G+ve Pyogenic cocci (Lecture 1) .

Gram-positive pyogenic cocci

The pyogenic cocci are spherical bacteria that cause various suppurative (pus-producing) infections. The major G + ve pathogens included are the following; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae. The major G-ve cocci include; Neisseria gonorrhoeae Neisseria meningitidis

Staphylococci
taphylococci are Gram-positive spherical bacteria that occur in microscopic clusters resembling grapes. !t is fre"uently found in the nose and s#in of a person. $bout %&' of the populations are long-term carriers of S. aureus.

Staphylococcus aureus
Morphology and identification. S. aureus is a facultatively anaerobic( Gram-positive coccus( which appears as grape-li#e clusters when viewed through a microscope and has large( round( golden-yellow colonies( often with hemolysis( when grown on blood agar plates. aureus means )golden) in *atin. S. aureus is catalase positive (meaning that it can produce the en+yme )catalase)) and able to convert hydrogen pero,ide (-%.%) to water and o,ygen( which ma#es the catalase test useful to distinguish staphylococci from enterococci and streptococci. $ large 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)) that causes clot formation while most other Staphylococcus species are coagulase-negative. -owever( while

G+ve Pyogenic cocci (Lecture 1) .

. the majority of S. aureus are coagulase-positive( some may be atypical in that they do not produce coagulase. Role in disease S. aureus may occur as a commensal on human s#in; it also occurs in the nose fre"uently and throat less commonly. The occurrence of S. aureus under these circumstances does not always indicate infection and therefore does not always re"uire treatment. !t can survive for some hours on dry environmental surfaces. !t can host phages that produce to,ins such as the 1anton-2alentine leu#ocidin( which increase its virulence. 1anton-2alentine leu#ocidin (12*) is one of many to,ins associated with S. aureus infection. 3ecause it can be found in virtually all 4$-56 $ (community-associated 5ethicillin-resistant taphylococcus aureus) strains that cause soft-tissue infections( it was long described as a #ey virulence factor( allowing the bacteria to target and #ill specific white blood cells #nown as neutrophils. 12* is produced from the genetic material of a bacteriophage which infects Staphylococcus aureus( ma#ing it more virulent. S. aureus can infect other tissues when normal barriers have been breached (e.g.( s#in or mucosal lining). This leads to furuncles (boils) and carbuncles (a collection of furuncles). !n infants S. aureus infection can cause a severe disease taphylococcal scalded s#in syndrome ( ) due to the production of 7,foliatin to,in (7T).

S. aureus infections can be spread through contact with pus from an infected wound( s#in-tos#in contact with an infected person by producing hyaluronidase that destroy tissues( and contact with objects such as towels( sheets( clothing( or athletic e"uipment used by an infected person. 8eeply situated S. aureus infections can be very severe. 1rosthetic joints put a person at particular ris# for septic arthritis( and staphylococcal endocarditis (infection of the heart valves) and pneumonia( which may be rapidly spread.

G+ve Pyogenic cocci (Lecture 1) .

. S. aureus also causes; -mastitis( -phlebitis( -meningitis(-urinary tract infectionsosteomyelitis(-hospital ac"uired (nosocomial) infection. -food poisoning by releasing enteroto,ins into food( and -to,ic shoc# syndrome by release of To,ic shoc# syndrome to,in (T T) (superantigens) into the blood stream.

Fig -1-Sites of infection and diseases caused by Staphylococcus aureus

Fig- -!irulence determinants of Staphylococcus aureus.

G+ve Pyogenic cocci (Lecture 1) .

"n#ymes and to$ins

taphylococci can produce disease through their ability to multiply and spread in tissue( and through the production of e,tracellular substances (en+ymes and to,ins).

"n#ymes
0. 4oagulase and clumping factor 4oagulase is an e,tracellular protein which binds to prothrombin. 4oagulase is a traditional mar#er for identifying S. aureus in the clinical microbiology laboratory. 3acteria could protect themselves from phagocytic and immune defenses by causing locali+ed clotting. 4lumping factor is a surface compound that is responsible for adherence of the organism to fibrinogen and fibrin. %. 4atalase; which converts hydrogen pero,ide into water and o,ygen. (8istinguishes them from Streptococci which are negative). 9. taphylo#inase% This factor lyses fibrin. $nd causes fibrinolysis which might aid in bacterial spreading. :. -yaluronidase; spreading factor. ;. 1roteinases. <. *ipases. =. >-lactamases

&o$ins.
0. -to,in (-hemolysin).causes membrane damage and partial hemolysis of blood on blood agar. %. ?-to,in is a sphingomyelinase which damages membranes rich in this lipid. 4auses complete hemolysis of blood on blood agar.
9. -to,in is a very small peptide to,in produced by most strains of S. aureus. !t is also

produced by S. epidermidis. 4auses no hemolysis of blood on blood agar

:

G+ve Pyogenic cocci (Lecture 1) .

