MICROBIOLOGY AND PARASITOLOGY

Differentiation of Medically Important Bacteria G (+) cocci - Staphylococcus - Streptococcus - Pneumococcus G (+) bacilli - Non spore former - Corynebacterium - Spore former - Bacillus - Clostridium

G (-) cocci Neisseria G (-) Bacill Proteus Klebsiella Escherichia Salmonella Shigella Haemophilus Pseudomones Vibrio Campylobacter - Helicobacter

Pyogenic cocci spherical bacteria that cause various supportive infection G (+): G(-):

S. aureus N. gonorrhaeae S. pyogenes N. meningitidis S. pneumonia ________________________________________________________________ S. aureus Gram + cocci in clusters - Fermentive utilization of glucose and manitol - Chemoorganotrohic heterotrophs - 8 Stereotypes: > serotypes 5(MRSA) & 8(TSS) - Cell Structure: > Protein A & Capsule antiphagocytic > Teichoic acid colonizing factor > Adhesins adheres to fibronectin, laminin, fibrin and collagen - Enzymes & Toxins secreted: > Hemolysins > Catalase > Coagulase

> Leucocidin > Erythrogenic toxin > Enterotoxin - Disease caused by toxins: > Enterotoxins > Exfoliation SSSS > TSS (Toxic shock syndrome) ____________________________________________________________ Streptococcus pyogene - cocci in chain gram + - Lancefield classification: > based on CW polysaccharide C or Lipoteichoic acid moiety - Virulence factors: > Hyaluronic Acid capsule > M-protein -> anti-phagocytic > Adhesins > C5a peptidase inactivates chemotactic activity of C5a - Secreted products: > Erythrogenic toxins > Streptolysin S: O2 stable, non-antigenic; lyses RBCs, lymphons > Streptolysin O: O2 labile, antigenic, very immunogenic; lyses RBCs, cardotoxic _____________________________________________________________ Streptococcus pneumoniae - Lancet / football shape G (+) diplococcic - present in normal flora - Virulence factors > Adhesins > IgA protease > Capsule - Disease > Pneumonia > Sepsis > Meningitis - Identification: > optochin (P) disc: sensitive > catalase: negative > Bile: soluble > BAP: a-hemolytic, glistening, mucoid - Vaccination it contains capsular polysacharride rom 23 serotypes - recommended for children <5 years and adults > = 65 years and other people that is at work.

_____________________________________________________________ Neisseria G (-) D-shape / Kidney bean shape intracellular diplococci catalase (+), oxidase (+) Non-motile Fastidious (Fe requiring) Needs 5-10 % CO2 Gonococcus Meningococcus Capsule none + Pili + + IgA protease + + CHO fermentation Glucose Glucose & maltose ______________________________________________________________ Corynebacterium diptheriae G (+) rod, aerobic Palisade / Chinese character like formation Non-sporulating w/ metanchromatic granules Virulence: > Cord factor > Exotoxin - Tellurite media black colonies w/ a surrounding brown halo - ID tests: > Catalase & Nitrate reduction (+) > Non-motile > Urease (-) - Shick Test ______________________________________________________________ Bacillus - Aerobic, spore-former - Square-ended, bamboo like w/ central spores - BAP (Blood Agar Plate): non or hemolytic or off white, ground glass
appearance, medusa-head colonies (filamentous edge)

- Catalase (+) A. Bacillus anthrasis - Virulence factors > poly-D-glutamic acid capsule > Exotoxin 3 components: 1. Edema Factor (EF) dec. phagocytosis 2. Lethal Factor (LF)

3. Protective antigen (PA) B. Bacillus cereus - Exotoxin (mostly boiled rice) - Virulence factors > poly-D-glutamic acid capsule > Exotoxin(enterotoxin) _____________________________________________________________ Clostridium G (+) rod, box-can shaped Anaerobic spore former Natural habitat: soil Exotoxins Clostridium tetani

- Terminal spores (drum stick appearance) - tetanospasmin block release of NT from presynaptic membranes of inhibitory Clostridium botulinum

- subterminal spores - Toxins A, B, C1, D, E, F & G - blocks release of the NT Ach from presynaptic stimulatory terminals Clostridium perfringens

- central spores - invasins & exotoxins gas gangrene and uterine infection - Hemolysins, protease, lipase, collagenase, hyaluronidase - enterotoxin _____________________________________________________________ Haemophilus influenza - G (-) coccobacillus - X(hemin) & V(NAD) factors - 6 serotypes: A-F > Type B invasive disease - Virulence factors > Capsule > outer membrane > Adherence > IgA protease

- Infection > Bacteremia, meningitis (endotoxin) > Unencapsulated strains -> OM, bronchitis, sinusitis Haemophilus ducreyi - school of fish - requires freshly clotted blood or a GC agar base medium _______________________________________________________________ Bordetella pertussis -(whooping cough) - Aerobic G (-) coccobacullus - not part of normal flora; exceptionally contagious - Obligatory parasite - Virulence factors > perussis toxin > hemagglutinins - MOA: pertussis covalently attaches an ADP-Ribose moistly to G protein involved in adenylate cyclase_>increases camp_>inhibits neutrophil function and also increases lymphocyte number, tracheal cytotoxin - LP> toxin- systemic effects of an infection: fever - Lab. Diagnosis: > Nasopharyngeal swab - Culture: Bordet-Gengon droplets of mercury, slightly b hemolytic - Non-motile - Catalase & oxidase (+) - Citrate & Urease (-) ________________________________________________________________ Pseudomonas aeruginosa G (-) rod, oxidase (+), glucose non fermenting pyocyanin & pyoverdin protease, amylase, pectinase, cellulose saprophytic, opportunistic or as plant pathogens Virulence factors > adhesins > flagella > pili > endotoxin > protease > lipase > exotoxin A ( inhibits protein synthesis ) - MDR(multi drug resistance) ________________________________________________________________

