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D T John

Publications and source records attributed to D T John.

At least 37 records · Page 2Linked to original sources
PubMed1985

Isolation of Naegleria australiensis from an Oklahoma Lake.

Eight isolates of Naegleria australiensis were obtained from a small lake in Tulsa, Oklahoma. The eight strains were isolated during the hot summer months of July through September, when water temperatures ranged from 27 to 33 degrees C. All eight isolates were pathogenic for mice. The mean time to death for mice was 10 days (range 6-13 days). This pathogenic free-living ameba has not before been reported from the United States or the Western Hemisphere.

Amoeba↗
PubMed1984

Sucker-like structures on the pathogenic amoeba Naegleria fowleri.

Using scanning electron microscopy, we observed sucker-like structures on amoebae of 13 human isolates of Naegleria fowleri. The number of suckers per amoeba seemed to vary according to the virulence of the strain. We propose the term amoebastome to describe this unique sucker-like structure of N. fowleri.

Amoeba↗
PubMed1983

Cytopathogenicity of Naegleria fowleri for rat neuroblastoma cell cultures: scanning electron microscopy study.

Neuroblastoma cells were inoculated with Naegleria fowleri Lee and examined for cytopathology at various periods post-inoculation by scanning electron microscopy. By 18 h post-inoculation, approximately 50% of neuroblastoma cells were nonviable, as evidenced by trypan blue exclusion and light microscopic examination. This cytopathology resulted from piecemeal consumption of target cells mediated by a sucker apparatus extending from the surface of N. fowleri.

Amoeba↗
PubMed1983

Naegleria fowleri infection acquired by mice through swimming in amebae-contaminated water.

Groups of mice were placed in water containing from 10(2) to 10(6) Naegleria fowleri amebae per ml and allowed to swim for 2.5 to 20 min. Mouse mortality ranged from 0 to 70% and was dependent upon the concentration of amebae per ml and the length of swimming exposure. That swimming mice can develop fatal naeglerial infection further confirms the mouse model for studying experimental primary amebic meningoencephalitis.

Amebiasis↗
PubMed1982

Serum agglutination and immunoglobulin levels of mice infected with Naegleria fowleri.

Agglutinins were not detected in sera from mice given one, two, or three intranasal (i.n.) inoculations or a single intravenous (i.v.) inoculation of trophozoites of Naegleria fowleri. However, agglutinins were produced following second and third i.v. inoculations. Serum immunoglobulin levels increased in both i.n.- and i.v.-inoculated mice. IgG and IgM increased substantially more for i.v.-inoculated mice. IgA levels increased more consistently for i.n.-inoculated mice.

Agglutination Tests↗
PubMed1982

Biological factors affecting enflagellation of Naegleria fowleri.

Naegleria fowleri is a pathogenic amoeboflagellate that can be evoked to transform from amoebae to flagellates by subculture to nonnutrient buffer. More than half of the amoebae of strains KUL, nN68, and Lovell became enflagellated 300 min after subculture to amoeba-saline, whereas no amoebae of strains NF66, NF69, and HB4 did. N. fowleri nN68 enflagellated best when grown at 32 or 37 degrees C and subcultured to amoeba-saline at 37 or 42 degrees C. Amoebae from the stationary phase of growth enflagellated more readily than did actively growing amoebae. Incubation in expended culture medium from stationary-phase cultures enhanced the capability of growing amoebae to enflagellate after subculture to amoebasaline. Enflagellation was more extensive when the population density in amoebasaline did not exceed 2 x 10(5) amoebae per ml. Cycloheximide at 1 microgram/ml and actinomycin D at 25 micrograms/ml inhibited growth of N. fowleri nN68. Cycloheximide at 0.5 microgram/ml and actinomycin D at 25 micrograms/ml completely prevented enflagellation when added at time zero. Cycloheximide at 0.5 microgram/ml, added 120 to 300 min after initiation of enflagellation, prevented further differentiation and caused existing flagellates to revert to amoeboid cells. Similarly, actinomycin D at 25 micrograms/ml, added 90 to 300 min after initiation of enflagellation, retarded differentiation and caused flagellates to revert. Radiolabeled precursors were incorporated into macromolecules during differentiation in nonnutrient buffer. Enflagellation of N. fowleri is a suitable model for studying regulation of a eucaryotic protist.

Amoeba↗
PubMed1982

Cytopathogenicity of Naegleria fowleri and Naegleria gruberi for established mammalian cell cultures.

Amebae of Naegleria fowleri and Naegleria gruberi were cytopathic for nine established mammalian cell cultures, including mouse and human fibroblasts, rabbit and monkey kidney cells, rat and mouse neuroblastoma cells, baby hamster kidney cells, and human epithelioma and carcinoma cells. Nine strains of N. fowleri were equally cytopathic for rodent neuroblastoma cells. As few as one ameba per million neuroblastoma cells destroyed the mammalian target cells after 9 days. The N. fowleri grew and destroyed rat neuroblastoma cells at 30 to 37 C whereas N. gruberi grew and destroyed the target cells at 25 to 30 C. Both N. fowleri and N. gruberi attached efficiently to the target cells at 30 to 37 C; N. gruberi but not N. fowleri attached efficiently at 25 C. Electron microscopic observations of mixed cultures of N. fowleri and neuroblastoma cells established that the amebae, after 12 hr, had ingested portions of the neuroblastoma target cells without causing cell lysis. Conversely, N. gruberi amebae, after attaching to target cells, disrupted the plasma membrane and cytoplasm of the target cells although the target cell nucleus remained intact. The amebae then ingested the target cell debris.

