FINAL EXAM Flashcards
- requires 15-21% oxygen for growth
- i.e., Mycobacteria
obligate aerobe
- can grow with or without oxygen
- i.e., Enterobacteriaceae
facultative anaerobe
- grow in reduced oxygen (< 5%)
- i.e., Campylobacter, Neisseria, Helicobacter
microaerophilic
cannot grow in the presence of oxygen
anaerobe
- grow in the presence of 5-10% carbon dioxide
- Haemophilus, Neisseria
capnophilic
- enriched media
- contains blood cells
- used to identify hemolysis
SBA
- enriched media
- contains lysed blood cells that release NAD
Chocolate agar
- enriched media
- selective for organisms that require cysteine for growth
- Legionella, Francisella
BCYE (Buffered Charcoal Yeast extract)
- differential media for gram-negatives
- contains lactose, bile salts, neutral red indicator, crystal violet
- lactose-fermenters appear pink
MAC (MacConkey Agar)
- selective media for gram-negative coliforms
- permit isolation of stool pathogens
- differential for lactose and sucrose fermentation
- orange (non-pathogens), green/blue (pathogens)
HE (Hektoen enteric) media
- selective media for gram-negative coliforms
- permit isolation of stool pathogens
- Citrobacter, E. coli are yellow on plate
- Shigella is colorless
- Salmonella is red with black centers
XLD (xylose-lysine-deoxycholate) agar
- antibiotic media
- chocolate agar
- vancomycin, colistin, nystatin and trimethoprim
MTM (Modified Thayer Martin)
What do antibiotics in MTM inhibit?
- Vancomycin inhibits gram-positives
- Colistin inhibits gram-negatives
- Nystatin inhibits yeasts
- Trimethoprim inhibits swarmers (Proteus)
Micrococcaceae General Characteristics
- Gram-positive cocci, catalase positive, singly, pairs or in clusters
- Colony morphology: cream-colored, buttery on BA or CNA, some produce Beta hemolysis
- aerobic or falcultative anaerobes, non motile, non spore forming, non encapsulated
- gram positive cocci
- catalase +, coagulase -
- Bacitracin S
Micrococcus luteus
- gram positive cocci
- beta hemolytic
- catalase +, coagulase +
Staphylococcus aureus
- gram positive cocci
- catalase +, coagulase -
- Novobiocin R, Bacitracin R
- UTI’s in young, sexually active women
Staphylococcus saprophyticus
S. aureus Virulence factors
- coagulase, hyaluronidase, lipase
- enterotoxins (A-E, G-I, exfoliative, cytolytic)
- Protein A
Infections/Syndromes caused by S. aureus
- gastric infections and food poisoning
- LITERALLY ANYTHING
- Toxic Shock Syndrome (tampons, DIC)
- Scalded Skin Syndrome (SSS)
- Toxic Epidermal Necrolysis (TEN)
- selective for E. coli O157:H7 (doesn’t ferment sorbitol)
- looks for sorbitol fermentation
SMAC plate
Plate that X and V strips are added to for Haemophilus differentiation
Mueller-Hinton agar
- contains lecithinase and lipase
- C. perfringens is lecithinase +
- used for anaerobes
Egg Yolk Agar (EYA)
differential media for gram positives
CNA
- selective media for Vibrios
- contains thiosulfate, citrate, bile salts and sucrose
- cholera and alginolyticus turn yellow
TCBS agar
selective media for Burkholderia cepacia
OFPBL
- shows Prevotella pigment
- B. fragilis grows on it
KVLB agar
- gram positive cocci
- catalase +, coagulase -
- Novobiocin S, Bacitracin R
Staphylococcus epidermidis
- gram positive cocci
- catalase -, beta hemolytic
- PYR +, CAMP -
- Optochin R, Bacitracin S
Group A Strep (S. pyogenes)
- gram positive cocci
- catalase -, alpha hemolytic
- Optochin S, sodium desoxycholate +
Streptococcus pneumoniae
- gram positive cocci
- alpha hemolysis
- bile esculin +, 6.5% NaCl -, PYR -
Group D Strep (S. bovis)
S. pyogenes Virulence factors
- Protein M, Protein F, DNase, hyaluronidase, Streptolysin
- capsule, toxins
Infections of Group A Strep
- Streptococcus Toxic Shock Syndrome
- skin infections, scarlet fever, necrotizing fasciatus
- Complications: acute rheumatic fever, acute glomerular nephritis
- gram positive cocci
- catalase -, beta hemolytic
- PYR -, CAMP +