Enterobacteriaceae - causes GIT disease - all are facultative anaerobes - soil, water, decaying matter, large bowel - G (-), small, non spore forming rods - facultative anaerobs - endotoxin, enterotoxins, capsules (K), hemolysins, fimbrae (colonize), flagella (H) - (+) glucose fermentation - Oxidase (-), Catalase (+), Nitrate (+) - Includes > Coliforms ferment lactose quickly * E. coli, klebsiella > Non coliforms non or slow lactose fermenters * Proteus, Salmonella, Shigella - Biochemical Test > H2S producers * Proteus mirabilis, Salmonella sp. > Urease (+) * Proteus, Klebsiella pneumoniae > Non motile at 37oC * Klebsiella, Shigella sp. ________________________________________________________________ Klebsiella pneumoniae - Faculative, G (-) rod, lactose (+) - Non motile - (+) large capsules - Endotoxins -> necrosis & abscess ________________________________________________________________ Escherichia coli - Most common inhabitants of I.T. non fastidious - Faculative, G (-) rod, lactose (+) - enterotoxins and cytotoxins > ETEC > EPEC enterotoxins > EAggEC > EIEC shiga like toxin - Adherence CFAs ( colonization factor antigens) ________________________________________________________________ Proteus mirabilis - Extremely motile - Splits urea to CO2 and NH3 - Pili

_______________________________________________________________ Salmonella typhi - Faculative, G (-) rod, lactose (-), motile, urease (-), H2S producer - Large inoculum required - Virulence > Faculative intracellular pathogen ( macrophages ) > smooth LPS for invasion > Flagella _______________________________________________________________ Shigella dysenteriae - Faculative, G(-) rod, lactose (-), non motile, non H 2S producer nor Urease - Low inoculum required - Virulence Host factors: > Acid resistant > intestinal motility and normal flora is very important Bacteria Factors >invasive capacity is important in virulence >non-invasive mutants are virulent > LPS with O antigen > shiga toxin is enterotoxic, cytotoxic ________________________________________________________________ Vibrio Cholera - G (-) rods w/ enteric like properties: > Glucose fermenters > Non fastidious > most are bile tolerant - Non enteric > vibrio means comma shaped > catalase & oxidase (+) > halophilic or halotolerant - water borne - Acid sensitive - Virulence > Cholera toxin are stimulants-> increase adenylate cyclase -> increase cAMP -> increase water and electrolyte secretion into gut ________________________________________________________________ Campylobacter jejuni - G (-) curved rods : gull winged - micro aerophilic - Source:

> birds , dogs, farm animals, transmitted via food & H2O - Virulence > Enterotoxin > cytotoxin - cholera like gastroenteritis - self limiting Treponema pallidum - cannot classify if gram or + because its little - Long, skinny organism > dark field microscopy - slow growing, microaerophilic - motile w/ axial filaments -> sluggish - Humans = reservoir - Virulence > Hyaluronidase > capsule like outer coat _______________________________________________________________ Leptospira interrogans Helicoidal, one or both ends hooked Obligate aerobes, Oxidase and Catalase (+) Zoonotic Virulence > hemolysin > endotoxin - Culture: Fletchers or Stuarts media containing 10% rabbit serum - Incubation: up to 6 wks. Aerobically at 30oC in the dark _______________________________________________________________ Helicobacter pylori - G (-), spiral shaped rods, microaerophilic - Urease: urea -> CO2 and ammonia _______________________________________________________________ Mycobacterium tuberculosis - long slender bacillus - high Cell Wall lipid content ( resistant to acids, alkali, drying, germicidals) - AFS- acid fast stain - very slow growing - intracellular parasite ( macrophages) - strict aerobe - no known toxins -Virulence: - Mycolic acids ~ acid fastness - WAX D delayed hypersensitivity

- cord factor inhibits PMN migration, induces granulomatous response - Sulfolipid evasion of phagolysosome formation ________________________________________________________________ Mycobacterium Leprae - unculturable - Reservoir: Humans & armada Mycoplasma pneumoniae - General Characteristics > Coccoid ; lack of a CW > smalles free living organism > fried-egg / mulberry colonies - Difference from viruses: > grow on cell free media in vitro > contain RNA & DNA > Intracellular and extracellular parasitism in vivo - Difference from bacteria: > (+) sterols in the CM > no DNA homology w/ known bacteria > low guanine & cytosine levels > LWM genome > no reversion to walled forms ______________________________________________________________ Chlamydia - G (-) obligate intracellular parasites - 4x as much RNA as DNA - rigid CW that resembles bacterial CW but (-) muramic acid, and has a high lipid content in its outer membrane - Life cycle stages: > EB ( elementary body) attaches and enters columnar epithelial cells > EB inhibits fusion w/ lysosome > EB transforms into RB( reticulate body) and divides by binary fission > many RBs are made and some reverts to EBs ( 100 1000 EBs / phagolysosome) that can infect more cells. ______________________________________________________________ Rickettsiae - G (-), small pleomorphic, obligate intracellular parasites - leaky CW w/o peptidoglycan

- Zoonotic ~ occur in the gut lining ticks, fleas, lic, and etc. are transmitted via anthropo bites - Virulence > endotoxins, hemolysin, phospholipase A - Rickettsiae multiply in endothelial cells of small blood vessels -> thrombosis and rupture vessel.