universe84a

Atlas of Fungi

The name ‘ Atlas of Fungi’ is given even due to the vast spectrum of mycology but puny collection and another thing is that only an epic center collection of my authentical performance. So, please if you have benefited from this atlas, let others know about it too and share them through social media.

List of Fungi

1. Candida albicans
2. Candida glabrata
3. Candida Krusei
4. Candida tropicalis
5. Cryptococcus neoformans
6. Geotrichum
7. Malassezia
8. Aspergillus flavus
9. Aspergillus fumigatus
10. Aspergillus niger
11. Bipolaris
12. Cladosporium
13. Curvularia
14. Epidermophyton floccosum
15. Trichophyton mentagrophyte
16. Trichophyton rubrum
17. Microsporum persicolor
18. Microsporum ferrugineum
19. Fonsecaea
20. Fusarium oxysporum
21. Fusarium solani
22. Mucor
23. Penicillium
24. Rhizopus
25. Scedosporium
26. Sporothrix schenckii
27. Syncephalastrum
28. Trichosporon
29. Acremonium
30. Ochroconis gallopava
31. Trichoderma species

Candida albicans

Candida albicans in a saline wet mount

To visualize Candida albicans in a saline wet mount, we would need to perform the following steps:

1. Obtain a sample of the material suspected to contain Candida albicans. This could be oral swabs or vaginal secretions, or  culture for example.
2. Add a small drop of sterile saline solution to a clean glass slide.
3. Using a sterile swab or pipette, transfer a small amount of the sample onto the drop of saline on the slide.
4. Mix the sample with the saline using a sterile applicator or pipette.
5. Place a coverslip over the sample and gently press down to spread the sample and remove any air bubbles.
6. Observe the sample under a microscope using the 10X or 40X objective lens.
7. Look for oval or round yeast cells, which may appear singly or in clusters. Candida albicans is typically identified by its characteristic budding morphology, in which smaller cells emerge from larger cells.
8. Other features to look for include the presence of pseudohyphae (elongated cells connected end-to-end), and the absence of true hyphae (long, branching cells).
9. Take note of the quantity of Candida albicans cells present, as well as any other microorganisms or cells that may be present in the sample.

C. albicans  in Gram stain

To visualize Candida albicans in a Gram stain, we would need to perform the following steps:

1. Prepare a Gram stain slide by placing a small drop of normal saline  on a clean glass slide.
2. Using a sterile loop or swab, transfer a small amount of the Candida albicans sample onto the drop of water on the slide.
3. Mix the sample with the water using the loop or swab.
4. Allow the slide to air dry completely.
5. Once the slide is dry, cover the sample with crystal violet stain for 1 minute.
6. Rinse the slide with distilled water to remove excess stain.
7. Cover the sample with iodine for 1 minute.
8. Rinse the slide again with distilled water.
9. Decolorize the sample with 95% ethanol for 10-20 seconds.
10. Rinse the slide with distilled water.
11. Counterstain the sample with safranin for 1 minute.
12. Rinse the slide with distilled water and blot dry with bibulous paper.
13. Observe the sample under a microscope using the oil immersion objective lens.
14. Look for oval or round yeast cells, which will appear purple or blue-violet in color due to the crystal violet stain.
15. Take note of any budding cells or pseudohyphae, which may be visible as smaller purple or blue-violet cells emerging from larger cells.
16. Record the findings and communicate them to the appropriate healthcare provider, who can make a diagnosis and recommend treatment if necessary.

Note that Gram staining is not always reliable for identifying Candida albicans, as it can be difficult to distinguish from other yeasts and fungal species. Other diagnostic tests may be needed to confirm the presence of Candida albicans. But the above picture of Known Gram stained smear of Candida albicans.

C. albicans growth/ colony characteristics on SDA

Candida albicans germ tube test positive

The germ tube test is a laboratory test used to identify Candida albicans, a type of yeast that can cause infections in humans. In this test, a sample of C. albicans is incubated in human serum or plasma at 37°C for two to three hours. The test is considered positive if the C. albicans cells produce germ tubes, which are elongated projections from the yeast cell.

If the germ tube test is positive for C. albicans, it indicates that the fungus is present and actively growing. This information can be useful in the diagnosis of a C. albicans infection, as well as in determining the most appropriate treatment.

However, it is important to note that not all strains of C. albicans will produce germ tubes, and other Candida species may also produce germ tubes, albeit less frequently. Therefore, the germ tube test should be used in conjunction with other laboratory tests and clinical observations to confirm a diagnosis of C. albicans infection.

Chlamydospores of Candida albicans

Candida albicans is a type of fungus that can exist as a commensal microorganism in the human body, particularly in the gastrointestinal and genitourinary tracts. Under certain conditions, such as a weakened immune system or prolonged antibiotic use, C. albicans can overgrow and cause infections.

