COINED THE TERM…ZOONOSES “Zoonoses” Stating that “Between animal and human medicine, there is no dividing line, nor should there be. The object is different but the experience gained constitute the basis of all medicine

1. DR. VEERENDRA MARAVI
JR-2 , DEPT OF MICROBIOLOGY

2.  Introduction
 Risk group
 Microsporum spp.,Trichophyton spp.
 Sporothrix schenckii ,Sporothix brasiliensis
 Mallasezia spp.
 Cryptococcus neoformans,Cryptococcus gattii
 Penicillium (Talaromyces) marneffei
 Lacazia loboi
 Emmonsia spp.
 Conidiobolus coronatus,Conidiobolus incongruus, Basidiobolus ranarum
 Histoplasma capsulatum
 Coccidioides immitis,Coccidioides posadasii
 Paracoccidioides brasiliensis, Paracoccidioides lutzii
 Blastomyces dermatitidis
 Pneumocystis carinii (mammals),Pneumocystis jirovecii (Human)
 conclusion

3. “Zoonoses” Stating that
“Between animal and human
medicine, there is no dividing
line, nor should there be. The
object is different but the
experience gained constitute
the basis of all medicine”
Rudolf Virchow
1821-1905

4. Diseases and infections
which are naturally
transmitted
between vertebrate
animals and humans
-WHO 1959

5. The society of Indian Human
And Animal Mycologist
(SIHAM) found in 1996 and
its 1st conference held in
Jabalpur MP

6. Infant
Children<5
Pregnant women
People undergoing chemotherapy
People with organ transplant
People with HIV / AIDS
Elderly

7. Farmers , Livestock owners and
occupational groups
1 Share air and space with animals
2 Frequent contact with domestic and wild animals

8. Disease causing - Dermatophytosis
Worldwide distribution
Transmission –Direct contact with infected animal or material
Animals –All domestic and wild animals
Clinical sign in animal -Classical ring lesion with central healing
and crusts at the peripheral area, degree of folliculitis
Clinical signs & disease in humans - Tenea capitis, Tinea barbae,
Tinea faciei, Tinea corporis, Tinea cruris, Tinea
unguium, Tinea pedis, Tinea manuum, Tinea

9. Specimen-
Skin Scraping
Nail Scraping
Hair plucking
1. Skin : from the margin of the lesion, with the scalpel.
2. Nail : deeper part is collected and superficial part is
discarded.
3. Hair : plucked by fine forceps

10. fluorescence when examined under Wood’s light

11. Microscopic examinations -KOH preparation of skin or nail :
branching hyphae or chains of arthospores in dermatophytes

12. Sabouraud dextrose agar (SDA)
White cottony fluffy growth normally
never green, blue and black
Dermatophyte Test Medium
If growth present turns medium red

14. Disease causing – Sporotrichosis
Worldwide distribution
Transmission- Work related trauma, scratches or Bite of animals
Animals- Cats , Dogs ,horses, Cow , Camel, Goat Mule Bird ,Pig,
Rat, Dolphin
Clinical sign in animals -Localized cutaneous, lymphocutaneous and
disseminated infection
clinical signs & disease in humans- Fixed cutaneous,
lymphocutaneous , osteoarticular and disseminated infection

15. Specimen - collected like Pus , exudate and aspirate from
nodules
Wet mount- Small, Elongated yeast cell , sensitivity is low
Gram stained Smear – Gram positive irregular stained
yeast cell
Tissue section- Organism appears as Cigar shaped bodies

16. Gold standerd
 takes 3 to 4 weeks
Specimen inoculated in two sets in SDA, blood agar,
,chocolate agar incubated at 25 ° C and 37° C .
Colony –Moist membranous , fine greyish velvety may
vary from cream to black

