A.

STATION: PICTURES

Penicillium Marnifeii- opportunistic mycoses

Fungus can be in the form of:

a) mold (hyphae structure)
b) yeast (budding structure)
Penicillium Marnifeii is a dimorphic organism (which means it can be in both mold and yeast forms)
Fungus can be stained with gram stained positive or by Indian ink look under light microscope
- in gram stain positive: appear as purple violet
Culture media for fungus: Sabbarouth Dextrose Agar (SDA)
Example of mold:
Example of yeast: Cryptococcus neoformans, Candida albicans
Who will get Candidiasis- immunocompromised person: HIV, elderly, uncontrolled DM, pregnant
lady, on steroid treatment, on radiotherapy or chemotherapy

Mycobacterium tuberculosis

Staining: ZiehlNeelsen stain

What to see: slender, acid fast bacilli

Culture media: Lowenstein-Jensen Medium

Clostridium perferinges

Clostridium perferinges has lecithinase activity (which is a toxigenicity property). It could be

demonstrated by Naglers reaction.
Naglers reaction in Clostridium perferinges: by using lactose egg yolk milk agar culture- with
antitoxin applied to the left half of the plate. So, the lecithinase activity will be inhibited (the area
appeared to be less opaque)

Corynebacterium diphteriae

Causes pharyngitis, with pseudomembranous characteristic.

Rotavirus
 What disease can it cause?
AGE
In what age group?
- Below 2 years old
How to prevent?
- Vaccination, by Rotarix

Herpes Simplex Virus

HSV is one of the herpes viruses.

What are the types of herpes virus?
The classification of Human Herpesviruses: (Medical microbiology, Jawets et al)
i. Human herpesvirus 1
Common name: Herpes simplex virus type 1
ii. Human herpesvirus 2
Common name: Herpes simplex virus type 2
iii. Human herpesvirus 3
Common name: Varicella-zoster virus
iv. Human herpesvirus 4
Common name: Epstein-Barr virus
v. Human herpesvirus 5
Common name: Cytomegalovirus
vi. Human herpesvirus 6
Common name: Human herpesvirus 6
vii. Human herpesvirus 7
Common name: Human herpesvirus 7
viii. Human herpesvirus 8
Common name: Kaposis sarcoma- associated herpesvirus

Herpes Simplex Virus: cytopathic effect (ballooning) on viral culture.

- To do viral culture, you need a cell (eg: monkeys cell, fibroblast cell). Introduce the virus into the
cell, the virus will grow inside it.

HSV clinical features(depend on types):

- cluster vesicles at border of lip (usually caused by HSV type 1)
- genital ulcer (usually caused by HSV type 2)

What are the diseases caused by varicella zoster virus?

- chicken pox
- shingles

What is shingles?
- It is caused by varicella zoster virus (VSV). The virus lives dormant in the dorsal root ganglion of
the spinal cord. Later in life, VSV can reactivate, causing vesicular eruption along the dermatomal
distribution.
 Who is at risk of getting shingles?
Immunocompromised patient- HIV, elderly, uncontrolled DM

Rubella
a virus, causing congenital infection
Maculopapular rash
to prevent rubella infection: vaccination (MMR) at 1 year old

Influenza

What are the types of influenza virus?

Type A, B and C.
Human can be infected by A and B. But the most common is by type A.
Genetically re- assortment
Avian flu (eg: H7N9)- usually would not affect human. Unless there is a direct contact and
intermediate host (eg: swine).

(http://www.niaid.nih.gov/topics/flu/understandingflu/pages/definitionsoverview.aspx)
Pigs can be infected by both bird (avian) influenza and the form of influenza that infects humans. In a
setting such as a farm where chickens, humans, and pigs live in close proximity, pigs act as an
influenza virus mixing bowl. If a pig is infected with avian and human flu simultaneously, the two
types of virus may exchange genes. Such a "reassorted" flu virus can sometimes spread from pigs to
people.

OTHER PICTURES:

- Malassezia furfur: hypopigmented lesions and skin biopsy

- Actinomycoses
- Histoplasma Capsulatum: on SDA fuzzy appearance mold colony
- Treponema pallidum
- Corynebacterium diphteriae
o Potassium Tellurite Agar- black
o Blood Agar- white and opaque
o Elek Test- to demonstrate toxin production by Corynebacterium diphteriae
- Rhabdovirus
- RSV
- Adenovirus
- Aspergillus fumigates: conidiophores
B. STATION: CULTURE MEDIUM

1. MacConkey agar
2. Blood agar
3. Chocolate ager
4. CLED
5. TCBS
6. Muller Hinton agar
7. Sabarouth Dextrose agar

1. MacConkey agar
culture growth for gram negative organism
lactose fermenter: turn to PINK
- Escherichia coli
- Klebsiella
non-lactose fermenter
- Salmonella typhii
- Pseudomonas aeruginosa

E.coli on MacConkey agar (lactose fermenterpink)

http://pharmicrosec3.blogspot.com/2011/04/urinary-tract-infections-urine-culture.html