- Bacitracin R, hippurate hydrolysis + (+ = purple)
Group B Strep (S. agalactiae)
Group B Strep Virulence factors
- capsule
- DNase, protease
Infections of Group B Strep
- invasive disease in newborns
- endometritis and endocarditis
S. pneumo Virulence Factors
- capsule
- IgA protease, hyaluronidase
Infections of S. pneumo
#1 pneumonia agent in adults #1 agent of adult meningitis
- gram positive cocci
- catalase -, bile esculin +
- 6.5% NaCl +, PYR +
Enterococcus
- gram negative diplococci
- oxidase +, catalase +, superoxol +
- grows on MTM
- utilizes glucose
Neisseria gonorrhoeae
- gram negative diplococci
- oxidase +, catalase +
- utilizes glucose and maltose
Neisseria meningitidis
- gram negative diplococci
- oxidase +, catalase +
- grows on MTM
- utilizes glucose, maltose and lactose
Neisseria lactamica
N. gonorrhoeae Virulence Factors
pili, capsule, IgA protease
Infections of N. gonorrhoeae
- Males: dysuria, discharge, prostatitis, urethral stricture
- Females: up to 80% asymptomatic, discharge, leads to PID
- gram negative cocci, non-motile
- oxidase +
- asaccharolytic
Moraxella catarrhalis
- gram positive bacilli
- catalase +, non-motile, bile esculin -
- Babe-Ernst granules (seen on Loeffler’s serum agar)
Corynebacterium diphtheriae
C. diphtheriae Virulence factors
- diphtheria exotoxin
- test for toxin with Elek test
Infections of C. diphtheriae
Respiratory infection spread by aerosols
- gram positive bacilli
- catalase +, CAMP + (block)
- growth at 4C, tumbling motility, umbrella motility
- bile esculin +
Listeria monocytogenes
- gram positive bacilli
- catalase -, non-motile
- TSI is H2S +
- mostly occupational exposure (animals)
Eresipelothrix rhusiopathiae
- gram positive bacilli (variable)
- resemble fungi
- weakly acid fast
Nocardia
- gram positive bacilli
- agent of anthrax
- medusa head or egg whites, non-motile
- lecithinase +, string of pearls morphology
Bacillus anthracis
Infections/Virulence of B. anthracis
- virulence: exotoxin
- cutaneous anthrax
- pulmonary anthrax
- GI anthrax
- injection anthrax
- gram positive bacilli
- catalase +
- motile (flagella)
Bacillus cereus
Enterobacteriaceae General Characteristics
- ferment glucose
- reduce nitrate to nitrites
- oxidase negative (except Plesiomonas)
- motile at body temperature (except Klebseilla, Shigella, Yersinia)
- gram negative bacilli
- lactose fermenter, beta hemolytic, A/A
- ferments glucose, lactose, trehalose, and xylose
- H2S -, DNase -, urease -, PAD -, and citrate -
- IMVC (+ + - -)
E. coli
Types of E. coli GI Infections
- Enteropathogenic: infantile diarrhea (no blood)
- Enterotoxigenic: Traveler’s diarrhea (large inoculum)
- Enteroinvasive: like Shigella (large inoculum), blood, WBCs
- Enterohemorrhagic: bloody diarrhea, no WBC’s, O157:H7, Shiga-like toxin
- Enteroadherent: DAEC and EAEC (UTI’s)
- gram negative bacilli, non-motile
- lactose fermenter, A/A
- citrate +, H2S -, PAD -, urease -, MR -
- Indole - (oxytoca is +)
- IMVC - - + +
Klebsiella
- gram negative bacilli
- lactose fermenter, A/A
- citrate +, H2S -, PAD -, urease -, MR -
- IMVC - - + +
- lysine + (except cloacae), ODC +
Enterobacter
- gram negative bacilli
- NLF, citrate +, H2S -, PAD -, urease -, MR -, A/A
- IMVC - - + +
- ONPG +, slow lactose, DNase +
Serratia
- gram negative bacilli
- swarming on lab media, NLF
- PAD +, K/A H2S +
- lactose -, urease +, indole -, ODC +, MR +
Proteus mirabilis
- gram negative bacilli
- swarming on lab media, NLF
- PAD +, A/A H2S +,
- lactose -, urease +, indole +, ODC -, MR +
Proteus vulgaris
- gram negative bacilli
- lactose fermenter
- MR +, VP -, LDC -
- citrate +
Citrobacter
- gram negative bacilli
- NLF, motile
- MR +, PAD +, H2S -, citrate -
Morganella
- gram negative bacilli
- nosocomial infections, NLF, K/A
- MR +, indole +, PAD +, H2S -, citrate +
Providencia
- gram negative bacilli
- NLF, motile, K/A H2S+
- LDC +, urea -, indole +, MR +, PAD -
Edwardsiella
- gram negative bacilli
- NLF
