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They role in human pathology. Principles of microbiological diagnosis, specific therapy. Lector Tvorko M. S. Classification. S. aureus, S. epidermidis, S. saprophyticus VIRULENCE FACTORS OF STAPHYLOCOCCUS AUREUS Staphylococcus epidermidis Growing on Blood Agar Staphylococcus aureus Growing on Blood Agar Staphylococcus saprophyticus Growing on Blood Agar Coagulase Test on Staphylococcus aureus Coagulase Test on Staphylococcus epidermidis Staphylococcus aureus Growing on Yelk-salt Agar Staphylococcus aureus Growing on Mannitol Salt Agar. Staphylococcus epidermidis Growing on Mannitol Salt Agar Staphylococcus aureus Growing on DNase Agar Staphylococcus epidermidis Growing on DNase Agar Staphylococcal Skin Infections   Localized infections  Folliculitis (sties, pimples, and carbuncles) result from S. aureus entering natural openings in the skin – hair follicle Impetigo of the newborn  highly contagious superficial skin infection caused by S. aureus. Toxemia occurs when toxins enter the bloodstream; • Scalded skin syndrome • Toxic shock syndrome Figure 21.4 Staphylococcus aureus enterotoxin Staphylococcal Food Poisoning Figure 25.6 Electron Micrograph of Streptococcus pyogenes Note gram-positive (purple) cocci in chains (arrows).    Streptococcus are gram-positive cocci classified according to their hemolytic enzymes and cell wall antigens. Group A beta-hemolytic streptococci (including Streptococcus pyogenes) are the pathogens most important to humans. Produce a number of virulence factors:  M protein,  erythrogenic toxin,  deoxyribonuclease,  streptokinases,  hyaluronidase. Figure 21.5 Streptococcus in chains (Gram stain) hemolysis reaction - sheep blood agar    (alpha)  partial hemolysis  green color (beta)  complete clearing  A and B (gamma)  no lysis White colonies Beta Hemolysis on Blood Agar (Indirect Lighting) Alpha Hemolysis on Blood Agar (Indirect Lighting) A Plate of Blood Agar showing Alpha, Beta, and Gamma Hemolysis (Indirect Lighting) Gamma Reaction on Blood Agar Streptococcal pharyngitis  Strep throat  Streptococcus pyogenes  Resistant to phagocytosis  Streptokinases lyse clots  Streptolysins are cytotoxic  Pharyngitis - Scarlet Fever Erythrogenic toxin produced by lysogenized S. pyogenes   Diagnosis by indirect agglutination Figure 24.3 Streptococcal Skin Infections  Erysipelas      infects the dermal layer reddish patches Can progress to local tissue destruction Enter the bloodstream Impetigo  isolated pustules Streptococcal toxic shock syndrome  Cellulitis, myositis and necrotizing fasciitis Figure 21.6, 7 Bacterial Infections of the Heart   The inner layer of the heart is the endocardium. Inflammation of the endocardium Endocarditis Subacute bacterial endocarditis –  from microbs in the mouth.( Arises from a focus of infection, such as a tooth extraction).  alpha-hemolytic streptococci  staphylococci  enterococci  Preexisting heart abnormalities are predisposing factors.  Signs include fever, anemia, and heart murmur.   Acute bacterial endocarditis   Staphylococcus aureus The bacteria cause rapid destruction of heart valves Rheumatic Fever  Rheumatic fever is an autoimmune complication of streptococcal infections. Rheumatic fever is expressed as arthritis or inflammation of the heart.  It can result in permanent heart damage.  Rheumatic fever can follow a streptococcal  infection, such as streptococcal sore throat.  Streptococci might not be present at the time of rheumatic fever.  Prompt treatment of streptococcal infections can reduce the incidence of rheumatic fever.  Penicillin is administered as a preventive measure against subsequent streptococcal infections.  Antibodies against group A beta-hemolytic streptococci react with streptococcal antigens deposited in joints or heart valves or cross-react with the heart muscle. Figure 23.5 Dental Caries  Streptococcus mutans, found in the mouth  uses sucrose       form dextran from glucose lactic acid from fructose. Bacteria adhere to teeth and produce sticky dextran, forming dental plaque. Acid produced during carbohydrate fermentation destroys tooth enamel at the site of the plaque. Gram-positive rods and filamentous bacteria can penetrate into dentin and pulp. Caries are prevented by restricting the ingestion of sucrose and by the physical removal of plaque. Figure 25.4 Pneumomoccal Pneumonia    Streptococcus pneumoniae:  Gram-positive encapsulated diplococci Diagnosis by culturing bacteria Penicillin is drug of choice Figure 24.13 Scanning Electron Micrograph of Streptococcus pneumoniae Streptococcus pneumoniae (diplococcus). Fluorescent stain Streptococcus pneumoniae on Blood Agar (Indirect Lighting) Enterococcus faecalis in a Blood Culture A bile esculin slant before inoculation. Enterococcus faecalis growing on a bile esculin slant. Note black color. Meningococci Meningococci Gram staining Identification of Neisseria meningitidis : Carbohydrate Utilization Colonies of Neisseria meningitidis on Blood agar Clinical symptoms of meningococcal infection Clinical symptoms of meningococcal infection Clinical symptoms of meningococcal infection Pathological changes of brain (meningococcal infection) Spinal fluid Diagnosis and Treatment of the Most Common Types of Bacterial Meningitis  Diagnosis is based on Gram stain and serological tests of the bacteria in CSF.  