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[Epizootiology and prevention of helminthiasis of game animals in Czechoslovakia and its relationship to the parasitic diseases of cattle].

Decades of surveying helminthic diseases in domestic and free-living ruminants in the CSR evidenced extensive infestation by lungworms, gastrointestinal nematodes, and liver flukes. In some years, parasitic invasion caused up to 50% of total losses of roe deer. Programmes to control systematically dictyocaulosis and fascioliasis were carried out with cattle and sheep in 1970-1980, with deer in 1981-1985. In the eighties, the incidence of F. hepatica in cattle and sheep dropped to less than 1%, of D. viviparus in cattle to 0.2%; no D. filaria is to be encountered in sheep presently. The share of parasitic diseases in total losses of roe deer decreased to 19% in 1983-1985.

Animals↗

Molluscicidal activity of Solanum aculeatum (family: Solanaceae) berries against Biomphalaria pfeifferi, Bulinus globosus and Lymnaea natalensis.

Aqueous suspensions of powder of sun- or freeze-dried berries of the plant Solanum aculeatum (Family Solanaceae), indigenous in Kenya, were tested for molluscicidal activity against Biomphalaria pfeifferi, Bulinus globosus and Lymnaea natalensis under laboratory conditions. One hundred or 50 mg powder L-1 of sun- or freeze-dried berries killed over 60% of the test B. pfeifferi, Bul. globosus and L. natalensis. Whereas 25 mg L-1 of the sun dried material killed less than 60% of the test snails, similar concentrations of the freeze dried molluscicide produced 60-80% mortality in the snails, under similar conditions. Using L. natalensis as the target snail, it was shown that the freeze dried material was more potent than the freeze-dried berries of S. incanum, S. nigrum or leaves of Polygonum senegalensis (Family Polygonaceae), all present in Kenya, and known to possess molluscicidal properties. The powdered material retained molluscicidal activity even after several months storage at room temperature. These findings suggest that S. aculeatum is a potent plant molluscicide, and has the potential for the control of vectors of schistosomiasis and fascioliasis in Kenya.

Animals↗

Application of biotechnology methods to the study of trematodes.

Advances in biotechnology over the past decade offer renewed hope for solutions to trematode parasitoses in Thailand. Trematode diseases have remained the same, but the tools (1) to exploit the innate ability of cells to replicate and produce biological products upon demand, (2) to manipulate the genetic makeup of an organism, (3) and to biologically or synthetically manufacture peptides have provided scientists with new reagents for diagnosing, treating, preventing and controlling trematode diseases. Although recent applications have been focused on schistosomiasis, they have potential application to other trematode diseases (paragonimiasis, clonorchiasis, opisthorchiasis, fasciolopsiasis, and fascioliasis) endemic to Thailand. The optimism which new technology has generated must be tempered with realism and continued support for basic parasitology. Progress is dependent on a combination of approaches involving techniques at the molecular, cellular, organism and population levels. The new tools must be used in conjunction with old knowledge. Most of all there must be a rational strategy for the use of products of new technology. Defined vaccines alone will not be the answer, even if they become a reality. They must be applied in a manner that optimizes their effectiveness. The silver bullets may be out there, but they are still in a roughly molded stage. They must be finished, loaded in an appropriate epidemiological gun, and fired by an expert at specific targets.

Animals↗

Immunoaffinity fractionation of Schistosoma mansoni worm antigens using human antibodies and its application for serodiagnosis.

A crude Schistosoma mansoni soluble worm antigen preparation (SWAP) was fractionated using an immunoaffinity column consisting of specific human anti-SWAP antibodies obtained from chronic S. mansoni-infected human sera and bound to CNBr-activated Sepharose 4B. The chromatographic separation resulted in three fractions: the unbound material (FW), and the eluted antigens with glycine-HCl (F1) and glycine-HCl-NaCl (F2). Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the purified antigens F1 and F2 consisted of several bands when stained with Coomassie blue and silver stain, with molecular weights between 20 X 10(3) and 200 X 10(3). The F1 and F2 fractions in addition to FW and SWAP were used in an enzyme-linked immunosorbent assay (ELISA) to measure antibody levels in sera from schistosomiasis patients. Each individual serum assessed with the purified F2 antigen gave 100% positivity and three to four times higher optical density in comparison to SWAP with only 88% positivity. No detectable cross-reactive antibodies against F2 were found when a limited number of sera from filariasis, fascioliasis and trichinellosis patients were screened. Furthermore, F2 was also used and found to be more sensitive generally in detecting anti-adult worm antibodies than SWAP in recently schistosomiasis-infected persons. Thus, F2 appears to be a highly sensitive and specific reagent for the serodiagnosis of schistosomiasis infection.

