Staphylococci

1. Staphylococci are spherical Gram-positive cocci.

2. They are arranged in irregular grape-like clusters, as they multiply in different planes.
3. All staphylococci produce catalase (streptococci do not produce catalase).
4. They are non-spore forming and non-motile.
5. Staphylococcus aureus produces coagulase (other staphylococci do not).
6. Staphylococcus aureus ferments mannitol & hemolyzed red blood cells.
7. True facultative anaerobe.
8. They are routinely cultured on enriched media containing broth and/or blood.
9. Some staphylococci produce ß-lactamase which are resistant to ß-lactam drugs such as penicillin
& cephalosporin .
10. Staphylococci rapidly develop resistance to many antimicrobial agents and present difficult
therapeutic problems.
11. Staphylococci are hardy, being resistant to heat and drying, and thus can persist for long periods
on fomites (inanimate objects), which can then serve as sources of infection.

Staphylococcus aureus
Same as Staphylococci

Staphylococcus epidermidis
1. Gram-positive cocci arranged in clusters.
2. Normal flora of skin and mucous membranes.
3. Staphylococcus epidermidis infections are almost always hospital-acquired (nosocomial).
4. It causes infections of intravenous catheter sites and prosthetic devices.
5. Most strains are aerobe and facultative anaerobe.
6. They grow in nutrient agar, blood agar.
7. They are highly antibiotic resistant (most strains produce beta lactamase).
8. Catalase positive but coagulase negative (also- phosphatase negative & DNAse negative).
9. It does not ferment mannitol.
10. Novobiocin sensitive.
11. Determinants of pathogenicity:
Glycocalyx: adheres to a variety of prosthetic devices (implant materials).
Highly resistant to antibiotics

Staphylococcus saprophyticus
1. Gram-positive cocci arranged in clusters.
2. Aerobe and facultative anaerobe.
3. They grow in nutrient agar, blood agar with formation of pigments.
4. Catalase positive but coagulase negative (also- phosphatase negative & DNA-ase negative).
5. They cannot ferment mannitol
6. Novobiocin resistant.
7. Causes community acquired infection.

Streptococcus agalactiae
1. Gram-positive cocci arranged in chains.
2. Aerobic and facultative anaerobes.
3. Grows in blood agar with clear zone of hemolysis (ie, B-hemolytic) and also grow in bile
containing media.
4. Habitat - They are present as vaginal commensal in 25% women. 5. It hydrolyses hippurate
 Streptococcus pyogenes
1. Parts of normal flora of skin & oropharynx.
2. Gram stain: Gram-positive cocci arranged in long chains or pairs.
3. Morphology:
a. Spherical in shape.
b. Non-motile, non-flagellated, non-spore bearing.
C. Pili present, so virulent.
d. Many strains have a hyaluronic acid capsule that is antiphagocytic.
4. Aerobic or facultative aerobic.
5. Catalase negative.
6. Grow in blood agar media with a clear zone of complete hemolysis.
7. The growth of S. pyogenes on agar plates in the laboratory is inhibited by the antibiotic bacitracin,
an important diagnostic criteria.

Streptococcus pneumoniae
1. Pneumococci are gram-positive lancet-shaped cocci arranged in pairs (diplococci) or short
chains. (The term "lancet-shaped" means that the diplococci are oval with somewhat pointed ends
rather than being round.)
2. Pneumococci possess polysaccharide capsules of more than 85 antigenically distinct types.
3. On blood agar they produce alpha-hemolysis (partial hemolysis of green-brown color).
4. They are lysed by bile or deoxycholate.
5. Optochin sensitive, ie, their growth is inhibited by optochin.
6. Aerobic and facultative anaerobe.
7. Growth is enhanced by 5-10% CO
8. Non-spore forming, non-flagellated and non-motile.

Neisseria
1.Morphology:
a. The typical neisseria is a gram-negative diplococcus.
b. Individual coccus is kidney-shaped, when the organisms occur in pairs, the flat or concave sides
are adjacent.
c. Nonmotile, non-spore-forming, and non-acid-fast.
2. Neisseria contain endotoxin in their outer membrane.
3. Growth Characteristics: The neisseriae grow best under aerobic conditions in an atmosphere
containing 5% CO2 (eg, candle jar).
4. Culture: In 48 hours on enriched media (eg, modified-Thayer Martin, New York City), gonococci
and meningococci form convex, glistening, elevated, mucoid colonies 1-5 mm in diameter. Colonies
are transparent or opaque, nonpigmented, and nonhemolytic.
5. Most neisseriae oxidize carbohydrates, producing acid but not gas.
6. Neisseriae are oxidase-positive, i.e., they possess the enzyme cytochrome c. This is an important
laboratory diagnostic test in which colonies exposed to phenylenediamine turn purple or black as a
result of oxidation of the reagent by the enzyme.
7. The organisms are rapidly killed by drying, sunlight, moist heat, and many disinfectants.
 Neisseria gonorrhoeae
1. Gram stain: Gram-negative diplococci (arranged in pairs).
2. Morphology:
a. They resemble paired kidney-shaped beans. (Each having a concave margin & a convex margin,
concave sides are adjacent.)
b. Non-flagellated, non-motile.
c. Possesses pili on the surface.
d. Non-capsulated.
3. Usually intracellular in neutrophils in lesions and extracellular in exudates.
4. Contain endotoxin in their outer membrane. Endotoxin of N. gonorrhoeae is lipo-oligosaccharide
(LOS) (Endotoxin of most of the Gram-negative bacteria is lipopolysaccharide (LPS).
5. Non-spore forming.
6. Non-acid fast.
7. More than 100 serotypes are known due to antigenic variation of pilus protein.