Chlamydospores are a type of specialized spore produced by some fungi, including C. albicans. These spores are typically round or oval in shape and have thick walls that help them resist harsh environmental conditions. Chlamydospores of C. albicans  is formed by the aggregation of cytoplasm and organelles within the cell, followed by the deposition of a thick cell wall around the mass.

Chlamydospores of C. albicans can be identified in the laboratory by their characteristic morphology and staining properties. They are often used as a diagnostic tool for identifying the presence of C. albicans in clinical specimens, such as vaginal swabs or skin scrapings.

While the function of chlamydospores in C. albicans is not well understood, it is thought that they may serve as a means of survival and dispersal for the fungus. Additionally, chlamydospores may play a role in the pathogenesis of C. albicans infections by allowing the fungus to persist in host tissues and resist antifungal treatments.

Carbohydrate Fermentation Test of Candida albicans-

Candida albicans on chromagar

CHROMagar Candida or HiCrome candida differential agar recommendation is for rapid isolation and identification of Candida species from mixed cultures in clinical and non-clinical samples.

Cultural characteristics of CHROMagar Candida
After incubation at 25-30°C for 40-48 hours.

Organism Growth      Color of Colony

Candida albicans  -TCC 10231 -good-luxuriant green

Candida glabrata ATCC 15126- good-luxuriant cream to white

Candida krusei ATCC 24408 -good-luxuriant purple, fuzzy

Candida tropicalis ATCC 750 -good-luxuriant blue to purple

Escherichia coli ATCC 25922- inhibited

Staphylococcus aureus ATCC 25923- inhibited

Candida glabrata on SDA

Candida glabrata Showing Germ Tube Test -Negative

Candida glabrata in Giemsa stained smear of sputum

Candida dubliniensis -White to cream-coloured smooth, glabrous yeast-like colonies on Potato dextrose agar (PDA)-

Candida dubliensis: Tease mount preparation of fungal culture (PDA) using LPCB showing spherical to subspherical budding blastoconidia, branched pseudohyphae and terminal chlamydospores too-

Carbohydrate Assimilation Test of Various Yeasts like Candida albicans, Cryptococcus neoformans, Candida species, Saccharomyces cerevisiae 

Cryptococcus neoformans

Nigrosin Stain Preparation of CSF Showing Capsule of Cryptococcus neoformans as shown below-

Cryptococcus neoformans growth on SDA is showing  Cream -coloured smooth, mucoid yeast-like colonies.

C. neoformans India Ink preparation from SDA colony-

Cryptococcus neoformans yeast cells and budding in Gram-stained smear-

Cryptococcus neoformans in LPCB Preparation-

Cryptococcus neoformans In Giemsa Stained Smear of CSF-

Cryptococcus neoformans showing Urease Test Positive-

Urease Test for Cryptococcus

Test medium used: Motility Indole

Urea (MIU) agar

1. Uninoculated

2. Inoculated MIU medium  after 5 days of incubation showing

positive result as shown in tube no 2.

Cryptococcus neoformans on Caffeic Acid Agar after 5 Days of Incubation-

Caffeic Acid Agar

Caffeic acid

⇓  Phenol oxidase (C. neoformans)

Brown pigment

Note: In the composition of the Bird SeedAgar, bird seed/ niger seed (Guizotia abyssinica seed) was replaced by the caffeic acid present on the coffee.

Coffee agar showing growth of  Cryptococcus neoformans (A-brown colony) and Candida albicans (B-white colony)

Carbohydrate Assimilation Test of Cryptococcus neoformans-

Geotrichum grow on SDA

LPCB Mount of Geotrichum candidum showing arthroconidia

Malassezia growth on SDA

Malassezia structures in a wet mount

LPCB mount of Malassezia from culture (SDA)

Aspergillus niger

Potassium hydroxide (KOH) preparation of aural discharge  showing septate hyphae

Gram staining showing plenty of pus cells with Gram-positive Conidia but lacking bacteria

Aspergillus has grown on CHOC, BAP, MAC and SDA media.

Fungal growth on Robertson’s Cooked Meat (RCM) medium

Aspergillus has grown even on anaerobically incubated 5%  sheep blood agar plate (BAP) as shown below-

Aspergillus niger spores on LPCB tease mount from
                      anaerobically incubated plate

Aspergillus niger growth  on CMA

Aspergillus niger growth on SDA showing dark brown to black colonies

Aspergillus niger conidiophores, vesicles, metulae, phialides 

        and conidia on LPCB preparation

Aspergillus fumigatus

Fungal spores ( conidia) on aural discharge Gram-stained smear of CSOM patient-

KOH mount of aural discharge: Potassium hydroxide (KOH) preparation showing septate hyphae with characteristic dichotomous branching  (at an angle of approximately 45°)-

Fungal growth on RCM-

Fungal Growth on Bacteria media:

Even fungal growth on bacterial media after a day incubation at 37.0°C as a shown image-

1. CHOC: Chocolate agar
2. BAP: 5% Sheep blood agar
3. MAC: MacConkey medium
4. SDA: Sabouraud dextrose agar ( for fungi)-

Aspergillus fumigatus Colony characteristics:

Colonies on SDA showing green surface pigmentation with a suede-like surface consisting of a dense felt of conidiophores.

