Absidia corymbifera. Absidia corymbifera showing a typical pyriform-shaped sporangium with a conical-shaped columella and pronounced apophysis.

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1 Absidia corymbifera Absidia corymbifera showing a typical pyriform-shaped sporangium with a conical-shaped columella and pronounced apophysis. Colonies are fast growing, floccose, white at first becoming pale grey with age, and up to 1.5 cm high. Sporangiophores are hyaline to faintly pigmented, simple or sometimes branched arising solitary from the stolons, in groups of three, or in whorls of up to seven. Rhizoids are very sparingly produced and may be difficult to find without the aid of a dissecting microscope to examine the colony on the agar surface. Sporangia are small (10-40 um in diameter) and are typically pyriform in shape with a characteristic conical-shaped columella and pronounced apophysis, often with a short projection at the top. Sporangiospores vary from subglobose to oblong-ellipsoidal ( x um), are hyaline to light grey and are smooth-walled. Temperature: optimum 35-37C; maximum 45C. The genus Absidia is characterized by a differentiation of the hyphae into arched stolons bearing more or less verticillate sporangiophores at the internode, and rhizoids formed at the point of contact with the substrate (at the node). This feature separates species of Absidia from the genus Rhizopus, where the sporangia arise from the nodes and are therefore found opposite the rhizoids. The sporangia are relatively small, globose, pyriform- or pear-shaped and are supported by a characteristic funnel-shaped apophysis. This distinguishes Absidia from the genera Mucor and Rhizomucor, which have large, globose sporangia without an apophysis. Absidia currently contains 21 mostly soil-borne species. A. corymbifera is the only species of Absidia known to cause disease in man and animals.

2 2 Clinical significance Absidia corymbifera is a common human pathogen, causing pulmonary, rhinocerebral, disseminated, CNS or cutaneous types of infection. It is also often associated with animal disease, especially mycotic abortion. A. corymbifera has a world-wide distribution mostly in association with soil and decaying plant debris. Clinical group: Opportunistic mycoses Mycosis: Zygomycosis Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

3 3 Acremonium sp. Acremonium sp. showing long awl-shaped phialides producing cylindrical, one-celled conidia mostly aggregated in slimy heads at the apex of each phialide. Colonies are usually slow growing, often compact and moist at first, becoming powdery, suede-like or floccose with age, and may be white, grey, pink, rose or orange in color. Hyphae are fine and hyaline and produce mostly simple awl-shaped erect phialides. Conidia are usually one-celled (ameroconidia), hyaline or pigmented, globose to cylindrical, and mostly aggregated in slimy heads at the apex of each phialide. This genus is distinguished from hyaline isolates of Phialophora by the absence or very limited development of a collarette on the phialide and the predominant formation of well differentiated, awl-shaped phialides with a basal septum. Microconidial Fusarium isolates may be confused with Acremonium, but they usually grow faster and have colonies with a characteristic fluffy appearance. For identification, potato dextrose agar and cornmeal agar are the most suitable media to use and exposure to daylight is recommended to maximize culture color characteristics. Clinical significance The genus Acremonium currently contains 100 species, of which most are saprophytic, being isolated from dead plant material and soil. A number of species are recognized as opportunistic pathogens of man and animals, causing mycetoma, onychomycosis, and hyalohyphomycosis, these include A. falciforme, A. kiliense and A. recifei, A. alabamensis, A. potroni, A. roseo-griseum and A. strictum. However, many reports only identify Acremonium species to genus level. Clinical manifestations of hyalohyphomycosis caused by Acremonium; include arthritis, osteomyelitis, peritonitis, endocarditis, pneumonia, cerebritis and subcutaneous infection.

4 4 Clinical group: Opportunistic mycoses Mycosis: Hyalohyphomycosis Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

5 5 Alternaria sp. Alternaria alternata showing branched acropetal chains and multicelled, obclavate to obpyriform conidia with short conical beaks. Colonies are fast growing, black to olivaceous-black or greyish, and are suede-like to floccose. Microscopically, branched acropetal chains (blastocatenate) of multicelled conidia (dictyoconidia) are produced sympodially from simple, sometimes branched, short or elongate conidiophores. Conidia are obclavate, obpyriform, sometimes ovoid or ellipsoidal, often with a short conical or cylindrical beak, pale brown, smooth-walled or verrucose. Alternaria species soon lose their ability to sporulate in culture. Potato dextrose agar and cornmeal agar are the most suitable media to use, and incubation under near ultra-violet light is recommended to maintain sporulation. Temperature: optimum 25-28C; maximum 31-32C. Clinical significance The genus contains 44 species of which most are plant parasites, but a few species are ubiquitous and are also frequently soil-borne. A. alternata is the commonest of these. Although usually seen as saprophytic contaminants, Alternaria species are recognized causative agents of mycotic keratitis and phaeohyphomycosis. Clinical manifestations include cutaneous infections, paranasal sinusitis, osteomyelitis and peritonitis in patients on continuous ambulatory peritoneal dialysis (CAPD). Clinical group: Opportunistic mycoses Mycosis: Phaeohyphomycosis Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

6 6 Apophysomyces elegans Culture of Apophysomyces elegans Colonies are fast growing, white becoming creamy white to buff with age, downy with no reverse pigment, and are composed of broad, sparsely septate (coenocytic) hyphae typical of a zygomycetous fungus. Sporangiophores are unbranched, straight or curved, slightly tapering towards the apex, up to 200 um long, 3-5 um in width near the apophysis, and hyaline when young but developing as the culture ages, a sepia to brown pigmentation and a conspicuous sub-apical thickening um below the apophysis with age. Sporangiophores arise at right angles from the aerial hyphae and often have a septate basal segment resembling the "foot cell" commonly seen in Aspergillus. Rhizoids are thin-walled, sub-hyaline and predominantly unbranched. Sporangia are multispored, small (20-50 in diameter), typically pyriform in shape, hyaline at first, sepia colored when mature, columellate and strongly apophysate. Columellae are hemispherical in shape and the apophyses are distinctively funnelor bell-shaped. Sporangiospores are smooth-walled, mostly oblong, occasionally subglobose, (3-4 x 5-6 um) and sub-hyaline to sepia in mass. Good growth at 26, 37 and 42C.

7 7 Young sporangium (left) and mature sporangium (right) of Apophysomyces elegans showing distinctive funnel-shaped apophyses and a conspicuous pigmented sub-apical thickening below the apophysis Apophysomyces elegans is readily distinguishable from other zygomycetes of medical importance, especially the morphologically similar, strongly apophysate pathogen Absidia corymbifera. A. elegans has sporangiophores with distinctive funnel- or bellshaped apophyses and hemispherical-shaped columellae, a conspicuous pigmented sub-apical thickening which constricts the lumen of the sporangiophore below the apophysis, and distinctive foot cells. Agar block method to induce sporulation of Apophysomyces elegans Laboratory identification of some zygomycetous fungi, especially Apophysomyces elegans and Saksenaea vasiformis may be difficult or delayed because of the mould's failure to sporulate on the primary isolation media or on subsequent subculture onto potato dextrose agar. Sporulation may be stimulated by the use of nutrient deficient media, like Czapek Dox agar, cornmeal-glucose-sucrose-yeast extract agar, or by using the agar block method described by Ellis and Ajello (1982).

8 8 Briefly, a small block of agar is cut from a well established culture grown on PDA and is placed in the centre of petri dish containing 1% agar in distilled water. After 21 days at 26C look for sporangium formation at the periphery of the petri dish. Clinical significance Apophysomyces elegans is a rare human pathogen usually associated with invasive lesions following traumatic implantation of the fungus through the skin. It is a soil fungus with a tropical to sub-tropical distribution. Clinical group: Opportunistic mycoses, Subcutaneous mycoses Mycosis: Zygomycosis Further reading Ellis, D.H Zygomycetes. Chapter 16 In Topley and Wilson's Microbiology and Microbial Infections. 9th edition Edward Arnold London pp Ellis, J.J., and L. Ajello An unusual source of Apophysomyces elegans and a method of stimulating sporulation of Saksenea vasiformis. Mycologia 74:

9 9 Aspergillus flavus Culture of Aspergillus flavus. On Czapek dox agar, colonies are granular, flat, often with radial grooves, yellow at first but quickly becoming bright to dark yellow-green with age. Conidial heads are typically radiate, mostly um in diameter, later splitting to form loose columns, biseriate but having some heads with phialides borne directly on the vesicle. Conidiophores are hyaline and coarsely roughened, the roughness often being more noticeable near the vesicle. Conidia are globose to subglobose (3-6 um in diameter), pale green and conspicuously echinulate. Some strains produce brownish sclerotia. Conidial head of A. flavus. Note: conidial heads with both uniseriate and biseriate arrangement of phialides may be present.

