4 Microbiology

AIIMS NOVEMBER 2017

1. Lipoarabinomannan urine assay is used for screening of?
a. Leprosy b. Tuberculosis
c. Corynebacterium d. Streptococcus
2. Replication cycle of which virus is shown in the Picture Below?

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a. HIV b. HBV
c. EBV d. Polio
184 Section I  •  Subject-wise MCQs and Answers with Explanations

3. Identify the parasite in fecal smear picture? 7. A lady presented with verrucous growth as show in the
picture. She gives history of thorn prick 6 months back.
What is the most likely causative agent?

PARASITO PLATE 13B

a. Entamoeba dyspar
b. Balantidium coli

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c. Cryptosporidium
d. Giardia

4. Best method of diagnosis of clostridium difficile is?
a. Toxin assay by ELISA
b. Pure strain Isolation by culture
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c. Gene detection by PCR
MICRO PLATE 12A
d. Colonoscopy
a. Mycetoma
5. Identify the Bacteria with structure shown in the picture?
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b. Sporotrichosis
c. Chromoblastomycosis
d. Tinea Nigra
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8. A female from Himachal pradesh presented with history

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of thorn prick a year back and now with verrucous lesion

on leg. Biopsy image is shown below. What is most likely
diagnosis?
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a. Vibrio
b. Leptospira
c. Helicobacter pylori
d. Campylobacter
6. All of following are dimorphic fungi EXCEPT: MICRO PLATE 13D
a. Histoplasma capsulatum a. Sporotrichosis
b. Penicillium marneffei b. Chromoblastomycosis
c. Pneumocystis jiroveci c. Actinomycetoma
d. Sporothrix schenckii d. Eumycetoma

AIIMS Nov 2013–May 2011

AIIMS (Nov 2017–May 2014) Questions with Explanations Covered in Volume II (Available Separately)
186 Section I  •  Subject-wise MCQs and Answers with Explanations

20. Which of the following fits to the life cycle picture given below? 22. Which of the following parasite’s life cycle is shown below?

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PARASITO PLATE 12I PARASITO PLATE 16D

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a. B. coli b. E. histolytica a. Fasciola buski b. Fasciola hepatica

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c. A. duodenale d. E. vermicularis c. Paragonimus westermani d. Clonorchis sinensis

21. Identify the organism from its egg in the picture? 23. Identify the organism in the slide?
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MICRO PLATE 1I
PARASITO PLATE 15D a. Nocardia sp b. Mycobacterium tuberculosis
a. Enterobius vermicularis c. Mycobacterium leprae d. Actinomyces sp
b. Trichuris trichiura 24. 1-3 b D glucan assay for fungi not used for?
c. Ascaris Lumbricoides a. Aspergillus b. Pneumocystis
d. Necator americanus c. Cryptococcus d. Pneumocystis jirovecii

AIIMS Nov 2013–May 2011

AIIMS (Nov 2017–May 2014) Questions with Explanations Covered in Volume II (Available Separately)