Antibodies↗

Simplification and standardization of dot-ELISA for human schistosomiasis mansoni.

Dot-ELISA, a technique that shares the same principles as the enzyme immunoassay, is useful for detection of anti-Schistosoma mansoni antibodies in the sera of patients with Schistosoma mansoni infections. The antigens were fixed to the nitrocellulose strips, blocked with 1% bovine serum albumin in 0.05% Tween 20. Patient sera (40) and normal laboratory personnel sera (9) were applied to the sheet directly, without cutting the strips into small discs. The nitrocellulose sheets are kept in a humid chamber for 30 min and then washed. After incubation with peroxidase-conjugated goat anti-human antibody, washing, and addition of substrate, positive reactions appear as brown dots against the white background. The room temperature assay takes about 2 hr. The optimum antigen concentration is 20-80 ng per dot and the optimum serum dilution is 1:100-1:400. The sensitivity and specificity of the assay are 90-95% and 90%, respectively. The level of positivity of the dot-ELISA by an arbitrary scale compares with standard micro-ELISA. The single positive reaction in a normal serum sample in dot-ELISA is also positive in micro-ELISA. Cross-reactivity between the S. mansoni antigen and human fascioliasis sera was noticed in 2 out of 8 patient sera. Good correlation between the arbitrary level of dot-ELISA and the absorbance of standardized micro-ELISA shows that the dot-ELISA is useful both for laboratory and field studies.

Antibodies↗

Interleukin 2 receptor in patients with localized and systemic parasitic diseases.

An enzyme-linked immunosorbent assay was used to quantify soluble interleukin 2 receptor (IL-2R) in the serum of patients with helminthic and protozoal infections. The results demonstrated that levels of IL-2R were normal in patients with helminthic infections limited to the intestinal tract (ascariasis, trichuriasis), but significantly elevated in patients with systemic or long-lasting infections (strongyloidiasis, schistosomiasis, fascioliasis, opisthorchiasis). In patients infected with Schistosoma mansoni levels of IL-2R were higher in those with the hepatosplenic than in those with the intestinal form of the disease. Patients with malaria also showed increased serum levels of IL-2R, irrespective whether the infection was caused by Plasmodium falciparum or P. vivax. No difference was observed between patients with acute or history of malaria. The highest levels of IL-2R were observed in patients with visceral leishmaniasis. Interestingly, in these patients the concentration of IL-2R correlated to specific antibody titre. The results are discussed in the context of preferential activation of T lymphocytes, B lymphocytes and/or macrophages during the course of the different parasitic infections investigated.

Enzyme-Linked Immunosorbent Assay↗

Hepatobiliary disorders presenting as fever of unknown origin in Cairo, Egypt: the role of diagnostic ultrasonography.

Among 130 patients with fever of unknown origin (FUO) studied from 1981 to 1985, 34 were diagnosed as having hepatobiliary disorders: amoebic liver abscess (11), pyogenic liver abscess (4), hepatic hydatid cysts (2), hepatic fascioliasis (2), tuberculous hepatic granulomas (1), chronic calcular cholecystitis with recurrent cholangitis (2), chronic active hepatitis (2), hepatocellular carcinoma (3), lymphoma involving the liver (4) and hepatic metastasis in (3) cases. Hepatobiliary disorders were the cause in 27% of FUO seen during 4 years.

Adolescent↗

[Interpretation of results of serologic analyses in large animal practices].

The backgrounds to and possibilities of serological studies for the purpose of establishing are reviewed, all this with regard to bovine disease. The serology of the virus infections infectious bovine rhinotracheitis (IBR), bovine viral diarrhoea (BVD), para-influenza virus 3 infection (PI3), bovine respiratory syncytial virus (BRS) and Chlamydia is discussed in greater detail. Likewise, the interpretation of serological findings in cases of brucellosis, leptospirosis and Johne's disease is dealt with. In addition, attention is paid to the results of serological testing in cases of lung-worm disease and fascioliasis.

