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Pavithra. G. Palan.

STAPHYLOCOCCUS. Pavithra. G. Palan. Staphylococci are gram positive cocci, Occur in grape like clusters, In Greek; staphyle - Bunch of grapes Kokkus - Berry. CLASSIFICATION: A) Based on coagulase production: 1. Coagulase positive: Eg- S. aureus

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Pavithra. G. Palan.

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  1. STAPHYLOCOCCUS Pavithra. G. Palan.

  2. Staphylococci are gram positive cocci, • Occur in grape like clusters, • In Greek; staphyle -Bunch of grapes Kokkus - Berry

  3. CLASSIFICATION: A) Based on coagulase production: 1. Coagulase positive: Eg- S. aureus 2. Coagulase negative: Eg- S. epidermidis S. saprophyticus B) Based on pathogenicity: 1. Common pathogen: Eg- S. aureus 2. Opportunistic pathogens: Eg- S. epidermidis S. saprophyticus 3. Non pathogen: Eg- S. homonis

  4. STAPHYLOCOCCUSAUREUS MORPHOLOGY: • These are spherical cocci. • Approximately 1μm in diameter. • Arranged characteristically in grape like clusters. • They are non motile and non sporing. • A few strains possess capsules.

  5. CULTURE: Media used :- i) Non selective media: Nutrient agar, Blood agar, MacConkey’s agar. ii) Selective media: Salt-milk agar, Ludlam’s medium

  6. Cultural Characteristics: i) On nutrient agar- The colonies are large, circular, convex, smooth, shiny, opaque and easily emulsifiable. Most strains produce golden yellow pigments.

  7. ii) On MacConkey’s agar- The colonies are small & pink in colour. iii) On blood agar- Most strains produce β- haemolytic colonies.

  8. Biochemical reactions: 1) Catalase test- Positive.

  9. 2) Coagulase test- i) Slide coagulase test- Positive. ii) Tube coagulase test- Positive. SLIDE COAGULASE TEST TUBE COAGULASE TEST

  10. 3) Reduces nitrate to nitrite. 4) Ferments mannitol anaerobically with acid only. 5) Urea hydrolysis test- Positive. 6) Gelatin liquefaction test- Positive. 7) Produces Lipase. 8) Produces Phosphatase. 9) Produces Thermostable nuclease.

  11. PATHOGENICITY: Source of infection: A) Exogenous: patients or carriers B) Endogenous: From colonized site Mode of transmission: A) Contact: direct or indirect( through fomites) B) Inhalation of air borne droplets

  12. Virulence factors: These include A) Cell associated factors B) Extracellular factors

  13. A) CELL ASSOCIATED FACTORS: a) Cell associated polymers b) Cell surface proteins a) CELL ASSOCIATED POLYMERS 1. Cell wall polysaccharide 2. Teichoic acid 3. Capsular polysaccharide b) CELL SURFACE PROTEINS: 1. Protein A 2. Clumping factor (bound coagulase)

  14. Structure of Staphylococcal cell wall

  15. B) EXTRACELLULAR FACTORS a) Enzymes b) Toxins

  16. a) Enzymes: 1. Free coagulase 2. Catalase 3. Lipase 4. Hyaluronidase 5. DNAase 6. Thermonuclease 7. Staphylokinase (Fibrinolysin) 8. Phosphatase

  17. b) Toxins: 1. Cytolytic toxins i) Haemolysins Alpha haemolysin Beta haemolysin Gamma haemolysin Delta haemolysin ii) Leucocidin (Panton-Valentine toxin) 2. Enterotoxin 3.Toxic shock syndrome toxin (TSST) 4. . Exfoliative (epidermolytic toxin)

  18. Disease: Diseases produced by Staphylococcus aureus is studied under 2 groups: A) Infections B) Intoxications

  19. A) INFECTIONS: Mechanism of pathogenesis: Cocci gain access to damaged skin, mucosal or tissue site Colonize by adhering to cells or extracellular matrix Evade the host defense mechanisms and multiply Cause tissue damage

  20. Common Staphylococcal infections are: 1) Skin and soft tissue: Folliculitis, furuncle (boil), carbuncle, styes, abscess, wound infections, impetigo, paronychia and less often cellulitis. Folliculitis

  21. Furuncle (boil) Carbuncle

  22. Styes Abscess

  23. Impetigo Paronychia Wound infection Cellulitis

  24. 2) Musculoskeletal: Osteomyelitis, arthritis, bursitis, pyomyositis. osteomyelitis 3) Respiratory: Tonsillitis, pharyngitis, sinusitis, otitis, bronchopneumonia, lung abscess, empyema, rarely pneumonia.

  25. 4) Central nervous system: Abscess, meningitis, intracranial thrombophlebitis. 5) Endovascular: Bacteremia, septicemia, pyemia, endocarditis. Endocarditis 6) Urinary: Urinary tract infection.