ANSWERS WITH EXPLANATIONS
AIIMS NOVEMBER 2017
1. Ans. (b)  Tuberculosis
Ref: Prescott’s Microbiology 9th ed page 744, Harrison 19th ed
page 1103, http://www.who.int/tb/publications/use-of-lf-lam-tb-
hiv/en/
Mycobacterial cell wall, lipoarabinomannan, is involved in the
pathogen-host interaction and facilitates the survival of M.
tuberculosis within macrophages. Tests based on the detection
of mycobacterial lipoarabinomannan (LAM) antigen in urine
have emerged as potential point-of-care tests for tuberculosis
(TB). LAM antigen is a lipopolysaccharide present in
mycobacterial cell walls, which is released from metabolically
active or degenerating bacterial cells and appears to be present
only in people with active TB disease. Urine-based testing
would have advantages over sputum-based testing because
urine is easy to collect and store, and lacks the infection control
risks associated with sputum collection. A low-cost point-
of-care urine assay for lipoarabinomannan (LAM) used for
screening patients prior to antiretroviral therapy (ART) rapidly
diagnoses a proportion of tuberculosis (TB) cases.
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Extra Edge
•• Phosphatidylinositol mannoside (PIM) and
lipoarabinomannan (LAM) are two lipid antigens presented
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by CD1b. LAM and PIM share a core region, where the
mannoses linked to PI are the stimulatory antigens.
•• Ethambutol is bacteriostatic against M. tuberculosis. Its
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primary mechanism of action is the inhibition of the
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arabinosyl transferases involved in cell wall synthesis, which
probably inhibits the formation of arabinogalactan and
lipoarabinomannan (LAM)
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•• LAM -B is a major antigen of M laprae which is distinct
from LAM of M tuberculosis.
2. Ans. (b)  HBV
Ref: Jawetz 24th ed page 496, 500
MICROBIOLOGY  •  Answers with Explanations 193
Two genera of viruses have Reverse transcriptase
1. Hepadna virus: Replicates through RNA intermediate. E.g.
Hepatitis B virus
2. Retrovirus: Replicates through DNA intermediate. E.G.
HIV virus
HBV Replication
The viral replication cycle are schematized. Blood borne
virion circulating through the liver attach to the basolateral
membranes through specific viral–host interactions. Pre-S1
protein on the envelope is believed to mediate viral attachment
to as yet uncharacterized cellular membrane proteins. Once
attached, the virus fuses with membrane protein in a slow
temperature dependent reaction. Upon cellular entry, viral core
is transported from the cytoplasm towards or into the nucleus
while the DNA within the capsid undergoes maturation. The
partially double stranded relaxed circular genome in the
capsid is converted into a covalently closed; fully double
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stranded circular DNA (ccc DNA). The maturation of the
genome is not dependent on any viral products but requires
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host machinery to remove covalently bound protein and RNA
primers and complete double stranded synthesis. The ccc DNA
amplifies viral infection, as it is the template for pregenomic
RNA production. It also serves as the template for production
of all the RNA species for the production of functional and
structural viral protein.
Viral assembly is a complex multi-step reaction in which
specific interactions between pregenomic RNA, P protein, host
chaperonins and viral core proteins result in capsid formation.
Once encapsidation occurs, the pregenomic RNA undergoes
reverse transcription with P protein. Reverse transcription
occurs within the capsid, which can recycle to the nucleus
or bud into the endoplasmic reticulum to acquire the
glycoprotein envelope. Enveloped virions are secreted through
the constitutive pathway of vascular transport.
HIV Replication
The seven stages of the HIV life cycle are: 1) binding, 2) fusion,
3) reverse transcription, 4) integration, 5) replication, 6)
assembly, and 7) budding.
MICROBIOLOGY
194 Section I  •  Subject-wise MCQs and Answers with Explanations

The HIV Life Cycle

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AIIMS (Nov 2017–May 2014)

 MICROBIOLOGY  •  Answers with Explanations 195

3. Ans. (b)  Balantidium coli Option (D)

Ref: Paniker Parasitology 7th ed page 107, CDC
See PLATE 13 B
The picture shown is of Trophozoite of Balantidium coli. Note
the cilli all around and the Bean/ Kidney shaped macronucleus.
Both Cyst and Trophozoite are seen in the Stool
4. Ans. (c)  Gene detection by PCR
Ref: Jawetz 24th ed page 187, Panikar 10th ed page 270, Harrison
19th ed page 857
Summary
Option (A)
Rapid, can differentiate toxigenic and nontoxigenic strain,
most used test practically
Option(B)
Slow, can not differentiate toxigenic and nontoxigenic strain,
Colonoscopy is positive only when pseudomembrane develops,
hence it is highly specific; but insensitive compared with other
tests. Negative result does not rule out C. difficile
Administration of antibiotics results in proliferation of
drug-resistant C difficile that produces two toxins. Toxin A, a
potent enterotoxin that also has some cytotoxic activity, binds
to the brush border membranes of the gut at receptor sites.
Toxin B is a potent cytotoxin. Both toxins are found in the
stools of patients with pseudomembranous colitis
The diagnosis of CDI is based on a combination of clinical
criteria:
•• Diarrhea (>3 unformed stools per 24 h for >2 days) with no
other recognized cause plus
•• Toxin A or B detected in the stool (by ELISA) or toxin
producing C. difficile detected in the stool by PCR or culture
or pseudomembranes seen in the colon.
Harrison’s 19th Ed says
Despite the array of tests available for C. difficile and its
toxins, no single traditional test has high sensitivity, high
specificity and rapid turnaround. Most laboratory tests for