10 10 Clinical significance Aspergillus flavus has a world-wide distribution and normally occurs as a saprophyte in soil and on many kinds of decaying organic matter. A. flavus is the second most common species (next to A. fumigatus) to be isolated from human infections, and it is often associated with invasive aspergillosis seen in immunosuppressed patients and in paranasal sinus infections. Clinical group: Opportunistic mycoses Mycosis: Aspergillosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

11 11 Aspergillus fumigatus Culture of Aspergillus fumigatus On Czapek dox agar, colonies show typical blue-green surface pigmentation with a suede-like surface consisting of a dense felt of conidiophores. Conidial heads are typically columnar (up to 400 x 50 um but often much shorter and smaller) and uniseriate. Conidiophores are short, smooth-walled and have conical-shaped terminal vesicles which support a single row of phialides on the upper two thirds of the vesicle. Conidia are produced in basipetal succession forming long chains and are globose to subglobose ( um in diameter), green and rough-walled. Note, this species is thermotolerant and grows at temperatures up to 55C. Conidial head of A. fumigatus (Note: uniseriate row of phialides on the upper two thirds of the vesicle) Clinical significance Aspergillus fumigatus is truly a cosmopolitan mould and has been found almost

12 12 everywhere on every conceivable type of substrate, especially soil and decaying organic debris. A. fumigatus is an important human pathogen and it is the most common cause of all forms of invasive and non-invasive aspergillosis. Clinical group: Opportunistic mycoses Mycosis: Aspergillosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

13 13 Teleomorph: Emericella nidulans Aspergillus nidulans Culture of Aspergillus nidulans On Czapek dox agar, colonies are typically plain green in color with dark redbrown cleistothecia developing within and upon the conidial layer. Reverse may be olive to drab-gray or purple-brown. Conidial heads are short, columnar (up to 70 x 30 um in diameter) and biseriate. Conidiophores are usually short, brownish and smooth-walled. Conidia are globose ( um in diameter) and rough-walled. Conidial head of A. nidulans. Note: conidial heads are short, columnar and biseriate. Conidiophores are usually short, brownish and smooth-walled. Conidia are globose and rough-walled.

14 14 Cleistothecium of Emericella nidulans (anamorph A. nidulans) showing numerous reddish-brown ascospores. Cleistothecia of A. nidulans are often surrounded by a mass of Hülle cells which are up to 25 um in diameter. Thick-walled Hülle cells of Emericella nidulans. Clinical significance Aspergillus nidulans is a typical soil fungus with a world-wide distribution. It has also been reported as a causative agent of aspergillosis in humans and animals. Clinical group: Opportunistic mycoses Mycosis: Aspergillosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

15 15 Aspergillus niger Culture of Aspergillus niger On Czapek dox agar, colonies consist of a compact white or yellow basal felt covered by a dense layer of dark-brown to black conidial heads. Conidial heads are large (up to 3 mm x um in diameter), globose, dark brown, becoming radiate and tending to split into several loose columns with age. Conidiophores are smooth-walled, hyaline or turning dark towards the vesicle. Conidial heads are biseriate with the phialides borne on brown, often septate metulae. Conidia are globose to subglobose ( um in diameter), dark brown to black and roughwalled. Conidial head of A. niger. Note: conidial heads are biseriate, large, globose, dark brown, becoming radiate with the phialides borne on metulae. Clinical significance Aspergillus niger is one of the most common and easily identifiable species of the genus Aspergillus, with its white to yellow mat later bearing black conidia. This is the third most common species associated with invasive pulmonary aspergillosis. It is also often a causative agent of aspergilloma and is the most frequently

16 16 encountered agent of otomycosis. A. niger may also be a common laboratory contaminant. Clinical group: Opportunistic mycoses Mycosis: Aspergillosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

17 17 Aspergillus terreus Culture of Aspergillus terreus On Czapek dox agar, colonies are typically suede-like and cinnamon-buff to sand brown in color with a yellow to deep dirty brown reverse. Conidial heads are compact, columnar (up to 500 x um in diameter) and biseriate. Conidiophores are hyaline and smooth-walled. Conidia are globose to ellipsoidal ( um in diameter), hyaline to slightly yellow and smooth-walled. Conidial head of A. terreus. Note: conidial heads are biseriate Clinical significance Aspergillus terreus occurs commonly in soil and is occasionally reported as a pathogen of humans and animals. Clinical group: Opportunistic mycoses Mycosis: Aspergillosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

18 18 Aureobasidium pullulans A. pullulans showing chains of 1- to 2-celled, darkly pigmented arthroconidia representing the Scytalidium anamorph of Aureobasidium and the presence of numerous hyaline, single-celled, ovoid-shaped conidia (ameroconidia) which are produced on short denticles. Colonies are fast growing, smooth, covered with slimy masses of conidia, cream or pink to brown or black. Hyphae are hyaline and septate, frequently becoming dark-brown with age and undergoing holothallic transformation to form chains of 1- to 2-celled, thick-walled, darkly pigmented arthroconidia commonly called chlamydoconidia. These arthroconidia actually represent the Scytalidium anamorph of Aureobasidium and are only of secondary importance in recognizing members of this genus. Conidia are produced synchronously in dense groups from indistinct scars or from short denticles on undifferentiated, hyaline to sub-hyaline hyphae. Conidia are hyaline, smooth-walled, single-celled (ameroconidia), ellipsoidal but of variable shape and size (8-12 x 4-6 um), often with an indistinct hilum (a mark or scar at the point of attachment). Temperature range for growth 2-35C; optimum 25C; maximum 35C (higher in some human pathogenic isolates). This species has two varieties: A. pullulans var. pullulans, with a colony which remains pink, light brown, or yellow for at least three weeks, and A. pullulans var. melanogenum which soon becomes black or greenish-black due to dark hyphae which often fall apart into separate cells. Clinical significance Aureobasidium pullulans has a world-wide distribution and it is usually isolated as a saprophyte, occasionally from skin and nails. However, it has also been reported as a rare causative agent of phaeohyphomycosis, mycotic keratitis and peritonitis in patients on continuous ambulatory peritoneal dialysis (CAPD). Clinical group: Opportunistic mycoses Mycosis: Phaeohyphomycosis Further reading De Hoog, G.S., and E.J. Hermanides-Nijhof The black yeasts and allied hyphomycetes. Studies in Mycology No. 15. Centraalbureau voor Schimmelcultures, Baarn, The Netherlands.

19 19 Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

20 20 Basidiobolus ranarum Synonym: Basidiobolus meristosporus; Basidiobolus haptosporus; Basidiobolus heterosporus Culture of Basidiololus ranarum. Colonies are moderately fast growing at 30C, flat, yellowish-grey to creamy-grey, glabrous, becoming radially folded and covered by a fine, powdery, white surface mycelium. Note, satellite colonies are often formed by germinating conidia ejected from the primary colony. Zygospores of Basidiobolus ranarum. Microscopic examination usually shows the presence of large vegetative hyphae (8-20 um in diameter) forming numerous round (20-50 um in diameter), smooth, thick-walled zygospores that have two closely appressed beak-like appendages. Two types of asexual spores are formed: primary and secondary. Primary spores are globose, one-celled, solitary and are forcibly discharged from a sporophore. The sporophore has a distinct swollen area just below the spore that actively participates in the discharge of the spore. Secondary spores are clavate, one-celled and are passively released from a sporophore. These sporophores are not swollen at their bases. The apex of the passively released spore has a knob-like adhesive tip. These spores may function as sporangia, producing several sporangiospores. Note, isolates often lose their sporulating ability with subculture and special media incorporating glucosamine hydrochloride and casein hydrolsate may need to be used to stimulate sporulation.

