1. Staphylococcus Dr. Jyotsna Agarwal Professor, Dept. of Microbiology K G Medical University, Lucknow

2. Family: Micrococcaceae Genus: –Staphylococcus - derived from Greek “stapyle” (bunch of grapes) –Include major human pathogen and skin commensals –Micrococcus - skin commensal

3. Staphylococcus: General Characteristics Gram-positive spherical cells (0.5-1.5  m) in singles, pairs, and clusters Appear as “bunches of grapes” Gram-stained smear of staphylococci from colony

4. Non motile Non–spore-forming Nonencapsulated Catalase-producing Oxidase: negative Glucose fermenters Primarily aerobic, some facultatively anaerobic Staphylococcus: General Characteristics

5. Approximately 33 species ~15 species associated with humans Staphylococcus divided into coagulase positive & coagulase negative categories Inhibited by high bile salt concentration Some are ß-hemolytic Colony morphology: buttery looking, cream or white colored Staphylococcus: General Characteristics

6. S. aureus S. intermedius S. hyicus S. delphini S. schleiferi Veterinary pathogens Animal-associated species Human pathogens Coagulase Positive Staphylococci

7. Primary pathogen Habitat: anterior nares (carriers) Colonization: axilla, perineum, pharynx Produce superficial to systemic infections Mode of transmission: Pus formation Staphylococcus aureus

8. Natural history of disease Usual sites - skin, nasopharynx, perineum Breach in mucosal barriers - can enter underlying tissue Characteristic abscesses with Pus Bacteria liberates toxins-

9. DISEASES Due to direct effect of organism –Local - skin –Deep abscesses –Systemic infections Toxin mediated –Food poisoning –toxic shock syndrome –Scalded skin syndrome

10. Virulence Factors of S. aureus Pathogen Factors ENZYMES – Catalase (counters host defences) –Coagulase –Hyaluronidase –Lipases –B lactasamase (antibiotic resistance) TOXINS- enterotoxin, TSST, epidermolytic toxin

11. SKIN LESIONS- superficial Boils Styes Furuncles Carbancles Wound infections

12. DEEP ABSCESSSES Direct / by blood Can be single / multiple Eg. - Breast abscess kidney, brain, Osteomyelitis, septic arthritis

13. TOXIN MEDIATED DISEASES 1. Staphylococcal food poisoning –Due to production of enterotoxins –preformed toxin, heat stable –short incubation period –Milk & milk products

14. 2. Toxic shock syndrome High fever, diarrhoea, shock and erythematous skin rash which desquamate Mediated via ‘toxic shock syndrome toxin’ 10% mortality rate

15. 3. Scalded skin syndrome Disease of young children Mediated through minor Staphylococcal infection by ‘epidermolytic toxin’ producing strains Good prognosis

16. Virulence Factors: Extracellular enzymes Cytolytic Toxins –Alpha hemolysin: lyses rbcs, damages plts, causes severe tissue damage –ß hemolysin: acts on sphingomyelin in the plasma membrane of rbcs

17. Virulence Factors: Extracellular enzymes Hyaluronidase: Hydrolyzes hyaluronic acid in connective tissue allowing spread of infection Staphylokinase: fibrinolysin which allows spread of infection Coagulase: virulence marker Lipase: allows colonization

18. Beta-lactamase or Penicillinase: confers resistance DNase: degrades DNA Protein A: in cell wall, it binds to F c part of IgG toblock phagocytosis Virulence Factors: Extracellular enzymes

19. Coagulase-Negative Staphylococci Habitat: skin and mucous membranes Common human isolates –S. epidermidis –S. saprophyticus

20. Coagulase-Negative Staphylococci: Staphylococcus epidermidis Virulence factor: “slime” Mode of infection: colonization of medical implants Infections are acquired nosocomially Serious infections among immunosuppressed patients or neonates may occur

21. Habitat: skin and mucosal membranes of the genitourinary tract Common cause of urinary tract infections in young, sexually active females Coagulase-Negative Staphylococci: Staphylococcus saprophyticus

22. Microscopic Examination 1.Gram-positive cocci 2.pairs and clusters 3.Numerous polymorphonuclear cells (PMNs) Laboratory Diagnosis: Direct Smear Examination

23. Laboratory Diagnosis: Cultural Characteristics Colony morphology –Smooth, butyrous, white to yellow, creamy –Grow well in 18-24 hours –S. aureus may produce hemolysis on blood agar S. aureus

24. Principle: tests for enzyme catalase Drop H 2 O 2 onto smear Bubbling = POS (Staph) No bubbling = Streptococci Identification Tests: Catalase

25. –Cell-bound “clumping factor” converts fibrinogen to fibrin which precipitates on cell causing agglutination –Extracellular enzyme “free coagulase” Identification Tests: Coagulase Test Two methods Slide test: screens for “clumping factor” Tube test

26. Novobiocin Susceptibility Test Test to differentiate coagulase-negative staphylococci from S.saprophyticus from urine samples –S. saprophyticus is resistant (top)

27. Beta-lactam group of antibiotics- (penicillins, cloxacillins, ampicillin, amoxycillin) Beta-lactamase producers treatment of choice Amoxyclavulinic acid or ampicillinn sulbactam combo or methicillin/ oxacillin For methicillin -resistant S. aureus (MRSA) treatment of choice- Vancomycin Antimicrobial Susceptibility

28. Summary Characters of Staphylococcus aureus Enzymes / Toxins Infections / Diseases Laboratory diagnosis Treatment / Abx resistance