Microbiology Flashcards
Gram stain limitations
Treponema (too thin to be visualized), Mycobacteria (high lipid content in cell wall detected by carbolfuchsin in acid-fast stain), Mycoplasma (no cell wall), Legionella pneumophila (primarily intracellular, silver stain), Rickettsia (ntracellular parasite), Chlamydia (intracellular parasite; lacks muramic acid in cell wall).
Giemsa stain
Chlamydia, Borrelia, Rickettsiae, Trypanosomes, Plasmodium
PAS (periodic acid-Schiff)
Stains glycogen, mucopolysaccharides; used to diagnose Whipple disease (Tropheryma whipplei).
Ziehl-Neelsen (carbol fuchsin)
Acid-fast organisms (Nocardia, Mycobacterium)
India ink
Cryptococcus neoformans (mucicarmine can also be used to stain thick polysacchardie capsule red).
Silver stain
Fungi (e.g., Pneumocystis), Legionella, H. pylori
Special culture requirements
H. influenzae (chocolate agar with factors V (NAD+) and X (hematin); N gonorrhoeae, N. meningitidis (Thayer-Martin (or VPN) media - Vancomycin (inhibits gramp-positive organisms), Polyymyxin (inhibits gram-negative organisms except Neisseria), and Nystatin (inhibits fungi); “to connect to Neisseria, please use your VPN client”; B. pertussis (Bordet-Gengou (potato) agar); C diphtheriae (Tellurite agar, Loffler medium); M tuberculosis (lowenstein-Jensen agar); M. pneumoniae (Eaton agar, requires cholesterol); Lactose-fermenting enterics (Pink colonies on MacConkey agar (fermentation produces acid, turning colony pink) E. coli is also grown on eosin-methylene blue (EMB) agar as colonies with green metallic sheen); Legionella (Charcoal yeast extract agar buffered with cysteine and iron); Fungi (Sabouradud agar)
Obligate aerobes
Nocardia, Pseudomonas aeruginosa, and Mycobacterium tuberculosis
Obligate anaerobes
Clostridium, Bacteroides, and Actinomyces
Obligate intracellular
Rickettsia, Chlamydia
Facultative intracellular
Salmonella, Neisseria, Brucella, Mycobacterium, Listeria, Francisella, Legionella, Yersinia pestis
Encapsulated bacteria
“SHiNE SKiS” Streptococcus pneumoniae, Haemophilus influenzae type B, Neisseria meningitidis, Eschericia coli, Salmonella, Klebsiella pneumoniae, and group B Strep
Catalase-positive organisms
Psuedomonas, Listeria, Aspergillus, Candida, E. coli, S. aureus, Serratia
Encapsulated bacteria vaccines
Pneumococcal vaccine, H. influenzae type B, Meningococcal
Urease-positive bugs
Cryptococcus, H. pylori, Proteus, Ureaplasma, Nocardia, Klebsiella, S. epidermidis, S. saprophyticus
Gram-positive lab algorithm
FA 129
Bugs with exotoxins
Corynebacterium diphtheriae (diptheria toxin), Pseudomonas aeruginosa (exotoxin A), Shigella (shiga toxin), EHEC (Shiga-like toxin), ETEC (Heat labile and heat-stablile toxin), Bacillus anthracis (edema factor), Vibrio cholerae (Cholera toxin), Bordetella pertussis (Pertussis toxin), Clostridium tetani (tetanospasmin), Clostridium botulinum (Botulinum toxin), Clostridium perfringens (Alpha toxin), Streptococcus pyogenes (Streptolysin O), Staph aureus (TSST-1), Strep pyogenes (Exotoxin A)
Endotoxin
An LPS found in outer membrane of gram-negative bacteria (both cocci and rods)- Edema, Nitric oxide, DIC/Death, Outer membrane, TNF-a, O-antigen, extremely heat stable, IL-1, Neutrophil chemotaxis
Staphylococcus aureus
Gram-positive cocci in clusters. Protein A (virulence factor) binds Fc-IgG, inhibiting complement activation and phagocytosis. Commonly colonizes the nose. Causes: Inflammatory disease- skin infections, organ abscesses, pneumonia (often after influenza virus infection), endocarditis, and osteomyelitis; Toxin-mediated disease - toxic shock syndrome (TSST-1), scalded skin syndrome (exfoliative toxin), rapid-onset food poisoning (enterotoxins); MRSA (methicillin-resistant S. aureus) infection - important cause of serious nosocomial and community-acquired infections, resistant to methicillin and nafcillin because of altered penicillin-binding protein
Staphylococcus epidermidis
Infects prosthetic devices and intravenous catheters by producing adherent biofilms. Component of normal skin flora; contaminates blood cultures. Novobiocin sensitive
Staphylococcus saprophyticus
Second most common cause of uncomplicated UTI in young women (first is E. coli). Novobiocin resistant
Streptococcus pneumoniae
Most common cause of (MOPS): Meningitis, Otitis media (children), Pneumonia, Sinusitis. Lancet-shaped, gram-positive diplococci. Encapsulated. IgA protease. Pneumococcus is associated with “rusty” sputum, sepsis in sickle cell anemia and splenectomy. No virulence without capsule.