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most sensitive if we already know its a toxic strain. Not of
much practical use toxins, including enzyme immunoassays, lack sensitivity.
However, testing of multiple additional stool specimens is not
Option (C)
Rapid, can differentiate toxigenic and nontoxigenic strain.
Newly approved for clinical testing, but appears to be more
Ea recommended. Nucleic acid amplification tests, including
PCR assays, have now been approved for diagnostic purposes
and appear to be both rapid and sensitive while retaining high
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sensitive than enzyme immunoassay toxin testing and at least specificity
as specific
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Relative Sensitivity and Specificity of Diagnostic Tests for Clostridium Difficile Infection
Type of test Relative Relative Comment
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sensitivitya specificitya
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Stool culture for C. difficile ++++ +++ Most sensitive test; specificity of + + + + if the C. difficile isolate
tests positive for toxin; with clinical data, is diagnostic of CDI. Can
not differentiate toxigenic from non toxigenic strain.
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turn around time too slow for practical use

Cell culture cytotoxin test +++ ++++ With clinical data, is diagnostic of CDI; highly specific but not as
on stool sensitive as stool culture; slow turnaround time
Enzyme immunoassay for + + to + + + +++ With clinical data, is diagnostic of CDI; rapid results, but not
toxin A or toxins A and B as sensitive as stool culture or cell culture cytotoxin test. Can
in stool differentiate toxigenic vs nontoxigenic strain.
Enzyme immunoassay for + + + to + + + + + + + Detects glutamate dehydrogenase found in toxigenic and
C. difficile common antigen nontoxigenic strains of C. difficile and other stool organisms; more
in stool sensitive and less specific than enzyme immunoassay for toxins;
rapid results
PCR for C. difficile toxin B ++++ ++++ Detects toxigenic C. difficile in stool; newly approved for
gene in stool clinical testing, but appears to be more sensitive than enzyme
immunoassay toxin testing and at least as specific
Colonoscopy or + ++++ Highly specific if pseudomembranes are seen; insensitive
sigmoidoscopy compared with other tests
aAccording to both clinical and test-based criteria.
Note: + + + +, >90%; + + +, 71–90%; + +, 51–70%; +, 50%

MICROBIOLOGY
196 Section I  •  Subject-wise MCQs and Answers with Explanations

5. Ans. (b)  Leptospira

Ref: Anantnarayan 9th ed page 377, Jawetz 24th ed ch 25
The spirochetes (Treponema, Borrelia, Leptospira) are long, for dimorphic fungi; HB PSC (use HB
slender, helically coiled, spiral or corkscrew-shaped, gram- pencil in PSC exam)
negative bacilli. They has an outer sheath or glycosaminoglycan •• Histoplasma capsulatum
coating. Inside the sheath is the outer membrane, which •• Blastomycoses sermatidis
contains peptidoglycan and maintains the structural integrity •• Paracoccidiodes brasiliense
of the organisms. Endoflagella (axial filaments) are the •• Penicillium marneffei
flagella-like organelles in the periplasmic space encased by •• Sporothrix schenkii
the outer membrane. The endoflagella begin at each end of the •• Coccidioides immitis
organism and wind around it, extending to and overlapping at •• Candida albicans
the midpoint. Inside the endoflagella is the inner membrane
(cytoplasmic membrane) that provides osmotic stability and 7. Ans. (b)  Sporotrichosis
covers the protoplasmic cylinder. Ref: Jawetz 24th ed page 630-632, Ananthanarayanan 8th ed
page 609
Similar Question was asked in AIIMS NOVEMBER 2012
Microbiology without Colour Plate

See MICRO PLATE 12

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8. Ans. (b)  Chromoblastomycosis

Ea Ref: Ananthanarayan Panikar 10th ed page 605

See MICRO PLATE 13

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Hints in Question
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•• Thorn Prick signifies “Traumatic inoculation” of Fungus.