18. Disease causing -Malassezia infection (pityriasis)
Worldwide distribution
Malassezia yeasts are commensal of human skin (part of the normal
microbiota)
Animals- Domestic animals such as dogs, cats, cows, sheep, pig,
horse, wild animals held in captivity, and animals from
wildlife
Disease in animals- Dermatitis, alopecia, stenosis, otitis externa
Clinical sign in humans- Chronic superficial disease of the skin
(pityriasis versicolor), folliculitis, seborrhoeic dermatitis and
dandruff, fungaemia

19. Sample-Skin from affected site , sponged with 70%
alcohol and after drying , edges of lesions scraped
with 15 no. Blade or glass slide
Wet Mount – large quantity and clusters of round yeast
cells ,2 -7μm in size ,occasional budding ,hyphae
are blunt ; short, stout may be curved with infrequent
branching (banana and grapes)
Albert Stain- Stains yeast cell and hyphae
purple ,clearly delineating the details of fungi against
a background of surrounding keratinocytes

21.  Malassezia are normal flora and
lipophilic so SDA used with olive oil
 fungus Dixon Agar is used now
SDA with chloramphenicol ,actidione
Tween 80 and film of olive oil-
small 3-6 mm yellowish colony,
slight rised with irregular edges in
7- 5 days at 32- 35° C

22. Disease causing - cryptococcosis
Distribution Worldwide
Transmission- Mainly by inhalation of fungus, occasionally through
breaks in the skin
Animals- Wide variety of mammals, birds, reptiles and amphibians
Clinical sign in animals- Focal or disseminated infection, affecting a
single organ system or many, central nervous system
involvement
Disease in humans- Cutaneous, Ocular, pulmonary and central
nervous system involvement

23. Sample –CSF, Serum or other body fluids site involved
Wet mount - with India ink or 10% Nigrosin with formalin-
Round to globular budding yeast μm size 5 to 20 μm with
distinct halo

24. Growth of cryptococcus is yeast like and highly mucoid
cream to buff-colored on SDA

25. Disease causing - Penicilliosis
Distribution in Southern China and South-East Asia
Transmission – Not clearly known
Animal- Bamboo rats, domestic animals such as dogs, cats
Clinical signs in animals- Skin dermatitidis, rhinitis, otitis externa
and disseminated infection
Clinical signs in Human - Non-specific clinical signs (generalized
lymphadenopathy, molluscum contagiosum like lesions of the
skin and mucosa) and disseminated infection

26. Sample – Impression smear if skin ,Lymph node biopsy,
Bone marrow aspirate
Stained with H and E – the yeast like
tissue multiply by trnsverse
fission rather then budding ,
have prominent central septum

27.  Gold standard
 On SDA and blood agar -
Greyish- white
colony with in two days
subsequently wooly-downy
to granuler, yellow orange
In center, bright brick red
in surrounding as
early as three days after
incubation in room temp.

28. Disease causing - Lobomycosis
Distribution- South and Central America, United States, Canada,
Europe,
Transmission – Trauma
Animal –Dolphin and aquatic animals
Clinical signs in animals- Granulomatous dermatitis
Disease in humans-Granulomatous dermatitis

29. Sample – Clinical curettage or
biopsy by surgical procedure
KOH wet mount – yeast cell with
Hyaline double refractile wall ,
size 6 to 12 micro meter;
Un branched 4 to 7 cells due to
Sequential budding linked to one
Another by tubular connection

30. Rhinosporodium seeberi ,Mycobacterium
leprae and Lacaziz loboi has not been
successfully grown till date.
One well documented case of experimental
inoculation in human volunteer who
developed single lesion 3 months after
intradermal injection.

