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Cryptococcus neoformans
Cryptococcus
Introduction
• Cryptococcus neoformans is an encapsulated
yeast and an obligate aerobe that can live in
both plants and animal
• C. neoformans usually infects the lungs or the
central nervous system (the brain and spinal
cord), but it can also affect other parts of the
body.
Habitat
• found in soil contaminated with pigeon
droppings or eucalyptus trees and decaying
wood.
• Cryptococcus neoformans grows easily in
pigeon feces, however the birds are not
naturally infected due to high body temp.
• Speciation:
• The genus Cryptococcus includes around 37
species.
• Among these, Cryptococcus neoformans is the
only species that is pathogenic to humans.
• Other species are Cryptococcus albidus,
Cryptococcus laurentii, Cryptococcus terreus,
and Cryptococcus gastricus.
Serotypes-Cryptococcus neoformans
• It has four serotypes (A to D).
• Serotyping is based on capsular agglutination
reactions.
• All types can cause human disease
• Cryptococcus neoformans serotype A is
responsible for the majority of human cases.
• Infection due to serotypes A and D common
• Serotypes A and D found in excreta of wild
and domesticated birds throughout world
• Serotypes B and C found in flowers Eucalyptus
camaldulensis
varieties
• It includes three varieties; var. gattii, var.
neoformans and var. grubii.
• The two varieties, neoformans and gattii are
morphologically similar, except that
basidiospores of var. neoformans are round
and those of var. gattii are more elliptical in
shape.
Morphology
• The typical vegetative form of C. neoformans is
the yeast form with a cell diameter of 2.5 μm to
10 μm.
• A characteristic polysaccharide capsule of
variable thickness (1-30μm) surrounds these
yeasts.
• In its natural environment the capsule is thinner
and the yeast smaller, while thicker capsules tend
to be found from infected tissues.
• The capsules stain pink by the Meyer’s
mucicarmine technique.
Life cycle
• Life cycle
– Asexual: yeast that reproduce by budding
• Human infections
– Sexual
• C. neoformans is heterothallic haploid yeasts that exist in two
mating types, MATαand MATa.
• MAT α strains are predominant in nature
• The haploid cells propagate asexually by budding
• In Sexual life cycle
– strains of opposite mating type come into contact on
substrates conducive for mating
– In the terminal swollen part of the basidium, the two
nuclei from opposite mating types fuse to form a single
transient diploidnucleus
– diploidnucleus undergoes one cycle of meiotic division
producing four haploid nuclei
Life cycle
– four haploid nuclei repeatedly undergo mitotic division
– basipetally produce bud of four long chains of
basidiospores (sexual spores) consisting of MATα and
MATa type spores in an equal ratio
• These basidiospores can cause infection
Life Cycle
Virulence Factors
• Virulence
– The polysaccharide capsule,
– melanin production,
– mannitol production,
– different enzymes :superoxide dismutase, proteases,
phospholipase B, and lysophospholipase the phenoloxidase
, urease
– the organism's ability to grow at 37°C
• these factors increase the degree of pathogenicity of a
microbe
Melanin Synthesis
• naturally occurring C. neoformans mutants
lacking melanin (Mel-) are less Virulent than
melanin-producing strains.
Cryptococcus
mannitol production
• high concentrations of D-mannitol in the CNS
may contribute to brain edema.
• D-mannitol protect the organism from
oxidative damage by hydroxyl radicals
• Mode of Infection:
– inhalation of the organism( basidiospores) from
environment.
– No person to person transmission
Disease
• Cryptococcosis
– subacute and chronic infection of lung
– earlier called European blastomycosis
– considered an opportunistic infections as it affects
mainly immunosuppressed individuals.
clinical types
• The clinical types of cryptococcosis are:
– pulmonary,
– CNS,
– visceral,
– osseous,
– cutaneous and mucocutaneous.
Pulmonary cryptococcosis
• In healthy individuals, the disease is primarily
pulmonary and is asymptomatic.
• Goes undiagnosed and resolves spontaneously
in majority of patients
• small proportion of cases ,haematogeneous
spread results in subacute or chronic
meningitis or meningoencephalitis
• All patients with pulmonary disease need a CSF examination
to r/o sub clinical meningitis
PATHOGENESIS
• Enters through lungs - inhalation of
Basidiospores of C neoformans
• Enters deep into lungs, Men acquires more
infections, and women less infected.
