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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5853_Библиотеки_им_академика_М_И_Перельмана
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111.4. Canavanine glycine bromothymol blue (CGB) agar
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For differentiation of C. neoformans and C. gattii (Kwon-Chung et al., 1982).
Solution A: Glycine (Univar) 10 g; KH
30 mg; distilled water 100 mL. Dissolve ingredients in small beaker and adjust pH to 5.6. Filter sterilise
solution using 0.22 μm filter. Store at 4
Solution B (aqueous bromothymol blue): Bromothymol blue 0.4 g; 0.01N NaOH 64 mL; distilled water
36 mL. Dissolve the bromothymol blue in the NaOH and add to the water.
1 g; MgSO4 1 g; thiamine HCl 1 mg; L-canavanine sulfate
2PO4
o
C.
Final mixture: To prepare a litre of the medium, mix 880 mL of distilled water, 20 mL of solution B, and
20 g of Bacto-agar (BD 214010) in a large flask. Autoclave to 121
o
C for 15 minutes and cool to 48oC. Add
100 mL of solution A and stir thoroughly before pouring into petri dishes or dispensing aseptically into tubes
to make slopes. A positive test with CGB medium indicates a change from pH from 5.8 (greenish-yellow) to
at least 7.0 (cobalt blue).
111.5. Cornmeal agar
For the routine cultivation and identification of fungi.
Ingredients: Cornmeal agar 8.5 g; distilled water 500 mL.
Method: Mix dry ingredients into 100 mL H
bring to boil. Dispense for slopes as required. Autoclave at 121
O, boil remaining water. Add boiling water to mixture and
2
o
C for 10 minutes, remove and slope or pour
for plates as required.
111.6. Cornmeal glucose sucrose yeast extract agar
To stimulate sporulation in mucormycetous fungi, especially Saksenaea and Apophysomyces.
Ingredients: Cornmeal agar 17 g; dextrose (glucose) 2 g; sucrose 3 g; yeast extract 1 g; distilled water
1000 mL.
Method: Soak dry ingredients in 100 mL H2O, then boil remaining water. Add boiling water to mixture and
bring to the boil. Dispense for slopes as required. Autoclave at 121
o
C for 10 minutes, remove and slope or
pour for plates as required.
111.7. Czapek dox agar
For the routine cultivation of fungi, especially Aspergillus, Penicillium and non-sporulating moulds.
Ingredients: Czapek dox agar (Oxoid CM97) 45.4 g; distilled water 1000 mL.
Method: Soak the ingredients in a small amount of water. Bring remaining water to boil, add to soaking
ingredients and bring to the boil again, stirring continuously. Dispense for slopes as required. Autoclave at
o
121
C for 10 minutes, remove and slope or pour for plates as required.
111.8. Modified Dixon’s agar
For primary isolation and cultivation of Malassezia species.
Ingredients: Malt extract 9 g; Bacto Tryptone 1.5 g; ox-bile desiccated 5 g; Tween 40 2.5 mL; oleic acid
0.5 g; glycerol 0.5 mL; Bacto-agar 3 g; distilled water 250 mL.
Method: Soak ingredients for 15 minutes in some water. Bring remaining water to boil, add to other ingredients. Bring to boil once more. Dispense for slopes. Autoclave at 121
o
C for 10 minutes and then slope.
Descriptions of Medical Fungi 306
111.9. Hair perforation test for dermatophytes
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To distinguish between dermatophytes species, particularly Trichophyton mentagrophytes and its variants.
Ingredients: Autoclaved blonde prepubertal hair that has been cut into 1 cm pieces. Sterile distilled water
5 mL in a suitable vial. Note: Perforations are easiest to see on blonde hair, and prepubertal hair has no fatty
acids that might inhibit fungal growth.
Method: Place hair in water in vial. Inoculate with small fragments of the test fungus. Incubate at room
temperature.
Individual hairs are removed at intervals up to 4 weeks and examined microscopically in lactophenol cot-
ton blue. Isolates of T. mentagrophytes produce marked localised areas of pitting and marked erosion,
whereas those of T. rubrum do not.
111.10. Lactritmel agar
For enhanced sporulation and the production of pigment by dermatophytes, especially Trichophyton rubrum.
Ingredients: Skimmed milk powder 7 g; honey 10 g; Cornmeal agar 17 g; Chloramphenicol 0.05 g; distilled
water 1000 mL.