Cultures are usually made on blood agar and incubated in an atmosphere containing reduced oxygen levels.  Cephalosporins may be administered before identification of the pathogen. Neisseria gonorrhoeae   Gram-, diplococcus  Gram stain pus, intracellular diplococcus Virulence factors  Pili  Initial attachment  Antigenic and phase variation  Opacity protein (Opa)  Tighter contact and invasion  Antigenic variation  LOS (lipooligosaccharide, lack OAg)  Inflammatory, major cause of symptom  IgA protease Affected Populations STI Rates World-Wide http://www.agi-usa.org/pubs/ib_std.html Gonococci (methylen blue staining) Gonococci Electron Micrograph of Neisseria gonorrhoeae Colonies of gonococci onto blood agar Identification of Neisseria : MTM Chocolate Agar with a Taxo (Oxidase) Disc Identification of Neisseria gonorrhoeae : Carbohydrate Utilization A Positive Direct Fluorescent Antibody Test for Neisseria gonorrhoeae Neisseria gonorrhoeae (3)  Pili: facilitate attachment, impede phagocytosis  Lipopolysaccharide: marked endotoxin activity; local cytopathic effect  IgA protease: cleaves IgA-1 subclass of immunoglobulins Neisseria gonorrhoeae (4)  Porin (“Por”, formerly protein I): may facilitate endocytosis  Opacity proteins (“Opa,” formerly protein II): contribute to attachment to human cells  Reduction-modifiable protein (“Rmp,” formerly protein III: stimulates blocking antibodies that reduce serum bactericidal activity GC Sexual Transmission  Efficiently transmitted by sexual contact  Greater efficiency of transmission from male to female  Male to female: 50 - 90%  Female to male: 20 - 80%   Vaginal & anal intercourse more efficient than oral Can be acquired from asymptomatic partner  Increases transmission and susceptibility to HIV 2-5 fold Gonorrhea         Many asymptomatic  Reason for spread Male- Urethritis, urethral discharge Female - Endocervicitis, discharge, dysuria, bleeding Pharyngitis Proctitis Disseminated gonococcal infection (DGI)  Pustular skin lesions  Septic arthritis Pelvic inflammatory disease (PID)  Endometritis, salpingitis, peritinitis  Infertility, ectopic pregnancy Ophthalmia neonatorum Gonococcal Infections in Women         Cervicitis  Pharyngitis Urethritis  DGI Proctitis Accessory gland infection (Skene, Bartholin) Pelvic inflammatory disease (PID) Peri-hepatitis (Fitz-Hugh-Curtis) Pregnancy morbidity Conjunctivitis Many infections asymptomatic Gonococcal Cervicitis Incubation 3-10 days Symptoms: Vaginal discharge Dysuria Vaginal bleeding Cervical signs : Erythema Friability Purulent exudate  Pelvic Inflammatory Disease Adhesions    Tube STD Atlas, 1997 Sx: lower abdominal pain Signs: CMT, uterine/ adnexal tenderness, +/fever Laparoscopy may show hydrosalpinx, inflammation, abscess, adhesions Gonococcal Infections in Men         Pharyngitis Urethritis  DGI Epididymitis  Urethral stricture Proctitis  Penile edema Conjunctivitis Abscess of Cowper’s/Tyson’s glands Seminal vesiculitis Prostatitis Many infections asymptomatic Gonococcal Urethritis     Incubation 2-7 days Abrupt onset of severe dysuria Purulent urethral discharge Most urethral infections symptomatic Epididymitis Epididymitis  Swollen painful epididymis  Urethritis  Epididymal tenderness or mass on exam Gonococcal Infections in Women & Men      Urethritis Proctitis Pharyngeal infections Conjunctivitis Disseminated Gonococcal Infection Blenorrhea Gonorrhea  Diagnosis     Intracellular Gram negative diplococci in discharge Growth on selective media, oxidase positive colonies Flourescent antibody Treatment  Cover for probable association with C. trachomatis GC Diagnostic Methods     Gram stain smear Culture Antigen Detection Tests: EIA & DFA Nucleic Acid Detection Tests    Probe Hybridization Nucleic Acid Amplification Tests (NAATs) Hybrid Capture Gonorrhea Diagnostic Tests Sensitivity Gram stain 90-95% (male urethra exudate) DNA probe Culture NAATs * 85-90% 80-95% 90-95% Specificity  95%  95%  99%  98% * Able to use URINE specimens Gram Stain for GC: Urethral Smear   Numerous PMNs Gram negative intracellular diplococci Gram Stain for GC: Cervical Smear  PMN with Gram negative intracellular diplococci GC Culture  Requires selective media with antibiotics to inhibit competing bacteria (Modified Thayer Martin Media, NYC Medium)  Sensitive to oxygen and cold temperature  Requires prompt placement in high-CO2 environment (candle jar, bag and pill, CO2 incubator)  In cases of suspected sexual abuse, culture is the only test accepted for legal purposes GC Culture Candle Jar GC Culture Specimen Streaking Cervical and Urethral GC Culture After 24 Hours Treatment http://www.cdc.gov/STD/treatment/4-2002TG.htm#Gonococcal Prevention      Abstinence Safe/”smart” sex Barrier contraceptives Educational programs Reduce misuse of antimicrobials
 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
									 
                                             
                                             
                                             
                                             
                                             
                                             
                                             
                                             
                                             
                                             
                                            