- K/A H2S +, indole -, VP -, PAD -, urease -, MR +, citrate + (typhi is -), lysine +
Salmonella
- gram negative bacilli
- NLF, non-motile
- K/A H2S -, ONPG +, ODC +, urease -, H2S -, LDC -, MR +
Shigella
Four Types of Shigella
- (A) S. dysenteriae: most serious
- (B) S. flexneri: 2nd most common in US
- (C) S. boydii: in developing countries
- (D) S. sonnei: most common in US
- gram negative bacilli, NLF
- K/A H2S -, non-motile at 37C
- grows at cold temps
- ODC +, MR +, PAD -
- enterocolita = target growth on CIN agar
Yersinia
Lysine decarboxylase (breakdown)
Lysine –> cadaverine + CO2
Ornithine decarboxylase (breakdown)
Ornithine –> putrescine
Arginine dihydrolase (breakdown)
arginine –> citrulline –> ornithine –> putrescine
Vibrio cholera infections/symptoms
- rice water stool
- extreme loss of electrolytes
- cholera toxin
- gram negative bacilli, curved rods
- catalase +, oxidase +, reduce nitrate
- Vibriostat (0/129) S, String test +
- yellow on TCBS, grows in 0% NaCl
Vibrio cholera
- gram negative bacilli, curved rods, oxidase +
- summer diarrhea in Japan
- associated with seafood
- some are urease +
- green on TCBS
Vibrio parahaemolyticus
- gram negative bacilli, curved rods, oxidase +
- associated with raw or undercooked seafood (oysters)
- lactose + vibrio
- green on TCBS
Vibrio vulnificus
- gram negative bacilli, curved rods, oxidase +
- mimics cholera, yellow on TCBS
Vibrio alginolyticus
- gram negative rods
- oxidase +, glucose fermenting, beta hemolytic
- straight gram-negative rods, indole +
- mesophilic and psychophilic groups
- infection associated with contaminated water
- string test -, Vibriostat disk R
- growth in 0% NaCl, inositol -
Aeromonas
- gram-negative rods
- similar to Shigella, but lower virulence
- oxidase +, Vibriostat S, growth in 0% NaCl
- NLF, motile, IBB agar, ODC +, LDC +, ADH +
Plesiomonas
- gram negative rod
- microaerophilic (5% O2)
- non-spore forming rods (seagull-wing shape)
- oxidase +, catalase +
- darting motility
- # 1 cause of diarrhea world-wide
- Campy BAP, Skirrow media
Campylobacter jejuni
- gram negative rod
- urease +
- causes ulcers, leads to gastric carcinoma
Helicobacter pylori
- gram negative bacilli
- blue/green hue on MAC , grape-like odor
- oxidase +, catalase +, glucose oxidizer
- motile, capsule, DNase, protease, Exotoxin A
- growth at 42C, ADH +, citrate +, K/K
Psuedomonas aeruginosa
Psuedomonas dyes
pyocyanin (green hue) and pyoverdin (fluorescence)
Infections/syndromes of Psuedomonas
- nosocomial infections
- pneumonia in CF patients
- Hot tub syndrome
- gram negative coccobacilli
- oxidase -, catalase -, non-motile, NLF
- purple on MAC, K/K
Acinetobacter baumanii
- gram negative bacilli, rough colonies
- lavender/green hue on MAC, NLF, ammonia odor
- oxidase -, catalase +, non-oxidizer of glucose
- DNase +, Esculin +, LDC +, Gelatin +, K/K
Stenotrophomonas maltophila
- gram negative bacilli, motile
- slow oxidase +, glucose oxidizer
- ONPG +, LDC +, K/K
- OFPBL, BCSA selective agars
Burkholderia cepacia
- gram negative long/thin bacilli, non-motile
- hospital contaminants
- DNase +, oxidase +, Gelatin +, indole +
- Doesn’t grow on MAC
- meningitis and septicemia in newborns
Elizabethkingia meningoseptica
- gram negative bacilli
- smooth, brown/tan colonies on SBA, grows on Mac
- motile (polar flagella), oxidase +, H2S + on TSI
Shewanella
- gram negative bacilli
- capsule, sattelitism on BAP with S. aureus, IgA protease
- Growth on chocolate
- oxidase +, catalase +, reduce nitrate, porphyrin -
H. influenzae
Special growth requirements of Haemophilus
- X & V factor: H. influenzae, H. aegyptius, H. hemolyticus (Quad I, IV)
- V factor: H. parainfluenzae (Quad I, III, IV)
- X factor: H. ducreyi
- Mueller-Hinton agar with X and V strips
H. influenzae infections
Hib is the #1 cause of meningitis is unvaccinated children
- gram negative bacilli
- fastidious, requires X factor, no growth on MAC