21 21 Asexual spores of B. ranarum. Note typical swollen sporophore in lower right corner Clinical significance Basidiobolus ranarum is commonly present in decaying fruit and vegetable matter, and as a commensal in the intestinal tract of frogs, toads and lizards. It has been reported from tropical Africa, India, Indonesia and South East Asia including the Northern Territory of Australia. Clinical group: Subcutaneous mycoses Mycosis: Zygomycosis Further reading Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

22 22 Beauveria sp. Beauvaria bassiana showing sympodial development of conidia on a geniculate or zig-zag rachis. Conidiogenous cells are flask-shaped, rachiform, proliferating sympodially and are often aggregated into sporodochia or synnemata. Conidia are hyaline and globose or ovoid in shape. Colonies are usually slow growing, downy, at first white but later often becoming yellow to pinkish. The genus Beauveria is characterized by the sympodial development of single-celled conidia (ameroconidia) on a geniculate or zig-zag rachis. Conidiogenous cells are flask-shaped, rachiform, proliferating sympodially and are often aggregated into sporodochia or synnemata. Conidia are hyaline and globose or ovoid in shape. Clinical significance Three species of Beauvaria are recognized, two of which are well known parasites of insects. B. bassiana is the most common species and is best known as the causal agent of the disastrous muscardine in silkworms. Beauveria species are occasionally isolated in the clinical laboratory as saprophytic contaminants. Clinical group: Saprophytes Mycosis: Hyaline hyphomycetes Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

23 23 Bipolaris sp. Teleomorph: Cochliobolus sp. Bipolaris australiensisshowing sympodial development of pale brown, fusiform to ellipsoidal, pseudoseptate, poroconidia on a geniculate or zig-zag rachis. Colonies are moderately fast growing, effuse, grey to blackish brown, suede-like to floccose with a black reverse. Microscopic morphology shows sympodial development of pale brown pigmented, pseudoseptate conidia on a geniculate or zig-zag rachis. Conidia are produced through pores in the conidiophore wall (poroconidia) and are straight, fusiform to ellipsoidal, rounded at both ends, smooth to finely roughened and germinating only from the ends (bipolar). Drechslera, Bipolaris, Curvularia and Exserohilum are all closely related and differentiation of the genera relies upon a combination of characters including conidial shape, the presence or absence of a protruding hilum, the contour of the basal portion of the conidium and its hilum, the point at which the germ tube originates from the basal cell and, to a lesser degree, the sequence and location of the first three conidial septa. The table below is modified from Domsch et al. (1980). Anamorph Main characters Teleomorph Drechslera Bipolaris Conidia cylindrical, germinating from any cell, hilum not protuberant. Conidia fusiformellipsoidal, central cells not much darker and broader than the distal Pyrenophora Cochliobolus

24 24 Curvularia Exserohilum ones, hilum not protuberant, germination bipolar. Conidia with 2-3 broader and darker central cells, often curved, with or without a prominent hilum, germination bipolar. Conidia fusiformcylindrical to obclavate, with a protuberant hilum, germination bipolar. Cochliobolus Setosphaeria Clinical significance The genus Bipolaris contains about 45 species which are mostly subtropical and tropical plant parasites. However, several species notably B. australiensis, B. hawaiiensis and B. spicifera, are well documented human pathogens. Clinical manifestations include mycotic keratitis, subcutaneous phaeohyphomycosis, sinusitis, peritonitis in patients on continuous ambulatory peritoneal dialysis (CAPD), and cerebral and disseminated infections. Phaeohyphomycosis caused by Bipolaris species has been reported in both normal and immunosuppressed patients. Clinical group: Opportunistic mycoses, Mycosis: Phaeohyphomycosis Subcutaneous mycoses Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London. McGinnis, M.R., M.G. Rinaldi and R.E. Winn Emerging agents of Phaeohyphomycosis: pathogenic species of Bipolaris and Exserohilum. J. Clin. Microbiol. 24:

25 25 Blastomyces dermatitidis Tissue section showing large, broad-base, unipolar budding yeast-like cells of Blastomyces dermatitidis. On Sabouraud's dextrose agar at 25C, colonies are variable in both morphology and rate of growth. They may grow rapidly, producing a fluffy white mycelium, or slowly as glabrous, tan, non-sporulating colonies. Growth and sporulation are enhanced by nitrogenous substances found in starling dung and yeast extract. Most strains become pleomorphic with age. Microscopically, hyaline, ovoid to pyriform, one-celled, smooth-walled conidia (2-10 um in diameter) of the Chrysosporium type, are borne on short lateral or terminal hyphal branches. On blood agar at 37C, colonies are wrinkled and folded, glabrous and yeast-like. Microscopically, the organism produces the characteristic yeast phase as seen in tissue pathology. B. dermatitidis can be described as a dimorphic fungus because it has both a mould and yeast phase. WARNING: Cultures of B. dermatiditis represent a severe biohazard to laboratory personnel and must be handled with extreme caution in an appropriate pathogen handling cabinet. In the past, conversion from the mould form to the yeast form was necessary to positively identify this dimorphic pathogen from species of Chrysosporium or Sepedonium. However, culture identification by the exoantigen test is now the method of choice. Clinical significance Blastomycosis is a chronic granulomatous and suppurative disease having a primary pulmonary stage that is frequently followed by dissemination to other body sites, chiefly the skin and bone. Although the disease was long thought to be restricted to the North American continent, in recent years autochthonous cases have been diagnosed in Africa, Asia and Europe. All available clinical and epidemiological evidence indicates that humans and lower animals contract blastomycosis from some source in nature. However, the natural habitat of B. dermatitidis has yet to be clearly delineated, despite some reports of its isolation from soil.

26 26 Clinical group: Dimorphic systemic mycoses Mycosis: Blastomycosis Further reading Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

27 27 Blastoschizomyces capitatis Synonym: Trichosporon capitatis On Sabouraud's dextrose agar colonies are white to cream colored, wrinkled and dull in appearance with a mycelial fringe. Microscopic morphology showing oval to long-oval blastoconidia x um in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: True mycelium and arthroconidia are abundant. The blastoconidia usually form clusters along or at the end of the hyphae. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Negative: Glucose; Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Galactose (Weak); L-Sorbose (Delayed); Citric acid; Glycerol; L-Arabinose; DL-Lactic acid; Succinic acid. Negative: Potassium nitrate; Lactose; Ribitol; Maltose; Cellobiose; D-Ribose; Raffinose; Sucrose; Melibiose; Galactitol; Trehalose; Erythritol; D-Mannitol; D- Xylose; Inositol; D-Glucitol; Soluble Starch; L-Rhamnose; Salicin; Melezitose; D- Arabinose. Blastoschizomyces capitatum is distinguished from Trichosporon beigelii by being lactose negative. Clinical significance Blastoschizomyces capitatis is a common environmental fungus and transient component of the normal skin flora, it is frequently isolated from sputum. Disseminated infections involving the spleen, liver, kidney, lung and brain have been reported in neutropenic patients with leukemia or from those undergoing bone marrow transplantation. Blood cultures are often positive and skin lesions similar to disseminated candidiasis may also be present.

