Bacteriology 4th Practical

MFSH MFSH 20032003

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Staphylococcus & StreptococcusStaphylococcus & StreptococcusStation 1: Staphylococcus VS. Streptococcus 3Staphylococcus (aureus & epidermidis)Staphylococcus (aureus & epidermidis) 44Station 2: Gram stain of Staphylococcus 5Station 3: Gram stain of staphylococcus (pus) 6Station 4: Staphylococcus growth on blood agar 7Station 5: Deoxyribonulease test 8Station 6: Mannitol sugar fermentation test 9Station 7: Coagulase test 10Station 8: Antibiotic sensitivity test, treatment & diseases 11Station 9: Summary and revision 12StreptococcusStreptococcus 1313Station 10: Gram stain of Streptococcus 14Station 11: Gram stain of Streptococcus (pus) 15Station 12: Gram stain of Streptococcus pneumoniae 16Station 13: Gram Stain of Streptococcus pneumoniae 17Station 14: Capsule stain of Streptococcus pneumoniae 18Station 15: Alpha heamolytic Streptococci 19Station 16: Beta heamolytic Streptococci 20Station 17: Gama or non-heamolytic Streptocooci 21Station 18: Optochin test 22Station 19: Bacitracin test 23Station 20: Capsule of Streptococcus Pyogens 24Station 21: Lancefield Grouping of beta heamolytic Streptococci 25Station 22: Treatment & diseases 26Station 23: Summary and revision 27

Staphylococcus StreptococcusSaline SalineH2O2 H2O2H2O + O2 (bubbles) H2O2

1) Bring 2 slides

2) Add saline

3) Put some of the bacteria

4) Add H2O2

5) See the difference

6) Take a look on the movie “Staph Vs Strept” included in the cd

Station 1Station 1

Station 2Station 2

Station 3Station 3

StaphylococciStaphylococci

Pus cellsPus cells

Station 4Station 4

اللون يكون اللون المفروض يكون المفروضللصفار للصفار مائل مائل

“ وقتها ” تأخذ لم لكن “ و وقتها ” تأخذ لم لكن و

يكون يكون المفروض المفروض F بياضا أكثر F اللون بياضا أكثر اللون

. الموضح .من الموضح من

تأخذ ” لم لكن تأخذ ” و لم لكن ووقتها“ وقتها“

Station 5Station 5

Station 6Station 6 MSA = Mannitol sugar agar

Color not changedColor not changedColor changed to Color changed to yellowyellow

Station 7Station 7

Liquid state Coagulated state

Station 8Station 8

Not all the members of the same Not all the members of the same species share all properties species share all properties together. For example, not all together. For example, not all staphylococcus aureus are staphylococcus aureus are sensitive to the same antibiotics. sensitive to the same antibiotics. (some of them are resistant to (some of them are resistant to penicillin and others are not).penicillin and others are not).

Bacteria

Control

When the distance between the control and the antibiotic is shorter than or equal to the distance between the antibiotic and the bacteria.

A =< B

The bacteria is called sensitive “to that anti-biotic”

When the distance between the control and the antibiotic is longer than the distance between the antibiotic and the bacteria.

A > B

The bacteria is called resistant “to that anti-biotic”

Sensitive

Sensitive Resistant

Sensitive

Some “tribes” of Staphylococcus areus are sensitive to Penicillin (also Cloxacilin, Methicillin…) while others are not.

Staph.aureus

Infections:-

1- Pyogenic infections and abscesses

2- Osteomylitis

3- Food poisoning “milk products” vomiting

4- Toxic shock syndrome in young women

5- SSSS “Staphylococcus Scalded Skin Syndrome”

Treatment:-

Drug of choice “Cloxacilin”

if MRSA, (beta-lactamase producers) then we use “Vancomycin”

Staph.epidermidis & Staph.saprophyticus

Infections:-

Epidermidis endocarditis with patients with catheters, cannulae & other artificial objects.

Saprophytics Urinary tract infections in young female

Treatment:-

“Vancomycin”

Station 9Station 9

Gram stainGram stain

Color on Blood Color on Blood agaragar

Mannitol Mannitol

fermentation testfermentation test

DNAse testDNAse test

Coagulase testCoagulase test

CatalaseCatalase

+ve +ve

Yellowish Whitish

+ve “yellow” -ve “red”

+ve -ve

+ve -ve +ve +ve

Station Station 1010

Station Station 1111

Station Station 1212

Station Station 1313

Capsule

(transparent)

Organism (red)

Station Station 1414

Streptococcus can be divided upon their Streptococcus can be divided upon their hemolytic activity into 3 groups:hemolytic activity into 3 groups:

1)1)αα Hemolytic (e.g. Streptococcus Hemolytic (e.g. Streptococcus pneumoniae)pneumoniae)

2)2)ββ Hemolytic (e.g. Streptococcus pyogenes) Hemolytic (e.g. Streptococcus pyogenes)

““These are also grouped by the These are also grouped by the LACEFIELDLACEFIELD grouping {from A-H, K-W} as you will see grouping {from A-H, K-W} as you will see later” later”

3)3)γγ or non Hemolytic (e.g. Streptococcus or non Hemolytic (e.g. Streptococcus mutans)mutans)

Station Station 1515

Station Station 1616

Station Station 1717

No colonies No colonies around the around the OPTOCHIN OPTOCHIN

There are There are colonies around colonies around the OPTOCHIN the OPTOCHIN

Station Station 1818

There will be some There will be some colonies around the colonies around the BACITRACIN strip.BACITRACIN strip.

Station Station 1919

Upon the Lancefield grouping

Station Station 2020

Station Station 2121

Station Station 2222

Alpha hemolytic streptococci (pnemococcus)

Infection:-

acute pnemonia

sinusitis

otitis media

meningitis

Treatment:-

“Penecillin”

Vaccination:

By the Polysaccharide capsule

Beta hemolytic streptococci gr.A (pyogens)

Infection:-

pharyngitis (sore throat)

tonsilitis

Treatment:-

“Penecillin”

Some notes:Some notes:

•All Staph. :- All Staph. :- VancomycinVancomycin But in But in non MRSAnon MRSA:- :- CloxacilinCloxacilin is better is better

•All Strept :- All Strept :- PenecilinPenecilin

Gram stainGram stain

Color on Blood agarColor on Blood agar

Color on ChocolateColor on Chocolate

Hemolytic activityHemolytic activity

Lancefiel GroupingLancefiel Grouping

Optochin testOptochin test

Bacitracin testBacitracin test

CatalaseCatalase

+ve +ve +ve Green Clear No Chg. Green No Chg. No Chg. Alpha Beta Non

+ve -ve -ve -ve +ve -ve -ve -ve -ve

Station Station 2323

NO Gp.A NO

يا يا دعواتكم دعواتكم

شبابشباب

A word can change the outcome of A word can change the outcome of

many things. So, if you either liked or many things. So, if you either liked or

didn’t like this project, why keep your didn’t like this project, why keep your

thoughts to yourself ?thoughts to yourself ?

Send us your comments and ideas Send us your comments and ideas

to to COORDINATOR1422@YAHOO.COM, It will

mean too much to us, “even if it was just “even if it was just

one word”one word”

THANXTHANX

M. F. ShaheenM. F. Shaheen 2003 2003