- pits agar, K/K, oxidase +, catalase -
- bleach-like odor
Eikenella
- gram negative bacilli, non-motile
- oxidase +, catalase -, A/A
- growth on MTM
- affects bones/joints of kids
Kingella
- gram negative coccobacilli, STI
- painful chancroid, buboes
- school of fish morphology
Haemophilus ducreyi
HACEK (organsims)
- Haemophilus (Aggregatibacter aphrophilus)
- Actinobacillus/Aggregatibacter actinomycetemcomitans
- Cardiobacterium hominis
- Eikenella corrodens
- Kingella species
HACEK (characteristics)
- Don’t grow on MAC, slow growers
- usually bite wounds (mouth flora)
- cause endocarditis
- gram negative bacilli, non-motile
- fastidious, no growth on MAC
- bipolar staining, zoonotic infections
- catalase +, oxidase +
Pasteurella multocida
- gram negative bacilli
- oxidase +, catalase +, H2S +
- urease +
- undulant fever, zoonotic infections
- specimen: bone marrow*, test with growth in dyes
Brucella
- gram negative bacilli
- requires cysteine for growth (BCYE agar)
- no growth on MAC, SBA
- zoonotic infections, rabbit fever, tularemia
Francisella tularensis
- gram negative bacilli (weakly staining)
- require cysteine for growth (BCYE agar)
- inhalation transmission, associated with AC units
- invades and survives in macrophages
- Pontiac fever
Legionella pneumophilia
Bordetella virulence factor
pertussis toxin
- gram negative bacilli
- smooth, silver pinpoint colonies (like mercury droplets)
- whooping cough
- specimen collected on Dacron swab
- use Bordet-Gengou agar
Bordetella pertussis
- can survive some oxygen exposure, but can’t perform metabolic processes unless in an anaerobic environment
- i.e., Bacteroides
Aerotolerant anaerobe
- requires strict anaerobic environment and are killed almost immediately in the presence of oxygen
- i.e., Clostridium
Obligate anaerobe
Exogeneous anaerobes
- exist outside of the body
- infection develops at site of entry
- i.e., Clostridium, Fusobacterium
Endogeneous anaerobes
- exist inside the body
- source of infection in/throughout body
- usually polymicrobial infections
- i.e., Bacteroides, Peptostreptococcus, Actinomyces
Skin anaerobes
Propionibacterium, Peptostreptococcus
Respiratory anaerobes
Prevotella, Porphyromonas, Fusobacterium
GI anaerobes
Bacteroides, Clostridium
Genitourinary tract anaerobes
Fusobacterium, Prevotella, Bacteroides, Lactobacillus
- gram positive bacilli, anaerobic
- spore forming, boxcar-shaped
- double zone of beta hemolysis
- lecithinase +, catalase -
- causes gas gangrene (alpha toxin)
Clostridium perfringens
- gram positive bacilli, anaerobic
- spore forming, heavily swarming
- terminal spores
Clostridium tetani
- gram positive bacilli, anaerobic
- spore forming, heavily swarming
- subterminal spores
Clostridium septicum
Clostridium botulinum infection
- causes botulism
- botulism toxins
- due to ingestion of toxin, ingestion of spores in infants
- antibiotic-associated diarrhea
- causes pseudomembranous colitis (blood, necrosis)
- nosocomial infection, CCFA agar
- fluoresces chartreuse
Clostridium difficile
- gram positive bacilli, anaerobic
- normal skin flora
- causes granulomatous disease (“sulfur granules”)
- molar tooth colonies, infected IUD’s, lumpy jaw
Actinomyces
- gram negative bacilli, anaerobic
- # 1 flora in colon
- grows on KVLB and BBE
- bile tolerant
- pilonidal cysts
Bacteroides fragilis
- gram negative bacilli, anaerobic
- # 1 gingiva flora
- brick-red fluorescence on KVLB (then turn black)
Prevotella melaninogenica
- gram negative bacilli, anaerobic
- slow grower
- brick-red fluorescence on KVLB (then turn black)
Porphyromonas
- gram negative bacilli, anaerobic
- long, thin rods with pointed ends
- fluoresces chartreuse
- nonhemolytic, indole +, lipase -
Fusobacterium nucleatum
- gram negative bacilli, anaerobic
- pleomorphic with rounded ends
- fluoresces chartreuse
- beta hemolytic, indole +, lipase +
Fusobacterium necrophorum
- gram positive cocci, anaerobic
- common contaminant, skin flora
