Microbiology
Systematic microbiology
Staphylococcus
• Introduction:
• Staphylococci are Gram positive cocci arranged in,grape-like clusters. They are
the commonest cause of suppuration.
• Sir Alexander Ogston (1880) a Scottish surgeon, established the pathogenic
role of coccus in abscesses and other suppurative lesions. He gave the name
Staphy lococcus (Staphyle, meaning a bunch of grapes,kokos, meaning a berry)
from the typical arrangement of the cocci in grape like clusters.
• He also observed that staphylococcal strains from pyogenic lesions produce
golden yellow colonies while those from the normal skin produce white
colonies on solid culture media.
• Rosenbach (1884) named them Staph. aureus and [Link] respectively.
• Passel (1885) described a third strain of Staphylococcus, that produces lemon
yellow colour and named it as Staph. citreus . Since pigment production
Staphylococcus
A. Staphylococcus aureus
Staphy lococcus aureus is a pathogenic staphylococcus that causes pyogenic infections in
man.
B. Staphylococcus epidermidis Staphylococcus epidermidis is a skin commensal.
C. Staphylococcus saprophyticus
Staphylococcus saprophyticus acts as an opportunistic pathogen.
Pathogenic strains (e.g. Staph. aureus) have certain characteristics like production of
enzymes coagulase,phosphatase, deoxyribonuclease and their ability to ferment mannitol.
Based on production of enzyme coagulase, staphylococci are classified as coagulase
positive
and coagulase negative.
Most of the coagulase positive strains produce golden yellow colonies and are known as
Staph. aureus (also named as Staph. pyogenes). These strains are toxigenic.
The coagulase negative strains are generally non-pathogenic, non-toxigenic and form
white
colonies, these are called Staph. epidermidis (formerly known as Staph. albus).
STAPHYLOCOCCUS AUREUS
• A. Morphology
• They are Gram positive cocci arranged in grape-like clusters,
non-motile, non-sporing.
• Size: ~1 μm in diameter .
• They may also be present singly, in pairs, tetrads or short chains
of three or four cells especially when examined from liquid
culture.
• A few strains may possess microscopically visible capsules,
especially in young cultures.
• Many apparently non-capsulated strains also have some amount
of capsular material on the surface.
B. Culture
They grow readily on ordinary culture media within a temperature range of
10-42°C,
The optimal temperature : 37°C and
pH 7.4-7.6.
They are aerobes and facultative anaerobes.
Culture media:
1. Nutrient Agar
After overnight or 24 hours incubation, the colonies are 2-4 mm in diameter,
circular, smooth, convex, opaque and easily emulsifiable.
Most of the strains produce golden yellow pigment. The pigment is not
diffusable into the medium.
Pigment production is best seen at 22- 25°C and only in aerobic cultures.
Pigment production is enhanced by incorporation of 1 % glycerol monoacetate
or milk in the medium. The pigment is believed to be a carotenoid.
• 2. Blood Agar
• Colonies are similar to those on nutrient agar and in Addition beta type of
haemolysis is seen.
• Haemolysis is best observed with sheep or rabbit blood.
• 3. Mac Conkey's Agar
• Colonies are very small and pink due to lactose fermenter.
• 4. Selective Media .
• These media are used for isolating Staph. aureus from samples such as faeces
containing other bacteria.
• Staphylococci can tolerate 8- 10% NaCl, lithium chloride,tellurite and polymyxin.
Incorporation of these reagents into media make them selective.
• Examples of selective media include salt milk agar (containing 8-10% Nacl)
and Ludlam's medium ( containing lithium chloride and tellurite).
• 5. Mannitol Salt Agar
• This is both a selective and an indiiator medium. It contains nutrient agar with
1% mannitol, 7.5% sodium chloride and phenol red as indicator. Yellow
coloured colonies are seen on this medium due to fermentation on
• of mannitol) by most strains of Staph. aureus.
• Mannitol fermentation leads to production of acid and lowers the pH of
medium (phenol red produces yellow colour in acidic pH).
6. Milk Agar
• This medium is prepared by mixing sterile nutrient agar and
• Sterilzed milk.
• Colonies of Staph. larger thanthose on nutriept agar and pigment production
is enhanced.
• 7. Liquid Medium:
• Uniform turbity is produced in peptone water or nutrient broth.
• C. Biochemical Reactions
• Staph. aureus is catalase positive (unlike
streptococci)and oxidase negative.
• It breaks down carbohydrates by fermentation
whereas micrococci break down aerobically.
• This property can be tested on Hugh-Leifson
O/F medium.