The fungi that cause subcutaneous mycoses normally reside
in soil or on vegetation. They enter the skin or subcutaneous
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tissue by traumatic inoculation with contaminated material.

All 4 options are Subcutaneous mycosis and can enter
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6. Ans. (c)  Pneumocystis jiroveci through Traumatic Inoculation

Ref: Ananthnarayan 9th Ed Page 593 •• Superficial mycoses is frequently seen in Sub-Himalayan
region including Himachal Pradesh.
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Dimorphic fungi: Fungi that have two growth forms, such as

•• Out of all Subcutaneous mycoses, Verrucous lesion is typical
a mold and a yeast. All dimorphic fungi are yeasts in humans
of Chromoblastomycosis.
(molds in dirt). Human infection is by spore inhalation, so no
Chromoblastomycosis (chromomycosis) is a subcutaneous
Person-Person transmission
mycotic infection caused by traumatic inoculation by any of
five recognized fungal agents that reside in soil and vegetation.

Sporothrix schenckii Round or cigar Branched septate hyphae w/ oval conidia Subcutaneous Sporotrichosis
shaped budding at tip of conidiophores (“daisies”) (gardener’s disease)
Yeast
Phialophora ver- None Brown septate hyphae Subcutaneous Chromoblastomycosis
rucosa
Fonsecaea pedrosoi None Brown septate hyphae and conidia Subcutaneous Chromoblastomycosis
Pseudallescheria None V-shaped septate hyphae with radiating Subcutaneous Mycetoma
boydii chains of conidia
Madurella myce- None Brown septate branched hyphae Subcutaneous Mycetoma
tomatis
Exophiala Bipolaris None Brown septate hyphae Subcutaneous Phaeohyphomycosis
Exserohilum None Brown septate hyphae Subcutaneous Phaeohyphomycosis

AIIMS (Nov 2017–May 2014)