Method: Weigh skimmed milk powder into stainless-steel jug. Slowly add some water, mixing milk into
smooth paste. Continue adding small quantities of water until powder is dissolved (about 150mL). Weigh
other ingredients into skimmed milk and allow to soak. Boil remaining water, and with it wash out honey
from beaker. Add to other ingredients and boil. Dispense for slopes if required. Autoclave for 10 minutes at
o
121
C. Dispense for plates or slopes as required. Note: Do not filter or adjust pH.
111.11. Littman oxgall agar
A confirmatory test for the identification of Trichophyton species.
Ingredients: Littman oxgall agar 27.5 g; distilled water 500 mL.
Method: Soak agar in 100 mL of water in stainless steel jug. Boil remaining 400 mL in a separate jug. When
water has boiled add to soaking agar and bring to the boil, stirring constantly. Dispense for slopes. Autoclave
for 10 minutes at 121
o
C, remove and slope.
111.12. Malt extract agar
For the routine cultivation and identification of fungi.
Ingredients: Malt extract 20 g; Bacto-agar 20 g; distilled water 1000 mL.
Method: Dissolve malt extract in a plastic beaker and adjust to pH 6.5 with NaOH. Soak agar in small
quantity of solution. Bring remaining solution to the boil, stirring constantly. Add to the soaking agar. Bring
to boil, stirring constantly. Dispense for slopes as required. Autoclave at 121
o
C for 10 minutes, remove and
dispense for slopes or pour for plates as required.
111.13. 1% Peptone agar
For the cultivation and differentiation of fungi.
Ingredients: Tryptone peptone 5 g; Bacto-agar 10 g; distilled water 500 mL.
Method: Soak agar and peptone in a small amount of water. Boil remaining water, add this to soaking ingre-
dients and bring to boil again. Dispense for slopes. Autoclave for 10 minutes at 121oC, then dispense for
slopes or plates as required.
Descriptions of Medical Fungi 307
111.14. Potato dextrose agar
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For the routine cultivation and identification of fungi.
Ingredients: Potato dextrose agar 39 g; distilled water 1000 mL.
Method: Soak potato dextrose agar in small amount of the water in a stainless-steel jug. Boil remaining water,
add to soaking ingredients, bring to the boil, stirring constantly. Dispense for slopes as required. Autoclave
o
at 121
C for 15 minutes. Remove and dispense for slopes or plates as required.
111.15. Rice grain slopes
To induce sporulation and for differentiation of M. audouinii and M. canis.
Method: Place approximately 1/2 teaspoon rice grains into wide neck 20 mL vials. Add 8 mL distilled water
to each vial. Cap the vials, then slope on racks ensuring rice grains are evenly distributed. Autoclave racks at
o
121
C for 15 minutes.
111.16. Sabouraud’s dextrose agar (SDA) with cycloheximide (0.05%),
chloramphenicol, gentamicin and yeast extract
For the primary isolation and cultivation of dermatophytes.
Ingredients: Sabouraud’s dextrose agar 65 g; cycloheximide (actidione) 0.5 g; chloramphenicol 0.05 g; yeast
extract 5 g; gentamicin (40 mg/mL) 0.56 mL; distilled water 1000 mL.
Method: Soak all ingredients, except gentamicin, in 100 mL water. Boil remaining water, add to soaking
ingredients, and bring to boil to dissolve, stirring well to prevent from charring. Add the gentamicin. Mix
well. Dispense for slopes if required. Autoclave at 121
plates as required.
o
C for 10 minutes. Remove and dispense for slopes or
111.17. Sabouraud’s dextrose agar with chloramphenicol and gentamicin
For the primary isolation and cultivation of yeasts and moulds.
Ingredients: Sabouraud’s dextrose agar 65 g; chloramphenicol 0.05 g; gentamicin (40 mg/mL) 0.56 mL;
distilled water 1000 mL.
Method: Soak all ingredients, except gentamicin, in 100 mL water. Boil remaining water, add to soaking
ingredients, and bring to boil to dissolve, stirring well to prevent from charring. Add the gentamicin. Mix
well. Dispense for slopes if required. Autoclave at 121
o
C for 10 minutes. Remove and dispense for slopes or
plates as required.
111.18. Sabouraud’s dextrose agar with 5% NaCl
For the cultivation and differentiation of dermatophytes especially T. rubrum from T. mentagrophytes.
Ingredients: Sabouraud’s dextrose agar 32.5 g; sodium chloride NaCl 25 g; Distilled distilled water 500 mL.
Method: Soak ingredients in approximately 100mL water. Bring remaining water to boil, add to soaking
ingredients, then bring back to the boil. Dispense for slopes. Autoclave at 121oC for 10 minutes, then dispense for slopes.