28 28 Clinical group: Opportunistic mycoses Mycosis: Miscellaneous Yeast Infections Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

29 29 Candida albicans Synonym: Candida stellatoidea Culture of Candida albicans. On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows spherical to subspherical budding yeast-like cells or blastoconidia, x um in size. Bubbing yeast cells of Candida albicans India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Pseudohyphae with blastoconidia and terminal vesicles (chlamydoconidia). Physiological Tests: Germ Tube test is Positive within 3 hours Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive

30 30 Germ-tubes of Candida albicans Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Maltose. Variable: Galactose; Trehalose. Negative: Sucrose (some strains positive); Lactose. Assimilation Tests: Positive: Glucose; Maltose; Galactose; Trehalose; Sucrose (some negative);d- Xylose; Soluble Starch; D-Mannitol; D-Glucitol (Delayed). Variable: Melezitose; Glycerol; Succinic acid; L-Arabinose; L-Sorbose; D-Ribose (some positive); Citric acid; DL-Lactic acid. Negative: Potassium nitrate; Lactose; Ribitol (some positive); Raffinose; Cellobiose; Melibiose; Erythritol; Inositol; L-Rhamnose; D-Arabinose; Galactitol; Salicin. Clinical significance Candida albicans occurs naturally as a commensal of mucous membranes and in the digestive tract of humans and animals. It accounts for up to 70% of Candida species isolated from sites of infection and has been reported as a causative agent of all types of candidiasis. Environmental isolations are usually from sources contaminated by human or animal excreta, such as polluted water, soil, air and plants. Clinical group: Cutaneous mycoses, Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands.

31 31 Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

32 32 Candida dubliniensis On Sabouraud's dextrose agar colonies are white to cream coloured, smooth, glabrous yeast-like in appearance [and are indistinguishable from those of C. albicans]. Microscopic morphology shows numerous ovoid, budding yeast-like cells or blastoconidia, x um in size. However, C. dubliniensis may be distinguished from C. albicans when grown on CHROMagar Candida. On this medium, C. albicans colonies are light blue-green in colour, while those of C. dubliniensis are a much darker green colour. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Pseudohyphae with blastoconidia and terminal vesicles (chlamydoconidia) are produced usually in abundance [similar to C. albicans]. Physiological Tests: Germ Tube test is Positive within 3 hours [as in C. albicans]. Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive [However, unlike C. albicans growth at 42C is very poor or absent]. Assimilation Tests: [Note the databases supplied by commercial systems like API 20C and ID32C do not contain profiles for C. dubliniensis, so often the first hint is a germ tube positive yeast with an unusual carbohydrate assimilation profile]. Positive: Glucose, Galactose, Maltose, Sucrose, D-Mannitol, Sorbitol, 2-Ketogluconate, N-Acetylglucosamine, Glucosamine. Variable: Trehalose, Palatinose. Negative: L-Arabinose, D-Xylose, D-Ribose, L-Sorbose, L-Rhamnose, Cellobiose, Lactose, Melibiose, Melezitose, Raffinose, Glycerol, Erythritol, Inositol, DL-Lactic acid, Glucuronate, DL-Lactate, Gluconate, Esculin hydrolysis. Clinical significance Candida dubliniensis has recently been recognised from the oral cavity of HIVinfected patients and are most frequently implicated in cases of recurrent infection following antifungal drug treatment. Phenotypically, isolates are very similar to C.

33 33 albicans in that isolates produce both germ-tubes and chlamydoconidia, however, they have unusual carbohydrate assimilation patterns and grow poorly or not at all at 42C. Clinical group: Cutaneous mycoses, Opportunistic mycoses Mycosis: Candidiasis Further reading Sullivan D. et al. Widespread geographic distribution of oral Candida dubliniensis strains in human immunodificiency virus-infected individuals. J. Clin. Micro : Sullivan D. and D. Coleman. Candida dubliniensis: an emerging opportunistic pathogen. Current Topics in medical Mycology. 1997; 8: Sullivan D and D. Coleman. Minireview. Candida dubliniensis: characteristics and identification. J. Clin. Micro. 1998; 36:

34 34 Candida famata Synonym: Torulopsis candida On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous yeast-like in appearance. Microscopic morphology shows numerous ovoid, budding yeast-like cells or blastoconidia, x um in size. No pseudohyphae produced. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ovoid, budding yeast-like cells only. No pseudohyphae produced. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Variable Growth at 37C is Variable Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Variable: Glucose; Sucrose; Maltose; Trehalose. Negative: Lactose; Galactose. Assimilation Tests: Positive: Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose; Melezitose; Glycerol; Raffinose; Cellobiose; L-Arabinose; D-Mannitol; D-Glucitol; Ribitol; Salicin; DL-Lactic acid; Succinic acid. Variable: Lactose; Melibiose; Erythritol; Soluble Starch; L-Rhamnose; D- Arabinose; L-Sorbose; D-Ribose; Galactitol; Citric acid. Negative: Potassium nitrate; Inositol. Clinical significance Candida famata is a common environmental isolate, however it is only rarely recovered from clinical specimens, usually associated with skin. Clinical group: Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands.

35 35 Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

36 36 Candida glabrata Synonym: Torulopsis glabrata On Sabouraud's dextrose agar colonies are white to cream colored, smooth and glabrous yeast-like in appearrance. Microscopic morphology shows numerous ovoid, budding yeast-like cells or blastoconidia, x um in size. No pse udohyphae produced. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Ovoid budding yeastlike cells only. No pseudohyphae produced. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Negative Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose. Variable: Trehalose. Negative: Latose; Maltose; Sucrose; Galactose. Assimilation Tests: Positive: Glucose; Trehalose. Variable: Glycerol; DL-Lactic acid. Negative: Potassium nitrate; Lactose; Galactose; Maltose; D-Xylose; Soluble Starch; Melezitose; Sucrose; Raffinose; Cellobiose; Erythritol; Inositol; L- Rhamnose; D-Arabinose; L-Arabinose; Ribitol; L-Sorbose; D-Ribose; Galactitol; D- Mannitol; D-Glucitol; Salicin; Citric acid; Succinic acid. Clinical significance Candida (Torulopsis) glabrata is one of the most common yeast species to be found on the body surface and is often isolated as an incidental finding from skin and urine. It has been implicated as an "opportunistic" cause of both superficial and systemic infections, especially in immunocompromised patients, and it has been isolated from patients with septicemia, pyelonephritis, pulmonary infections, endocarditis and hyperalimentation.

37 37 Clinical group: Cutaneous mycoses, Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

38 38 Candida guilliermondii On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows spherical to subspherical budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Branched pseudohyphae with dense verticils of blastoconidia. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Sucrose; Variable: Galactose. Negative: Lactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Sucrose; Galactose; Maltose; Trehalose; D-Xylose; Melezitose; Glycerol; Raffinose; Cellobiose; Melibiose; L-Rhamnose; D-Arabinose (delayed); L- Arabinose; Ribitol; L-Sorbose (delayed); Galactitol (delayed); D-Mannitol; D- Glucitol; Salicin; Citric acid; Succinic acid. Variable: D-Ribose; DL-Lactic acid. Negative: Potassium nitrate; Lactose; Erythritol ; Inositol; Soluble Starch. Clinical significance Candida guilliermondii has been isolated from numerous human infections, mostly of cutaneous origin. Systemic infections are rare, although one has been reported in a patient with aplastic anemia. C. guilliermondii has also been isolated from normal skin and in sea water, faeces of animals, fig wasps, buttermilk, leather, fish and beer. Clinical group: Cutaneous mycoses Mycosis: Candidiasis

39 39 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

40 40 Candida haemulonii Synonym: Torulopsis haemulonii On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows numerous ovoid to globose, budding yeast-like cells or blastoconidia, x um in size. No pseudohyphae produced. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ovoid, budding yeast-like cells only. No pseudohyphae produced. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Sucrose; Trehalose. Variable: Maltose. Negative: Lactose; Galactose. Assimilation Tests: Positive: Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose; Melezitose; Glycerol; Raffinose (delayed); L-Arabinose (delayed); Ribitol (delayed); Galactitol (delayed); D-Mannitol; L-Rhamnose; D-Glucitol; Citric acid; Succinic acid. Negative: Potassium nitrate; Lactose; L-Sorbose; Cellobiose; D-Ribose; Melibiose; Erythritol; Inositol; Soluble Starch; Salicin; D-Arabinose; DL-Lactic acid. Clinical significance Candida haemulonii has been reported from a few cases of fungemia but clinical isolations remain rare. It has also been isolated from fish and a dolphin. C. haemulonii may be difficult to distinguish from C. famata using some commercial yeast identification systems due to data base limitations. Clinical group: Opportunistic mycoses Mycosis: Candidiasis