Viridans group streptococci
Viridans strep are a-hemolytic. They are normal flora of the oropharynx and cause dental caries (strep mutans) and subacute bacterial endocarditis at damaged valves (S. sanguinis). Resistant to optochin, differentiating them from S. pneumoniae.
Streptococcus pyogenes (group A streptococci)
Causes: Pyogenic - pharyngitis, cellulitis, impetigo; Toxigenic - scarelt fever, toxic shock-like syndrome, necrotizing fascitis; Immunologic - rheumatic fever, acute glomerulonephritis. Bacitracin sensitive. Antibodies to M protein enhance host defenses against S. pyogenes but can give rise to rheumatic fever. ASO titer detects recent S. pyogenes infection. JONES criteria for rheumatic fever: Joints (polyarthritis), O (carditis), Nodules (subcutaneous), Erythema marginatum, Sydenham chorea. Pharyngitis can result in rheumatic fever and glomerulonephritis. Impetigo more commonly precedes glomerulonephritis than pharyngits. Scarlet fever: scarlet rash with sandpaper-like texture, strawberry tongue, circumoral pallor.
Streptococcus agalactiae (GBS)
Bacitracin resistant, b-hemolytic, colonizes vagina; causes pneumonia, meningitis, and sepsis, mainly in babies. Produces CAMP (authors of test) factor, which enlarges the area of hemolysis formed by S. aureus. Hippurate test +. Screen pregnant women at 35-37 weeks. Patients with + culture receive intrapartum penicillin prophylaxis.
Enterococci (group D strep)
Enterococci (E. faecalis and E. faecium) are normal colonic flora that are penicillin G resistant and cause UTI, biliary tract infections, and subacute endocarditis (following GI/GU procedures). Lancefield group D includes the enterococci and the nonenterococcal group D streptococci. Lancefield grouping is based on differences in the C carbohydrate on the bacterial cell wall. Variable hemolysis. VRE are an important cause of nosocomial infection.
Streptococcus bovis (group D strept)
Colonizes the gut. Can cause bactermia and subacute endocarditis in colon cancer patients.
Corynebacterium diphtheriae
Causes diphtheria via exotoxin encoded by B-prophage. Potent exotoxin inhibits protein synthesis via ADP-ribosylation of EF-2. Symptoms include pseudomembranous pharyngitis (grayish-white membrane) with lymphadenopathy, myocarditis, and arrythmias. Lab diagnosis based on gram-positive rods with metachromatic (blue and red) granules and Elek test for toxin. Toxoid vaccine prevents diphtheria. Black colonies on cystine-tellurite agar.
Spores: bacterial
Some bacteria can form spores at the end of the stationary phase when nutrients are limited. Spores are highly resistant to heat and chemicals. Have dipicoloinic acid in their core. Have no metabolic activity. Must autoclave to kill spores (as is done to surgical equipment) by steaming at 121 C for 15 minutes. Spore-forming gram-positive bacteria found in soil: Bacillus anthracis, Clostridium perfringens, C. tetani. Other spore formers include B. cereus, C. botulinum, Coxiella burnetti
Actinomyces vs Nocardia
Both form long, branching filaments resembling fungi.
Actinomyces (Gram-postive anaerobe, not acid fast, normal oral flora, causes oral/facial abscesses that drain through sinus tracts, form yellow “sulfur granules”, treat with penicillin)
Nocardia (Gram-positive aerobe, Acid fast (weak), found in soil, causes pulmonary infections in immunocompromised and cutaneous infections after trauma in immunocompetent, treat with sulfonamides)
Leprosy (Hansen disease)
Caused by Mycobacterium leprae, an acid-fast bacillus that likes cool temperatures (infects skin and superficial nerves- “glove and stocking” loss of sensation) and cannot be grown in vitro. Reservoir in United states: armadillos. Hansen disease has 2 forms: Lepromatous - presents diffusely over the skin, with leonine (lion-like) facies, and is communicable; characterized by low cell-mediated immunit with a humoral Th2 response. Tuberculoid - limited to a few hypoesthetic, hairless skin plaques; characterized by high cell-mediated immunity with a largely Th1-type immune response. Treatment: multidrug therapy consisting of dapsone and rifampin for 6 months for tuberculoid form; and dapsone, rifampin, and clofazimine for 2-5 years for lepromatous form.
Gram-negative algorithm
FA 136
Lactose-fermenting enteric bacteria
Grow pink colonies on MacConkey agar. Citrobacter, Klebsiella, E. coli, Enterobacter, and Serratia. E. coli produces B-galactosidase, which breakes down lactose into glucose and galactose. EMB agar - lactose fermenters grow as purple/black colonies. E. coli grows purple colonies with a green sheen.