Aspergillus fumigatus: Tease mount preparation of fungal culture using LPCB Conidial head morphology showing stipe, conidiophore, vesicle, metulae, phialides, and conidia

Note- a uniseriate row of phialides on the upper two-thirds of the vesicle

Robertson’s Cooked Meat (RCM) medium :

This medium is usually used to cultivate anaerobic bacteria but also facilitate the growth of fungus as shown in this image.

Later confirmed growth of Aspergillus flavus.

Aspergillus flavus Colony Morphology-

Colonies on SDA are granular, flat often with radial grooves, initially

yellow but quickly changes bright to dark yellow-green with time.

Aspergillus flavus:  Tease mount preparation of fungal culture using LPCB showing conidial head morphology showing rough-walled stipe near the vesicle, conidiophore (septate), vesicle, metulae, phialides, and conidia

Note- Both uniseriate and biseriate rows of phialides may be present.

Bipolaris growth on SDA

Bipolaris on LPCB preparation showing sympodial development of  pale brown, fusiform, to ellipsoidal, pseudoseptate, proconidia on a geniculate 

Cladosporium growth on SDA

Tease mount preparation of Cladosporium culture using LPCB  showing Conidia of Cladosporium

KOH preparation of a skin scale showing ringworm fungi   (dermatophytes) with branching septate  hyphae

Trichophyton mentagrophytes Colony characteristics on SDA-

Colonies are flat, white to cream in colour,  with a powdery to the granular surface (A).

Reverse pigmentation-

Yellow to brown (B)

 LPCB Preparation from Trichophyton mentagrophytes culture showing hyphae (1),   microconidia (2), macroconidia (3), chlamydospores(4), and spiral hyphae(5)

Trichophyton mentagrophytes Colony morphology on SDA-

Colonies are flat, white to cream in colour,  with a powdery to the granular surface (A).

Reverse pigmentation- Yellow to brown (B)

Epidermophyton floccosum-

Young culture  of Epidermophyton  floccosum on SDA:

-Slow grower

-Generally, acquire greenish-brown or khaki colour

Reverse pigmentation: Deep yellowish-brown

LPCB Preparation from the young culture of Epidermophyton floccosum

showing blastospores and pseudomycellium but lacking chlamydospores

Microsporum ferrugineum Colony Morphology-

Microsporum ferrugineum colonies on SDA are waxy, glabrous, convoluted

thallus with a cream to buff coloured surface(A). Reverse Pigmentation-

Lacking (B)-

Microsporum ferrugineum LPCB Preparation

LPCB preparation from the culture of Microsporum ferrugineum showing

bamboo hyphae (1) and the absence of both microconidia and macroconidia

Microsporum persicolor growth on Dermasel agar and LPCB preparation

Cladosporium growth on SDA

Conidiophores(1) and conidia (2) of Cladosporium

Fonsecaea colony morphology on SDA Plate and Tube

Conidiophores (1) and conidia(2) of Fonsecaea

Fusarium oxysporum growth on SDA after 5 days of incubation-

Microconidia, phialides and macroconidia of Fusarium oxysporum-

Microconidia, phialides and macroconidia of Fusarium oxysporum-

Fusarium solani growth on SDA after 5 days of incubation-

Fusarium solani in LPCB preparation showing microconidia, long phialides and chlamydospores-

Sporangia and sporangiospores of Mucor

Mucor structures under the microscope Showing sporangium, sporangiospores, columella, sporangiophore or aerial hyphae

Paecilomyces in LPCB preparation showing conidiophores, phialides, conidia and conidia in divergent chains-

Paecilomyces marquandii growth on CMA

Paecilomyes marquandii in LPCB preparation showing conidiophores, phialides and conidia 

Penicillium growth on SDA after 4 days of incubation

Penicillium in LPCB preparation showing conidiophores, flask-shaped phialides arranged in groups from branched metulae forming a penicillus and conidia

Penicillium cheresanum–In LPCB Preparation showing long chains of single–celled conidia

Penicillium marneffei

Penicillium marneffei exhibits thermal dimorphism by growing in living tissue or in the culture at 37°C as a yeast-like fungus, and in the culture at below 30°C as a mould (as in Figure).

