Biomedical subjects
Publications and source records attributed to L Cerva.
The cellular division of trophozoites of Giardia intestinalis group was followed microscopically in stained preparations from axenic cultures. Eleven successive stages were described, documented by photomicrographs and their survival times calculated.
Histological sections of 414 appendices were examined parasitologically. Enterobius vermicularis was found in 8.7%, eggs of Ascaris lumbricoides in 0.5%, trophozites of Dientamoeba fragilis in 4.8%, Endolimax nana in 2.2%, Entamoeba coli in 1% and cysts of Giardia intestinalis in 1.9% of cases. Appendicopathies associated with Enterobius were most frequent in the age group from 6 to 10 years (24.3%) and from 21 to 25 years (12.2%). Patients older than 15 years were practically women only. Dientamoeba was most frequent in the age group from 11 to 15 years (11.3%). In women D. fragilis was three times more frequent than in men. The coincidence of D. fragilis and E. vermicularis infections was 50%. No interactions were seen between the protozoans in the contents of the appendix and its mucous membrane. Statistical evaluation indicates possible etiologic role of E. vermicularis in the occurrence of acute appendicities. D. fragilis appears to be the most common intestinal protozoan parasite in Bohemia.
From nutritive media produced in the CSSR a modification of Diamond's medium TYI-S-33 was prepared which ensures the long-term axenic cultivation of Entamoeba histolytica and Giardia intestinalis. Details of the cultivation technique are also presented.
Serum samples of 1,054 inhabitants of Bohemia (Czechoslovakia) were examined by means of indirect haemagglutination test with antigens from Naegleria fowleri and Acanthamoeba culbertsoni. With N. fowleri antigen the frequency of positive reactions did not exceed 3.5 per cent in lowest serum dilutions only. N. fowleri could not be identified as a possible causative agent in any chronic form of disease in man. Significant accumulation of positive findings with A. culbertsoni antigen was observed in hepatitis A patients and convalescents (52 per cent). The potential interpretations of this phenomenon are discussed.
The growth of Naegleria fowleri cultures in a BCS medium was not affected either by trimethoprim at 400 micrograms/ml or by aminopterine, 3,5-diaminopterine and methotrexate at 500 micrograms/ml. N. lovaniensis propagation in the same medium was inhibited with 10 micrograms/ml of trimethoprim, 50 micrograms/ml methotrexate and 100 micrograms/ml 3,5-diaminopteridine. Aminopterine was ineffective at a concentration of 500 micrograms/ml. The inhibitory effect of trimethoprim on N. lovaniensis cultures depended on the medium composition and could be neutralized by an addition of folic or tetrahydrofolic acids and a suspension of heat-killed Enterobacter aerogenes. Thymine, thymidine, hypoxantine and 2-amino-4-hydroxy-6-(tetrahydroxybutyl)-pteridine did not have an adverse effect. Trimethoprim activity in N. fowleri cultures could not be enhanced by the addition of Triton X-100 and Polymyxine B. Cryolyzate of N. fowleri amoebae did not influence the trimethoprim inhibition of N. lovaniensis cultures. Deviation in dihydrofolatereductase chemical structure or thymine dependency seems to be the probable explanation for N. fowleri antifolate resistance.
A hydrophobic, galactose-specific lectin was isolated by means of affinity chromatography from Sarcocystis gigantea. Adsorbents with different spacer lengths were tested. S. gigantea lectin differs from sheep muscle lectin in the spacer length needed for adsorption. Sodium dodecyl sulfate-gel electrophoresis of the S. gigantea lectin revealed a subunit size about 19 kDa and the presence of disulfide cross-linked dimers. The lectin is present in high concentration in cystozoites, cyst fluid and cyst wall material.
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Axenic cultures of Tetrahymena pyriformis, strain I MT IV, grown in a defined medium at room temperature, were used to study interactions of these protozoa with vaccination strain L Sc 2ab of poliovirus type 1, vaccination strain P 712 of poliovirus type 2 and with type 30 echovirus, strain 480/78. T. pyriformis cultures in media containing 10(3.0) TCD50/1 ml of type poliovirus, 10(3.0) TCD50/1 ml of type 2 poliovirus or 10(2.5) TCD50/1 ml echovirus 30 and in virus-free medium did not differ one from another in their growth and die-away kinetics during the 21 days of observation. Two-day T. pyriformis cultures were infected with poliovirus 1 (initial concentration 10(3.2) TCD50/1 ml), and poliovirus 2 and echovirus 30 (initial concentrations 10(3.0) TCD50/1 ml). Viruses were titrated in test tube cultures of BGM cells. The supernatant fluid, standardized sediment and samples of control virus suspension free of protozoa were titrated after 0, 2, 6, 10, 13, 18, 28 and 30 days. Most of the virus in culture was found associated with the sediment, both in the period of active growth and during the die-away phase of T. pyriformis protozoa. The virus in sediment was present at higher titres and its survival time was longer than in virus in liquid phase. Thirteen days after the first contact between T. pyriformis and virus the sediment and supernatant fluid of the old protozoan culture and the T. pyriformis-free control viral suspension were taken and used as inocula for new two-day T. pyriformis cultures.(ABSTRACT TRUNCATED AT 250 WORDS)
The water extract from cryolyzed whole muscle cysts of Sarcocystis gigantea from sheep, in spite of the high lectin content, is a suitable antigen for the detection of specific antibodies by means of indirect haemagglutination reaction (IHA). The agglutinating effect of lectin from parasitic cysts can be eliminated with a 0.5% concentration of lactose dissolved in all solutions used for IHA. In sera of slaughterhouse sheep, positive titres ranging from 1:80 to 1:1 280 were registered. Positive reactions in lower titres were observed also with antibodies against S. dispersa, S. cuniculi and Sarcocystis sp. from pigs. Sensibilized erythrocytes can be stored in refrigerator at least for 1 week.