  26. B) INTOXICATIOINS: The disease is caused by the bacterial exotoxins, which are produced either in the infected host or preformed in vitro. There are 3 types- • Food poisoning • Toxic shock syndrome • Staphylococcal scalded skin syndrome

  27. 1) Food poisoning: • Enterotoxin is responsible for manifestations of staphylococcal food poisoning. • Eight types of enterotoxin are currently known, named A, B, C1-3, D, E, and H. • It usually occurs when preformed toxin is ingested with contaminated food. • The toxin acts directly on the autonomic nervous system to cause the illness, rather than gut mucosa.

  28. The common food items responsible are - milk and milk products, meat, fish and ice cream. • Source of infection- food handler who is a carrier. • Incubation period- 2 to 6 hours. • Clinical symptoms- nausea, vomiting and diarrhoea. • The illness is usually self limited, with recovery in a day or so.

  29. 2) Staphylococcal Toxic shock syndrome (STSS): • STSS is associated with infection of mucosal or sequestered sites by TSST( formerly known as enterotoxin type F) producing S.aureus. • It is fatal multisystem disease presenting with fever, hypotension, myalgia, vomiting, diarrhoea, mucosal hyperemia and erythematous rash which desquamates subsequently.

  30. 2 types of STSS known: i) Menstrual associated STSS: Here colonization of S.aureus occurs in the vagina of menstruating woman who uses highly absorbent vaginal tampons. ii) Non menstrual associated STSS: Here colonization of S.aureus occurs in other sites like surgical wound.

  31. 3) Staphylococcal scalded skin syndrome (SSSS): • Exfoliative toxin produced by S.aureus is responsible for this. • It is a skin disease in which outer layer of epidermis gets separated from the underlying tissues.

  32. Types of SSSS: Severe formMilder form In new born - Ritter’s disease - Pemphigus neonatorum In older patients - Toxic epidermal - Bullous necrolysis impetigo

  33. Pemphigus neonatorum Ritter’s disease Toxic epidermal necrolysis Bullous impetigo

  34. LAB DIAGNOSIS: Specimens collected: Depends on the type of infection. • Suppurative lesion- Pus, • Respiratory infection- Sputum, • Bacteremia & septicemia- Blood, • Food poisoning- Feces, vomit & the remains of suspected food, • For the detection of carriers- Nasal swab.

  35. Methods of examination: I) Direct microscopy: • Direct microscopy with Gram stained smear is useful in case of pus, where cocci in clusters are seen. • This is of no value for specimen like sputum where mixed flora are normally present.

  36. II) Culture: a) Media used: b) Cultural Characteristics: c) Gram staining: Smears are examined from the culture plate and reveals Gram positive cocci(1μm in diameter) arranged in grape like clusters.

  37. d) Biochemical reactions: III) Antibiotic sensitivity tests done as a guide to treatment. IV) Bacteriophage typing is done for epidemiological purposes. V) Serological tests are not useful.

  38. TREATMENT: • Drug resistance is common. • Benzyl penicillin is the most effective antibiotic, if the strain is sensitive. • Cloxacillin or Methicillin is used against beta-lactamase producing strains. • Methicillin Resistant Staphylococcus aureus (MRSA) strains have become common. • Vancomycin is used in treatment of infections with MRSA strains.

  39. EPIDEMIOLOGY: • Staphylococci are primary parasites of human beings and animals. • Hospital infections caused by staphylococci deserve special attention because of their frequency & they are caused by strains resistant to various antibiotics. • Staphylococci are the common cause of postoperative wound infection and other hospital cross infections.

  40. PREVENTION: • Isolation & treatment of MRSA patients. • Detection of carriers among hospital staff, their isolation & treatment. • Avoid indiscriminate usage of antibiotics.

  41. Coagulase Negative Staphylococci( CoNS ): Two species of coagulase negative Staphylococci can cause human infections- 1. Staphylococcus epidermidis 2. Staphylococcus saprophyticus

  42. S. Epidermidis: • It is a common cause of stitch abscesses. • It has predilection for growth on implanted foreign bodies such as artificial valves, shunts, intravascular catheters and prosthetic appliances leading to bacteraemia. • In persons with structural abnormalities of urinary tract, it can cause cystitis. • Endocarditis may be caused, particularly in drug addicts.

  43. S.saprophyticus: • It causes urinary tract infections, mostly in sexually active young women. • The infection is symptomatic and may involve the upper urinary tract also. • Men are infected much less often. • It is one of the few frequently isolated CoNS that is resistant to Novobiocin.

  44. Distinguishing features of the major species of staphylococcus

  45. Novobiocin sensitivity test

  46. THANK YOU

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