- catalase -, indole -, urease -, nitrate -, SPS S
Peptostreptococcus anaerobius
- gram positive cocci, anaerobic
- common contaminant, skin flora
- catalase -, indole -, urease -, nitrate -, SPS R
Peptostreptococcus asaccharolyticus
Anaerobic specimen collection
- aspirates collected and transported on PRAS media
- use methanol as fixative for gram staining instead of heat
Vancomycin disk susceptibility
- S: Porphyromonas, Clostridium
- R: Bacteroides, Prevotella, Fusobacterium
Kanamycin disk susceptibility
- S: Bacteroides, Fusobacterium
- R: B. fragilis, Prevotella, Porphyromonas
Colistin disk susceptibility
- S: Prevotella, Fusobacterium, Bacteroides
- R: B. fragilis, Porphyromonas, Clostridium
- helical shapted
- Borrelia, Treponema, Leptospira
- mostly identified by serology
- Fletcher’s media, Stuart’s media, Kelly medium
Spirochetes
- obligate aerobic helical rods
- tightly coiled, thin, flexible
- 20 serovars
- reduced phagocytosis, hemolysis and endotoxins
- disease associated with contact with infected urine
- zoonotic disease
Leptospires
- loosely coiled
- arthropod borne (tick)
- microaerophilic
- causes relapsing fever
- can get Jarisch-Herxheimer reaction (death of spirochetes can cause sudden endotoxin release)
Borrelia recurrentis
- Lyme disease
- transmitted via tick bite (Ixodes)
- complement evasion (binds factor H)
Borrelia burgdorferi
Treponema virulence
- penetrate intact mucous membranes
- crosses placenta
- dissemination
- antigenic variation
Syphilis infection
- Primary: PAINLESS chancre at site of infection
- Secondary: widespread macular rash
- Latent: non-infectious, detect with serology
- Late: CNS disease, congenital syphilis
Congenital syphilis (symptoms)
- non-immune hydrops (placental disease causing fetal death)
- bone lesions, hepatosplenomegaly
- visible deformities (tibias, Hutchinson’s teeth)
- obligate intracellular parasite
- EB and RB stages of life cycle
- most common STI in the US
- can cause Reiter syndrome, Lymphogranuloma venereum
Chlamydia trachomatis
Chlamydia infection in men
- Nongonococcal urethritis (NGU), Prostatitis, Epididymitis
- urethral discharge, dysuria, hemospermia, conjunctivitis
- usually asymptomatic
Chlamydia infection in women
- Cervicitis, endometritis, salpingitis, PID, perihepatitis, conjunctivitis
- vaginal discharge, dysuria, pain bleeding
- usually asymptomatic
- infertility and sterility
Chlamydia infection in neonates
- conjunctivitis, nasopharyngeal infection, pneumonia
- otitis media (less common)
- erythromycin eyedrops at birth to take care of infections
- bird chlamydia
- parakeets, turkey
- PARROT FEVER (psittacosis)
- pneumonia in humans
- survive in macrophages or epithelial cells
- bird-to-bird transmission
Chlamydia psittaci
- ROCKY MOUNTAIN SPOTTED FEVER
- transmitted via ticks
- flulike symptoms
- rash spread over ankles, wrists, hands and soles of feet
- NO RASH ON THE FACE
Rickettsia rickettsii
- ticks and dogs
- rash over palms, soles of feet, body and ON THE FACE
- taches noires (black spots at primary site of infection)
Mediterranean spotted fever (Boutonneuse Fever)
- endemic typhus (murine typhus)
- oriental rat flea, cat flea
- infection when flea defecates on skin and scratching infects the bite
Rickettsia typhi
- epidemic louse-borne typhus
- known as BRILL-ZINSSER DISEASE
- areas of sanitation disruption
- defecates in bite wound via scratching
- rash over the body and FACE
Rickettsia prowazekii
- transmitted via mouse mite
- rash on face, trunk and extremities, but NOT ON PALMS OR SOLES
Rickettsia akari
- scrub typhus
- transmitted via chigger, reservoir is rat
- tache noire at site of infection
- rash not on palms, soles of feet or the face
Orientia tsutsugamushi
- dogs infected with brown dog ticks
- rickettsial-like inclusions IN MONOCYTES (HME)
- Lone star tick (A. americanum)
Ehrlichia chaffeensis
- Q FEVER (Query fever)
- potential bioterror agent
