Cell surface differences of Naegleria fowleri and Naegleria lovaniensis exposed with surface markers (original) (raw)
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Infection and immunity, 1975
Antigens prepared from each of five strains (CA, CJ, HB-1, HB-3, and TY) of pathogenic Naegleria and the EG strain of nonpathogenic Naegleria gruberi were compared by the gel diffusion and immunoelectrophoresis techniques. Axenically grown amoebae were used as sources of antigens. Antisera were produced in individual rabbits against three strains (CA, CJ, and HB-1) of pathogenic Naegleria and the EG strain of N. gruberi. In the gel diffusion experiment each of the six antigens was reacted with each of the four antisera in agar gel. The results of these experiments revealed that the antigens of N. gruberi reacted strongly with the homologous antiserum but minimally with each of the three heterologous antisera. The antigens of all five pathogenic strains reacted extensively with the anti-CA, anti-CJ, and anti-HB-1 sera and moderately with the anti-EG serum. In the immunoelectrophoresis test each of the six antigens was separated electrophoretically in agar gel and reacted with each of...
Morphological response of cultured cells to Naegleria amoeba cytopathogenic material
Journal of Cell Science, 1985
Naegleria amoebae contain cytopathogenic material (NACM). The morphological response of cultured cells to this material follows a number of characteristics in common with those resulting from infectious agents. The cytopathologic changes varied depending on the strain of the cultured cells. Among those from 17 different vertebrate sources, both primary and continuous cell lines, some were destroyed completely by dilutions of NACM up to 10~8 while others appeared unaffected by NACM at any concentration. The response had no apparent relationship to species, organ source, or passage level of the cells. The reaction was typified by a long latent period (4-10 days) during which the number of cells in the culture increased up to 10-fold, followed abruptly by a short period (less than 24 h) during which all of the cells were destroyed. The latent period was prolonged when the culture conditions were adverse, or when the amount of NACM in the inoculum was minimal. A high multiplicity of NACM in the inoculum lysed the entire culture, while dilutions near the endpoint caused generalized or only focal changes of rounded cytopathic cells. The cytopathic effect could be maintained in cultured cells by serial passage, such that the total activity greatly exceeded what could be attributed to the original inoculum. These findings are consistent with the concept that NACM has properties of an infectious agent and that its quantity is enhanced and spread through the culture by cell-to-cell contact and by cell division.
Microbiology, 1980
The behaviour of Naegleria gruberi amoebae on coverslips coated with concanavalin A was followed by reflection interference microscopy. Locomotion continued briefly on the protein and much material was shed before the cells halted. The effect was overcome by perfusion with hapten sugars. These carbohydrates dissociated those trails formed in water from the substrate save for areas of focal contact, whereas trails generated in 10 mM-KC1 were unaffected by hapten. These results argue for direct cell-substrate contact over a limited portion of the cell surface during this type of locomotion.
Parasitology Research, 2010
Naegleria fowleri is the etiologic agent of primary amoebic meningoencephalitis, a rapidly fatal parasitic disease of humans. The adherence of Naegleria trophozoites to the host cell is one of the most important steps in the establishment and invasiveness of this infectious disease. Currently, little is known about the surface molecules that may participate in the interaction of N. fowleri with their target cells. In the present study, we investigated the composition of glycoconjugates present on the surface of trophozoites of the pathogenic N. fowleri and the nonpathogenic Naegleria gruberi. With the use of biotinylated lectins in western blot and flow cytometric analysis, we showed that N. fowleri trophozoites present high levels of surface glycoconjugates that contain α-D-mannose, α-D-glucose, and terminal α-L-fucose residues. A significant difference in the expression of these glycoconjugates was observed between N. fowleri and the nonpathogenic N. gruberi. Furthermore, we suggest that glycoconjugates that contain D-mannose and L-fucose residues participate in the adhesion of N. fowleri and subsequent damage to MDCK cells. I
Journal of clinical microbiology, 1993
Monoclonal antibodies (MAbs) reactive to the pathogenic amoeba Naegleria fowleri were analyzed by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay, Western blotting (immunoblotting), and radioimmunoprecipitation assay (RIPA). Two MAbs (3A4 and 5D12) showed reactivity by ELISA with all N. fowleri strains tested and no reactivity with the five other Naegleria species, N. lovaniensis, N. gruberi, N. australiensis, N. jadini, and N. andersoni. These MAbs reacted with the three morphological forms of N. fowleri (trophozoites, cysts, and flagellates). The reactivity on Western blots was suppressed by treatment with metaperiodate, suggesting a carbohydrate epitope. Differences in reactivity patterns between trophozoites and cysts observed with radioimmunoprecipitation assay might reflect differences in biological properties. The formalin stability of the epitope may be useful in detecting N. fowleri in fixed biopsies and in investigating the pathological process.