Salmonella typhii on MacConkey agar (non-lactose fermentercolourless)

 http://www.cdc.gov/meningitis/lab-manual/chpt06-culture-id.html

2. Blood agar
Staphylococci
Staphylococcus aureus- golden, yellow colony
Streptococci
HAEMOLYSIS
a) (partial)
- Streptococcus pneumonia (SENSITIVE TO OPTOCIN)
- Streptococcus viridans (resistance to optocin)
b) (clear)
- Streptococcus pyogenes (SENTITIVE TO BACITRACIN)
c) gamma (non- haemolytic)
 Staph aureus on blood agar- golden yellow colony

http://abcnews.go.com/Health/mrsa-outbreak-hits-students-mexicos-belen-high-school/story?id=15338881
 Strep pneumonia on blood agar, sensitive to optocin

http://old.mgm.ufl.edu/~gulig/mmid/mmid-lab/labimage/imagky.html
 Strep pyogene on blood agar, beta haemolysis, sensitive to bacitracin

http://www.haloarchaea.com/teaching/molec_prac_course/classical.html

3. Chocolate agar
Haemophilus influenza microscopically: gram negative coccobacilli
Neisseria meningitides microscopically: gram negative diplococci
 Haemophilus influenza on chololate agar - greyish

http://www.microbiologyinpictures.com/bacteria%20photos/haemophilus%20influenzae%20photos/HAIN1.html
4. CLED (Cystine Lactose Electrolyte Deficient) agar
Escherichia coli yellow

E.coli on CLED agar- yellow colony

http://www.flickriver.com/photos/nathanreading/6141285541/

5. TCBS (Thiosulfate-Citrate-Bile salts-Sucrose) agar

Yellow colony: Vibrio cholera
Green colony: Vibrio parahaemolyticus

Vibrio cholera on TCBS agar- yellow colony

http://commons.wikimedia.org/wiki/File:Vibrio_cholerae_on_TCBS_agar.jpg
6. Mueller- Hinton agar
Haemophilus influenza requires both factors (X and V) to grow
 Haemophius ducreii only needs factor V
Parainfluenza only needs factor X.

XV V X
Haemophilus influenza Haemophilus ducreii Parainfluenza

Haemophilus influenza on Mueller- Hinton agar with factors X and V

http://t1.gstatic.com/images?q=tbn:ANd9GcSjKOq9iFJ54_k9EfdE0MA03QXRlhhn6yHfEHP9SuGJJFHa-RdG

7. Sabouraud Dextrose agar (SDA)

For fungal growth, and some other organisms. eg: for candida albicans, Cryptococcus neoformans

Candida albicans on SDA

http://thunderhouse4-yuri.blogspot.com/2009/12/candida-albicans.html

C. STATION: TSI (Triple Sugar Iron)

 Some idea on TSI:

http://biology.clc.uc.edu/fankhauser/labs/microbiology/Triple_Sugar_Iron/TSI_Use.htm

Triple Sugar Iron medium is a differential medium that can distinguish between a number of Gram-
negative enteric bacteria based on their physiological ability (or lack thereof) to:

a. metabolize lactose and/or sucrose

b. conduct fermentation to produce acid
c. produce gas during fermentation
d. generate H2S.

The medium contains 1.0% each of sucrose and lactose and 0.1% glucose. If only glucose is fermented,
acid produced in the butt will turn it yellow, but insufficient acid products are formed to affect the methyl
red in the slant. However, if either sucrose or lactose are fermented, sufficient fermentation products
will be formed to turn both the butt and the slant yellow. If gas is formed during the fermentation, it
will show in the butt either as bubbles or as cracking of the agar. If no fermentation occurs (as for an
obligate aerobe), the slant and butt will remain red. The medium also contains ferrous sulfate. If the
bacterium forms H2S, this chemical will react with the iron to form ferrous sulfide, which is seen as a
black precipitate in the butt (a black butt). The following table summarizes these reactions:

TRIPLE SUGAR
IRON FUNCTION RESULT/INTERPRETATION
INGREDIENTS
phenol red a pH indicator: Phenol red turns yellow in an acid environment. It
below 6.8 it is thus indicates whether the acids of fermentation
yellow have been produced. Failure to turn the butt
above 82., it is red yellow indicates that no fermentation has occured,
and that the bacterium is an obligate aerobe.
0.1 % glucose if only glucose is If only glucose is fermented, only enough acid is
fermented, only a produced to turn the butt yellow. The slant will
small amount of acid remain red.
is produced
1.0 % lactose if the culture can a large amount of acid turns both butt and slant
1.0% sucrose ferment either yellow, thus indicating the ability of the culture to
lactose (lac+) and/or ferment either lactose or sucrose
sucrose (suc+), a
large amount of acid
is produced
FeSO4 A source of iron and A few bacteria are capable of reducing the SO4=
(ferrous sulfate) sulfur to H2S (hydrogen sulfide).
The iron combines with the H2S to form FeS
(ferrous sulfide) a black compound. This will turn
the butt black. Thus, a black butt indicates H2S
production.

Lactose fermenter (SLANT YELLOW):

 - E. coli
- Klebsiella

E. coli Salmonella typhii Pseudomonas aeruginosa

Slant Yellow: Lactose Red: alkaline Red: alkali
fermenter
Gas Presence of gas (gas is Black (H2S- hydrogen No gas
formed during sulphide)
fermentation)
But Yellow Yellow: glucose fermenter Red: alkali

TSI

http://quizlet.com/19153953/familiarize/embedv2?&m

D. STATION: Robertson Cook Meat Medium

- for Clostridium perfringes (causing gas gangrene)

Clostridium perfringes in Robertson Cook Meat Medium

 catalog.hardydiagnostics.com

E. STATION: Microscope