Descriptions of Medical Fungi 308
111.19. Tap water agar
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For the stimulation of sporulation in Apophysomyces and Saksenaea isolates.
Ingredients: Bacto-agar 15 g; distilled water 1000 mL.
Method: Add agar to water in stainless-steel jug, allow to soak. Autoclave at 121
and dispense for slopes or plates as required.
111.20. Urea agar slopes with 0.5% glucose
For the differentiation of urease producing organisms.
o
C for 10 minutes, remove
Urea glucose broth base: Urea, broth base 0.9 g; glucose 5 g; distilled water 450 mL. Add the urea broth base
and glucose to the distilled water in a 500mL beaker. Dispense in 5 x 90 mL amounts. Autoclave at 121
for 20 mins. When cool store at 4
o
C.
o
C
Method to make slopes: 40% Urea solution 10 mL; Bacto-agar 3 g; distilled water 100 mL. Add agar to
100 mL of distilled water in a 250 mL pyrex bottle. Autoclave at 121
o
C for 15 minutes and cool to 50oC in
a water bath. Add 90 mL of the urea glucose broth base and 10 mL of 40% urea solution to the autoclaved
agar solution and dispense in 3 mL aliquots. Angle the vials on racks to form slopes.
111.21. Vitamin Free Agar (Trichophyton Agar No. 1)
For the differentiation of Trichophyton species.
Ingredients: Trichophyton agar No. 1 11.8 g; distilled water 200 mL.
Method: Add agar to water in stainless-steel jug, allow to soak. Bring to boil to dissolve, stirring constantly.
Once boiled remove immediately to avoid discolouration. Dispense for slopes. Autoclave at 121oC for
10 minutes, remove and dispense for slopes.
DOI: 10.1079/9781800622340.0111
Descriptions of Medical Fungi 309
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Glossary
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For additional definitions of mycological terminology, the reader is referred to Ainsworth and Bisby’s
Dictionary of the Fungi (Kirk et al., 2008).
Acerose: Needle-like and stiff, like a pine needle.
Acervulus (pl. acervuli): A small, saucer or disk-shaped conidial fruiting body (embedded in host plant
tissue), conidiophores are formed on a cushion-like structure at the bottom of the cavity.
Acicular: Slender and pointed, needle-shaped.
Acrogenous: Development at the apex. Conidia born at the tip of the conidiophore.
Acropetal: Describes a chain of conidia in which the youngest conidium is at the tip and the oldest is at the
base.
Acropleurogenous: Formed at the end and on the sides. Conidia developing at the tip and along the sides of
the conidiophore.
Aculaete: Bearing narrow spines.
Acuminate: Gradually narrowing to a point.
Adelophialide: Reduced form of phialide lacking a basal septum. Seen in the general Phialemonium and
Lecythophora.
Adiaspore: A large spherical chlamydospore produced in the lungs of humans and animals by the enlargement of an inhaled conidium of Blastomyces (Emmonsia) parvus.
Aerial mycelium: Hyphal elements growing above the agar surface.
Allantoid: Slightly curved with rounded ends; sausage-like in form.
Ameroconidium (pl. ameroconidia): A one-celled conidium.
Ampulliform: Flask-like in form.
Anamorph: An asexual state of a fungus.
Annellide: A specialised conidiogenous cell producing conidia in basipetal succession by a series of short
percurrent proliferations (annellations). The tip of an annellide increases in length and becomes narrower as
each subsequent conidium is formed.
Annelloconidium (pl. annelloconidia): A conidium produced by an annellide.
Anthropophilic: Of dermatophytes, that are primarily parasitic on man. They are unable to colonise other
animals and they have no other environmental sources.
Apiculus: A short, sharp projection from a conidium.
Apophysis: A swelling. The term is primarily applied to the funnel-shaped swelling of a sporangiophore,
immediately below the columella, seen in Mucorales.
Apothecium (pl. apothecia): A cup-like or saucer-like ascoma in which the hymenium is exposed at maturity.
© CAB International 2023. Descriptions of Medical Fungi (eds S. Kidd, C.Halliday and D. Ellis)
3 11
Applanate: Flattened.
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Appressorium (pl. appressoria): A swelling on a germ tube or hypha, especially for attachment in an early
stage of infection (by many fungal plant pathogens), whether or not morphologically differentiated from
vegetative hyphae, as long as the structure adheres to and penetrates the host.
Arthric: Thallic conidial ontogeny involving the conversion and subsequent disarticulation of a determinant
conidiogenous hypha.
Arthro- (prefix): Jointed.
