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The effect of mouse strain, age, sex, and the size of infective dose on the susceptibility to infection with the coccidium Cryptosporidium parvum Tyzzer, 1912 was determined using several murine models. Mice were infected with C. parvum oocysts originally of cervine origin, maintained by repeat passage in calves. All mice in the experimental groups proved susceptible to infection, though this resulted asymptomatic in all cases. C. parvum infection in BALB/c and Porton mice exhibited some variation. BALB/c mice demonstrated a longer prepatent period than Porton mice. They also produced a greater oocyst output over the patent period, though the differences were not statistically significant. Differences were observed between mice infected at either 3 or 4 weeks of age. Prepatent period was shorter in those mice infected at 3 weeks of age, reaching 100% infection rate by day 7 post-inoculation. The patent period was longer in younger mice showing that age at time of infection can modify the oocyst shedding profile. However, no sex related differences in the course of infection were observed. The effect of different infective doses of oocysts was analysed. The three doses used (10⁴, 10⁵, 10⁶) proved infective for all mice, there were no statistical differences in either prepatent or patent periods, or in the oocyst shedding profiles. Experimental cryptosporidiosis was also induced in cyclophosphamide-immunosuppressed mice. Cyclophosphamide was orally administered by stomach tube at a dose of 50 mg/kg/day starting 10 days before the intragastric inoculation of 10⁶ oocysts of C. parvum per mouse and continuing until the end of the experiment. Immunosuppressed mice had a shorter prepatent period, remained infected longer and shed more oocysts than immunocompetent mice. Immunosuppression produced high mortality rates; during the course of the experiment 44% of immunosuppressed-infected and 30% of immunosuppressed-uninfected mice died. There were no deaths in the untreated groups. Differences in the clinical course of the infection were also observed between immunosuppressed and immunocompetent mice; however, some mice recovered without immunosuppression withdrawal.

Plasmodia of a Henneguya species measuring 70-900 µm and exhibiting season-dependent stages of development were detected throughout a three-year study on gill myxosporosis of Lake Balaton pikeperch (Stizostedion lucioperca (L.)). Sixty-five out of 160 fish (41%) examined in the period of study were infected by the parasite. Infection was the most prevalent (48%) among pikeperch specimens exceeding 40 cm in length. The highest prevalence of infection (58%) was recorded in 1995-1996 while the lowest (30%) in 1996-1997. The youngest plasmodia appeared in April, and started to develop within the capillaries of the secondary lamellae of the gill filaments. The round or ellipsoidal plasmodia which continued their gradual growth in the subsequent months of the year achieved a size of 800-900 µm by the late autumn months, but remained in intralamellar location throughout the developmental cycle. Mature spores developed in the plasmodia by the end of winter. On the basis of their shape and size, the spores were identified as Henneguya creplini (Gurley, 1894). However, because of the uncertain taxonomy of species assigned to the genus Henneguya the taxonomic position of the parasite requires further study. The host reaction consisting of epithelial proliferation and granulation tissue formation starts around the infected secondary lamella only after the maturation of spores and the disruption of plasmodia.

Microsporidia are eukaryotic, obligate intracellular organisms defined by their small spores containing a single polar tube that coils around the interior of the spore. After appropriate stimuli the germination of spores occurs. Conditions that promote germination vary widely among species, presumably reflecting the organism's adaptation to their host and external environment as well as preventing accidental discharge in the environment. It appears that calcium may be a key ion in this process. Regardless of the stimuli required for activation, all microsporidia exhibit the same response to the stimuli, that is, increasing the intrasporal osmotic pressure. This results in an influx of water into the spore accompanied by swelling of the polaroplasts and posterior vacuole. The polar tube then discharges from the anterior pole of the spore in an explosive reaction and is thought to form a hollow tube by a process of eversion. If the polar tube is discharged next to a cell, it can pierce the cell and transfer the sporoplasm into this cell. Polar tubes resist dissociation in detergents and acids but dissociate in dithiothreitol. We have developed a method for the purification of polar tube proteins (PTPs) using differential extraction followed by reverse phase high performance liquid chromatography (HPLC). This method was used to purify for subsequent characterization PTPs from Glugea americanus, Encephalitozoon cuniculi, E. hellem and E. intestinalis. These proteins appear to be members of a protein family that demonstrate conserved characteristics in solubility, hydrophobicity, mass, proline content and immunologic epitopes. These characteristics are probably important in the function of this protein in its self assembly during the eversion of the polar tube and in providing elasticity and resiliency for sporoplasm passage.