41 41 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

42 42 Candida kefyr Synonym: Candida pseudotropicalis On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows numerous short-ovoid to long-ovoid, budding yeast-like cells or blastoconidia, x um, sometimes becoming elongate (up to 16.0 um) in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant, long, wavy, branched pseudohyphae is usually formed, with ovoid blastoconidia, budding off singly, in pairs or chains, often in a verticillated position. Note, in some strains pseudohyphae may be scarce or almost absent. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Variable Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Galactose (delayed). Variable: Glucose; Sucrose; Lactose; Maltose. Negative: Trehalose. Assimilation Tests: Positive: Glucose; Galactose; Sucrose; Lactose; Raffinose; DL-Lactic acid. Variable: Trehalose; D-Xylose; Glycerol; L-Arabinose; Cellobiose; L-Sorbose; D- Ribose; D-Mannitol; Salicin; Citric acid; Succinic acid. Negative: Potassium nitrate; Ribito; Maltose; Melibiose; Galactitol; Erythritol; Inositol; D-Glucitol; Soluble Starch; L-Rhamnose; Melezitose; D-Arabinose. Clinical significance Candida kefyr is a rare cause of candidiasis and is usually associated with superficial cutaneous manifestations rather than systemic disease. It has been isolated from nails and lung infections. Environmental isolations have been made from cheese and dairy products. Clinical group: Cutaneous mycoses, Mycosis: Candidiasis

43 43 Opportunistic mycoses Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

44 44 Candida krusei On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous yeast-like colonies. Microscopic morphology shows predominantly small, elongated to ovoid budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant long, wavy, branched pseudohyphae with elongated to ovoid blastoconidia, budding off in verticillate branches. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Variable Growth on Cycloheximide medium is Negative Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose. Negative: Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Glycerol (delayed); DL-Lactic acid; Succinic acid. Variable: L-Sorbose; Citric acid. Negative: Potassium nitrate; Lactose; Ribitol; Galactose; Raffinose; Maltose; Cellobiose; D-Ribose (some positive); Sucrose; Melibiose; Galactitol; Trehalose; Erythritol; D-Mannitol; D-Xylose; Inositol; D-Glucitol; Soluble Starch; L- Rhamnose; Salicin; Melezitose; D-Arabinose; Arabinose. Clinical significance Candida krusei is regularly associated with some forms of infant diarrhoea and occasionally with systemic disease. It has also been reported to colonize the gastrointestinal, respiratory and urinary tracts of patients with granulocytopenia. Environmental isolations have been made from beer, milk products, skin, faeces of animals and birds and pickle brine Clinical group: Opportunistic mycoses Mycosis: Candidiasis

45 45 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

46 46 Candida lusitaniae On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows numerous subglobose, ovoid, or elliptical budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant pseudohyphae with short chains of blastoconidia. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Negative Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Sucrose (delayed); Trehalose (delayed). Variable: Galactose; Maltose. Negative: Lactose. Assimilation Tests: Positive: Glucose; Maltose; Sucrose; Trehalose; D-Xylose; Glycerol; Cellobiose; L- Rhamnose; D-Ribose (delayed); D-Mannitol; Ribitol; D-Glucitol; Salicin; DL- Lactic acid; Succinic acid. Variable: Galactose; Melezitose; D-Arabinose; L-Arabinose; L-Sorbose; Citric acid. Negative: Potassium nitrate; Lactose; Raffinose; Melibiose; Galactitol; Erythritol; Inositol; Soluble Starch. Clinical significance Candida lusitaniae has been isolated from several cases of disseminated candidiasis, including septicemia and pyelonephritis. It has also been reported to colonize the human respiratory, gastrointestinal and urinary tracts and many strains have proven to be resistant to amphotericin B. C. lusitaniae was first isolated from the alimentary tract of warm blooded animals and environmental isolations have been made from cornmeal, citrus peel, fruit juices, and milk from cows with mastitis. C. lusitaniae may also be difficult to distinguish from C. tropicalis using some yeast identification systems.

47 47 Clinical group: Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

48 48 Candida norvegensis Synonym: Torulopsis norvegica On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows numerous ovoid, budding yeast-like cells or blastoconidia, x um in size. No pseudohyphae produced. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ovoid, budding yeast-like cells only. No pseudohyphae produced. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Negative Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose (delayed). Negative: Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Cellobiose; Citric acid; Succinic acid. Variable: Soluble Starch. Negative: Potassium nitrate; Lactose; Ribitol; Galactose; Raffinose; L-Sorbose; Maltose; D-Ribose; Sucrose; Melibiose; Galactitol; Trehalose; Erythritol; D- Mannitol; D-Xylose; Inositol; D-Glucitol; L-Rhamnose; Salicin; Melezitose; D- Arabinose; Glycerol; L-Arabinose; DL-Lactic acid. Clinical significance Candida norvegensis is a very rare clinical isolate that has been reported as a causative agent of peritonitis and disseminated candidiasis in a patient on CAPD. Clinical group: Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands.

49 49 Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

50 50 Candida parapsilosis On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows predominantly small, globose to ovoid budding yeast-like cells or blastoconidia, x um in size, with some larger elongated forms present. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant, muchbranched pseudohyphae in a delicate tree-like pattern with 2-3 blastoconidia in small clusters at intervals along the pseudohyphae. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Negative Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose. Variable: Galactose. Negative: Lactose; Sucrose (some positive); Maltose (some positive); Trehalose (some positive). Assimilation Tests: Positive: Glucose; Galactose; Maltose; Trehalose; D-Xylose; Glycerol; L-Arabinose; Ribitol (delayed); D-Mannitol; Sucrose; D-Glucitol. Variable: D-Ribose; L-Sorbose; Succinic acid; Citric acid; DL-Lactic acid. Negative: Potassium nitrate; Cellobiose; Lactose; Raffinose; Melibiose; Galactitol; Erythritol; Inositol; Soluble Starch; L-Rhamnose; Salicin; Melezitose; D-Arabinose. Clinical significance Candida parapsilosis is an opportunistic human pathogen which may cause both superficial cutaneous infections, especially of the nail and systemic disease, especially endocarditis. Other clinical manifestations include endophthalmitis and fungemia. Environmental isolations have been made from intertidal and oceanic waters, pickle brine, cured meats, olives and normal skin, and faeces. Clinical group: Opportunistic mycoses Mycosis: Candidiasis

51 51 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

52 52 Candida tropicalis On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows spherical to subspherical budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Negative - no capsules present. Cornmeal and Tween 80 Agar Plate: Abundant long, wavy, branched pseudohyphae with numerous ovoid blastoconidia budding off. Terminal vesicles (chlamydoconidia) are not produced. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Maltose; Galactose; Trehalose (delayed). Variable: Sucrose. Negative: Lactose. Assimilation Tests: Positive: Glucose; Galactose; Maltose; Trehalose; D-Xylose; Soluble Starch; Succinic acid; D-Mannitol; L-Arabinose (weak); D-Glucitol. Variable: Sucrose; Salicin; Melezitose; Glycerol; Cellobiose; D-Ribose; Ribitol; L- Sorbose; Citric acid; DL-Lactic acid. Negative: Potassium nitrate; L-Rhamnose; Lactose; Raffinose; Melibiose; Galactitol; Erythritol; Inositol; D-Arabinose. Clinical significance Candida tropicalis is a major cause of septicemia and disseminated candidiasis, especially in patients with lymphoma, leukemia and diabetes. It is the second most frequently encountered medical pathogen, next to C. albicans, and is also found as part of the normal human mucocutaneous flora. Sucrose negative variants of C. tropicalis have also been increasingly found in cases of disseminated candidiasis. Environmental isolations have been made from faeces, shrimp, kefir, and soil.

