VMC 311: Veterinary Bacteriology Compiled by Dr.

Gaurav Singhal

Staphylococcus (from the Greek: staphyle, "bunch of grapes" and kokkos, "granule") is a genus of
Gram-positive bacteria. Under the microscope they appear round (cocci) or oval, and form grape-like
clusters. They are Gram positive, non motile, non spore producing and do not form capsules. They are
aerobic and facultative, liquefy Gelatin and also ferment carbohydrates to acid. Most of the
Staphylococcus are harmless and reside normally on the skin and mucous membranes of humans and
other organisms and in soil. They are found in suppurative conditions where they produce variety of
toxic substances.

History

• Micrococci were discovered by Ogston in pus in 1881 and he divided them into Streptococci
and Staphylococci.
• Rosenbach in 1884 grew them on artificial media and differentiated into two species: S.
pyogenes aureus and S. pyogenes albus.
• Passet in 1885 discovered the third species which he named S. pyogenes citreus.
• Gordon in 1905 concluded that S. pyogenes aureus is the golden form, S. pyogenes albus is the
non pigmented variety of golden form and S. pyogenes citreus is the intermediate one.
• Classification by Shaw, Stitt and Cowan in 1951:

Group I: Staphylococcus; Group II: Micrococcus

Staphylococcus Toxins:

S. aureus is important because of the variety of toxins it produces causing reactions in the tissues of
host.

1 Coagulase: Some Staphylococci produces coagulase enzyme that coagulate plasma and
causes blood clot formation which help S. to multiply and protect them from immediate
phagocytosis. Rabbit and human plasma are mostly readily coagulated. Coagulase enzyme is
resistant to heat and its antigenicity is doubtful. The active principle in coagulase is
precipitated by alcohol, acetic acid or half saturation with ammonium sulfate.

The main classification of staphylococci is based on their ability to produce coagulase enzyme:

Coagulase-positive

• S. aureus is coagulase-positive, meaning that they can produce coagulase enzyme. However,
while the majority of S. aureus are coagulase-positive, some may be atypical in that they do not
produce coagulase. S aureus is also catalase-positive (meaning that it can produce the enzyme
catalase) and able to convert hydrogen peroxide (H2O2) to water and oxygen, which makes the
catalase test useful to distinguish Staphylococci from Enterococci and Streptococci.
• S. pseudintermedius inhabits and sometimes infects the skin of domestic dogs and cats.

Coagulase-negative

1
VMC 311: Veterinary Bacteriology Compiled by Dr. Gaurav Singhal

• S. epidermidis, a coagulase-negative staphylococcus species, is a commensal of the skin, but

can cause severe infections in immune-suppressed patients.
• S. saprophyticus, another coagulase-negative species that is part of the normal vaginal flora, is
predominantly implicated in genitourinary tract infections in sexually-active young women.