Three-spined stickleback (Gasterosteus aculeatus L.), nine-spined stickleback (Pungitius pungitius (L.)) and flounder (Platichthys flesus (L.)) are widespread teleosts, which all have behaviours involving migration between freshwater and brackish/sea water environments. Their importance in dispersal of the freshwater monogenean Gyrodactylus salaris Malmberg, 1957, which causes heavy losses of Atlantic salmon (Salmo salar L.) parr in infected Norwegian rivers, was tested indirectly by their susceptibility and resistance to the parasite in laboratory experiments. Gyrodactylus salaris attached to the three fish species, but no parasite reproduction was observed. The infections were eliminated after a maximum of 3 days on flounder, 6 days on nine-spined stickleback, and 8 days on three-spined stickleback. Thus these fishes are innately resistant to G. salaris, and are therefore of no importance concerning the population dynamics of G. salaris in freshwater systems. However, attachment of parasites indicates that these fish species may function as transport hosts, and theoretically play a part in the dispersal of G. salaris in nature.

Spermatozoa of the monogenean Heterocotyle capricornensis Chisholm et Whittington, 1996 are long and filiform, comprising an elongate nucelus, probably a single elongate mitochondrion and two incorporated axonemes, one of which is shifted with respect to the other. The shift results in a region at each end of the sperm where only one axoneme is present, accompanied by the nucleus and mitochondrion at one end and the nucleus and/or mitochondrion at the other. By taking note of the direction of dynein arms on the axonemal doublet microtubules, each axoneme is identified and followed from beginning to end. No basal bodies remain in mature sperm but the main nuclear end is interpreted as proximal/anterior based on the final stages of spermiogenesis. A group of four or five cortical microtubules from the spermatid zone of differentiation persists in mature sperm, but is not closely associated with a region of extracellular matrix, as it is in other monocotylids. The sperm structure is compared with that of other monocotylids and the phylogenetic implications are discussed. Aberrant sperms in one individual were folded and fused along much of their length.

Relatively few effective compounds are available for treating microsporidiosis in humans. In this study, several compounds were assayed for activity against Encephalitozoon intestinalis (Cali, Kotler et Orenstein, 1993) and Vittaforma corneae Shadduck, Meccoli, Davis et Font, 1990 in vitro. Of the benzimidazoles tested, albendazole was most effective and the MIC50 values were 8.0 ng/ml and 55.0 ng/ml for E. intestinalis and V. corneae, respectively. Fumagillin and its analogue, TNP-470 were nearly equally effective against both E. intestinalis and V. corneae. The MIC50 values of fumagillin were 0.52 ng/ml and 0.81 ng/ml, and the MIC50 values of TNP-470 were 0.35 ng/ml and 0.38 ng/ml for E. intestinalis and V. corneae, respectively. In addition, 12 of 44 purines and pteridines with putative tubulin binding activity that were synthesized at Southern Research Institute (SRI), inhibited microsporidial replication by more than 50% at concentrations that were not toxic to the host cells. Several chitin synthesis/assembly inhibitors inhibited growth of the microsporidia in vitro but were toxic for the host cells making it difficult to interpret the results. One exception was lufenuron, which caused no significant toxicity to the host cells and expressed approximate MIC50 values of 2.95 µg/ml and 6.3 µg/ml against E. intestinalis and V. corneae, respectively. These results warrant further studies on albendazole, fumagillin, TNP-470, lufenuron, and the selected SRI purines and pteridines for developing therapeutic strategies for microsporidiosis.