“R. seeberi had first been regarded as a sporozoon by Malbran, its
discoverer, in 1892, as a protozoan by Seeber who first published
a description of the pathogen and then, as a phycomycete by
Ashworth in 1923. Through molecular biological analysis of the
organism’s ribosomal DNA, Herr et al. classified the organism
in a new clade which was named the Mesomycetozoa, which
includes fish and amphibian pathogens in the former DRIP
clade (Dermocystidium, the rossette agent, Ichthyophonus
and Psorospermium). It is of interest that the histopathology
of these fish and amphibian diseases closely resembles that of
rhinosporidiosis. In addition, morphological similarities were
noted between R. seeberi and these pathogens. Indeed it was
speculated by Herr et al. that some of these pathogens could be
classified in the genus Rhinosporidium with the suggestion that
Rhinosporidium is a monotypic genus. An independent group of
workers supported this conclusion concerning taxonomy, in that
their analysis of R. seeberi 18S rRNA from infected tissue showed
that this organism is a protist “from a novel clade of parasites
that infect fish and amphibians”. These studies finally resolve the
debate on the taxonomy of R. seeberi, particularly that it is not a
classic fungus “but rather the first known human pathogen from
the DRIPs clade, a novel clade of aquatic protistan parasites”.
20. Ans. (b)  E. histolytica
Ref: Paniker parasitology 6th ed page 24, Sternberg Pathology
5th ed page 1537
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See PARASITO PLATE 12 KEY
Entamoeba histolytica exists in three morphological forms
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Trophozoite, Precyst and Cyst. Tetranucleated cyst is the
infective form and it divides into 8 uninucleated cysts as shown
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in the picture.
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21. Ans. (d)  Necator americanus
Ref: Parasitology: A Conceptual Approach by Eric Loker, Bruce
Hofkin- Page 48, CDC
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See PARASITO PLATE 15
Recall Bias........................................................
Two hookworms Nectar americanus and Ancylostoma duodenale eggs are
similar and cannot be differentiate. Ascaris Lumbricoides was the option
not Ancylostoma duodenale. Hence the answer will be Necator Americanus
Note
Necator Americanus egg is non Bile stained. But
the picture in the exam was yellowish due to iodine
mount (in iodine wet mounts, both bile-stained
and nonbile-stained helminthic ova are stained
brown) and hence many students got confused
and marked other options.
MICROBIOLOGY  •  Answers with Explanations 201
22. Ans. (b)  Fasciola hepatica
Ref: BRS Microbiology and Immunology 4th ed page 208,
Textbook of Parasitology by Chatterjee Page 187.
See PARASITO PLATE 16
23. Ans. (a)  Nocardia sp
Ref: Jawetz 24th ed chapter 15, BRS Microbiology 4th ed page 39-41
Indirect repeat Microbiology AIIMS Nov 2016 (Different
options, plus stain was also asked).
See MICRO PLATE 1 KEY
Nocardia
These bacteria are gram-positive, catalase-positive, and partially
acid-fast bacilli. Nocardiae form extensive branching substrates
and aerial filaments that fragment after formation, breaking
into coccobacillary cells. The cell walls contain mycolic acids
that are shorter chained than those of Mycobacteria. They are
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considered to be weakly acid-fast, but if they are stained with
the routine acid-fast reagent (carbol-fuchsin) but decolorized
with 1–4% sulfuric acid instead of the stronger acid-alcohol
Ea decolorant, most isolates will stain acid-fast. Gram-stained
smears reveal gram-positive bacilli, coccobacillary cells, and
branching filaments. With the modified acid-fast stain, most
isolates will be acid-fast.
24. Ans. (c)  Cryptococcus
Ref: Toxicology of 1 - 3-Beta-Glucans: Glucans as a Marker
for Fungal Exposure edited by Shih-Houng Young, Vincent
Castranova page 203, 204, J. Clin. Microbiol. July 2014 vol. 52
no. 7 2328-2333
Indirect repeat AIIMS NOV 2017 Microbiology (See for details
of 1- 3 β D glucan)
25. Ans. (a)  Heterophile antibody test
Ref: Ananthanarayan 9th ed 96-97, Textbook of Microbiology
& Immunology By Subhash Chandra Parija 2nd ed page 483,
Dorland’s Medical Dictionary 32nd ed page 1896
Patient with primary EBV infection develops IgM antibodies
that binds on antigen on RBC of other species particularly
sheep and horse but not guinea pig kidney cells. Heterophile
antibodies also occur in normal sera but these also binds to
Guinea pig kidney cells. This forms the basis of Heterophile
antibody test.
Paul–Bunnell test: Originally called as Heterophile Antibody
test was first described by Paul and Bunnell for demonstration
of heterophile antibodies in patients with infectious
mononucleosis in 1932. It determined the highest dilution of
the patient’s serum that was capable of agglutinating sheep red
blood cells by Hemagglutination. The Paul-Bunnell test is a
useful test to screen for the presence of heterophile antibodies
because it is simple and inexpensive.
MICROBIOLOGY
 Microbiology

MICRO PLATE 1

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A B C

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D E F
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G H I

MICRO PLATE 1 KEY

A. Gram positive (blue/purple) cocci (round) in clusters (grape like), C. Gram positive cocci in group of two (diplococcus/Lancet shaped),
for example, Staphylococcus although some very short chains may be seen: Streptococcus
B. Gram positive cocci in chains. For example, Streptococci Pneumoneae
D. Gram positive bacillus: Listeria, Cornybacterium

E. Gram positive bacillus in chain: Bacillus spp (Anthrax, cereus)
F. Gram negative cocci (Diplococci), insat shows intracellular Gram
negative diplococci and polymorphonuclear leukocytes in ure-
thral exudate; Neisseria sp., Moraxella catarrhalis, Acineto-
bacter, and Brucella.
G. Gram negative bacillus: E. coli, Pseudomonas, Haemophilius,
Klebsiella, Salmonella
MICROBIOLOGY  • Color Plates 1027
H. Nocardia Gram Stain: (Blue in pink background). Showing fila-
mentous, branching gram positive bacilli. (you must know how to
differentiate chains from filaments; chains are more beaded and
filaments are ore branched)
I. Nocardia, partially acid fast staining: (Pink in blue background).
Partial Acid-Fast staining employed: Carbol fuscin stain (3 min),
decolourize with 1% H2SO4 (until colour no longer comes off
~1 min) and counterstain with methyene blue (30 sec).

