Investigations of Entomophthora muscae (Cohn) (Entomophthorales: Entomophthoraceae) conidial infection on housefly Musca domestica (Linn.) by scanning electron microscopy

1993 ◽  
Vol 39 (4) ◽  
pp. 363-366
Author(s):  
C. M. Tu ◽  
B. L. Singh

An entomogenous fungus, Entomophthora muscae, was studied morphologically with respect to the formation of primary and secondary conidia and germ tubes. The fungus penetrated the insect cuticle by germ tubes that were produced at the base of each conidium that penetrated directly through the cuticle. Fungal germ-tube formation and penetration of host integument were observed. The tough germ-tube penetration point seemed to provide abundant energy for the penetration of the host integument. Conidia not directly on the integument formed secondary conidia but were never observed to form germ tubes. Neither appressorium nor infection cushion was observed on the germ tubes.Key words: Entomophthora muscae, entomogenous fungus, insect mycosis, insect pathology, conidia.


1990 ◽  
Vol 36 (4) ◽  
pp. 249-253 ◽  
Author(s):  
Ruth C. Mock ◽  
Jordan H. Pollack ◽  
Tadayo Hashimoto

Candida albicans formed germ tubes when exposed to air containing 5 to 15% carbon dioxide (CO2). The CO2-mediated germ tube formation occurred optimally at 37 °C in a pH range of 5.5 to 6.5. No germ tubes were produced at 25 °C, even when the optimal concentration of CO2 (10%) was present in the environment. The requirement of CO2 for germ tube formation could be partially substituted by sodium bicarbonate but not by N2. Carbon dioxide was required to be present throughout the entire course of germ tube emergence suggesting that its role is not limited to an initial triggering of morphogenic change. We suggest that carbon dioxide may be a common effector responsible for the germ tube promoting activity of certain chemical inducers for C. albicans. Key words: Candida albican germ tubes, CO2-induced germ tube formation, endotrophic germ tube formation.



1970 ◽  
Vol 48 (9) ◽  
pp. 1692-1692 ◽  
Author(s):  
Y. Hiratsuka

Germ tubes of Cronartium coleosporioides Arth. (= Peridermium stalactiforme Arth. and Kern) emerged between processes through short irregular slits. Germ tube walls were folded when they emerged and expanded after the emergence.



1991 ◽  
Vol 69 (9) ◽  
pp. 2044-2054 ◽  
Author(s):  
Rajendra Chaubal ◽  
V. A. Wilmot ◽  
Willard K. Wynn

Adherence of germinating urediniospores of the common maize rust fungus (Puccinia sorghi Schw.) to substrata was studied by ultrastructural and cytochemical examination of extracellular matrix produced by germ tubes in conjunction with measurements of adhesion to plastic and glass surfaces. Copious amounts of extracellular matrix on germ tubes could consistently be visualized by scanning and transmission electron microscopy only when (i) a cationic detergent (cetylpyridinium chloride, polydiallyldimethylammonium chloride) or a cationic stain (ruthenium red, alcian blue, cuprolinic blue) was added to the fixation solutions, (ii) germ tubes were fixed by rapid-freezing and freeze-substitution and observed with a scanning electron microscope, or when (iii) germ tubes were observed in a frozen-hydrated state by low-temperature scanning electron microscopy. Incubation of germinated spores with dilute alkalies (NaOH, KOH), pronase E (nonspecific protease), and laminarinase (β-1,3 (1,3; 1,4-glucanase) removed the extracellular matrix and detached germ tubes from surfaces. Treatments with water, dilute acids, ionic and neutral detergents, organic solvents, hydrocarbons, and several polysaccharide-degrading enzymes did not remove the extracellular matrix and also did not detach germ tubes. These results, together with staining patterns obtained with lectins and other polysaccharide-specific reagents, indicate that the extracellular matrix is composed mainly of glycoproteins rich in acidic amino acids and β-1,3-glucan polymers, and that it is probably responsible for the adhesion of the rust germ tubes to the host leaf surfaces. Key words: Puccinia sorghi, germ tube adhesion, extracellular matrix, cytochemistry.