Animals↗

[Parasite egg development during methane fermentation].

Investigations were carried out on the development of the most frequently encountered parasitic diseases in farm animals raised under industrial conditions. The parasite eggs were obtained from great numbers of animals with fascioliasis, dicrocoeliasis, ascaridiasis, and paramphistomum infection, etc., and were mixed with fecal homogenate in a lagoon of a fermentor in the conditions of methane fermentation. It was found that the parasite eggs in such conditions retained their vitality in a state of anabiosis. After the fermentation procedure was over and the manure compost mass was spread in the field the parasite eggs could resume their biologic development and become infective. This made it necessary to compost the manure liquid and use it after subsidiary thermal treatment.

Animals↗

Elimination of Biomphalaria pfeifferi, Bulinus tropicus and Lymnaea natalensis by the ampullarid snail, Marisa cornuarietis, in a man-made dam in northern Tanzania.

Marisa cornuarietis is a well known ampullarid competitor/predator of Biomphalaria glabrata in Puerto Rico. For the first time in Africa a flourishing population of Marisa has been established in a small, permanent, man-made dam at Kisangara, near Moshi, Tanzania. Prior to the release of M. cornuarietis in June 1977, this dam supported thriving populations of the pulmonate snail hosts Biomphalaria pfeifferi and Lymnaea natalensis; Bulinus tropicus and the melaniid Melanoides tuberculata were also common. Some 24 months after the establishment of Marisa the three pulmonate species had been eliminated; only M. tuberculata remained at about the same population density as originally recorded. Marisa has not caused any obvious adverse environmental impact in the dam. There is at present no valid evidence that this ampullarid would be a threat to local rice production, which is the only crop at risk, but carefully designed field trials should be undertaken to confirm or refute this view. In view of the vast number of permanent, lentic habitats throughout the Afrotropical region, which act as important transmission sites of schistosomiasis and fascioliasis, the role of Marisa cornuarietis as a cost-effective biological control agent in integrated control operations deserves henceforth to be energetically explored.

Animals↗

[Additional data on superficial argentophilic structures of the miracidium and cercaria of Fasciola gigantica (Cobbold, 1855) (Trematoda:Fasciolidae) and on the epidemiology of this parasite].

The life cycle of a senegalese strain of Fasciola gigantica was carried out in France from adults recovered in the liver of naturally infected cattle (Bos taurus). Laboratory-reared Lymnaea natalensis experimentally infected shed numerous cercariae. Metacercariae were used for successful infection of sheep. In the miracidium, variations are observed in the disposition of the medio-ventral epithelial cell of the second tier. The distribution of the superficial argentophilic structures is similar to that of the miracidium of F. hepatica. The chaetotaxy of the cercaria is very closely related to that of the cercaria of a strain of F. gigantica from Madagascar shed by Lymnaea hovarum but it differs clearly from that of the cercaria of F. hepatica. The encystment of a great number of cercariae (25 %) on the surface of water suggests that this is an important factor in the extension of the foci of contamination, and therefore increases greatly the risks of infection not only for animal but also for man. It is possible that human fascioliasis caused by F. gigantica is a disease, the importance of which is under-estimated.

Animals↗

[Detection of antibodies against ovum antigens in Schistosoma mansoni bilharziasis].

Fifty-five sera from bilharziasis patients from West Indies and thirty-five control sera (from normal subjects and patients with hydatidosis and fascioliasis) were examined by ELISA for antibodies reacting with S. mansoni soluble egg antigen MSA1 . MSA1 antigen was prepared according to Pelley and coupled to isothiocyanate substituted plastic discs. Sensitivity and specificity of reaction were good. Moreover, in experimental infection in mice, sera of mice cured with praziquantel became negative. The sera were also examined by a whole egg antigen in counter immunoelectrophoresis. The results of the two different techniques correspond well although ELISA using purified antigen prove to be more specific and sensitive.

Animals↗

Application of different Paragonimus antigens to immunodiagnosis of human lung fluke infection.

