Isolation of Histoplasma capsulatum from a texas cave

Summary Sixty-three soil samples collected from caves in Texas and the Republic of Mexico were studied to determine the incidence of pathogenic fungi by the use of the flotation technique in mice.Histoplasma capsulatum was isolated from the Kickapoo cave in Texas, and there was some histopathological evidence of its presence in the soil of Devil' cave (Cueva del Diablo) in Mexico. In the latter case, an incidence of human contact with the cave which resulted in an illness is presented.     

Isolation of a New Soluble Antigen from the Yeast Phase of Histoplasma capsulatum

A method is described by which a soluble antigen was prepared from the yeast phase of Histoplasma capsulatum. This soluble preparation had a specificity greater than that of whole-cell yeast-phase antigens. In complement fixation tests with sera from human cases of histoplasmosis, blastomycosis, and coccidioidomycosis, the soluble antigen reacted in 12.1% of 141 tests with heterologous sera, whereas conventional whole-cell yeast antigens reacted in 47.3% of 91 tests with heterologous sera. The reactivities of the two types of antigens with homologous sera were essentially the same.     

Antigens from Histoplasma capsulatum and Blastomyces dermatitidis

Certain groups of fungi share chemical structures which makes difficult the isolation and differentiation of specific antigens by the usual methods of extraction and purification. Therefore, we have oriented our studies to the immunological and biochemical characterization of differences and similarities of molecular structures from fungi, etiologic agents of systemic mycoses, hoping to establish criteria for the utilization and handling of these antigens.A deproteinized polysaccharide-protein complex (D-PPC) was isolated from Histoplasma capsulatum and Blastomyces dermatitidis. The immunological studies with humoral tests indicate a variable cross reaction between antigens of both species. In immunodiffussion systems, the reaction was specific for each species using saline solution or phosphate buffer solution, while using an agarose veronal system, the cross reactions were very evident. In addition, differences in cross reactions were obtained with immunoelectrophoresis, haemagglutination and complement fixation microtest. This variation in cross reaction responses suggest that these antigens (D-PPC) share common structures but at the same time must have some different component owned by each one of the fungi which makes them more specific than crude antigens.     

Selenocystine-resistant mutants of Histoplasma capsulatum

Cysteine metabolism has been thought to be important to the phenomenon of dimorphism inHistoplasma capsulatum. We sought mutants with genetic blocks in the metabolism of cysteine by selection of colonies resistant to the toxic analogue, selenocystine. The 22 resistant strains thus obtained were all deficient in uptake of cystine from the surrounding medium but were normally able to convert from mycelium to yeast and back again. Furthermore, they had normal quantities of NADH-dependent cystine reductase when this enzyme was measured. We conclude that mutants defective in cystine uptake can be readily obtained by selection of colonies resistant to selenocystine, and that a lesion in cystine-uptake does not appear to affect the phenomenon of dimorphism in this organism.Preliminary reports of this work were presented at the Second International Congress of Mycology, Tampa, 1977 and at the first International Conference on Histoplasmosis, Atlanta, 1978.     

Chemical composition of Histoplasma capsulatum

The chemical composition of yeast and mycelial cells of three strains ofHistoplasma capsulatum was analyzed and is expressed as per cent dry weight. Cultures were grown in a liquid synthetic medium, mycelial cells at 25°C and yeast at 37°C on gyrotory shakers. After 7 days, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and protein were higher in the yeast cells while mycelial cells contained more lipid and carbohydrate. The components of one strain were also studied at different stages of growth. The DNA in both yeast and mycelial cells remained relatively constant, but other components varied with the age of culture. In yeast cells the RNA level was 6.8 % at 2 days and then declined sharply remaining constant around 3.5 %. A protein content of 29 % on day 2 decreased gradually to 19 % on day 14. An initial lipid content of 21 % rose to 33 % on day 5 and then decreased. Similarly, an initial carbohydrate level of 17 % rose to 25 % on day 7 and then declined. The mycelial cells contained 4 % RNA up to 10 days followed by a slight decline to 3 % on day 14. A protein content of 20 % on day 5 increased to 24 % on day 10 and then decreased to 15 % on day 28. The lipid content of 33 % on day 5 rose to 38 % on day 7 and then decreased gradually. The carbohydrate level of 20 % at 5 days increased to 38 % on day 10 and declined gradually to 27 % after 28 days.

---

Résumé La composition chimique des cellules levuriformes et mycéliennes de trois souches deHistoplasma capsulatum a été déterminée. Le champignon a été cultivé dans un milieu synthétique liquide secoué à 25° C pour la phase mycélienne et à 37° C pour la phase levuriforme. Après 7 jours d'incubation, les cellules levuriformes étaient plus riches en acides nucléiques et en protéines que les cellules mycéliennes qui étaient par contre plus riches en lipides et en hydrates de carbone. La composition d'une des souches fut étudiée à différentes étapes de la croissance. La teneur en ADN des deux phases resta relativement constante mais des variations furent observées dans le cas des autres constituants chimiques. Pour ce qui est des levures, l'ARN qui constituait 6,8 % du poids des cellules sèches à deux jours, tomba rapidement à 3,5 % et resta constant. Les proteines passèrent de 29 % au deuxième jour à 19 % au quatorzième jour. Au contraire, la teneur en lipides passa d'un valeur initiale de 21 % à 33 % au cinquième jour, pour diminuer de nouveau par la suite. De même, une teneur initiale en hydrates de carbone de 17 % passa à 25 % au septième jour puis diminua par la suite. Dans les cas des cellules mycéliennes contenaient 4 % de ARN jusqu'au dizième jours, puis cette valeur tomba légèrement jusqu'à 3 % au quatorzième jour. Les protéines qui représentaient 20 % au cinquième jour augmentèrent jusqu'à 24 % au dizième jour pour tomber à 15 % au vingthuitième jour. La teneur en lipides de 33 % au cinquième jour augmenta jusqu'à 38 % au septième jour pour diminuer graduellement. De même les taux en hydrates de carbones qui représentaient 20 % au cinquième jour augmentèrent jusqu'à 38 % au dixième jour et diminuèrent graduellement jusqu'à 27 % au vingt-huitième jour.