1982 ◽  
Vol 60 (11) ◽  
pp. 2313-2324 ◽  
Author(s):  
Michael T. Higham ◽  
Kathleen M. Cole

Spore development was studied in Choanephora cucurbitarum by using transmission and scanning electron microscopy. Sporangioles are produced by expansion of the ampulla wall. A two-layered spore wall is then constructed within the spine-covered sporangiole wall. The outer spore wall layer is longitudinally grooved and is devoid of spines or appendages. The inner wall layer is thinner and electron transparent. During wall production, dark-staining granular vesicles were observed in the spore cytoplasm. Their contents stained similarly to the material of the outer wall layer. Mature spores possessed a third, innermost wall layer. This was identified as a new wall layer, which was continuous with the germ-tube wall of germinated spores. Released spores were observed to be contained within the sporangiole during dispersal and germination.



1971 ◽  
Vol 17 (7) ◽  
pp. 851-856 ◽  
Author(s):  
D. N. Mardon ◽  
I. S. K. Hurst ◽  
E. Balish

Candida albicans formed germ tubes within 3 h at 37C in a glucose–salts–biotin (GSB) medium containing L-alpha-amino-n-butyric acid as the nitrogen source. Optimal germ-tube production was obtained when the inoculum was grown on Sabouraud dextrose agar. The GSB medium containing L-alpha-amino-n-butyric acid promoted germ-tube formation more effectively than GSB medium plus gamma-amino-butyric acid or Sabouraud dextrose broth.Carbon-14 incorporation studies revealed that during germ-tube formation (0–4 h) the 3 carbon of alpha-amino-n-butyric acid was incorporated intracellularly to a greater extent than the 1 carbon. However, during blastospore formation (5–16 h), this difference was less pronounced.When six other Candida species were grown in GSB plus L-alpha-amino-n-butyric acid medium, few germ tubes were observed with the exception of one Candida stellatoidea strain. However, even this strain of C. stellatoidea produced far fewer germ tubes in this minimal culture medium than any strain of C. albicans tested.



2009 ◽  
Vol 77 (4) ◽  
pp. 1596-1605 ◽  
Author(s):  
Suman Ghosh ◽  
Dhammika H. M. L. P. Navarathna ◽  
David D. Roberts ◽  
Jake T. Cooper ◽  
Audrey L. Atkin ◽  
...  

ABSTRACT The opportunistic fungal pathogen Candida albicans is a part of the normal flora but it also causes systemic candidiasis if it reaches the bloodstream. Upon being phagocytized by macrophages, an important component of innate immunity, C. albicans rapidly upregulates a set of arginine biosynthetic genes. Arginine, urea, and CO2 induced hyphae in a density-dependent manner in wild-type, cph1/cph1, and rim101/rim101 strains but not in efg1/efg1 or cph1/cph1 efg1/efg1 strains. Arginase (Car1p) converts arginine to urea, which in turn is degraded by urea amidolyase (Dur1,2p) to produce CO2, a signal for hyphal switching. We used a dur1,2/dur1,2 mutant (KWN6) and the complemented strain, KWN8 (dur1,2/dur1,2::DUR1,2/DUR1,2) to study germ tube formation. KWN6 could not make germ tubes in the presence of arginine or urea but did in the presence of 5% CO2, which bypasses Dur1,2p. We also tested the effect of arginine on the interaction between the macrophage line RAW 264.7 and several strains of C. albicans. Arginine activated an Efg1p-dependent yeast-to-hypha switch, enabling wild-type C. albicans and KWN8 to escape from macrophages within 6 h, whereas KWN6 was defective in this regard. Additionally, two mutants that cannot synthesize arginine, BWP17 and SN152, were defective in making hyphae inside the macrophages, whereas the corresponding arginine prototrophs, DAY286 and SN87, formed germ tubes and escaped from macrophages. Therefore, metabolism of arginine by C. albicans controls hyphal switching and provides an important mechanism for escaping host defense.



2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Tapiwa Matare ◽  
Pasipanodya Nziramasanga ◽  
Lovemore Gwanzura ◽  
Valerie Robertson

Objective. The potential of NaHCO3 versus human serum to induce germ tube formation in Candida albicans was investigated. Specimens. A total of 100 isolates were obtained from oral swabs of patients presenting with thrush. Approval for the study was granted by the Joint Research Ethics Committee (JREC/23/08). Method. Confirmed C. albicans isolates by routine methods were tested for germ tube induction using 5 different concentrations of Tris-maleate buffered NaHCO3 and Tris-maleate buffer control. Standard control strains included were C. albicans (ATCC 10231) and C. krusei (ATCC 6258). Microculture was done in 20 μL inoculums on microscope slides for 3 hours at 37°C. The rate of germ tube formation at 10-minute intervals was determined on 100 isolates using the optimum 20 mM Tris-maleate buffered NaHCO3 concentration. Parallel germ tube formation using human serum was done in test tubes. Results. The optimum concentration of NaHCO3 in Tris-maleate buffer for germ tube induction was 20 mM for 67% of isolates. Only 21% of isolates formed germ tubes in Tris-maleate buffer control. There was no significant difference in induction between human serum and Tris-maleate buffered NaHCO3. Conclusion. Tris-maleate buffered NaHCO3 induced germ tube formation in C. albicans isolates at rates similar to human serum.