Neisseria meningitidis
#Morphology:
1. Gram negative, oval diplococci with flattened apposed surfaces (Plano-convex).
2. Arranged in pairs or may be tetrads.
3. Has a predominant polysaccharide capsule (it enhances virulence by anti-phagocytic action).
4. Have endotoxin composed of lipopolysaccharide (LPS).
5. Intracellular or extracellular.
6. Non-spore forming, non-flagellated and non-motile.
7. Non-acid fast.
8. It has 13 serotypes
#Properties of polysaccharide capsule of the meningococcus:
1. It enhances virulence by its antiphagocytic action.
2. It is the antigen that defines the serologic groups.
3. It is the antigen detected in the spinal fluid of patients with meningitis.
4. It is the antigen in the vaccine.
#Cultural characteristics of meningococcus:
1. Require enriched media for growth. Chocolate agar medium is satisfactory.
2. Optimum temperature: 37°C
3. Optimum pH: 7.4-7.6
4. Aerobic but growth facilitated by presence of 10% CO2
5. On solid media like blood agar, colonies are 1-2 mm in diameter, smooth, translucent and round
with regular edge without haemolysis.
6. In liquid media like serum broth, the growth produces slight turbidity with little granular deposit
at the bottom.

Corynebacterium
Morphology of C. diphtheriae:
1. Corynebacteria (from the Greek koryne, club) are gram-positive rods that appear club-shaped
(wider at one end).
2. The cells are arranged in singly, in pairs resembling 'V, 'L' and in-groups resembling "Chinese
letters".
3. The rods have a beaded appearance. The beads consist of metachromatic granules.
4. Show pleomorphism in microscopic and colonial morphology.
5. Have metachromatic granules. 6. Non-spore forming and non-motile.
Other properties:
1. Aerobic and facultative anaerobes.
2. Cultural characteristics of C. diphtheriae:
Grows best at 37°C.
Grows in media enriched with blood, serum or egg e.g. Loeffler's serum slope & blood tellurite agar.
Blood agar: Small, granular and gray colony, with irregular edges and small zones of hemolysis.
Blood tellurite agar: Brown to black colony with a brown-black halo because the tellurite is
reduced intracellularly.
3. Lysogenic conversion: Nontoxigenic diphtheria organisms undergo lysogenic conversion and
become lysogenic and toxigenic. These lysogenic strains produce diphtheria toxins and produce
pathogenicity.

Clostridium
1. Gram-positive bacilli and non-acid-fast.
2. Rod shaped with rounded ends.
3. Spore forming: Possesses spherical and terminal spores. Spores are larger than the bacillary body.
4. Obligate anaerobes.
5. Motile with peritrichous flagella.
6. Produce exotoxins.
7. Non-capsulated.

Clostridium tetani
1. C tetani is a Gram-positive rod, which forms spores readily in nature and in culture, yielding a
round terminal spore that gives the organism a "drumstick" or "tennis racket" appearance.
2. Non-capsulated and non-acid-fast.
3. Motile with peritrichous flagella (slow and steady movement).
4. Strict anaerobe
5. Produces a neurotoxic exotoxin, tetanospasmin or tetanus toxin, a metalloproteinase.

Clostridium botulinum
1. Gram-positive bacilli. 2. Non-capsulated. 3. Anaerobic and spore forming.
4. Spores are oval, sub-terminal and slightly bulging. 5. Motile with peritrichous flagella. 6. Produce
toxins (protein in nature)

Clostridium perfringens
1.Morphology:
a)Gram-positive bacilli. b)Non-motile c)Capsulated.
d)Oval, sub-terminal/central spore
2. Saccharolytic
3. It grows overnight under anaerobic conditions, producing hemolytic colonies on blood agar.
4. In broth containing fermentable carbohydrate, growth of C perfringens is accompanied by the
production of large amounts of hydrogen and carbon dioxide gas, which can also be produced in
necrotic tissues, hence the term gas gangrene.
5. Transmission: Spores are located in the soil; vegetative cells are members of the normal flora of
the colon and vagina. Gas gangrene is associated with war wounds, automobile and motorcycle
accidents, and septic abortions (endometritis)
 Bacillus anthracis
1. Gram positive, spore forming rods with square ends.
2. Rectangular in shape and arranged in very long chains (box car appearance).
3. Possess antiphagocytic capsule, which is composed of polypeptide (polymer of D-glutamate).
4. Non-motile, non-acid-fast, nonhemolytic.
5. Colonies are characterized by a rough, uneven surface with multiple curled extensions at the ed
resembling a "Medusa head."
6. B.anthracis endospores are usually oval, sub-terminal or central in position. They are extremely
hardy and survive in the environment for decades.
7. Secretes an exotoxin, called anthrax toxin, which is made up of three proteins: protective antigen
(PA). edema factor (EF), and lethal factor (LF).
8. Disease: B. anthracis causes anthrax, which is common in animals but rare in human.