Utilizing enzyme immunoassay, Paragonimus africanus, P. uterobilateralis, P. ecuadoriensis and P. westermani antigens were tested with regard to their reactivity to sera from patients infected with Paragonimus africanus, P. uterobilateralis, P. ecuadoriensis, Fasciola hepatica, Opisthorchis viverrini, different schistosomes, Cysticercus cellulosae, nematodes and from patients with nonparasitic diseases as well as from healthy blood donors. Simultaneous application of Paragonimus antigens to each serum led to a pattern of reactivity which rendered possible species-specific diagnosis. Most distinct cross reactivity occurred with fascioliasis sera, followed by opisthorchiasis, schistosomiasis and sera from patients with cysticercosis. Cross-reactivity was observed to least extent when P. westermani antigen was applied. A follow-up study of 42 paragonimiasis patients treated with praziquantel (2-cyclohexylcarbonyl-1,2,3,6,7, 11b-hexahydro-4H-pyrazino[2,1-a]isoquinolin-4-one, EMBAY 8440, Biltricide) demonstrated that substantial decrease of IgG antibodies occurred between the first and sixth months after therapy, whereas change of IgE antibody levels was less conclusive.

Antigens, Helminth↗

[The environment of the Moscow megalopolis and the problems of parasitic contamination].

Studying parasitic contamination is regarded as an individual research trend for large-scale land and water areas whose natural habitat is greatly modified by man-made influences. On the one hand, parasitic contamination as its particular form acts as a factor that destabilizing the environment, on the other, the man-made deformation of natural biocenoses, which is induced by socioeconomic influences promote its occurrence and manifestations. The investigations of parasitic contamination are beyond the scope of the routine parasitological goals, though they involve their whole package and should be considered as an ecological objective of environmental protection. The cenotic communications of the links of the parasitic systems with the physicochemical and biological ingredients of the habitat predetermine qualitative and quantitative differences of parasitic contamination in the large urban areas and, finally, the dissimilar course of epidemiological, epizootic, and phytototic processes in parasitic diseases in their various sectors. The major factors influencing the levels of the parasitic pollution of the Moscow megalopolis were analyzed. These included various lines of economic activities, the population's living conditions, procedures for manufacturing foodstuffs, demographic changes. In the recent years, the situation in many parasitic infections in man (trichinosis, cryptobiasis, enterobiasis, toxocariasis, etc.), animals (fascioliasis, trichinosis, etc.) and plants (viral, fungal, and nematode diseases). There is an increase in the population of mosquitos, lice, fleas, and other insects which are vectors of dangerous parasitic diseases of man and animals.

Animals↗

Purification and characterization of a specific Fasciola antigen.

Specific Fasciola antigen was prepared from homogenates of Fasciola hepatica adult worms. The homogenate was ultracentrifuged and the supernatant containing crude Fasciola antigen was then passed over a cyanogen bromide activated sepharose 4B column coupled with antiserum against Schistosoma mansoni adult worm surface antigen. The specific, Schistosoma-free Fasciola antigen was tested for its specificity by immunodiffusion. Characterization of the specific Fasciola antigen was done by gradient poly-acrylamide gel electrophoresis and immunoblotting technique. The electrophoresis migration pattern of specific Fasciola antigen, stained with Coomassie blue, showed 7 bands in the 12-54 kDa regions. Using the immunoblotting technique, a batch of positive fascioliasis sera recognized two specific bands at the 33 and 54 kDa regions.

Animals↗

[Evaluation of ELISA-IgG test using a purified antigen in the diagnosis of human hydatidosis].

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of IgG antibodies to purified sheep hydatic cyst fluid antigen in 56 sera of confirmed cases of hydatidosis. The cut-off value was determined using serum samples from 80 healthy persons, employing two serum dilutions (1:100 and 1:500) with two and three standard deviations (SD). This assay was compared with the indirect hemagglutination test (IHAT). The sensitivity of ELISA-IgG was 94.3% for hepatic cysts and 92.9% for pulmonary cysts, whereas the values for IHAT were 77.1 and 64.3% respectively. According to Mac Nemar test, both method presented statistical significance (p < 0.05). In order to find out the specificity, additional 70 serum samples from individuals with other parasitoses, such as cysticercosis (30), trichinosis (26) and fascioliasis (14) were also tested. IHAT presented a specificity of 92.7% and for ELISA-IgG the specificity using a cut-off of mean + 3 SD was 99.3 and 100.0% with sera dilution of 1:100 and 1:500 respectively when a cut-off of mean + 2 SD was considered, we found a specificity of 91.3 and 97.3% for 1:100 and 1:500 dilutions. The use of ELISA-IgG and purified antigen in the diagnosis of human hydatidosis is discussed.

