Animals↗

Treatment of gastrointestinal infections.

During the past year, we have seen the continuing spread of antimicrobial drug resistance in important gastrointestinal pathogens. Typhoid fever seen in the United States was multidrug resistant, but still susceptible to quinolones. The mechanism of quinolone resistant typhoid in Vietnam was better elucidated. Treatment failures in enterobacteriaceae treated with quinolones suggest that the minimum inhibitory concentration break point for resistance should be lowered. Evidence is mounting that ciprofloxacin and ofloxacin may be safely used to treat serious infections in children. Cefixime showed some promise in treating shigellosis in an open labeled trial. Decreased gastric acid secretion was associated with cholera but not dysentery. Phase 1 trials of vaccines for cholera and enterotoxigenic Escherichia coli showed promise. The antifungal drug, clotrimazole, demonstrated ability to inhibit secretory diarrhea in laboratory studies. Nitazoxanide demonstrated efficacy in both protozoan and helminthic infections in humans, including fascioliasis.

Journal Article↗

Kinetic disposition of triclabendazole in buffalo compared to cattle.

Concentrations of triclabendazole sulfoxide and its sulfone metabolite in plasma were measured in buffalo and cross-bred cattle after single intraruminal administration of triclabendazole at two different doses. Plasma concentrations of both metabolites were significantly lower in buffalo than cattle at both doses, which resulted in a smaller area under the concentration-time curve, a lower concentration maximum and a lower relative bioavailability. Thus, the recommended doses of 12 mg/kg body weight for the treatment of bubaline fascioliasis may not be valid for buffalo because of the substantially lower uptake of the drug in this species.

Animals↗

Intrahepatic bile duct changes in human hepatosplenic schistosomiasis mansoni.

Wedge liver biopsies of 132 patients with hepatosplenic mansonian schistosomiasis were studied and divided in two groups according to the presence (Group I - 69 cases) or absence (Group II - 63 cases) of markers of the actual presence of the parasite in the liver tissue. Histological variables indicating bile duct injury were analysed in each case: periductal fibrosis, hyperplasia of the bile duct epithelium, bile duct degeneration, and marginal ductular proliferation. The presence of one or more of these variables defined two sub-groups: A - bile duct lesions present (73 cases), and B - bile duct lesions absent (59 cases). The variables "bile duct degeneration" and "ductular proliferation" were related to the actual presence of the parasite in the host. In 55.3% of all cases of human mansonian schistosomiasis a spectrum of injuries to the bile ducts was present. Epithelial hyperplasia alone or associated with patterns of mucopolysaccharide production was observed in 87.6% cases of the Sub-group A. The bile ducts changes in mansonian schistosomiasis are close to those described in liver fluke infestations such as clonorchiasis, fascioliasis and opistorchiasis. Statistical analysis revealed that high mucopolysaccharide production was associated with epithelial hyperplasia. The pathogenesis of the bile duct changes in human mansonian schistosomiasis and its relation to the parasitic infestations and their antigens is discussed.

Bile Ducts, Intrahepatic↗

Development and evaluation of a Western blot kit for diagnosis of human trichinellosis.

We evaluated industrially prepared Western blot strips designed to avoid the cross-reactions observed with indirect immunofluorescence and enzyme-linked immunosorbent assays used for the serodiagnosis of trichinellosis. The antigen preparations were crude extracts of Trichinella spiralis. The Western blot profile characteristic of trichinellosis was characterized by comparing 60 sera from patients infected by Trichinella to 11 sera from healthy subjects, 51 sera from patients with other proven parasitic diseases (cysticercosis, schistosomiasis, strongyloidosis, fascioliasis, toxocariasis, liver amebiasis, anisakiasis, filariasis, toxoplasmosis, hydatidosis, or malaria), and 23 sera from patients with autoantibodies. Specific 43- to 44-kDa and 64-kDa bands were obtained with all of the sera from 51 patients with acute trichinellosis, in 4 out of 9 patients at the early stages of the disease, and in only 1 control patient, who had suspected anisakiasis and in whom trichinellosis could not be ruled out by muscle biopsy.

Animals↗

Nitazoxanide: a new broad spectrum antiparasitic agent.