2013 ◽  
Vol 59 (3) ◽  
pp. 197-203 ◽  
Author(s):  
Ola M. Gomaa ◽  
Abeer S. Gaweesh

Adhesion of Candida cells to surfaces is considered the first step in colonization. Some natural products, such as propolis, could be used to block cell adhesion and therefore preventing colonization. In this study, Egyptian propolis ethanol extract concentrations in the range of 25 to 125 ng/μL were used to inhibit the adhesion of oral Candida. The exopolysaccharides showed a 2.5-fold decrease, while the surface-bound exopolysaccharides showed only about 1.15-fold decrease. On the other hand, surface-bound proteins decreased by 7.5-fold upon the addition of 75 ng/μL propolis. The inhibition of adhesion was detected by scanning electron microscopy. The non-slip incubation assay showed a significant decrease in germ tube formation (GTF) compared with an increase upon the addition of fetal bovine serum and corn meal, both of which had a positive effect on GTF compared with a negative GTF effect when using propolis, suggesting that propolis could be considered a quorum-sensing molecule. The use of propolis would help in maintaining the cleanliness of dental fixtures and (or) treating recurrent candidiasis as a complementary and alternative treatment, especially in elders and immunocompromised patients.



1977 ◽  
Vol 55 (9) ◽  
pp. 1137-1142 ◽  
Author(s):  
I. Chet ◽  
D. Timar ◽  
Y. Henis

Sclerotia of Sclerotium rolfsii germinated without the addition of an exogenous nutrient source. The germinating sclerotia excreted sugars and amino acids to the surrounding medium. Continuous washing of sclerotia prevented germination. In germinating sclerotia, the rate of uptake and incorporation into macromolecules of [3H]uridine and[3H]leucine was very low in the first 6 h but increased rapidly during the next 6 h. Electron microscopy of germinating sclerotial slices revealed that only the granulated medullar cells were involved in germ-tube formation. The germination tubes were very electron-dense and had a large number of mitochondria near their tips. The huge vesicles of the cortex cells appeared almost empty in the germinating sclerotium. It is suggested that these cells serve as an external nutrient reservoir for the germination tubes.



1982 ◽  
Vol 152 (2) ◽  
pp. 555-562
Author(s):  
E Mattia ◽  
G Carruba ◽  
L Angiolella ◽  
A Cassone

A number of strains of Candida albicans were tested for germ tube formation after induction by N-acetyl-D-glucosamine (GlcNAc) and other simple (proline, glucose plus glutamine) or complex (serum) compounds. A proportion of strains (high responders) were induced to form germ tubes evolving to true hyphae by GlcNAc alone or by proline or glucose plus glutamine mixture. The majority of strains were low responders because they could be induced only by serum or GlcNAc-serum medium. Two strains were found to be nonresponders: they grew as pseudohyphae in serum. Despite minor quantitative differences, all strains efficiently utilized GlcNAc for growth under the yeast form at 28 degrees C. They also had comparable active, inducible, and constitutive uptake systems for GlcNAc. During germ tube formation in GlcNAc, the inducible uptake system was modulated, as expected from induction and decay of GlcNAc kinase. Uranyl acetate, at a concentration of 0.01 mM, inhibited both GlcNAc uptake and germ tube formation and was reversed by phosphates. Germinating and nongerminating cells differed in the rapidity and extent of GlcNAc incorporation into acid-insoluble and alkali-acid-insoluble cell fractions. During germ tube formation induced by proline, GlcNAc was almost totally incorporated into the acid-insoluble fraction after 60 min. Moreover, hyphal development on induction by either GlcNAc or proline was characterized by an apparent "uncoupling" between protein and polysaccharide metabolism, the ratio between the two main cellular constituents falling from more than 1 to less than 0.5 after 270 min of development. The data suggest that utilization of the inducer for wall synthesis is a determinant of germ tube formation C. albicans but that the nature and extent of inducer uptake is not a key event for this phenomenon to occur.



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