Ectoparasitic Prototransversotrema steeri Angel infected Acanthopagrus australis (Günther) in western (W. Bay) and southern (S. Bay) Moreton Bay with greatest abundance in winter (June - August), and none in summer (December - February). This confirms previous observations on seasonality of P. steeri from a small estuary in New South Wales. Transversotrema licinum Manter, which is a new host and geographical record, infects A. australis in W. Bay but not S. Bay, with large abundances in summer and winter, lowest abundance in spring. Increased size of P. steeri in winter in S. Bay could be explained by increased size of fish sampled since fish length and size of each parasite species were correlated positively. Transversotrema licinum increased in size from summer to winter irrespective of fish size. Number of eggs in utero and parasite size were correlated for both species; worms in mid-size range had most eggs, and P. steeri had more eggs (average = 19.2, 0-80) than T. licinum (3.3, 0-21); eggs were similar in length. Number of eggs per parasite, adjusted for parasite size, was greatest in autumn (P. steeri) or summer (T. licinum), i.e. early in infection period. Spawning migration to eastern Moreton Bay did not influence number of eggs per parasite.

Histochemical studies of enzyme activities and structural elements in Gyrodactylus derjavini Mikailov, 1975 parasitizing fins of Oncorhynchus mykiss Walbaum were conducted. Marked activities of non-specific esterase, acid phosphatase, alkaline phosphatase and amino-peptidase were found in the intestinal caeca of the parasite. A strong activity of acetylcholinesterase was seen in the nervous system. Extraintestinal non-specific and eserine-sulphate resistant esterase was localized in the distal part of the hamulus sheath. Activities of peroxidase and glucuronidase were not detected. In the embryo, developing hamuli were enclosed in a sheath rich in phospholipids. Deposits of neutral lipids were sparse. The fully developed ventral and dorsal hamulus bars stained strongly for calcium. Lectin binding assays showed a mannose rich region in the cephalic duct openings, strong reactions for galactose in the glycocalyx whereas reactions for lactose were weak. These findings are discussed with respect to the parasite-host relationship.

Sheep scab caused by the mite Psoroptes ovis (Hering) is a highly contagious disease of sheep. As a first step in developing a mite-derived vaccine for controlling the disease, the soluble antigens in mite extracts which induce an immune response in sheep were identified by electrophoretic and immunoblotting techniques. At least 22 proteins were present in P. ovis extracts as revealed by Coomassie Blue staining. Mite-infested sheep serum recognised six antigenic bands in the extracts with approximate relative molecular weights ranging from 12 to 183 kDa. A deeply staining band at 31.2 kDa and another at 41.8 kDa are of particular diagnostic value. Immunoblotting studies showed that there was no cross reactivity between P. ovis and two other ectoparasites of sheep in the UK, the sheep louse Bovicola ovis (Schrank) and the sheep tick Ixodes ricinus L.

The role of microtubules in the secretory processes in the tegument of adult trematode Fasciola hepatica L. is studied by estimating the effects of colchicine, a substance known to disrupt microtubules, on the number of T2 vesicles. Tissue slices of the worm are incubated in Hedon-Fleig medium with or without 5 x 10(-4)M colchicine. The dynamics of the colchicine-provoked secretory block is examined by morphometry on samples processed for electron microscopy. T2 vesicles are estimated as a total number or separately within three levels (apical, sub-apical and central) of the distal tegument. The secretory block is demonstrated as reduction in the total number of T2 vesicles. The separate counting within three levels of the distal tegument demonstrates in control samples a trend of sub-apical condensation of T2 vesicles. This pattern of T2 distribution remains unchanged in colchicine-treated samples in spite of the reduction of the mean T2 counts within each of the levels examined. The data illustrate the role of microtubules in both the tegumental transport of secretory vesicles and the stratification of the organelles within the tegument.

The sera of 67 HIV-infected persons without clinical signs of Toxoplasma gondii infection and sera of 777 immunocompetent persons from three distinct regions of Czechoslovakia were examined for the presence of toxoplasmic antibodies using the complement-fixation test (CFT). Additionally Toxoplasma positive HIV+ individuals were re-examined for the presence of IgG and IgM toxoplasmic antibodies by ELISA methods. Results show that overall prevalence of toxoplasmic antibodies is not significantly greater in HIV-positive subjects (29.8 %) than in the general population (26.1 %). Similarities between these two tested groups were also documented by a close correlation of their geometrical means of titres (13.9 versus 14.5). All 20 HIV-infected patients who were positive in CFT were positive in ELISA IgG reaction, and none in ELISA IgM reaction. The detected antibody levels were suggestive of a latent Toxoplasma infection only. But because of the risk of the infection reactivation all of these patients should be attended to on a systematic basis.