     

Isolation of Histoplasma capsulatum from an oil bird (steatornis caripensis) cave in Venezuela

Summary 1.Histoplasma capsulatum was recovered from two of fifteen soil specimens collected in a Venezuelan cave. This cave, located near Caripe in the state of Monagas, harbors a large colony of the oil bird,Steatornis caripensis.2. This finding confirms the previous discovery in Tingo Maria, Peru, of the association ofH. capsulatum with oil birds.3. The relationship ofH. capsulatum with animal habitats is discussed along with the public health importance of this association.

---

Zusammenfassung 1.Histoplasma capsulatum ist in zwei Bodenproben aus fünfzehn, die einer Höhle in Venezuela entnommen wurden, gefunden worden. Diese Höhle, die sich in der Nähe von Caripe, im Staate Monagas befindet, herbergt eine grosse Kolonie von Fettvögeln,Steatornis caripensis.2. Dieser Befund bestätigt die Vergesellschaftung vonH. capsulatum mit den Fettvögeln, wie es vorher bereits in Tingo Maria, Peru, entdeckt worden ist.3. Die Beziehung vonH. capsulatum zum tierischen Habitat wird diskutiert unter Betonung der Wichtigkeit dieser Vergesellschaftung vom Standpunkte des öffentlichen Gesundheitswesens.

     

A prototrophic yeast-strain of Histoplasma capsulatum

A newly derived strain of Histoplasma capsulatum can be grown stably as yeast in a minimal medium containing glucose, biotin, tartrate and inorganic salts.     

Inhibition of Histoplasma capsulatum by Garlic

The mycelial phase ofHistoplasma capsulatum was inhibited by both the volatile and water soluble components of garlic,Allium sativum L. Garlic extract at a concentration of 254 parts per billion (ppb) was inhibitory, while 8.1 parts per million (ppm) were lethal to pure cultures ofH. capsulatum. The role of garlic as an eradicent is discussed.The work was conducted while the author was a graduate student at the University of Kentucky, Lexington, Kentucky.     

DNA polymerases from a dimorphic fungus,Histoplasma capsulatum

DNA polymerases from the yeast and mycelial phases of a fungus,Histoplasma capsulatum, were partially purified and analyzed. Two classes of enzymes were identified in each morphological form. No significant differences were found among polymerases Ia and Ib from mycelia and I from yeast; similarly, polymerases II from both phases were almost identical. (Numbering refers to the order of elution from phosphocellulose.) However, all of the class I polymerases differed significantly from the class II enzymes. DNA polymerases I (M_r 122,000) were unaffected by 0.2m KCl and resistant to 10 mmN-ethylmaleimide; their levels fluctuated significantly in relation to cell growth, and they showed no nuclease activity. All of the enzymes had similar template dependence and Mg²⁺ requirements. These results are compared to data on other eucaryotic DNA polymerases. Possible roles for class I and class II polymerases are discussed.     

Histoplasma capsulatum at the host-pathogen interface

Histoplasma capsulatum is the most common cause of invasive fungal pulmonary disease worldwide. The interaction of H. capsulatum with a host is a complex, dynamic process. Severe disease most commonly occurs in individuals with compromised immunity, and the increasing utilization of immunomodulators in medicine has revealed significant risks for reactivation disease in patients with latent histoplasmosis. Fortunately, there are well developed molecular tools and excellent animal models for studying H. capsulatum virulence and numerous recent advances have been made regarding the pathogenesis of this fungus that will improve our capacity to combat disease.     

Hydroxamic Acid from Histoplasma capsulatum That Displays Growth Factor Activity

Growth factor(s) present in a spent liquid medium after culture of the yeast form of Histoplasma capsulatum enhanced both yeast and mycelial growth of nine isolates tested. Hydroxamic acid extracted from the culture fluid displayed growth factor activity.     

Cysteine-independent and cysteine-requiring yeast-strains of Histoplasma capsulatum

Recently we described a strain of Histoplasma capsulatum, designated H-35, which is able to grow as yeast on a minimal medium consisting of inorganic salts, glucose and a trace of biotin. Using this strain as a prototrophic wild type we sought auxotrophic mutants. Mutagenized yeast-cells were starved for inorganic sulfate in sulfur-free minimal medium. Sulfate was then added, and growing prototrophic cells were killed by addition of amphotericin B. After 24 hours non-growing auxotrophs were rescued by removal of amphotericin and addition of yeast extract. This mutant enrichment cycle was repeated two additional times, after which the cells were plated on blood agar and 800 yeast-colonies were picked. Seventeen of these yeast-strains required cysteine for growth, as compared with strain H-35, which grew as yeast on minimal medium.