Gram and Acid-Fast Staining Methods

Most bacteria are classified as gram-positive or gram-negative according to their response to the Gram staining procedure. This procedure
was named for the histologist Hans Christian Gram, who developed this differential staining procedure in an attempt to stain bacteria
in infected tissues. The Gram stain depends on the ability of certain bacteria (the gram-positive bacteria) to retain a complex of crystal
violet (a purple dye) and iodine after a brief wash with alcohol or acetone. Gram-negative bacteria do not retain the dye-iodine complex
and become translucent, but they can then be counter stained with safranin (a red dye). Thus, gram-positive bacteria look purple under
the microscope, and gram-negative bacteria look red. The distinction between these two groups turns out to reflect fundamental
differences in their cell envelopes
Gram stain steps
•• Fix smear by heat. •• Wash with water. Do not blot.

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•• Cover with crystal violet. •• Cover for 10–30 seconds with safranin (2.5% solution in 95%
•• Wash with water. Do not blot. alcohol).
••
••
••
Cover with Gram’s iodine.
Wash with water. Do not blot.
Decolorize for 10–30 seconds with gentle agitation in acetone
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•• Wash with water and let dry.
ƒƒ School of fish appearance is seen in — H. ducrei
ƒƒ Fish in stream pattern is seen in — V. cholera
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(30 mL) and alcohol (70 mL).

Ziehl-Neelsen Acid-Fast Stain

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•• Fix smear by heat.

•• Cover with carbolfuchsin, steam gently for 5 minutes over direct flame (or for 20 minutes over a water bath).
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•• Wash with water.

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•• Decolorize in acid-alcohol until only a faint pink color remains.

•• Wash with water.
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•• Counterstain for 10–30 seconds with Loeffler’s methylene blue.

•• Wash with water and let dry.
Kinyoun carbolfuchsin acid-fast stain
•• Formula: 4 g basic fuchsin, 8 g phenol, 20 mL 95% alcohol, 100 mL distilled water.
•• Stain fixed smear for 3 minutes (no heat necessary) and continue as with Ziehl-Neelsen stain.
Acid Fast Organisms/Structures
Organisms All mycobacteria; M. tuberculosis, M. leprae, Atypical mycobacteria
Actinomycetes including Nocardia (week+) and Rhodococcus (Except Actinomyces and Streptomyces)
Legionella
Oocycts Cryptococcus parvum
Isospora belli
Cyclospora cayetanensis
Parasites Sarcocystis
Taenia saginata eggs (Taenis solium eggs do not stain well, can be used to diff)
Hydatid cyst especially hooklets
Others Bacterial spoers
Head of sperm

MICROBIOLOGY
1028 Section II  •  Subject-wise Color Plates

Classification of Bacteria
Listeria
Gram + Cornybacterium
Bacillus spp
E coli
Pseudomonas
Haemophilius
Klebsiella
Bacilli (Rods) Bordetella B
Yersinia
Gram – Pasteurella
Franciscella
AEROBES

Brucella
Salmonella
Proteus
Campylobacter
Actinobacter
Staphylococcus

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Gram + Streptococcus
C
Cocci Enterococcus
Neisseria

Filamentous
Gram –

Gram +
Moraxella
Nocardia
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Gram – Microthrix parvicella
Clostridia
Gram + Actinomyces (agar culture)
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Bacilli (Rods) Lactobacillus

Bacteriods D
Gram –
ANAEROBES

Fusobacterium
Streptococcus viridians
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Cocci
Gram + Peptococcus MICRO PLATE 2 KEY
Peptostreptococcus
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Gram – Veillonella
Microscopy of STD's
Gram + Actinomyces (broth culture)
Filamentous Trick: In genital ulcer smear, check if pathogen is intracellular or
Gram – – extracellular. If intracellular then check the lobes of the nucleus
(Monocytes or Neutrophils)
MICRO PLATE 2 A. Haemophilus ducreyi on gram stain appearance of ulcer shows
characteristic “Extracellular schools of fish” appearance.