53 53 Clinical group: Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

54 54 Candida viswanathii On Sabouraud's dextrose agar colonies are white to cream colored, smooth, glabrous and yeast-like in appearance. Microscopic morphology shows numerous globose, ovoid, or cylindrical budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant wavy pseudohyphae with verticillated branched chains of blastoconidia. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Negative Growth on Cycloheximide medium is Positive Growth at 37C is Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Positive: Glucose; Galactose; Maltose; Trehalos (delayed). Variable: Sucrose; Negative: Lactose. Assimilation Tests: Positive: Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose; Soluble Starch; Melezitose; Glycerol; L-Arabinose (delayed); D-Mannitol; Ribitol; Salicin; Citric acid; Succinic acid. Negative: Potassium nitrate; Lactose; Raffinose; L-Sorbose; Cellobiose; D-Ribose; Melibiose; Galactitol; Erythritol; Inositol; D-Glucitol; L-Rhamnose; DL-Lactic acid; D-Arabinose. Clinical significance Candida viswanathii is only rarely isolated from clinical specimens. Clinical group: Opportunistic mycoses Mycosis: Candidiasis Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands.

55 55 Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

56 56 Chrysosporium tropicum Culture of Chrysosporium tropicum. Colonies are moderately fast growing, flat, white to tan to beige in colour, often with a powdery or granular surface texture. Reverse pigment absent or pale brownish-yellow with age. Hyaline, one-celled (ameroconidia) are produced directly on vegetative hyphae by non-specialized conidiogenous cells. Conidia are typically pyriform to clavate with truncate bases (6 to 7 by 3.5 to 4 um) and are formed either intercalary (arthroconidia), laterally (often on pedicels) or terminally. No macroconidia or hyphal spirals are seen. Chrysosporium tropicum showing typical pyriform to clavate shaped conidia with truncated bases which may be formed either intercalary, laterally or terminally. Clinical significance Species of Chrysosporium are occasionally isolated from skin and nail scrapings, especially from feet, but because they are common soil saprophytes they are usually considered as contaminants. There are about 22 species of Chrysosporium, several are keratinophilic with some also being thermotolerant, and cultures may closely resemble some dermatophytes, especially Trichophyton mentagrophytes, and some strains may also resemble cultures of Histoplasma and Blastomyces. Clinical group: Saprophytes Mycosis: Hyaline hyphomycetes Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK.

57 57 McGinnis, M.R Laboratory handbook of medical mycology. Academic Press, London, UK.

58 58 Cladophialophora bantiana Synonym: Xylohyphae bantiana; Cladosporium bantianum; Cladosporium trichoides. Conidiophore and conidia of Cladophialophora bantiana. Colonies are moderately fast growing, olivaceous-grey, suede-like to floccose and grow at temperatures up to 42-43C. Conidia are formed in long, sparsely branched, flexuous, acropetal chains from undifferentiated conidiophores. Conidia are one-celled (very occasionally two-celled), pale brown, smooth-walled, ellipsoid to oblong-ellipsoid and are 2-3 x 4-7 um in size. Cladophialophora bantiana may be distinguished from Cladosporium species by the absence of conidia with distinctly pigmented hila, the absence of characteristic shield cells and by growth at 42C (compared with Cladophialophora carrionii which has a maximum growth temperature of 35-36C. Clinical significance Cladophialophora bantianahas been isolated from soil and is a recognized agent of cerebral phaeohyphomycosis. The fungus is neurotropic and may cause brain abscess in both normal and immunosuppressed patients. Clinical group: Opportunistic mycoses Mycosis: Phaeohyphomycosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

59 59 Synonym: Cladosporium carrionii Cladophialophora carrionii Culture of Cladophialophora carrionii On Sabouraud's dextrose agar, colonies are slow growing, reaching 3-4 cm in diameter after one month, with a compact suede-like to downy surface. Colonies are olivaceous-black in color and have well defined margins. Microscopic morphology shows the typical Cladosporium type conidial ontogeny with elongate conidiophores producing branched acropetal chains of smooth-walled conidia, x um in size. Maximum growth temperature 35-37C. Conidiophores and conidia of C. carrionii Clinical significance Cladophialophora carrionii is a recognized agent of chromoblastomycosis and it has been isolated from soil and fence posts made from Eucalyptus sp. Cases of chromoblastomycosis caused by C. carrionii are commonly found in Australia, Venezuela, Madagascar and South America. Isolates from phaeomycotic cysts and opportunistic infections have also been reported. Clinical group: Subcutaneous mycoses Mycosis: Chromoblastomycosis Further reading De Hoog G.S. and J Guarro Atlas of clinical fungi. Centraalbureau voor Schimmelcultures, Baarn and Delft, The Netherlands.

60 60 Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London.

61 61 Cladosporium sp. Culture of Cladsporium cladosporioides. Colonies are rather slow growing, mostly olivaceous-brown to blackish brown but also sometimes grey, buff or brown, suede-like to floccose, often becoming powdery due to the production of abundant conidia. Vegetative hyphae, conidiophores and conidia are equally pigmented. Conidiophores are more or less distinct from the vegetative hyphae, are erect, straight or flexuous, unbranched or branched only in the apical region, with geniculate sympodial elongation in some species. Conidia are 1- to 4-celled, smooth, verrucose or echinulate, with a distinct dark hilum and are produced in branched acropetal chains. The term blastocatenate is often used to describe chains of conidia where the youngest conidium is at the apical or distal end of the chain. Note, the conidia closest to the conidiophore and where the chains branch, are usually "shield-shaped". The presence of shield-shaped conidia, a distinct hilum, and chains of conidia that readily disarticulate, are diagnostic for the genus Cladosporium. Conidiophores and conidia of Cladosporium cladosporioides For identification, isolates are best grown on potato dextrose agar or 2% malt extract agar at 20-25C. Microscopic mounts are best made using a cellotape flag or slide culture preparation mounted in lactophenol cotton blue. A drop of alcohol is usually needed to detach the cellotape flag from the stick, and to act as a wetting agent. Clinical significance Cladosporium species have a world-wide distribution and are among the most

62 62 common air-borne fungi. Some 500 species have been described. Cladosporium species are frequently isolated as contaminants. Clinical group: Saprophytes Mycosis: Dematiaceous hyphomycetes Further reading Domsch, K.H., W. Gams, and T.H. Anderson Compendium of soil fungi. Volume 1. Academic Press, London, UK.

63 63 Coccidioides immitis Culture of Coccidioides immitis On Sabouraud's dextrose agar at 25C, colonies are initially moist and glabrous, but rapidly become suede-like to downy, greyish white in color with a tan to brown reverse. However, considerable variation in both growth rate and culture morphology has been noted (Rippon, 1988). Microscopic morphology shows typical single-celled, hyaline, rectangular to barrel-shaped, alternate arthroconidia, x um in size, separated from each other by a disjunctor cell. This arthroconidial state has been classified in the genus Malbranchea and is similar to that produced by many non-pathogenic soil fungi such as Gymnoascus species. WARNING: Cultures of Coccidioides immitis represent a severe biohazard to laboratory personnel and must be handled with extreme caution in an appropriate pathogen handling cabinet. C. immitis is a dimorphic fungus, existing in living tissue as spherules and endospores, and in soil or culture in a mycelial form. Culture identification by the exoantigen test is now the method of choice. Arthroconidia of C. immitis. Clinical significance Coccidioidomycosis is initially, a respiratory infection, resulting from the inhalation of conidia, that typically resolves rapidly leaving the patient with a strong specific immunity to re-infection. However, in some individuals the disease may progress to a chronic pulmonary condition or to a systemic disease involving the meninges, bones, joints and subcutaneous and cutaneous tissues. Coccidioides

64 64 immitis is a soil inhabiting fungus endemic in south-western U.S.A., northern Mexico and various centers in South America. Several cases have now been diagnosed in Australia, all in patients with a history of travel to endemic areas. Clinical group: Dimorphic systemic Mycosis: Coccidioidomycosis mycoses Further reading Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

65 65 Colletotrichum coccodes Culture of Colletotrichum coccodes. Colonies usually darkly pigmented with white aerial mycelium, consisting of numerous black sclerotia and light brown coloured conidial masses, reverse is dark brown. Sclerotia are usually abundant, setose, spherical and are often confluent. Conidia are straight, fusiform, attenuated at the ends, X 3-4 um. Appressoria are common, clavate, brown, X Sclerotia with setae of C. coccodes.