- goats, cattle, sheep
- influenza-like illness
Coxiella burnetii
- don’t have cells walls (CWD)
- slow growing, fastidious
- Fried Egg appearance
Mycoplasma
- adhere to epithelium of mucosal surfaces in oropharynx
- bronchitis, pharyngitis
- WALKING PNEUMONIA
- isolation indicates pathogenicity
- spread via close contact (dorms, military, prison)
- epidemics can occur
Mycoplasma pneumoniae
- adhere to epithelium of urogenital tract
- normal flora, opportunistic
- salpingitis, pyelonephritis, PID, postpartum fevers
- requires arginine (pink)
- plate to A8 agar
Mycoplasma hominis
- urogenital tract infection
- normal flora of lower urinary tract in women
- upper urinary tract infection
- significant infection to fetus: chorioamnionitis (infection of placental membrane), congenital pneumonia, chronic lung disease, meningitis
- 10% NGU in men
- requires urea (pink)
Ureaplasma urealyticum
- slender, slightly curved or straight rods
- non-motile, non-spore-forming
- cell wall has high lipid content (mycolic acid)
- acid-fast (resist gram stain), SPUTUM SPECIMEN IS BEST
- aerobic
- require complex media
- slow growers (2-6 weeks)
Mycobacteria
Two major Mycobacteria groups
- M. tuberculosis complex
- Nontuberculous mycobacteria (NTMs)
- transmitted by airborne droplet
- bacteria phagocytosed and multiply intracellularly
- slow growing
- raised, dry, rough colonies
- nonpigmented, non-chromogen
- cord factor (inhibits PMN migration)
- grow best at 35-37C
- positive niacin accumulation, reduces nitrate
Mycobacterium tuberculosis
TB treatment
- 9 month course of therapy
- isoniazid and rifampin
- maybe streptomycin or ethambutol
- drug combo if resistant
- most common NTM causing TB in the US
- slow growing, nonpigmented, non-chromogen
- soil and water sources
- disease in swine and poultry
- large increase in number of isolates due to HIV patients developing AIDS (most common systemic disease)
- produces heat-stable catalase
M. avium complex (MAC complex)
M. avium and M. intracellulare
- Johne’s disease
- niacin -, nitrate -, tween 80 -
M. paratuberculosis
- 2nd most common NTM causing lung disease in US
- CHRONIC PULMONARY DISEASE
- photochromogenic, slow grower
- catalase +, Tween 80 +
- reduces nitrate
- pyrazinamidase production
M. kansasii
- photoreactivity
- carotene pigment upon exposure to light
photochromogens
- photoreactivity
- produce a pale yellow to orange color in dark or light
scotochromogens
- photoreactivity
- buff or lack of color
non-chromogens
- disseminated AIDS infection
- enteritis, genital, soft tissue
- SQ and heat-stable catalase +
- pyrazinamidase +, urease +
- slow grower, non-chromogen
M. genavense
- cutaneous infections
- due to contact with saltwater or inadequately chlorinated freshwater
- “swimming pool granuloma”
- slow grower, photochromogen
M. marinum
- cervical lymphadenitis in kids
- smooth colonies with dense centers
- light yellow to deep orange pigment
- Tween 80 -, don’t reduce nitrate
- urease +, high SQ catalase
- slow grower, scotochromogen
M. scrofulaceum
- pulmonary disease
- phochromogenic, slow grower
- niacin +
M. simiae
- pulmonary disease
- similar to TB
- slow grower, scotochromogen
- Tween 80 +, reduces nitrate
- inhibited by NaCl
M. szulgai
- nodule developing into a severe shallow ulcer
- nonchromogenic buff colonies, slow grower
- heat-stable catalase
M. ulcerans
- hot and cold-water taps, birds
- pulmonary infection in patients with predisposing condition
- scotochromogenic, slow grower
- niacin - , reduce nitrate
M. xenopi
- tap water and soil sources
- “tap water” bacillus
- slow grower, scotochromogen
M. gordonae
- rapid growing, non-chromogen
- M. abscessus group
- opportunistic
- more drug resistance than M. fortuitum
- positive 3-day arylsulfatase test
- growth on Mac without crystal violet
- doesn’t reduce nitrate
M. chelonae
- rapid growing, non-chromogen
- localized infections of skin and soft tissues