31. Disease causing - Adiaspiromycosis
Case reported from- Case reports from Asia , Australia,
Europe, North America
Transmission- Inhalation of the fungus
Animal - Wild rodents ,Rabbits , Squirrels
Clinical sign in animals- Deep mycoses
Disease in humans- Lung and disseminated disease

32. Sample – by fine neddle aspiration or biopsy tissue
sputum exam, and bronchial washing is usually negative
H & E stain- Shows large thick walled adiaconidia
surrounded by granulomatous reaction.
 symmetrical round to oval
Neither budding nor endosporulation seen in adiaconidia
Size 50 to 500 micro meter thickness 20 to 70 µm
Trilaminar wall

33. (A)Histopathological section of a weasel showing numerous granulomatous
lesions. A central thick walled spore, (<50μm in size) surrounded by
macrophages and eosinophils is present in a single granulomatous lesion.
H&E stain.
(B)(B) Histopathological section of an otter’s lung showing an adiaspore (about
300μm in size) surrounded by epithelioid macrophages and multinucleate
giant cells. H&E stain

34. SDA and BHI blood agar but without actidione at 25° C -
Grows moderately rapid initially velvety later on produces
aerial mycelia colonies are pale white, buff to pale brown
Colony microscopically - Branching septate hyphae with
diameter 0.,5 to 2 µm
,branch at right angles from vegetative hyphae

35. Disease causing -Entomophthoromycosis
Case reported from- Tropical countries of Africa, Asia, United
States and Europe
Transmission- Traumatic implantation or inhalation of the fungus
Animal -Horses, dogs, sheep
Clinical signs in animals- Cutaneous and disseminated infection
Disease in humans - Chronic subcutaneous and invasive infection

36. Sample – Biopsy taken from affected site
KOH wet mount- Coenocytic hyphae ,
short,6-15µm
wide with
cross-walls seprating empty hyphal
fragments from actively growing part
H & E staining – Non septate hyphae
with surrounding eosinophilic sleeve
H &E

37. SDA – AT 25 ºC - Rapidly growing waxy creamy colonies
adherent to surface with pale reverse
 At 37ºC – Furrows and folds appear ‘
Indue course ,waxy colony become powdery from center
due to production of short ,white aerial mycelia

38. Growth in SDA

39. Disease - Histoplasmosis
Distribution - Worldwide (endemic in Mississippi and Ohio River
valleys in USA)
Transmission - Inhalation of the fungus
Animal- Cattle, sheep, horses
Clinical sign in animals - Non-specific signs (chronic gastrointestinal
infection) and disseminated infection
clinical sign in humans - Chronic progressive lung disease, chronic
cutaneous or systemic disease or an acute fulminating fatal
systemic disease

40. Specimen- Sputum, bone marrow and lymph node
aspiration/ biopsy and peripheral blood film , biopsy from
skin lesion
KOH wet mount- Tiny yeast cell
Thik and thin smears are prepared from peripheral blood
or bone marrow, stained with geimsa stain,- Fungus
appear as smalloval yeast cells 2-4 µm, within
mononuclear or polymorphonuclear cells ,occasionally in
giant cells,

41.  Multiple yeast cell of histoplasma capsulatum with in histiocytes of alveoli
in renal transplant recipient( H&E * 400)

42. SDA with antibacterial antibiotic –Incubated at 25ºC to
37ºC – the fungal culture for demonstration for yeast
phase is not beneficial for primary isolation but conversion
from mycelial to yeast to confirm identity of isolates

43. Disease – Coccidioidomycosis
Distribution- Southwestern USA, northern Mexico, Central and
South America
Transmission - Inhalation of arthroconidia and skin trauma
Animals - Dogs, llamas, nonhuman primates, cats, horses,
domesticated or wild mammals, snakes
Clinical signs in Animals - Asymptomatic to severe and fatal
infection
Disease in humans- Cutaneous, pulmonary, disseminated infection

44. Specimen- Sputum Gastric
contents , CSF exudate or
pus
KOH wet mount- Refractile
thick walled globular
spherules of about 20
Immature spherules are
smaller and without
endospore

45. Gold Standard
 In SDA cultivated in well
stopped narrow neck bottles
or culture tube at ºC as well
as 37ºC – Appear with in 3-5
days , initially moist and
smooth then become downy
greyish white with tan to
brown underside.