Amoeba↗
PubMed1982

Intravenous infection of mice with Naegleria fowleri.

Naegleria fowleri produced fatal meningoencephalitis in mice following intravenous (i.v.) inoculation. Amebae were present in the peripheral circulation for 120 minutes after i.v. inoculation with a dose of 10(7) trophozoites per mouse. Amebae were cultured from and observed in brain (days 1-21), lung (days 1-12), and liver and kidney (days 1-5). Infected mice exhibited weight loss, leukocytosis, reduced lymphocyte/neutrophil ratio, neurologic symptoms, and mortality. Histologically, the disease was characterized by an acute, hemorrhagic, necrotizing meningoencephalitis. Although amebae were detected in tissues other than brain, pathologic involvement of these tissues was minimal.

Amebiasis↗
PubMed1980

Growth of Naegleria fowleri in several axenic media.

Growth of Naegleria fowleri under agitated and unagitated culture conditions was compared using axenic media developed by Balamuth, Cerva, Chang and Nelson. Under agitated culture conditions, the best growth was achieved with Nelson medium. Using unagitated cultures. Cerva medium produced the greatest cell yields. The media of Cerva and Nelson are considerably lower in organic nutrients and salt concentration than are the media of Balamuth and Chang. The pH of the media rose during ameba growth with the increases proportional to the amount of growth, i.e. the greater the cell yield the greater the rise in pH. Maximum cell yields varied by 2.5 fold for 10 human isolates of N. fowleri cultured in Nelson medium.

Culture Media↗
PubMed1978

Innate resistance of mice to experimental infection with Naegleria fowleri.

The mouse system provides an excellent model for studying host resistance to Naegleria fowleri, the agent of primary amoebic meningoencephalitis. Innate resistance to infection with N. fowleri was examined with respect to infecting dose and the age, sex, and strain of mice. Intravenous inoculation with 10(7) amoebae per mouse produced 100% mortality in 9 days, whereas inoculation with fewer amoebae reduced the cumulative mortality. Male and female DUB/ICR mice of varying ages were inoculated intravenously with 2.5 X 10(5) N. fowleri per g of body weight. The youngest mice died first, with 100% mortality for both males and females, and mortality decreased with increasing age. Female mice were significantly more resistant to infection than males. Five strains of mice weighing approximately 20 g were inoculated intravenously with weight-adjusted doses; mortality ranged from 10% in C57BL/6 mice to 95% in A/HeCr mice.

Age Factors↗
PubMed1977

Cell size, macromolecular composition, and O2 consumption during agitated cultivation of Naegleria gruberi.

Cell size, macromolecular composition, carbohydrate utilization patterns, and O2 concentrations were measured throughout the growth stages of Naegleria gruberi in agitated culture in a complex medium. Biphasic logarithmic growth occurred during the intial 83 hr of growth and the mean generation time was 7.0 hr and 19 hr during initial and secondary log growth stages, respectively. The maximum yield was 5 X 10(6) amebae/ml. The pH rose rapidly (1 pH unit) during the secondary log growth phase (52-83 hr) and continued into the stationary growth phase (83-120 hr). Dry weight, total protein, carbohydrate, and RNA per ameba increased just before the secondary log growth phase. RNA increase 31% to 35% per ameba at the end of each phase of log growth. DNA increased approximately 2-fold throughout the different growth phases. Average cell size increased 90% during biphasic log growth then decreased during stationary phase. O2 tension decreased from 100% to 18% of saturation during the biphasic growth phase, then increased during stationary growth to near 100% saturation. Glucose and total carbohydrate assays showed little utilization of those substrates throughout the growth stages. Naegleria gruberi presumably has a predominantly aerobic metabolism, also its metabolism may change during the different growth phases.

Amoeba↗
PubMed1977

Immunization of mice against Naegleria fowleri infection.

Naegleria fowleri produces fatal meningoencephalitis in humans and in experimentally infected laboratory animals. The course of the disease in mice is dependent upon the infecting dose of amoebae, route of inoculation, and prior exposure to Naegleria antigens. DUB/ICR mice were immunized by various routes and antigen preparations, held for 21 days, and, together with noninfected control mice, challenged intravenously (i.v.) or intranasally (i.n.) with 10(7) or 10(6) N. fowleri per mouse, respectively. Mice immunized with liver or formalinized N. fowleri or live N. gruberi subcutaneously, intraperitoneally, i.v., or i.n. were significantly protected against a subsequent lethal challenge with N. fowleri i.v. or i.n. In general, i.v. inoculation afforded greated protection than other routes of immunization, intact cells immunized mice better than did cell fragments, and N. gruberi appeared to be a better immunogen than N. fowleri.

Administration, Intranasal↗
PubMed1977

Concanavalin A-induced agglutination of Naegleria.

Concanavalin A (Con A) agglutinated all Naegleria gruberi strains tested but did not agglutinate any N. fowleri strains tested. Agglutination was time and temperature dependent and Con A concentration and ameba concentration dependent over certain ranges. Agglutination increased to maximum up to 1 h incubation with Con A. At least 1 X 10(6) amebae/ml were needed for maximum agglutination, and Con A concentrations higher than 100 microgram/ml did not appreciably increase agglutination. Incubation of 4 degrees C or with 10 mM alpha-methyl-D-mannoside inhibited agglutination of N. gruberi. These data indicate a difference in polysaccharide structure of cell membranes of N. fowleri and N. gruberi.

Agglutination↗