. :. *eu#ocidin is a multicomponent protein to,in produced as separate components which act together to damage membranes. The bicomponent to,in 1anton-2alentine leu#ocidin (12*) is associated with severe necroti+ing pneumonia in children. ;. 7,foliatin to,in (7T). 4auses the scalded s#in syndrome in neonates( which results in widespread blistering and loss of the epidermis. The protease activity of the e,foliative to,ins causes peeling of the s#in observed with <. To,ic shoc# syndrome to,in (T multisystem involvement( including des"uamative s#in rash. =. 7nteroto,ins $-7( G-!( @-5; cause emesis (vomiting) when ingested and the bacterium is a leading cause of food poisoning. The last three to,ins act as superantigens. uperantigens stimulate T cells nonspecifically without normal antigenic recognition. Ap to one in five T cells may be activated( whereas only 0 in 0&(&&& are stimulated during a usual antigen presentation. Role of pigment in virulence ome strains of S. aureus are capable of producing staphylo,antin - a pigment that acts as a virulence factor. !ts has an antio,idant action that helps the microbe to evade #illing with reactive o,ygen used by the host immune system. !t is thought that staphylo,antin is responsible for . aureusB characteristic golden colour. 1igment is viewed in the laboratory by culturing the bacterium on mil# agar to monitor the golden yellow pigment of S. aureus. 'iagnosis 8epending upon the type of infection present( an appropriate specimen is obtained accordingly and sent to the laboratory for definitive identification by using biochemical or en+yme-based tests. $ Gram stain is first performed to guide the way( which should show typical gram-positive bacteria( cocci( in clusters. econdly( culture the organism in 5annitol alt $gar( which is a selective medium with =CD' Ea4l that allows S. aureus to grow producing yellow-colored colonies as a result of salt utili+ation and subse"uent drop in the mediumBs p-. Growth on 5il# agar facilitates the observation of the golden pigment of
;

T); this to,in is associated with fever( shoc# and

G+ve Pyogenic cocci (Lecture 1) .

. S. aureus. Furthermore( for differentiation on the species level( catalase (positive for all species)( coagulase (fibrin clot formation)( 8E$se (+one of clearance on nutrient agar)( lipase (a yellow color and rancid odor smell)( and phosphatase (a pin# color) tests are all done.

Rapid 'iagnosis and &yping

8iagnostic microbiology laboratories and reference laboratories are #eys for identifying outbrea#s and new strains of S. aureus. 6ecent genetic advances have enabled reliable and rapid techni"ues for the identification and characteri+ation of clinical isolates of S. aureus in real-time. These tools support infection control strategies to limit bacterial spread and ensure the appropriate use of antibiotics. These techni"ues include 6eal-time 146 and Guantitative 146 and are increasingly being employed in clinical laboratories.

Sensitivity to antimicrobial agents%

taphylococci are variably sensitive to many antimicrobial drugs. 6esistance falls into several classes; 6esistant to penicillin; production of >-lactamase under plasmid control. 6esistance to naficillin( methicillin and o,acillin. The appearance of vancomycin resistance strains. 1lasmid mediated resistant to tetracycline( erythromycin and aminoglycosides.

&reatment
*ocal antiseptics are used to control furunculosis; tetracyclines are used for long-term treatment of acne. urgical drainage and removal of dead bone is accompanied by longterm administration of appropriate drugs is used in chronic osteomyelitis. 3acteremia( endocarditis( pneumonia re"uire prolonged intravenous therapy with >-lactamase resistant penicillin.
<

G+ve Pyogenic cocci (Lecture 1) .

"pidemiology and control

The chief source of infection is shedding human lesions( fomites contaminated from such lesions( and the human respiratory tracts and s#in. !n hospitals the area for severe staphylococcal infections are the newborn nursery( intensive care units( operating rooms and cancer chemotherapy wards. The application of topical antiseptics to nasal or perineal carriage sites may diminish shedding of organism. 6ifampin coupled with a second oral antistaphylococcal drug provides a long-term cure of nasal carriageHalthough staphylococci can rapidly develop resistant to rifampin.

(ther medically important species of staphylococcus

S.epidermidis( a coagulase-negative staphylococcus species( is a commensal of the s#in( but can cause severe infections in immune-suppressed patients and those with central venous catheters. S. saprophyticus( another coagulase-negative species that is part of the normal vaginal flora( is predominantly implicated in genitourinary tract infections in se,ually-active young women.

Al-Mustansiriya University College of medicine Microbiology Dept. rd grade !-""-#$$% Dr. Shama