Arthroconidium (pl. arthroconidia): A thallic conidium released by either the splitting of a double septum or
by the fragmentation or lysis of a disjunctor cell.
Ascocarp: A fruiting body containing asci and ascospores.
Ascomycota: A group of fungi that reproduce sexually by the endogenous formation of ascospores in an ascus.
Ascomycetous: Referring to the Ascomycota.
Asco- (prefix): Pertaining to an ascus.
Ascoma pl. ascomata: An ascus-containing structure, ascocarp.
Ascospore: A spore produced in an ascus by “free cell formation”.
Ascus (pl. asci): A sac-like cell containing ascospores. Asci are characteristic of the Ascomycota.
Aseptate: Lacking septa, often pertaining to the hyphae seen in zygomycetous fungi (also see coenocytic).
Avellaneous: Hazel, wood-brown, pale brown in colour.
Awl-shaped: Tapering to slender stiff point.
Ballistoconidium (pl. ballistoconidia): A conidium that is forcibly discharged; as in Basidiobolus and
Conidiobolus.
Bamboo-hyphae: Straight hypha with darkened septa at regular intervals.
Basidioma: A basidium-producing organ; basidiocarp.
Basidiomycota: A group of fungi that reproduce sexually by the exogenous formation of basidiospores from
a basidium.
Basidiospore: A propagative cell (typically a ballistospore) containing one or two haploid nuclei produced,
after meiosis, on a basidium. The colour, form and ornamentation of the basidiospore are fundamental to
basidiomycete classification and an essential part of any specific description.
Basidium (pl. basidia): The cell or organ, diagnostic for basidiomycetes, from which, after karyogamy and
meiosis, basidiospores (generally 4) are produced externally each on an extension (sterigma) of its wall.
Basipetal: Describes a chain of conidia having the youngest conidia at the base of the chain, the oldest
conidium is at the apex.
Basocatenulate: A chain of conidia having the youngest conidia at the base of the chain, the oldest conidium
is at the apex.
Bipolar budding: Blastoconidia developing at the opposite poles of a parent cell.
Biseriate: In two series, i.e. phialides arising from metulae as in the genus Aspergillus.
Biverticillate: (of a penicillus), having branching at two levels, i.e. having metulae bearing phialides as in the
genus Penicillium.
312 Glossary
Blastic: A form of conidial development where there is a recognisable enlargement or "blowing out" of a
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conidial initial before being delimited by a septum.
Blastocatenate: A chain of conidia having the youngest cell at the tip.
Blastoconidium (pl. blastoconidia): An asexual conidium that forms by a blowing out or budding process.
Bud: A young conidium. Usually used to denote the young blastoconidia of yeasts.
Budding: Asexual multiplication by the production of a small outgrowth or bud from a parent cell.
Butyrous: Buttery.
Capsule: A hyaline mucopolysaccharide sheath around the cell wall of certain yeasts e.g. Cryptococcus and
Rhodotorula.
Catenulate: Conidia arranged in chains.
Cerebriform: Resembling the brain in texture or structure.
Chlamydospore: A thick-walled, thallic conidium formed within the vegetative hyphae. Chlamydospores
function as organs of perennation (survival from one year to the next by vegetative means) rather than
dissemination.
Cicatrised: (of conidiogenous cells and conidia), having thickened scars.
Cigar shaped: Cylindrical but gradually becoming narrower towards both ends.
Circinate: Twisted round, coiled.
Circumscissile: Opening or cracking along a circle.
Clamp connection: A specialised hyphal bridge over a septum in the basidiomycetous fungi.
Clavate: Club-like; narrowing in the direction of the base.
Cleistothecium (pl. cleistothecia): An enclosed ascocarp containing randomly dispersed asci.
Coenocytic: Infrequently septate, multi-nucleate hyphae as in the zygomycetous fungi.
Collarette: A small collar. Usually, a remnant of a cell wall present at the tip of a phialide, or around a
sporangiophore.
Columella (pl. columellae): A sterile dome-like structure at the tip of a sporangiophore or within a
sporangium.
Columnar: Forming a column.
Conidiogenous cell: A cell that forms conidia.
Conidiophore: A specialised hypha upon which conidia develop.
Conidium (pl. conidia): An asexual reproductive propagule formed in any manner that does not involve
cytoplasmic cleavage. Conidia function as organs of dissemination.
Cottony: Having a loose and coarse texture.
Cylindrical: Cylindric, having parallel walls and circular cross-section.
Dematiaceous: A dark brown, greenish-grey or black colour.
Dermatophyte: A fungus with the ability to utilise keratin to infect hair, nail and skin of animals and humans.