46. Disease - Paracoccidioidomycosis
Distribution - South America
Transmission - Inhalation of the fungus, injuries of the skin and
mucosal membranes
Animal - Dogs, domesticated and wild animals (armadillos and
monkeys)
Clinical signs in Animals - Non-specific clinical signs depending on
the organ ( lymphadenomegaly, apathy, and
hepatosplenomegaly)
Clinical signs in humans - Mucocutaneous, pulmonary or
disseminated infection

47. Specimen – sputum, BAL fluid, CSF, Pus , crust from
granulomatous lesion, biopsy material
10% KOH wet mount- Round refractile yeast cell, varying
in size from 2-10 to 30 µm or more. Present as single or
short chains of cells. Multipolar budding is seen

48. paracoccidioidomycosis. (A) Wet mount of a fresh sputum examination
from a patient after clarification with potassium hydroxide. (B) Grocot
silver methenamine staining of a smear from lymph node aspirate.

49. Grow on SDA with antibacterial antibiotics and actidione
– Growth of both yeast and mycelial are slow on primery
isolation. Growth at 37ºC in week that is yeast like soft
,off-white to cream colored wrinkled and rough to pasty in
appearance.
Colonies at 25ºC are flat to wrinkled, leathery ,color of
mycelial colony varies from white to tan with yellowish –
brown reverse

50. Paracoccidioides brasiliensis (BAT strain). Aspect of the colony growing on
Sabouraud-agar at 25°C and at 37°C .

51. Disease - Blastomycosis
Distribution - Worldwide (endemic in North American continent,
Distribution -autochthonous in Africa, South America and Asia
Transmission - Inhalation of airborne conidia
Animals - Dogs, cats, horses, marine mammals
Clinical signs in animals - Cutaneous, pulmonary, systemic
infection
Clinical signs in humans - Cutaneous, pulmonary, disseminated
infection (granulomatous and suppurative lesions in lung skin
and bones

52. Specimen- Sputum, BAL,
pus from abscess, biopsy ,
urine after prostatic
massage
KOH wet mount-Yeast cell
,double-contoured , thick
walled, multi nucleated
yeast with single broad-
based budding daughter
cells
Blastomyces dermatitidis broad-based budding yeast in the aspirate of a
chest wall abscess. Note the presence of multiple nuclei, the thickened cell
wall, and the broad-based bud.

53.  Fungal culture in SDA at
25°C and at 37°C ,growth is
slow takes two to four weeks
 Colonies look yeast like at
early then hyphal projections
develops on surface and
finally enier surface become
downy/ fluffy white.
 Old culture become tan
colored

54.  LPCB - a low power view of hyphae
growing away from the point of
inoculation of a slide culture (100X)
Blastomyces dermatitidis - Conidia seen
growing along hyphae. (400X)

55. Disease - Pneumocystosis
Distribution – Worldwide
Transmission - Inhalation of airborne conidia
Animals - Rodents, dogs, cats, cattle
Clinical signs in animals - Lethal pneumonia in immune
debilitated animals
Clinical signs in humans - Asymptomatic, interstitial
pneumonia, progressive pneumonia (in
immunocompromised hosts)

56. Specimen- Sputum ,lung biopsy ,BAL fluid
BAL is ideal sample for cytological staining
H&E stains- Shows frothy oedema fluids in alveoli of
patient
Selective staining method is Giemsa – stains the
trophozoites as deep blue with no stains of cyst walls

58. Gold standard
Not use full in routine diagnosis
Yet to be grown in vitro
Grown in continuous cell line derived from
adenocarcinoma
Yield is low

60. The important of fungal zoonotic infection has been
demonstrated. There is no doubt that these types of fungi
need to be controlled.
 Control of human exposure to animal reservoirs can
protect susceptible populations .
Raise awareness of the scale of the problem for zoonotic
fungi in order to better define the burden, distribution,
mortality and socio-economic consequences, and also
provide proper platform for prevention and control
strategies.

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62. THANKYOU