Enterobacteriaceae
1. They are gram-negative rods, either motile with peritrichous flagella or nonmotile.
2. They are found primarily in the colon of humans and other animals, many as part of the normal
flora.
3.They are all facultative anaerobes.
4.They all ferment glucose (fermentation of other sugars varies).
5.None have cytochrome oxidase (i.e. they are oxidase negative).
6.They reduce nitrates to nitrites as part of their energy-generating processes.
7. They are catalase-positive.
8. They are capable of growing on ordinary media.

Escherichia coli
1.Gram staining property: E. coli is a straight gram-negative rod.
2. Morphology:
a. Motile with peritrichous flagella.
b. Most strains are fimbriated/piliated.
c. It has 3 antigens: O (cell wall antigen), H (flagellar antigen), K (capsular antigen).
3.Facultative anaerobe.
4.E. coli ferments lactose, (a property that distinguishes it from the two major intestinal pathogens,
Shigella and Salmonella.)
5. Catalase positive
6. Oxidase negative
7. Because there are more than 150 O, 50 H. and 90 K antigens, the various combinations result in
more than 1000 antigenic types of E. coli.
8. Specific serotypes are associated with certain diseases, e.g.O55 and O111 cause outbreaks of
neonatal diarrhea.

Salmonella
1. Salmonellae are gram-negative rods that do not ferment lactose but do produce H2S features that
are used in their laboratory identification.
2. They form colorless colonies on MacConkey agar media.
3. Most strains are motile with peritrichous flagella
4. Fimbriae present. 5. Non-spore forming 6. Catalase positive 7. Oxidase negative
8.Survive freezing in water for long periods.
9. Antigens: O antigens, H antigens, Vi antigen.
10. Salmonellae are resistant to certain chemicals (eg, brilliant green, sodium tetrathionate, sodium
deoxycholate) that inhibit other enteric bacteria.
 Shigella
1. Gram-negative rods.2. Non-lactose fermenter.3. Form colorless colonies on MacConkey agar
4. Fimbriae present (In case of Sh. flexneri).
5. Non-spore forming and non-capsulated.
6. Shigella infections are almost always limited to the GIT; bloodstream invasion is quite rare.
7. Culture: Shigellae are facultative anaerobes but grow best aerobically. Convex, circular,
transparent colonies with intact edges reach a diameter of about 2 mm in 24 hours.

Proteus
1. Gram-negative rods.
2.Lactose non-fermenter, oxidase-negative (do not have the enzyme 'cytochrome oxidase).
3. Distinguished from other members of the Enterobacteriaceae by their ability to produce the
enzyme phenylalanine deaminase.
4. Produce the enzyme 'urease', which cleaves urea to form NH, & CO₂ (character by fishy odour),
5. Certain species are very motile and produce a striking swarming effect on blood agar,
characterized by expanding rings (waves) of organisms over the surface of the agar.
6. The cell wall O antigens of certain strains of Proteus, such as OX-2, OX-19, and OX-K, cross-react
with antigens of several species of rickettsiae.

Klebsiella
1.Gram negative rods 2. Lactose fermenter.
3. It has a very large polysaccharide capsule, which gives its-colonies a striking mucoid appearance.
4. Frequently found in the large intestine but also present in soil and water.

Vibrio Cholerae
1. Vibrios are curved, comma-shaped Gram-negative rods.
2. Actively motile by means of a single polar flagellum.
3. In hanging drop preparation, they have a 'fish-in-stream' appearance.
4. Vibrios are oxidase-positive, which differentiates them from enteric gram-negative bacteria.
5. Non-spore forming and non-capsulated
6.Antigenic structures- O & H antigen

Helicobacter pylori
1. Curved Gram-negative rods. 2. Strongly urease-positive. 3. It has multiple flagella at one pole and
is actively motile.
4. Microaerophilic (ie, requires less O2 concentration for survival).
5. Oxidase positive, catalase positive.

Campylobacter jejuni
1. Campylobacters are gram-negative rods with comma, S or "gull wing"
2. They are motile with a single polar flagellum. 3.Do not form spores
4. They have lipopolysaccharides with endotoxic activity.
5. Habitat: Normal intestinal flora of many warm-blooded animals (e.g. cattle, chickens, dogs, birds
6. Culture: a. Selective media are needed, and incubation must be in an atmosphere with reduced O 2
( 5% O2 ) with added CO2 (10% CO2)
C. jejuni grows well at 42° C whereas Campylobacter intestinalis does not.
c. The colonies tend to be colorless or gray. They may be watery and spreading or round and convex,
and both colony types may appear on one agar plate.
7. Biochemical characteristics:
a.Oxidase- and catalase-positive.
b. Do not oxidize or ferment carbohydrates