• Self limiting in most cases,
• Pulmonary infections can occur.
• Present as discrete nodules - Cryptococcoma.
Disseminated cryptococcosis
• CNS cryptococcosis
• Cutaneous and mucocutaneous cryptococcosis
• In osseous cryptococcosis cranial bones and vertebrae are
commonly affected.
• Visceral cryptococcosis -most commonly heart, testis,
prostate and eye.
Cryptococcus
Cryptococcus
Route of infection
Route of cryptococcal
meningoencephalitis
risk group of cryptococcosis
• Patients with:
– AIDS,
– carcinoma,
– leukemia,
– collagen vascular disease,
– Hodgkin’s disease,
– sarcoidosis and
– undergoing immunosuppressive therapy
Laboratory diagnosis:
• The specimens collected according to site of
infection
• Pulmonary : are sputum, blood, lung biopsy
• CNS: CSF, Serum CRAG
• Cutaneous and mucocutaneous: skin biopsy,
Serum CRAG
Wet mount Preparation
• 10% KOH mount:
– Globose yeast cells are seen in most clinical
materials, such as CSF and pulmonary tissue
Negative staining:
India ink preparation and Nigrosin
Cryptococcus in Lung
PAS staining
Silver Stain
Mucicaramine staining
Culture
• Aspirates and tissues are inoculated onto
Sabouraud dextrose agar.
• Colonies of C.neoformans are fast growing(
takes 48 to 72 hours.)
• Colonies-soft, glistening, smooth, usually
mucoid, and cream to yellowish brown in
color.
• It grows well at 25°C as well as 37°C.
Cryptococcus
Bird seed agar
Biochemical test
• phenoloxidase enzyme test:
– phenoloxidase enzyme that results in production
of melanin and thus a brown to black
discoloration of the colony when it is grown on
caffeic acid agar or bird seed agar.
• Urease test:
– Cryptococcus neoformans differs from Candida by
hydrolysing urea and not forming pseudohyphae.
Cryptococcus
serological
• Detection of cryptococcal antigens,
particularly polysaccharide capsular antigen
by latex-agglutination in CSF, blood or urine.
• Detection of Serum antibodies by
agglutination and immunofluorescence
Treatment
• Amphotericin B combined with 5-
fluorocytosine
• Fluconazole
Thank you

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Cryptococcus

  • 3. Introduction • Cryptococcus neoformans is an encapsulated yeast and an obligate aerobe that can live in both plants and animal • C. neoformans usually infects the lungs or the central nervous system (the brain and spinal cord), but it can also affect other parts of the body.
  • 4. Habitat • found in soil contaminated with pigeon droppings or eucalyptus trees and decaying wood. • Cryptococcus neoformans grows easily in pigeon feces, however the birds are not naturally infected due to high body temp.
  • 5. • Speciation: • The genus Cryptococcus includes around 37 species. • Among these, Cryptococcus neoformans is the only species that is pathogenic to humans. • Other species are Cryptococcus albidus, Cryptococcus laurentii, Cryptococcus terreus, and Cryptococcus gastricus.
  • 6. Serotypes-Cryptococcus neoformans • It has four serotypes (A to D). • Serotyping is based on capsular agglutination reactions. • All types can cause human disease • Cryptococcus neoformans serotype A is responsible for the majority of human cases.
  • 7. • Infection due to serotypes A and D common • Serotypes A and D found in excreta of wild and domesticated birds throughout world • Serotypes B and C found in flowers Eucalyptus camaldulensis
  • 8. varieties • It includes three varieties; var. gattii, var. neoformans and var. grubii. • The two varieties, neoformans and gattii are morphologically similar, except that basidiospores of var. neoformans are round and those of var. gattii are more elliptical in shape.
  • 9. Morphology • The typical vegetative form of C. neoformans is the yeast form with a cell diameter of 2.5 μm to 10 μm. • A characteristic polysaccharide capsule of variable thickness (1-30μm) surrounds these yeasts. • In its natural environment the capsule is thinner and the yeast smaller, while thicker capsules tend to be found from infected tissues. • The capsules stain pink by the Meyer’s mucicarmine technique.