2. Fibrinolysin: Many S. produces a substance fibrinolysin which lyses the fibrin and hence
interferes with clot formation. This helps in the spread of S. It is non antigenic and precipitated
by alcohol and acetone. Fibrinolysis is inhibited by beta-hemolysin.
3. Leucocidin: this toxin was described by Van de Velde in 1874. It helps S. to kill leucocytes
and hence it also helps in its spread. It has little or no effect on erythrocytes. It is antigenic.
4. Hyaluronidase: It is also called a spreading factor. The exact role of this substance in S.
infection is not known. This enzyme is normally responsible for the degradation of Hyaluronic
acid and as such it is assumed that the substance is most active during early stages of S.
infection.
5. Dermonecrotoxin: This toxin causes necrosis of skin. It occurs in natural infections in the
pustular dermatitis of the dog.
6. Lethal toxin: this toxin is responsible for the lethal effects in rabbits, horses, cats, guinea pigs
and mice. The animal becomes unsteady along with paralysis of hind legs, the respiration
becomes irregular and gasping, in-coordination of movements, pupils of the eye are initially
contracted followed by dilatation. This is followed by violent convulsions and death.
7. Hemotoxin: S. produces hemolyses of erythrocytes on blood agar plates. There are three
antigenically distinct hemolysins – Alpha Hemolysins, Beta hemolysins and Gama hemolysins.
The S. producing Alpha and Beta hemolysins form double-zone effect on the agar plate. The
Alpha hemolysin produces a clear zone surrounding the colony while the Beta hemolysin
produces a wider hemolytic zone. This double zone hemolysis is augmented by incubating the
colonies at 37O C initially followed by incubation at 10O C. hence it is termed as “hot-cold”
hemolysis.
8. Enterotoxin: Some strains of S. produces enterotoxin responsible for causing gastroenteritis
leading to food poisoning (Toxic Shock Syndrome). The most common symptoms are nausea,
dizziness, prostration, severe abdominal pain and diarrhea. The organisms are isolated from the
udder of cow, bakery products, raw milk, cream and ice-cream. Enterotoxin is not destroyed by
100O C for 30 minutes. N/100 NaOH and N/100 HCl destroy this toxin. The “Kitten test” is
used for the experimental studies of this substance. All enterotoxin producing strains are
hemolytic and coagulase positive but not vice versa.

Biochemical identification

Staphylococcus species can be differentiated from other aerobic and facultative anaerobic gram
positive cocci by several simple tests. Staphylococcus species are facultative anaerobes. Facultative
anaerobes are capable of growth both aerobically and anaerobically. All species grow in the presence
of bile salts and are catalase positive. Growth also occurs in a 6.5% NaCl solution. On Baird Parker
Medium Staphylococcus spp. are fermentative, except for S. saprophyticus which is oxidative.
Staphylococcus species are resistant to Bacitracin and susceptible to Furazolidone.

2
VMC 311: Veterinary Bacteriology Compiled by Dr. Gaurav Singhal

Staphylococcus can cause a wide variety of infections in humans and other animals through either
toxin production or invasion. Staphylococcal toxins are a common cause of food poisoning, as it can
grow in improperly-stored food.

Staphylococcus aureus ("golden cluster seed" or "the seed gold" and also known as golden staph)

It is the most common cause of staph infections. It is a spherical bacterium, frequently part of the skin
flora found in the nose and on skin. About 20% of the populations are long-term carriers of S. aureus.
S. aureus was discovered in Aberdeen, Scotland in 1880 by the surgeon Sir Alexander Ogston in pus
from surgical abscesses. It was first cultivated by Rosenbach in 1884.

Distribution and transmission: It is distributed through out the world and found on skin and mucous
membranes of man and animals. It is present in animal products such as eggs, milk and meat. These
bacteria can survive on dry surfaces, increasing the chance of transmission. Infected persons, who
handle the food, help in the transmission of S. aureus. Like streptococcus, S. is also an opportunist
organism and waits for suitable conditions for invasion.

S. aureus infections can be spread through contact with pus from an infected wound, skin-to-skin
contact with an infected person by producing hyaluronidase that destroy tissues, and contact with
objects such as towels, sheets, clothing, or athletic equipment used by an infected person.

Morphology and Staining: S. aureus is a facultatively anaerobic, Gram-positive coccus, which

appears as grape-like clusters when viewed through a microscope and has large, round, golden-yellow
colonies, often with hemolysis, when grown on blood agar plates. The golden appearance is the
etymological root of the bacteria's name “aureus” which means "golden" in Latin. It does not produce
spores, capsules and flagella.

Growth Requirements and Characteristics: It grows well in pus, all common laboratory media,
blood and serum. The optimum temperature for growth is 37O C and the optimum pH is 7.2. S. aureus
is catalase positive (meaning that it can produce the enzyme "catalase") and able to convert hydrogen
peroxide (H2O2) to water and oxygen, which makes the catalase test useful to distinguish staphylococci
from enterococci and streptococci. A small percentage of S. aureus can be differentiated from most
other staphylococci by the coagulase test: S. aureus is primarily coagulase-positive (meaning that it
can produce "coagulase", a protein product, which is an enzyme) that causes clot formation while most
other Staphylococcus species are coagulase-negative.