Denticle: A small projection or peg on which conidia are produced.
Glossary 313
Determinate conidiophore: Growth ceasing with the production of terminal conidia. The conidiophore does
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not alter in length after the formation of conidia.
Deuteromycetes: An artificial subdivision to accommodate those fungi where only the asexual state is
known.
Dichotomous: A type of hyphal branching, frequently successively, into two more or less equal arms.
Dictyoconidium (pl. dictyoconidia): A conidium with both longitudinal and transverse septa; a muriform
conidium.
Didmoconidium (pl. didymoconidia): A two-celled conidium.
Dimorphic: Having two different morphological forms.
Disjunctor cell: An empty cell that fragments and/or undergoes lysis to release a conidium.
Distoseptate: (of conidial septation), having the individual cells each surrounded by a sac-like wall distinct
from the outer wall, as in Drechslera, cf. euseptate.
Divergent: Tending to develop in different directions.
Dolipore septum: A characteristic septum found in the Basidiomycetes that flares out near the pore to form
an elongate channel.
Double septum: A two layered septum that may undergo centripetal separation (schizolysis) to release a
conidium.
Downy: Having a short and dense mycelial texture.
Dysgonic: A slow growing variant.
Echinulate: Having sharply pointed spines; spinose.
Ectothrix: Natural hair invasion by a dermatophyte characterised by arthroconidia on the outside of the hair
shaft.
Effuse: Spread out, radiate.
Elliptical: Oval, with a symmetric curve.
Elongate: Lengthened.
Endospore: A spore produced within a spherule.
Endothrix: Natural hair invasion by a dermatophyte characterised by the development of arthroconidia
within the hair shaft only.
Enteroblastic: A mode of blastic conidium ontogeny in which inner cell wall layers of the conidiogenous cell
are involved in conidium development.
Erect: Upright.
Euseptate: Of conidial septation, having cells separated by multilayered walls of similar structure to lateral
walls, cf. distoseptate.
Evanescent: Having a short existence; fugacious.
Exudate: Droplets of fluid formed on the surface of a colony.
Falcate: Curved like a sickle.
Farinose: Covered with a sort of white, mealy powder.
314 Glossary
Flexuous: Wavy.
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Floccose: Fluffy or cottony.
Foot cell: A basal cell of a conidiophore as seen in Aspergillus and Fusarium.
Funiculose: (of hyphae), aggregated into rope-like strands.
Fusiform: Spindle-shaped, tapering toward the end.
Geniculate: Bent like a knee.
Geophillic: Of dermatophytes, that normally inhabit the soil where they are believed to decompose kerati-
naceous debris. Some species may cause infections in animals and man following contact with soil.
Germ tube: The initial hypha that develops from a conidium or spore.
Glabrous: Smooth.
Globose: Spherical or almost so.
Glioconidia: Conidia aggregated in slimy heads at the tip of an annellide or phialide.
Guttulate: Containing one or more oil droplets.
Gymnothecium (pl. gymnothecia): A non-ostiolate ascocarp composed of loosely interwoven hyphae and
containing randomly dispersed asci; this arrangement allows many open spaces for the release of ascospores
formed within the fruiting structure.
Heterothallic: A fungus that requires mating between two compatible strains for sexual reproduction to
occur.
Hilum: A mark or scar, especially that on a spore at the point of attachment to a conidiogenous cell or
Holoblastic: A mode of blastic conidium ontogeny in which all the cell wall layers of the conidiogenous cell
are involved in conidium development.
Holothallic: A mode of thallic conidium ontogeny in which all the cell wall layers of the conidiogenous cell
are involved in conidium development.
Homothallic: A fungus capable of sexual reproduction on a single thallus.
Hülle cells: Thick-walled cells with characteristic thin-walled pores, usually associated with cleistothecia of
Aspergillus nidulans.
Hyaline: Transparent or nearly so; translucent; frequently used in the sense of colourless.
Hyalo-: A prefix (of conidia) meaning hyaline or brightly coloured, especially for groups of anamorphic
fungi.
Hyalohyphomycosis: A mycotic infection of humans or animals caused by a non-dematiaceous fungus. cf.
phaeohyphomycosis.
Hypha (pl. hyphae): A single filament of a fungus.
Hyphomycetes: A class of mycelial moulds which reproduce asexually by conidia on hyphae or aggregations
of hyphae.
Hymenium (pl. hymenia): A layer of cells producing basidiospores.
Indeterminate: (1) Having the edge not well-defined, especially of fruit-bodies and leaf-spots; (2)
(of conidiophores), continuing growth indefinitely.
Glossary 315
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