= DUcreyi = Do You Cry = painful ulcer

Haemophilus ducreyi causes chancroid (soft chancre), a sexually
transmitted disease. Chancroid consists of a ragged ulcer on the
genitalia, with marked swelling and tenderness. The regional
lymph nodes are enlarged and painful. The disease must be dif-
ferentiated from syphilis, herpes simplex infection, and lympho-
granuloma venereum.
The small gram-negative rods occur in strands in the lesions,
usually in association with other pyogenic microorganisms.
H. ducreyi requires X factor but not V factor. It is grown best
from scrapings of the ulcer base on chocolate agar containing 1%
A IsoVitaleX and vancomycin, 3 g/mL, and incubated in 10% CO2

AIIMS

at 33°C. There is no permanent immunity following chancroid
infection. Treatment with intramuscular ceftriaxone, oral tri-
methoprim-sulfamethoxazole, or oral erythromycin often results
in healing in 2 weeks.
B. Donovanosis (granuloma inguinale) causes genital ulceration.
The causative organism, calymmatobacterium granulomatis
reclassified as Klebsiella granulomatis. Donovan body (safety pin
like inclusion bodies inside the monocyte).
MICROBIOLOGY  • Color Plates 1029
MICRO PLATE 3 KEY
A. Actinomyces HandE stain: High magnification micrograph of
a sulfur granule formed by actinomyces in the mandible. His-
topathology—For most purposes, recognition is based on the
appearances of sulphur granules using the hematoxylin and eosin
(HandE) stain. These granules actually represent colonies of
A. israelii, a gram-positive, anaerobic filamentous bacteria.
B. Actinomyces Grams stain: Showing tangled mass of branching
filaments (black arrow), surrounded by a hypocellular artifactual
cleft (yellow arrow) surrounded by neutrophils and macrophages.

N ote
Specific fungal stains such as the methenamine silver and periodic
acid Schiff (PAS) stains are useful to ensure that fungi are present,
but are seldom helpful for specific diagnosis. Actinomycete

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C. Neisseria is an intracellular gram-negative diplococci seen with filaments also take up silver based stains
polymorphonuclear leukocytes (see for multiple lobes of the
nucleus) in urethral exudate.
D. Chlamydia is gram-negative, obligate intracellular parasites.
They must grow and reproduce within host cells. Intracytoplas-
Ea Actinomycosis versus Nocardia
Actinomycosis Nocardia (Aerobic)
mic basophilic inclusion containing clamps of elementary bodies
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of chlamydia-basophilic inclusions. •• Gram stain Gram +ve filamentous Gram +ve filamentous
branching branching
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MICRO PLATE 3 •• Acid Not AFB +ve weak (but N.

fastness madurae AFB –ve)
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•• Morpho •• Non-motile •• Non-motile

•• Non-sporing •• Non-sporing
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•• Non-capsulated •• Non-capsulated
•• Infection in In immunocompitant In immunocompro-
mised (HIV/AIDS)
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•• Clinical •• Oro-cervicofacial •• Airborn inhalation

features (MC type)- woody/ –thick sputum
lump jaw •• CXR-Lt. lower
•• Appendix in GIT lobe nodule with
•• PID in IUCD users central cavitation.
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(A. israelii)
•• Microscopy Spidery colony and Paraffin bait
Sun-ray appearance- technique
Ray fungus
Sulphur granules
•• Treatment Penicillin TMP-SMX/
Sulfonamides

B
By Nephron - Own work, CC BY-SA 3.0,
https://commons.wikimedia.org/w/index.php?curid=18555026

MICROBIOLOGY