Colonies on agar plates are round, smooth, glistening, opaque, convex, amorphous, entire edge and of
a golden yellow color. The growth is butyrous in consistency. A uniform turbidity with powdery
sediment is produced in broth. Gelatin liquefaction is saccate in type.

Resistance: They are the most resistant of all cocci. A temperature of 60O C for half an hour kills
them. They are resistant to chemicals but killed in 1% phenol in 15 minutes and 10% formaldehyde in
10 minutes. They remain alive in dried pus for months. They are sensitive to many antibiotics. It can
survive on domesticated animals such as dogs, cats and horses, and can cause bumblefoot in chickens.
S. aureus can infect other tissues when barriers have been breached (e.g., skin or mucosal lining). This
leads to furuncles (boils) and carbuncles (a collection of furuncles). Penicillin is the standard antibiotic
3
VMC 311: Veterinary Bacteriology Compiled by Dr. Gaurav Singhal

used in the treatment of S. With continuous use, S. are getting resistant to Penicillin. As such, first-line
therapy is most commonly penicillinase-resistant penicillin (for example, oxacillin or flucloxacillin).

Biochemical Properties: S. aureus produces acid from glucose, maltose, mannitol, lactose, sucrose
and glycerol. It acidifies and coagulates litmus milk and produces hemolysis on blood agar. It is indol
negative, ammonia positive, methyl red positive, reduces nitrates to nitrites, hydrolyse gelatin and
coagulate serum.

Antigenic structure: S. possesses carbohydrate substance which is antigenic. Agglutination tests have
been used to distinguish strains.

Pathogenicity: S. aureus is associated with suppurative wound infection in man and animals. It can
cause a range of illnesses from minor skin infections, such as pimples, impetigo (may also be caused
by Streptococcus pyogenes), boils (furuncles), cellulitis folliculitis, carbuncles, sinusitis, tonsillitis,
ulcerative keratitis, scalded skin syndrome (a cutaneous reaction to exotoxin absorbed into the
bloodstream) and abscesses, to life-threatening diseases such as pneumonia, meningitis, osteomyelitis,
endocarditis, toxic shock syndrome (TSS) and septicemia. Its incidence is from skin, soft tissue,
respiratory, bone, joint, endovascular to wound infections. It is still one of the four most common
causes of nasocomial infections, often causing postsurgical wound infections.

S. aureus is one of the causal agents of mastitis in dairy cows. Its large capsule protects the organism
from attack by the cow's immunological defenses. It also causes pustular dermatitis in dogs and
abscess formation in all animals including fowls. It is the secondary invader in small pox, diphtheria,
septic sore throat, scarlet fever, TB and pneumonia. The severity of the infections in man and animals
is determined by the toxins produced by S. aureus. Wound infections and boils are produced by
dermonecrotoxin and leucocidin. Enterotoxin causes gastroenteritis and food poisoning.

Immunity: It is not produced to subsequent attacks. There is a temporary rise in the phagocytic
activity due to opsonin but this is soon lost. Most satisfactory immunity would be produced by toxoids
and antitoxins for various toxic substances. It may be used with success in cases of septicemia and
pustular dermatitis.

Diagnosis: Demonstration of S. on direct smears from a lesion (Gram-positive cocci in clusters), by

the production of hemolysin (Beta hemolysis), Catalase and Coagulase, production of enterotoxin
during food poisoning cases, Hotis test of milk sample (adherent greenish-brown colonies with white
centres after the milk sample has been incubated for 16-20 hrs.). For staphylococcal food poisoning,
phage typing can be to determine the source of infection.