Both nonvirulent (15 strains) and virulent (1 strain) variants of Naegleria fowleri were isolated from the cooling circuits in seven of ten examined plants in North Moravia. No amoebae were found in waters with a high salinity (electric conductivity more than 614 microseconds) and with a low content of phosphates (less than 0.37 mg . 1(-1)) and oxygen (less than 7.4 mg . 1(-1)). Other results of the physical, chemical and bacteriological examinations showed no direct bearing on the occurrence of N. fowleri.
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A description is given of the preparation of antigen from Sarcocystis dispersa cystozoites and the procedure of the indirect haemagglutination test (IHA). The antibodies against this antigen were detected in experimentally infected mice from day 20 p.i. (1: 640). In the following weeks the antibody titres reached the value of 1: 40,960. The sera of pigs, sheep and horses spontaneously infected with other Sarcocystis species reacted with this antigen in low titres only. The bovine sera gave negative reactions even in cases when Sarcocystis cysts were present in the muscles of the examined animals. A possible application of IHA for the research and diagnostic purposes is discussed.
Six strains of nonvirulent and three strains of virulent variants of Naegleria fowleri amoebae were isolated from the examined cooling water samples from 9 power plants. The virulent variants were obtained solely from effluents discharged from power plants with a closed-circuit cooling N. fowleri was not detected outside the reach of the thermal pollution. A disinfection of out-flowing cooling water seems to be an unnecessary investment in our climate. Warm discharge water should under no conditions be used directly for sports and recreational purposes.
Two complexes of lakes and canals supplying water for two electric power plants, their steam condensors and an adjoining river were investigated by means of culture methods for the presence of Naegleria fowleri in Poland in the period from 1974 to 1980. Sixty-four strains of N. fowleri were isolated, 13 isolates being virulent for mice when instilled intranasally. These strains were found in the steam condensor of the power station A and in waters polluted with warm water of this plant. Pathogenic N. fowleri strains occurred also in an adjoining river connected with the water system of the power plant. The results show the possible role of the steam condensor A as an incubator and regular source of pollution with pathogenic amoebae for its own system of cooling waters and even the adjoining river.
Cultures of Acanthamoeba castellanii, kept at room temperature in sterile Bacto-Casitone (Difco) medium, were artificially infected with vaccination poliovirus strains type 1 and type 3 and with echovirus type 4 and echovirus type 30. No remarkable virus cumulation was observed on the surface or inside amoeba cells during the observation period of 21 days. Amoeba-adsorbed viruses were impossible to remove by repeated washings. Virus neutralization with specific antisera showed that enteroviruses were most probably present only on amoeba surfaces. In contrast to amoeba-free virus suspension, echoviruses bound to amoebae and their cell pulp persisted even after 52 to 75 days. However, the tested amoeba species played only the role of a solids-like carrier in this survival of echoviruses.
Sera of 1 218 animals were examined using indirect haemagglutination reaction with antigens of Naegleria fowleri and Acanthamoeba culbertsoni. Rabbit sera (214 samples) gave negative reaction with both antigens. Bovine sera (593 samples) reacted positively in 1.5%, particularly with the antigen of A. culbertsoni. Out of the 411 pigs examined 12% were positive with one or both of the antigens. In one of the pig farms of a modern type the positivity with one or both of the antigens. In one of the pig farms of a modern type the positivity was as much as 32.2%. The titres of positive animal sera have never been higher than 1 : 320. The amoebae of the limax group do not seem to be involved in the pathogenesis of diseases of cattle and rabbits. A sporadic occurrence of peracute cases is possible. In case of a sudden death, animals with symptoms of encephalitis or pneumonia should be examined for the presence of amoebae. Hypersensitivity should be considered as a possible cause of pig diseases.
Trimethoprim in a concentration of 4 micrograms per milliter of Bacto-Casitone (Difco) medium inhibits the growth of nonvirulent Naegleria fowleri isolates. The growth of virulent strains is unaffected even with 400 micrograms of the drug per milliliter of medium. Differences in sensitivity constitute the possibility of a simple selection of environmental isolates. The pathogenicity and virulence of Naegleria species may be connected with the metabolism of folic acid.