ELECTROPHORETIC KARYOTYPE AND LINKAGE GROUPS OF THE AMEBOFLAGELLATE NAEGLERIA-GRUBERI
1990
fectivity test-a simple test which may serve to distinguish Trypanosoma brucei from T. rhodesiense. Bull. Wld. Health Org., 42:650-65 1. 16. Rickman, L. R. & Robson, J. 1970. The testing of proven T. brucei and T. rhodesiense strains by the blood incubation infectivity test. Bull. Wld. Health Org., 42:9 11-916. 17. Riflcin, M. R. 1978. Identification of the trypanocidal factor in normal human serum: high density lipoprotein. Proc. Natl. Acad. Sci., 75:3450-3454. 18. Rifkin, M. R. 1983. Interaction ofhigh-density lipoprotein with Trypanosoma brucei: effect of membrane stabilizers. 1973. Experimental infections with African trypanosomes. V. Preliminary parasitological, clinical, hematological, serological, and pathological observations in rhesus monkeys infected with Trypanosoma rhodesiense. Amer.
Studies on the rhizoplast from Naegleria gruberi
Journal of Cell Science
A procedure, utilizing homogenization and centrifugation in a low ionic strength buffer containing Triton X-100, has been used to facilitate the isolation of the rhizoplast from flagellates of Naegleria gruberi. This has enabled a study to be made of the physical and biochemical properties of this organelle. The rhizoplast is shown to be a proteinaceous structure with chemical properties similar to those of the molluscan gill ciliary rootlet. Polyacrylamide gel electrophoresis gives a possible subunit molecular weight of approximately 240000 Daltons. Studies with antisera raised against the rhizoplast fraction demonstrated the absence of rhizoplast antigens in amoeboid forms of Naegleria gruberi and is taken as evidence that the organelle is synthesized de novo during transformation of the amoeba to the flagellate form. Results of optical diffraction studies on isolated rhizoplasts are also presented.
Southeast Asian Journal of Tropical Medicine and Public Health, 2011
Seven stains were studied to determine the best color and contrast for staining the developmental stages of free living pathogenic Acanthamoeba and Naegleria species. The acid-fast bacilli stain (AFB) produced a blue color without contrast; trichrome-eosin and modified Field's showed various color contrasts; Giemsa, iron-hematoxylin, modified AFB and Gram produced only one color which distinguished the nucleus, nucleolus, cytoplasm, food- and water-vacuoles. The motile organs (acanthopodia, pseudopodia, lobopodia and flagella) were also clearly differentiated but produced a similar color as the cytoplasm. These motile organelles were first induced by incubating at 37 C for at least 15 minutes and then fixing with methanol in order to preserve the protruding morphology prior to staining. The trichrome-eosin and iron-hematoxylin stains showed good color contrast for detecting all three stages, the trophozoite, cyst and flagellate; Giemsa and Gram stained the trophozoite and flagellate stages; the modified Field's and modified AFB stains stained only the trophozoite stage. Depending on the purpose, all these stains (except the AFB stain) can be used to identify the developmental stages of Acanthanweba and Naegleria for clinical, epidemiological or public health use.
Naegleria fowleri: Light and electron microscopy study of mitosis
Experimental Parasitology, 2009
a b s t r a c t DAPI and Feulgen stains were used as specific DNA markers for studying the mitosis process in Naegleria fowleri. Both DAPI and Feulgen stains reacted with DNA in the nuclei of the amoebae. Representative figures of N. fowleri mitotic nuclei with a defined arrangement according to the phase of the cell cycle were observed. A notable characteristic is that the nucleolus is present throughout the stages of mitosis. During metaphase, several deeply stained DNA condensations following an elongated pattern were observed, corresponding almost certainly to tightly grouped chromosomes. Ultrastructural observations demonstrated that the nucleus divides by cryptomitosis, a process in which the nuclear membrane does not disappear during the mitosis. Centrioles were not found, and a spindle of microtubules was observed running the length of the nucleus from pole to pole however, they did not come to a focal point.