Nitazoxanide (Alinia, Romark Laboratories) was synthesized based on the structure of niclosamide. In vitro studies have demonstrated activity against a broad range of parasites as well as some bacteria. Three controlled trials demonstrated efficacy in cryptosporidiosis, however, the efficacy in advanced AIDS patients (CD4 cell counts = 50) at approved doses was limited. Trials have also demonstrated efficacy comparable to metronidazole (Flagyl, GD Searle and Co.) in giardiasis with fewer side effects. Nitazoxanide is also effective versus intestinal helminths and tapeworms as well as in chronic fascioliasis. Side effects in clinical trials have been similar to placebo. Nitazoxanide is the first agent proven to be effective in cryptosporidiosis. It has also proven efficacy in giardiasis. Nitazoxanide is efficacious again intestinal helminths. Additional indications may be developed in the future.

Animals↗

Hepatobiliary alterations in massive biliary ascariasis. Histopathological aspects of an autopsy case.

Hepatobiliary alterations found in an autopsy case of massive Biliary Ascariasis, are reported on histological grounds. Severe cholangitis was the main finding, but other changes were also detected, such as pyloric and intestinal metaplasia, hyperplasia of the epithelial lining, with intraductal papillomas and adenomatous proliferation. Remnants of the worm were observed tightly adhered to the epithelium, forming microscopic intrahepatic calculi. Mucopolysaccharides, especially acid, showed to be strongly positive on the luminal border, and in proliferated glands around the ducts. The authors discuss the similarity between such findings and Oriental Cholangio-hepatitis, and suggest that inflammation and the presence of the parasitic remnants are responsible for the hyperplastic and metaplastic changes, similarly with what occurs in chlonorchiasis, fascioliasis and schistosomiasis.

Ascariasis↗

[Molluscacide activity of piquerol A isolated from Piqueria trinervia (Compositae) on 8 species of pulmonate snails].

In laboratory trials an aqueous solution of Piquerol A from Piqueria trinervia, collected in several regions of Mexico, showed a molluscicide action on the adults of eight different pulmonates snails species: Fossaria (Fossaria) humilis, F. (Bakerilymnaea) sp., Pseudosuccinea columella and Stagnicola attenuata from Mexico; F. (B.) cubensis and Physa cubensis from Cuba; P. columella and Biomphalaria glabrata from Brazil; B. glabrata from Puerto Rico; and S. elodes from U.S.A. The solution was tested at 50, 25 and 5 ppm concentration, for two periods of 6 and 24 hours, at room temperature (20-22 degrees C). A 100% mortality was obtained for all species at 50 ppm concentration after 6 hours of exposure; the same percentage at 25 ppm after 24 hours; and 60 to 100% mortality at 5 ppm concentration during 24 hours of exposure. No recovery was observed among any of the treated snails. Piquerol A is a sesquiterpene with low stability in nature and has previously only been tested as an insecticide and as an inhibitor of metabolism in cell cultures: no field trails have been made on its toxicity to other aquatic fauna as yet, but it is believed Piquerol A could be an excellent molluscicide for use in areas where focal transmission of schistosomiasis and fascioliasis are taking place. This is the first time experiments on molluscicides have been carried out in Mexico.

Animals↗

Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR.

From complete mitochondrial DNA sequence of Fasciola hepatica available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella by F. hepatica during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS) region of rDNA from L. columella in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica miracidia in L. columella during the pre-patent period and identification of transmission areas.

Animals↗

Notes on human cases of cystic echinococcosis in Peru.

Cystic echinococcosis (CE) is a high prevalent zoonosis in the central and southern Peruvian Andes. Serum samples (n50)from patients presenting presumptive clinical and radiological diagnosis of CE (group 1), were tested for antibodies against Echinococcus granulosus metacestode using Arc-5 double diffusion assay (DD5), immunoelectrophoresis (IEF), and immunoelectrotransfer blot (EITB) techniques. Serum samples (n18) from patients presenting other parasite infections (paragonomiasis, cysticercosis, and fascioliasis) or healthy blood donors (n15), were designated as control groups. The overall sensitivity of the tests was of 94% (DD5 and IEF tests) or 96% (EITB test). Only patients from group 1 were seropositive for CE. Polypeptides of 21, 31, and 48 kDa were considered positive for CE. Based on these results, this study demonstrates that CE also occurs in other coastal departments (Piura, Ancash, Ica, Arequipa, and Tacna) besides Lima.