- positive 3-day arylsulfatase test
- reduces nitrate
M. fortuitum
- rapid growing, scotochromogen
- rare cases of pulmonary, skin, soft tissue and bone infections
- nonpigmented colonies
- negative arylsulfatase test
- reduces nitrate
- grows on Mac w/o crystal violet
M. smegmatis
- Hansen’s disease
- person-to-person transmission
- low infection rate
- can’t be cultured in vitro
- armadillos and foot pads of white mice
M. leprae
Digestion/decontamination of specimens
- liquefy sample through digestion of proteinaceous material
- allow chemical decontaminating agent to contact and kill the non-mycobacterial organisms (sodium hydroxide and N-acetyl-L-cysteine)
- mycobacteria survive chemical treatment because of high lipid content in cell walls
- liquefying mucin allows mycobacteria to utilize nutrients
Specimens that need digestion and decontamination
sputum, gastric washing, BAL, bronchial washings, transtracheal aspirate
Specimens that only need decontamination
urine, autopsy tissue, fluids
Specimens that don’t need digestion and decontamination
CSF, pleural fluid, joint fluid
Acid-fast staining
- Ziehl-Neelsen (uses heat)
- KINYOUN***(no heating)
- red organism with blue background
Mycobacteria culture media
- Lowenstein-Jensen (LJ) media: eggs, salts, glycerol, potato flour, MALACHITE GREEN
- Middlebrook media
- all media contain malachite green (suppress gram-positive)
BACTEC system principle
amount of labeled CO2 liberated is detected by instrument and is interpreted as “growth index”
Mycobacteria ID
- acid fast organism
- colony morphology (smooth and soft or rough)
- growth rates: rapid growers (7 days)
- photoreactivity
- niacin accumulation (most +)
- nitrate test (M. kansasii and tuberculosis +)
- catalase (M. tb, bovis are -)
- Tween 80 hydrolysis
- arylsulfatase test
- pyrazinamidase (distinguishes M. marinum (+) from kansasii (-) and M. bovis (-) from tb (+))
- Tellurite reduction (MAC +)
- urease (M. scrofulaceum (+) and gordonae (-))
MTB ID
MTB is only non-pigmented producer that is nitrate + and niacin + in culture media
Colony count using 0.001 loop
75 colonies seen =
75,000 CFU/mL
100,000 is significant; ID and perform sensitivity
Common causes of UTI’s
- S. saprophyticus (young, sexually active females)
- Enterococcus, Enterobacteriaceae, S. aureus, S. epidermidis
Upper vs. Lower UTI’s
- Upper: infection of renal parenchyma or ureters
- Lower: infection of bladder, urethra or prostate (males)
Complicated vs. Uncomplicated UTI’s
- Complicated: UTI’s due to one or more structural or functional abnormalities
- Uncomplicated: UTI’s in sexually active women with normal GU tracts; no procedures
UTI Risk factors
- AGE (kids with long-term medical problems, adults with GU abnormalities, elderly because of catheterization, prostate problems, bladder prolapse)
- INSTITUTIONALIZE CARE (catheters, asymptomatic bacteruria)
- PREGNANCY (increased ASB)
- RENAL TRANSPLANT (leads to septicemia)
- BLADDER CATHETERIZATION
UTI Routes of Infection
- Ascending route: most common route, migration from bladder, acquired UTI’s
- Descending route: infection via blood, < 5% of UTI’s
Infections in urine cultures that aren’t UTI’s
- Herpes simplex (HSV)
- Chlamydia trachomatis
- Neisseria gonorrhoeae
Specimen type/collection and processing
- Voided midstream specimen collection (CCMS)
- Catheterized specimen collection (best for kids)
- Suprapubic aspiration (for anaerobes)
- 24hr old specimen is unacceptable
- first morning urine is best
- processed within 2 hours
- REFRIGERATE if not processed immediately
Culture with 3 or more uropathogens
contamination
Reportable results
- a pure culture of S. aureus is significant
- yeast in any culture is significant
- unusual organisms are reportable
- only report significant UTI antibiotics
Acceptable sputum smear
< 10 epithelial cells
> 25 PMN’s
Pharyngitis pathogens
- LOOK FOR GROUP A STREP
- viral with rhinorrhea
- C. diphtheriae