Neisseria
Gram-negative diploccoci. Both ferment glucose (but meningococci also ferments maltose) and produce IgA proteases. N. gonorrhoeae is often intracellular
N. gonococci
No polysaccharide capsul, no maltose fermentation, no vaccine (due to rapid antigenic variation of pilus proteins), Sexually transmitted, causes gonorrhea, septic arthritis, neonatal conjunctivitis, pelvic inflammatory disease (PID), and FItz-Hugh-Curtis syndrome. Condoms prevent sexual transmission. Erythromycin oinment prevents neonatal transmission. Treatment: ceftriaxone + (azithromycin or doxycycline) for possible chlamydia coinfection
N. meningococci
Polysaccharid capsule, maltose fermentation, vaccine (none for type B), respiratory and oral secretions, causes meningococcemia and meningitis, Waterhouse-Friderichsen syndomre, Rifampin, cirprofloxacin, or ceftriaxone prophylaxis in close contacts. Treatment: ceftriaxone or penicillin G
Haemophilus influenzae
Small gram-negative (coccobacillary) rod. Aerosol transmission. Most invasive disease caused by capsular type B. Nontypeable strains cause mucosal infections (otitis media, conjunctivitis, bronchitis). Produces IgA protease. Culture on chocolate agar requies factors V (NAD) and X (hematin) for growth; can also be grown with S. aureus, which provides factor V. Haemophilus causes epiglotitis, meningitis, otitis media, and pneumonia. Treat mucosal infections with amoxicillin +/- clavulanate. Treat meningits with ceftriaxone. Rifampin prophylaxis in close contacts. Does not cause the flu. Vaccine contains type B capsular polysaccharide conjugated to diphtheria toxoid or other protein. Given between 2 and 18 months of age.
Legionella pneumophila
Gram-negative rod. Gram stains poorly - use silver stain. Grow on charcoal yeast extract culture with iron and cysteine. Detected clinically by presence of antigen in urine. Aerosol transmission from environmental water source habitat. No person-to-person transmission. Treatment: macrolide or quinolone.
Legionnaires’ disease - sever pneumonia, fever, GI and CNS symptoms.
Pontiac fever - mild flu-like syndrome
Pseudomonas aeruginosa
Aerobic gram-negative rod. Non-lactose fermenting, oxidase +. Produces pyocyanin (blue-green pigment); has a grape-like odor. Water source. Produces endotoxin (fever, schock) and exotxoin A (inactivates EF-2). Pseudomonas is assocated with wound and burn infections, Pneumonia (especially cystic fibrosis), Sepsis, External otitis (swimmer’s ear), UTI, Drug use and Diabetic Osteomyelitis, and hot tub folliculitis. Malignant otitis externa in diabetics. Echthyma gangrenosum - rapidly progressive, necrotic cutaneous lesions caused by Pseudomonas bacteremia. Typically seen in immunocompromised patients. Treatment: aminoglycoside plus extended-spectrum penicillin (e.g. piperacillin, ticarcillin, cefepime, imipenem, meropenem).
Klebsiella
An intestinal flora that causes lobar pneumonia in alcoholics and diabetics when aspirated. Very mucoid colonies caused by abundant polysaccharide capsules. Red “currant jelly” sputum. Also cause of nosocomial UTIs.
Salmonella vs Shigella
Salmonella (have flagella, can disseminate hematogenously, have many animal reservoirs, produce hydrogen sulfide, antibiotics may prolong fecal excretion of organism, invades intestinal mucosa and causes a monocytic response, can cause bloody diarrhea, does not ferment lactose)
Shigella (no flagella, cell to cell transmission; no hematogenous spread, only reservoirs are humans and primates, does not produce hydrogen sulfide, antibiotics shorten duration of fecal excretion of organism, invades intestinal mucosa and cuases PMN infiltration, often causes bloody diearrhea, does not ferment lactose)
Salmonella typhi
Causes typhoid fever. Found only in humans. Characterized by rose spots on the abdomen, fever, headache, and diarrhea. Can remain in gallbladder and cause a carrier state.
Campylobacter jejuni
Major cause of bloody diarrhea, expecially in children. Fecal-oral transmission through foods such as poultry, meat, unpasteurized milk. Comma or S-shaped, oxidase +, grows at 42 C. Common antecedent to Guillain-Barre syndrome and reactive arthritis.
Vibrio cholerae
Produces rice-water diarrhea via entertoxin that permantly activates Gs, inc cAMP. Comma shaped, oxidase +, grows in alkaline media. Endemic to developing countries. Prompt oral rehydration is necessary.
Yersinia enterocolitica
Usually transmitted from pet feces (e.g. puppies), contaminated milk, or pork. Causes mesenteric adenitis that can mimic Crohn disease or appendicitis.