The pathogenicity of the coccidium Isospora suis was studied in 42 conventional and 26 gnotobiotic piglets at 1-13 days post infection (DPI). The prepatent period of the coccidium I. suis was 4.5-5 days. Only in one experiment in conventional piglets the prepatent period of 3.5 days was recorded. At least 9 days pathogenesis of I. suis was recorded during which double alteration of the small intestine occurred. The first alteration was found at 3-4 DPI. The second phase of alterative changes was observed at 8-9 DPI. In comparison with gnotobiotic piglets in conventional piglets clinical signs and pathological changes used to appear one day sooner but reparation and regeneration of alterative changes was quicker. In the first phase of the infection (3-7 or 8 DPI) the heaviest pathological changes were in the posterior jejunum. In the second phase of the infection (8 or 9-12 DPI) the heaviest changes were in the middle jejunum. In short period repeated lesions of mucosal barrier of the small intestine multiply the pathogenicity of I. suis and rank I. suis in front place among other enteropathogens of nursing piglets.

In the period between autumn 1983 and winter 1985, 33 digestive tracts and 317 coprological samples of hares were examined. The prevalence of individual coccidian species was reported on the basis of the examination of both the digestive tracts and coprological samples. The following coccidian species were found: Eimeria babatica (with 26.4% prevalence in the season 1983/1984 and 31.1% in 1984/1985), E. europaea (21.2% and 30.3%), E. hungarica (6.5% and 7.6%), E. leporis (24.2% and 45.4%), E. macrosculpta (1.3% and 2.5%), E. robertsoni (36.8% and 29.4%), E. sculpta (6.9% and 13.4%), E. semisculpta (15.6% and 10.0%), and E. townsendi (67.1% and 74.8%), respectively. The species E. babatica, E. macrosculpta, and E. sculpta were recorded in Czechoslovakia for the first time. The oocyst morphology and location of coccidia in the digestive tracts of hares were studied.

A total of 2,883 foreign students at the age of 18-30 years were examined for amoebiasis after their arrival to Czechoslovakia. Stool examinations revealed the presence of Entamoeba histolytica in 112 of them (3.9%). Students from 38 countries were found to be infected with this parasite. In a set of 2,064 students from these countries E. histolytica prevalence in stool was 5.4%. There were greater differences in the prevalence between individual countries inside a geographical region than between individual geographical regions. The highest E. histolytica prevalence in stool was found in students from tropical and southern Africa (6.7% of 745 examined) and the lowest in students from South-eastern Asia (3.1% of 321 examined). In a simple cross-section study, antibodies against E. histolytica were detected by enzyme-linked immunosorbent assay (ELISA) in the sera of 1,001 persons. Antibodies were detected in 7.9% of students at the following titres: 1:200 in 4.5%, 1:600 in 1.5%, 1:1,800 in 1.9%. Antibodies occurred more frequently in students carrying E. histolytica cysts (X2 = 14.9). Titre of ELISA antibodies in patients with confirmed liver abscess was higher than 1:1,800. counterimmunoelectrophoresis (CIEP) test was used for serum examinations of patients who had been demonstrated by ELISA to be seropositive and of those carrying E. histolytica cysts. In a set of 170 patients CIEP antibodies were also more frequent in those carrying E. histolytica cysts (X2 = 26.95). A comparison of the results of ELISA and CIEP tests in the same patients revealed that CIEP antibodies were more dependent on the actual parasitization with E. histolytica than ELISA antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

A total of 26,478 ixodid ticks (935 pools) were examined by intracerebral inoculation of suckling mice. Six species of ticks were tested: Ixodes ricinus (23,470 individuals), I. trianguliceps (12), Haemaphysalis punctata (831), H. concinna (39), Dermacentor reticulatus (69) and D. marginatus (2,057). The ticks were collected largely by flagging vegetation, a substantial minority (4%) from animals. Three strains of Francisella tularensis were isolated, one each from I. ricinus (males, district Breclav, southern Moravia), D. reticulatus (males, district Breclav) and D. marginatus (engorged females collected from sheep in Roznava district, eastern Slovakia). D. marginatus and D. reticulatus represent new vector species for Czechoslovakia.