Adolescent↗

[The technology for manufacturing antiparasitic preparations. 1. The development of a technology for producing the anthelmintic triklazan in finely dispersed form and the evaluation of its antinematodal, anticestodal and antitrematodal activities].

The authors describe the technology of preparing a new anthelmintic agent triclazan, N-(3,4-dichlorophenyl)-2-[(benzo-2,1,3-thiadiazole-4-sulfonyl)amino]-5- chlorobenzamide, finely dispersed (the particles size 4-8 mu). Triclazan is highly effective not only in hymenolepiasis and trichocephaliasis, as was shown previously, but in fascioliasis, monieziasis and intestinal nematodiasis of sheep as well.

Animals↗

[Cryptosporidia and other endoparasites in heifers imported into the Czech Republic].

Totally 887 heifers of Holstein-Friesian breed mostly in late pregnancy imported to the Czech Republic from France (597), Germany (89), Denmark (181) and Holland (20) were examined coprologically from September 1993 to March 1995 in the parasitological laboratory of the National Veterinary Institute (NVI). Prague. Feces were sampled individually, rectally, always on days 1-3 following importation from heifers housed in particular quarantine sheds. In compliance with presently valid veterinary regulations, all animals were examined for liver fluke disease (fascioliasis) and lungworm. Moreover, 634 heifers were submitted to qualitative coprological examination aimed at revealing the presence of cysts and oocysts of protozoa, eggs of taenias and nematodes of gastrointestinal tract. The method according to Pavlásek (1991), especially designed for proving oocysts of the genus Cryptosporidium, was applied in all fecal specimens delivered to the SVI from animals in quarantine (N = 887). From trematodes, 12 heifers imported from France were positive for eggs of Fasciola hepatica and in other two animals eggs of the genus Paramphistomum were found. None of the imported heifers showed lungworm disease. Summary of data on occurrence of endoparasites gained during qualitative examination of samples of feces taken from heifers imported from France, Germany and Denmark is presented in Tab. I. Parasitologically, 91.2 to 100% of imported animals were positive. Taeniasis (the genus Moniezia) was detected in 2.8% of heifers imported from France and in 9.8% animals from Denmark. Protozoal parasites were found in 58.8% (Denmark) and 92.8% (Germany) heifers. Coccidial oocysts most frequently observed represented the genus Eimeria (E. bovis, E. auburnensis and E. zuernii). Gastrointestinal nematodes of nine genera were found in 72.5 to 80.8% of heifers. The most frequent findings were genera Ostertagia, Haemonchus and Trichostrongylus. Oocysts, morphologically identical with Cryptosporidium muris Tyzzer (1907), 1910, were detected in 4.5% of heifers imported into the Czech Republic from France and in 7.9% of those from Germany. In view of the fact the imported heifers were sampled always on days 1-3 of their quarantine following their importation it is quite impossible, considering the development of the protozoon, they could become infected just in the territory of the Czech Republic. Therefore, with the highest probability, our findings of C. muris-like oocysts in heifers are of priority importance for France and Germany because in the literature these countries do not report cattle as a host of this protozoon. We have found out 57.9% out of 19 animals positive for C. muris on one farm of a private cattle keeper. On the basis of a long-term monitoring of three dairy cows and one bull, the duration of the patent period is longer than 18 months, while we do not know precise onset of shedding oocysts of the protozoon in these naturally infected animals. Furthermore, the paper discusses the need of future studies of C. muris from the point of view of spread, pathogenicity, specificity and host spectrum. The author proposes and recommends obligatory examinations of imported animals with special attention paid to presence of coccidia of the genus Cryptosporidium in order to maintain, with respect to their zoonotic character, these protozoal infections under proper control. At present the parasitological laboratory of the NVI in Prague has a bank of oocyst isolates of the C. muris type from cattle (Bos taurus), from desert hamsters (Phodopus roborovskii Satunin, 1903) and camels (Camelus bactrianus). Experimental infections is permanently kept in laboratory mice following successful transmission from desert hamsters.

Animals↗