Adolescent↗

Cloning and expression of a cysteine proteinase gene from Paragonimus westermani adult worms.

A gene encoding a cysteine proteinase from Paragonimus westermani has been cloned and expressed in Escherichia coli. The cysteine proteinase cDNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from the conserved active site of the cysteine proteinase. The 5' and 3' regions of the gene were amplified using a PCR technique for the rapid amplification of cDNA ends. The cloned gene has an open reading frame of 687 bp and deduced amino acid sequence of 229. Sequence analysis and alignment showed significant homologies with the eukaryotic cysteine proteinases and conservation of the Cys, His, and Asp residues that form a catalytic triad. Analysis of the expressed protein on sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that the molecular weight of the protein was approximately 28.5 kDa. The expressed protein reacted with the sera of patients with paragonimiasis but not with the sera of fascioliasis and clonorchiasis. These results suggest that the expressed protein may be valuable as a specific diagnostic material for the immunodiagnosis of paragonimiasis.

Amino Acid Sequence↗

Expression of cysteine proteinase of Clonorchis sinensis and its use in serodiagnosis of clonorchiasis.

A gene encoding cysteine proteinase from Clonorchis sinensis has been cloned and expressed in Escherichia coli. The cysteine proteinase cDNA fragment was amplified by reverse transcription-polymerase chain reaction using degenerate oligonucleotide primers derived from conserved active site of cysteine proteinases. The 5' and 3' regions of the gene were amplified using rapid amplification of cDNA ends. The cloned gene has an open reading frame of 696 bp and deduced amino acid sequence of 232. Sequence analysis and alignment showed significant homologies with the eukaryotic cysteine proteinases and conservation of the Cys, His, and Asp residues that form the catalytic triad. Analysis of the expressed protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the molecular weight of the protein was approximately 28.5 kDa. Proteolytic activity of the expressed protein was inhibited by cysteine proteinase inhibitors such as L-trans-epoxysuccinyl-leucylamide-(4-guanidino)-butane, iodoacetic acid, and leupeptin. The expressed protein showed biochemical properties similar to those of cysteine proteinases of other parasites. The expressed protein strongly reacted with the sera from patients with clonorchiasis but not with the sera from patients with paragonimiasis, fascioliasis, cysticercosis, and sparganosis, or with sera from normal human controls. These results suggest that the expressed protein may be valuable as a specific diagnostic material for the immunodiagnosis of clonorchiasis.

Amino Acid Sequence↗

Tissue parasitic diseases in Korea.

Parasitic disease is still important subject in the field of infectious diseases in Korea considering it's number and morbidity. Recently there was conspicuous reduction of parasitic disease in terms of soil-transmitted nematodiasis, but parasitism affecting organs other than intestinal tract is still a considerable problem. This survey covers the parasitic diseases cross-sectioned at a pathology laboratory of a referral hospital, trying to elucidate the significance of its relative frequency and also to describe some histopathological changes made by different parasites. Entire pathological materials of parasitic diseases, that were referred, examined and confirmed at the Department of Pathology, Seoul National University Hospital from 1968 to 1987, were used for the study. There was a total of 594 cases of tissue parasitic diseases. This number accounted for 0.33 per cent of total accessions of surgical pathology. There occurred average 30 cases of tissue parasitic disease each year at this Hospital. Protozoal diseases were constituted of 15 cases of amebiasis, 7 cases of leishmaniasis (imported) and 5 cases of Pneumocystis carinii infections. Among helminthic infections cysticercosis was the most common (425 cases), and was followed by paragonimiasis (35 cases), sparganosis (31 cases), clonorchiasis (32 cases) and ascariasis (16 cases). In addition there were 4 cases of anisakiasis, 2 cases of fascioliasis, 2 cases of echinococcosis (imported) and a case of strongyloidiasis and a case of metagonimiasis respectively. It is emphasized that imported parasitic diseases such as leishmaniasis and hydatid disease become steadily found nowadays. Schistosomiasis is another possible imported disease, but not found in this series. And certain cestodiasis particularly cysticercosis and sparganosis should be the subjects of epidemiologic re-evaluation in view of steady occurrence of their morbidity.

Adult↗