  • 10. Life cycle • Life cycle – Asexual: yeast that reproduce by budding • Human infections – Sexual • C. neoformans is heterothallic haploid yeasts that exist in two mating types, MATαand MATa. • MAT α strains are predominant in nature
  • 11. • The haploid cells propagate asexually by budding • In Sexual life cycle – strains of opposite mating type come into contact on substrates conducive for mating – In the terminal swollen part of the basidium, the two nuclei from opposite mating types fuse to form a single transient diploidnucleus – diploidnucleus undergoes one cycle of meiotic division producing four haploid nuclei
  • 12. Life cycle – four haploid nuclei repeatedly undergo mitotic division – basipetally produce bud of four long chains of basidiospores (sexual spores) consisting of MATα and MATa type spores in an equal ratio • These basidiospores can cause infection
  • 14. Virulence Factors • Virulence – The polysaccharide capsule, – melanin production, – mannitol production, – different enzymes :superoxide dismutase, proteases, phospholipase B, and lysophospholipase the phenoloxidase , urease – the organism's ability to grow at 37°C • these factors increase the degree of pathogenicity of a microbe
  • 15. Melanin Synthesis • naturally occurring C. neoformans mutants lacking melanin (Mel-) are less Virulent than melanin-producing strains.
  • 17. mannitol production • high concentrations of D-mannitol in the CNS may contribute to brain edema. • D-mannitol protect the organism from oxidative damage by hydroxyl radicals
  • 18. • Mode of Infection: – inhalation of the organism( basidiospores) from environment. – No person to person transmission
  • 19. Disease • Cryptococcosis – subacute and chronic infection of lung – earlier called European blastomycosis – considered an opportunistic infections as it affects mainly immunosuppressed individuals.
  • 20. clinical types • The clinical types of cryptococcosis are: – pulmonary, – CNS, – visceral, – osseous, – cutaneous and mucocutaneous.
  • 21. Pulmonary cryptococcosis • In healthy individuals, the disease is primarily pulmonary and is asymptomatic. • Goes undiagnosed and resolves spontaneously in majority of patients • small proportion of cases ,haematogeneous spread results in subacute or chronic meningitis or meningoencephalitis • All patients with pulmonary disease need a CSF examination to r/o sub clinical meningitis
  • 22. PATHOGENESIS • Enters through lungs - inhalation of Basidiospores of C neoformans • Enters deep into lungs, Men acquires more infections, and women less infected. • Self limiting in most cases, • Pulmonary infections can occur. • Present as discrete nodules - Cryptococcoma.
  • 23. Disseminated cryptococcosis • CNS cryptococcosis • Cutaneous and mucocutaneous cryptococcosis • In osseous cryptococcosis cranial bones and vertebrae are commonly affected. • Visceral cryptococcosis -most commonly heart, testis, prostate and eye.
  • 28. risk group of cryptococcosis • Patients with: – AIDS, – carcinoma, – leukemia, – collagen vascular disease, – Hodgkin’s disease, – sarcoidosis and – undergoing immunosuppressive therapy
  • 29. Laboratory diagnosis: • The specimens collected according to site of infection • Pulmonary : are sputum, blood, lung biopsy • CNS: CSF, Serum CRAG • Cutaneous and mucocutaneous: skin biopsy, Serum CRAG
  • 30. Wet mount Preparation • 10% KOH mount: – Globose yeast cells are seen in most clinical materials, such as CSF and pulmonary tissue
  • 31. Negative staining: India ink preparation and Nigrosin
  • 34. Culture • Aspirates and tissues are inoculated onto Sabouraud dextrose agar. • Colonies of C.neoformans are fast growing( takes 48 to 72 hours.) • Colonies-soft, glistening, smooth, usually mucoid, and cream to yellowish brown in color. • It grows well at 25°C as well as 37°C.
  • 37. Biochemical test • phenoloxidase enzyme test: – phenoloxidase enzyme that results in production of melanin and thus a brown to black discoloration of the colony when it is grown on caffeic acid agar or bird seed agar. • Urease test: – Cryptococcus neoformans differs from Candida by hydrolysing urea and not forming pseudohyphae.
  • 39. serological • Detection of cryptococcal antigens, particularly polysaccharide capsular antigen by latex-agglutination in CSF, blood or urine. • Detection of Serum antibodies by agglutination and immunofluorescence
  • 40. Treatment • Amphotericin B combined with 5- fluorocytosine • Fluconazole