Penicillium marneffei in LPCB Preparation showing conidiophores, phialides and conidia

Rhizopus in LPCB mount showing sporangiophores (A), columellae (B) and rhizoids (C)

Scedosporium growth on SDA

Conidiophores and conidia of Scedosporium in LPCB Preparation

Growth of Sporothrix schenckii

Conidiophores and conidia of Sporothrix schenckii in LPCB from SDA growth

Syncephalastrum growth on SDA after 3 days of incubation 

LPCB mount showing sporangiophores, merosporangia and merospores

of Syncephlastrum-

Trichosporon growth on PDA

LPCB Mount of Trichosporon

Acremonium on SDA after 13 days of incubation showing powdery colonies

Acremonium on SDA after 13 days of incubation showing powdery colonies-

Acremonium in LPCB preparation showing long awl-shaped phialides producing cylindrical, one-celled conidia mostly aggregated in slimy heads at the apex of each phialide-

Aural discharge in KOH mount showing hyphae and conidia-

Ochroconis gallopava Growth on SDA has Smooth to suede-like, dry, flat, tobacco brown colonies with dark brown diffusible pigment-

Ochroconis gallopava fungal elements in 0.05% Tween 80 wet mount from a culture showing conidia, conidiophores, hyphae and hyphae are brown with relatively thick walls.

Hyphae (A), Conidiophores (B) and conidia (C) of Ochroconis gallopava

Antifungal Susceptibility Testing (AFST)

–              Disk Diffusion Method

Approved guideline M-44 A, CLSI, USA

–              only for testing Candida species

-Medium: MHA + 2% glucose and 0.5 μg methylene blue dye (GMB); pH 7.2-7.4.

-There should be no excess moisture on plates.

-Turbidity standard for inoculum preparation – 0.5 McFarland standard

Fungus in Ziehl-Neelsen stained smear of sputum showing yeast cells, budding and paseudohyphae

Fungal elements in KOH mount of Sputum showing yeast cells, budding and pseudohyphae

Conidium of Exserohilum in KOH Mount of Skin Scrapping

Conidium and Hypha of Exserohilum in KOH Mount of Skin Scrapping

Dematiaceous fungi or black mould on SDA

Conidia and Hyphae of Dematiaceous Fungi in LPCB Preparation

Trichoderma growth on SDA

Conidia, phialides and conidiophores of Trichoderma-

Fungus, Trichoderma growth on SDA, Conidia, phialides and conidiophores in LPCB Preparation-

Conidia, phialides and conidiophores of Trichoderma species-

Dematiaceous Fungus on SDA and its conidia and hyphae in LPCB preparation-

Septate hyphae and conidia of fungi-

Cylindrocarpon growth on SDA-

Hyphae, chlamydospores and conidia of Cylindrocarpon in LPCB Preparation-

Cylindrocarpon growth on SDA and its structures in LPCB mount-

Scopulariopsis on SDA and CMA-

Conidiophores (annelids) and conidia of Scopulariopsis in LPCB Preparation-

Scopulariopsis growth on SDA and CMA and its Conidiophores (annelids) and conidia in LPCB preparation-

Sporothrix schenckii growth on SDA at 37°C-

Yeast cells of Sporothrix schenckii in LPCB preparation of culture incubated at 37° C-

Aspergillus niger growth on SDA-

Aspergillus niger growth on CMA-

A structure resembling Candida ciferrii in KOH mount of sputum in a COPD patient –

Structures resembling Malbranchea-

Arthroconidia of Malbranchea

Scopulariopsis growth on CMA and SDA agar-

Various forms of conidia, globose, pyriform to truncate of Scopulariopsis in LPCB tease mount-

Mycelium of Scopulariopsis in LPCB tease mount-

Conidia and conidiophores (annellides) of Scopulariopsis in LPCB tease mount-

Fungal hyphae in KOH Mount of Sputum specimen-

Fungal hyphae and conidia in KOH mount of Sputum-

Trichoderma with various structures, Conidiophores, phialides, conidia, septate hyphae-

Fungal hyphae in the urine of diabetic patient-

Curvularia in LPCB showing conidia and hyphae-

Curvularia growth on SDA-

Fungus encountered during PBS comment

You may even grow Cryptococcus on Blood agar as well MacConkey medium if there is no SDA in your laboratory-

Fungal Elements-Hyphae, yeast cells and Conidia in KOH Mount of Skin Scrapping – 

Pseudohypha of Candida, bacteria, pus cell and epithelial cell in Gram-stained smear of sputum-

Curvularia colony on SDA

Curvuaria reverse pigment expression on SDA-

Melanized fungus, Curvuaria colony on SDA-

Fungal septate hyphae in the v KOH mount of sputum-

KOH mount of sputum under the Microscope showing fungal elements, yeast cells, budding and fungal hyphae-

Yeast cells in Giemsa stained smear of sputum-

Contd…