66 66 Conidia of C. coccodes. Appressoria of C. coccodes. Clinical significance Plant fungus reported from a case of mycotic keratitis. Clinical group: Opportunistic mycoses Mycosis: Mycotic Keratitis Further reading

67 67 Synonym: Entomophthora coronata Conidiobolus coronatus Culture of Conidiobolus coronatus. Colonies grow rapidly and are flat, cream colored, glabrous, becoming radially folded and covered by a fine, powdery, white surface mycelium and conidiophores. The lid of the petri dish soon becomes covered with conidia, which are forcibly discharged by the conidiophores. The color of the colony may become tan to brown with age. Conidiophores are simple, forming solitary, terminal conidia which are spherical, um in diameter, single-celled and have a prominent papilla. Conidia may also produce hair-like appendages called villae. Conidia germinate to produce either, (1) single or multiple hyphal tubes that may also become conidiophores which bear secondary conidia, or (2) produce multiple short conidiophores, each bearing a small secondary conidium. Spherical conidia with hair-like appendages (villae) and prominent papillae characteristic of Conidiobolus coronatus. Clinical significance C. coronatus is commonly present in soil and decaying leaves. It has a world-wide distribution, but it is mainly found in tropical rain forests of Africa. Clinical group: Subcutaneous mycoses Mycosis: Zygomycosis

68 68 Further reading Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

69 69 Cryptococcus albidus On Sabouraud's dextrose agar colonies are cream colored, smooth, mucoid and yeast-like in appearance. Microscopic morphology shows globose to ovoid budding yeast-like cells or blastoconidia, x um in size. India Ink Preparation: Positive - distinct capsules surrounding the yeast cells are present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding yeast cells only. No pseudohyphae present. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Positive Growth on Cycloheximide medium is Negative Growth at 37C is Negative (Positive) Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Negative: Glucose; Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Potassium nitrate; Glucose; Maltose (weak); Sucrose; Trehalose (weak); D-Xylose (weak); Melezitose; Raffinose (weak); Cellobiose; Inositol; L-Arabinose (weak); D-Mannitol (weak); Citric acid (weak); Succinic acid (weak); Salicin; D- Glucitol. Variable: Soluble Starch; Lactose; Galactose; Melibiose; L-Rhamnose; D- Arabinose; Ribitol; L-Sorbose; D-Ribose; Galactitol; DL-Lactic acid. Negative: Erythritol; Glycerol (some positive). Clinical significance Cryptococcus albidus is a common transient on human skin and it has also been reported as a rare cause of pulmonary, CNS and vaginal infection. However, these reports must be viewed with considerable caution, as the culture identifications have not been verified. Clinical group: Cutaneous mycoses, Opportunistic mycoses Mycosis: Cryptococcosis

70 70 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

71 71 Cryptococcus laurentii On Sabouraud's dextrose agar colonies are cream colored, often becoming a deeper orange-yellow with age, with a smooth mucoid texture. Microscopic morphology : Spherical and elongated budding yeast-like cells or blastoconidia, x um in size. No pseudohyphae present. India Ink Preparation: Positive - narrow but distinct capsules surrounding the yeast cells are present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding yeast cells only. No pseudohyphae present. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Positive Growth on Cycloheximide medium is Variable Growth at 37C is Negative (weak growth in some strains) Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Negative: Glucose; Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose (weak); Melezitose; Lactose; Raffinose; Cellobiose; Melibiose; Inositol (delayed); L-Rhamnose; D-Arabinose; L-Arabinose; D-Mannitol; Ribitol; D-Ribose (delayed); Galactitol; Salicin. Variable: Erythritol; Soluble Starch; D-Glucitol; Glycerol; Citric acid; DL-Lactic acid; Succinic acid. Negative: Potassium nitrate; L-Sorbose (some positive). Clinical significance Cryptococcus laurentii has been reported as a rare cause of pulmonary and cutaneous infection and CAPD associated peritonitis in humans. It may also be occasionally recovered as a saprophyte from skin. Clinical group: Opportunistic mycoses Mycosis: Cryptococcosis

72 72 Further reading Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA

73 73 Cryptococcus neoformans var. gattii Teleomorph: Filobasidiella neoformans var. bacillispora Cryptococcus neoformans is an encapsulated basidiomycete yeast-like fungus with a predilection for the respiratory and nervous system of humans and animals. Two varieties, C. neoformans var. neoformans and C. neoformans var. gattii are distinguishable biochemically and by molecular techniques. CGB agar turns blue for Cryptococcus neoformans var. gattii. Canavanine-glycine-bromthymol blue (CGB) agar is now the medium of choice to determine the varietal status of C. neoformans isolates. This simple biotype test is based on the ability of C. neoformans var. gattii isolates to grow in the presence of L- canavanine and to assimilate glycine as a sole carbon source. On Sabouraud's dextrose agar colonies are cream colored, smooth, mucoid yeastlike in appearance. Microscopic morphology shows globose to ovoid budding yeast-like cells or blastoconidia, x um in size. India ink preparation showing capsules of C. neoformans India Ink Preparation: Positive - distinct, wide gelatinous capsules surrounding the yeast cells are present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding yeast cells only. No pseudohyphae present.

74 74 Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Positive Growth on Cycloheximide medium is Negative Growth at 37C is Weakly Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Negative: Glucose; Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests: Positive: Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose; Melezitose; Raffinose (weak); Cellobiose (weak); Inositol; L-Rhamnose; D-Arabinose; L- Arabinose (delayed); D-Ribose (weak); Galactitol (delayed); D-Mannitol; D- Glucitol. Variable: Soluble Starch; Glycerol; Succinic acid; Ribitol; L-Sorbose; Salicin; Citric acid; DL-Lactic acid. Negative: Potassium nitrate; Lactose; Melibiose; Erythritol. Bird seed agar plate showing brown colonies of C. neoformans and white colonies of Candida albicans. Bird seed agar or caffeic acid agar: Colonies of both varieties turn dark brown in color; C. neoformans selectively absorbs a brown pigment from these media. Supplementary description of Cryptococcus neoformans var. gattii. Canavanine-glycine-bromthymol blue (CGB) agar turns blue within 2-5 days; Cryptococcus neoformans var. neoformans leaves this medium unchanged [see above].

75 75 Growth of C. neoformans var gattii on Eucalypt agar Clinical significance The distribution of cryptococcosis due to Cryptococcus neoformans var. gattii is geographically restricted, non-immunocompromised hosts are usually affected, large mass lesions in lung and/or brain (cryptococcomas) are characteristic and morbidity from neurological disease is high. Human disease is endemic in Australia, Papua New Guinea, parts of Africa, the Mediterranean region, India, south-east Asia, Mexico, Brazil, Paraguay and Southern California. Environmental isolations, initially from the Barossa Valley in South Australia have established that C. neoformans var. gattii has a specific ecological association with Eucalyptus camaldulensis, a species of red gum widely distributed in mainland Australia. Subsequently, another species of red gum E. tereticornis was confirmed as a natural habitat. This species has a more restricted distribution occurring along the eastern coastal seaboard of Australia, extending to Papua New Guinea. More recently, high concentrations of C. neoformans var. gattii have been isolated from single specimens of three additional eucalypts, Eucalyptus rudis (flooded gum), E. gomphocephala (tuart) and E. blakelyi (Blakely's red gum). Three of these species (E. camaldulensis, E. tereticornis and E. gomphocephala) have been exported to several countries in which human disease due to C. neoformans var. gattii has been reported though the association is not exact. Outside of Australia limited

76 76 isolations of C. neoformans var. gattii have been made from E. camaldulensis trees growing in California, Apulia, Italy and in northern India. Evidence for an epidemiological association between this cryptococcal habitat and human infection is circumstantial. There is correlation between the global distribution of human infection with C. neoformans var. gattii and the five species of eucalypts and environmental searches conducted in Australia and elsewhere have so far failed to identified any other natural source. In Australia, 92% of human isolates and all of those from koalas (a native animal which live in close association with eucalypts) and from all five host eucalypts identified to date (ie E. camaldulensis, E. tereticornis, E. rudis and E. gomphocephala), exhibit the same genetic fingerprint (VGI) when identified by random amplification of polymorphic DNA (RAPD) and PCR fingerprinting. This finding is consistent with an epidemiological association between mammalian disease and exposure to host eucalypts. However, the occurrence of human C. neoformans var. gattii infection in other countries which lack the host trees, and our own observations of a distinct genetic type (VGII) in certain locations in Australia suggest that additional environmental niches for this fungus are yet to be discovered. Clinical group: Opportunistic mycoses Mycosis: Cryptococcosis Further reading Cryptococcosis Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London. Kwon-Chung K.J., Polacheck I., and Bennett J.E Improved diagnostic medium for separation of Cryptococcus neoformans var. neoformans (Serotypes A and D) and Cryptococcus neoformans var. gattii (Serotypes B and C). J. Clin. Microbiol. 15(3):

77 77 Cryptococcus neoformans var. neoformans Teleomorph: Filobasidiella neoformans var. neoformans Cryptococcus neoformans is an encapsulated basidiomycete yeast-like fungus with a predilection for the respiratory and nervous system of humans and animals. Two varieties, C. neoformans var. neoformans and C. neoformans var. gattii are distinguishable biochemically and by molecular techniques. CGB agar turns blue for Cryptococcus neoformans var. gattii. Canavanine-glycine-bromthymol blue (CGB) agar is now the medium of choice to determine the varietal status of C. neoformans isolates. This simple biotype test is based on the ability of C. neoformans var. gattii isolates to grow in the presence of L- canavanine and to assimilate glycine as a sole carbon source. Basidiospores of C. neoformans var. neoformans. Mating experiments have shown that the two varieties of C. neoformans exhibit different sexual states: Filobasidiella neoformans var. neoformans is the teleomorph of C. neoformans var. neoformans and Filobasidiella neoformans var. bacillispora is the teleomorph of C. neoformansvar. gatti.

78 78 Culture of C. neoformans On Sabouraud's dextrose agar colonies are cream colored, smooth, mucoid yeastlike in appearance. Microscopic morphology shows globose to ovoid budding yeast-like cells or blastoconidia, x um in size. India ink preparation showing capsules of C. neoformans India Ink Preparation: Positive - distinct, wide gelatinous capsules surrounding the yeast cells are present. Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding yeast cells only. No pseudohyphae present. Physiological Tests: Germ Tube test is Negative Hydrolysis of Urea is Positive Growth on Cycloheximide medium is Negative Growth at 37C is Weakly Positive Fermentation Reactions: Where fermentation means the production of gas and is independent of ph changes. Negative: Glucose; Sucrose; Lactose; Galactose; Maltose; Trehalose. Assimilation Tests:

79 79 Positive: Glucose; Galactose; Maltose; Sucrose; Trehalose; D-Xylose; Melezitose; Raffinose (weak); Cellobiose (weak); Inositol; L-Rhamnose; D-Arabinose; L- Arabinose (delayed); D-Ribose (weak); Galactitol (delayed); D-Mannitol; D- Glucitol. Variable: Soluble Starch; Glycerol; Succinic acid; Ribitol; L-Sorbose; Salicin; Citric acid; DL-Lactic acid. Negative: Potassium nitrate; Lactose; Melibiose; Erythritol. Bird seed agar plate showing brown colonies of C. neoformans and white colonies of Candida albicans. Bird seed agar or caffeic acid agar: Colonies of both varieties turn dark brown in color; C. neoformans selectively absorbs a brown pigment from these media. Supplementary description of Cryptococcus neoformans var. gattii. Canavanine-glycine-bromthymol blue (CGB) agar turns blue within 2-5 days; Cryptococcus neoformans var. neoformans leaves this medium unchanged [see above]. Growth of C. neoformans var gattii on Eucalypt agar Clinical significance

80 80 In humans, C. neoformans var. neoformans affects immunocompromised hosts predominantly and is the commonest cause of fungal meningitis; worldwide, 7-10% of patients with AIDS are affected. AIDS associated cryptococcosis accounts for 50% of all cryptococcal infections reported annually and usually occurs in HIV patients when their CD4 lymphocyte count is below 200/mm3. Meningitis is the predominant clinical presentation with fever and headache as the most common symptoms. Secondary cutaneous infections occur in up to 15% of patients with disseminated cryptococcosis and often indicate a poor prognosis. Lesions usually begin as small papules that subsequently ulcerate, but may also present as abscesses, erythematous nodules, or cellulitis. This variety is found in nature in avian excreta, especially weathered pigeon droppings, which are believed to be the source of infection. Clinical group: Opportunistic mycoses Mycosis: Cryptococcosis Further reading Cryptococcosis Kreger-Van Rij, N.J.W. (ed) The Yeasts: a taxonomic study. 3rd Edition. Elsevier Science Publishers B.V., Amsterdam, The Netherlands. Kwon-Chung, K.J. and J.E. Bennett Medical Mycology. Lea & Febiger, Philadelphia and London. Kwon-Chung K.J., Polacheck I., and Bennett J.E Improved diagnostic medium for separation of Cryptococcus neoformans var. neoformans (Serotypes A and D) and Cryptococcus neoformans var. gattii (Serotypes B and C). J. Clin. Microbiol. 15(3):

81 81 Cunninghamella bertholletiae Synonym: Cunninghamella elegans; Cunninghamella echinulata var. elegans. Cunninghamella bertholletiaeshowing simple sporangiophores forming a swollen, terminal vesicle around which single-celled, globose to ovoid sporangiola develop on swollen denticles. Colonies on Sabouraud's agar are very fast growing, white at first, but becoming rather dark grey and powdery with development. Sporangiophores to 20 um wide, straight, with verticillate or solitary branches. Vesicles subglobose to pyriform with the terminal ones up to 40 um and the lateral ones 10 to 30 um in diameter. Sporangiola are globose (7-11 um diameter), ellipsoidal (9-13 x 6-10 um), verrucose or short-echinulate, hyaline singly but brownish in mass. Temperature: optimum 25-30C; maximum up to 50C. The genus Cunninghamella is characterized by white to grey, rapidly growing colonies, producing erect, straight, branching sporangiophores. These sporangiophores end in globose or pyriform-shaped vesicles from which several one-celled, globose to ovoid, echinulate or smooth-walled develop on swollen denticles. Chlamydoconidia and zygospores may also be present. Cunninghamella species are mainly soil fungi of the Mediterranean and subtropical zones, and they are only rarely isolated in temperate regions. The genus now contains seven species, with C. bertholletiae the only species known to cause disease in man and animals. Clinical significance Cunninghamella bertholletiae is a rare cause of zygomycosis in humans often associated with trauma and immunosuppression. It is a common soil fungus found throughout the temperate regions of the world. Clinical group: Opportunistic mycoses Mycosis: Zygomycosis

82 82 Further reading Rippon, J.W Medical Mycology. 3rd Edition. W.B. Saunders Co., Philadelphia, USA.

83 83 Curvularia sp. Teleomorph: Cochliobolus sp. Colonies are fast growing, suede-like to downy, brown to blackish brown with a black reverse. Conidia are pale brown, with three or more transverse septa (phragmoconidia) and are formed apically through a pore (poroconidia) in a sympodially elongating geniculate conidiophore similar to Drechslera. Conidia are cylindrical or slightly curved, with one of the central cells being larger and darker. Germination is bipolar and some species may have a prominent hilum. Culture of Curvularia lunata. Conidia of Curvularia lunata. Clinical significance The genus Curvularia contains some 35 species which are mostly subtropical and tropical plant parasites. However, three ubiquitous species have been recovered from human infections, principally from cases of mycotic keratitis; C. lunata, C.

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