Otitis media pathogens
Streptococcus pneumoniae, Haemophilus influenzae
Epiglottitis pathogens
Streptococcus, Staphylococcus, Hib***
Pertussis pathogens
Bordetella
Health-care-acquired pneumonia pathogens
Pseudomonas, Kleb. pneumoniae, Acinetobacter, MRSA GNR’s, S.pneumo, H. influenzae, Legionella
Causes of pneumonia in CF patients
Pseudomonas aeruginosa, Burkholderia cepacia
Main Respiratory Tract Pathogens
Pseudomonas aeruginosa, Burkholderia cepacia, S. pneumoniae, Klebsiella pneumoniae, S. pyogenes, H. influenzae
Bronchitis pathogens
viruses, H. influenzae
Community-acquired pneumonia pathogens
- Kids: viruses, H. parainfluenzae, Mycoplasma pneumoniae
- Adults: S. pneumoniae, Hib, M. pneumoniae
Lower Respiratory Tract pathogens
- neonates: Chlamydia trachomatis
- infants: viruses
- 5-18 months: S. pneumo, H. influenzae
- 3-19 years: viruses, S. aureus, M. pneumoniae
- 18-45 years: M. pneumoniae
- older: S. pneumoniae, Legionella
- hospitalized: GNR’s, S. pneumoniae, S. aureus
Respiratory Tract pathogens with capsules
S. pneumoniae, H. influenzae, Pseudomonas aeruginosa, Klebsiella pneumoniae
Sinusitis pathogens
- complication of URT infection
- viruses (> 90%)
- S. pneumo, H. flu, S. pyogenes, M. catarrhalis, S. aureus
Empyema
- collection of purulent fluid in pleural cavity
- S. aureus, S. pneumo, S. pyogenes
- rapid onset, lack of fever
- absence of blood or pus
- large number of watery stools (> 20/day)
- Enterotoxigenic E. coli, cholera, S. aureus, C. perfringens, Bacillus cereus, viral or parasitic
Enterotoxin-mediated diarrhea
- fecal leukocytes, RBC’s, fever
- Salmonella, Campy, Shigella, E. coli, Entamoeba
Diarrhea mediated by invasion of bowel mucosal surface
- S. typhi, Y. enterocolitica
- presents with constipation
- fecal leukocytes and RBC’s
Diarrhea mediated by invasion of full thickness of bowel with lymphatic spread
- presence of viable bacteria in the blood
- one of the most serious infections
Bacteremia
bacteremia present with the absence of physical signs/symptoms
Occult Bacteremia
Bacteremia plus clinical presentation of signs/symptoms of bacterial invasion and toxin production
Septicemia
Systemic response to infection sometimes accompanied by organ involvement
Sepsis
Sepsis accompanied by hypotension
Septic shock
Primary vs. Secondary Bacteremia
- Primary: arises from endovascular source
- Secondary: arises from exovascular source
Common blood culture contaminants
Propionibacterium, S. epidermidis, Corynebacterium
Transient vs. Intermittent vs. Continuous Bacteremia
- Transient: result of procedure, caused by flora
- Intermittent: result of abscess (meningococcemia, gonococcemia)
- Continuous: intravascular source, continuously present, i.e., prosthetic heart valve
Common Bacteremia causes
Candida albicans, Malassesia furfur, E. coli, P. aeruginosa, S. aureus, CoNS, Enterococcus, S. pyogenes, Aeromonas hydrophila, Acinetobacter baumanii, Kleb pneumoniae
- anticoagulant in blood culture bottles
- neutralizes bactericidal activity (complement)
- prevention of phagocytosis
- inactivation of certain antimicrobial agents (streptomycin, gentamycin)
- may inhibit Peptostreptoccus, N. gonorrhoeae, N. meningitidis, Gardnerella)
Sodium polyanethol sulfonate (SPS)
BacT/ALERT principle
measures CO2 derived pH changes by colorimetric sensor
Contamination vs. Pathogen
- S. aureus, E. coli, Enterics, S. pneumo, P. aeruginosa, Candida are almost always true pathogens
- CoNS, diptheroids, skin flora should be questioned
- more than one bottle growing the same thing usually indicates significance
- growth of skin biota in single bottle usually indicates contaminant
- infection in/around hair follicles
- S. aureus is most common
- Enterobacteriaceae
- Pseudomonas aeruginosa associated with hot tubs
folliculitis, furuncolosis, carbuncles
- dermatophyte fungi, Trichophyton, Epidermophyton, Microsporum
- ringworm
- tinea pedis: athlete’s foot
- tinea versicolor: Malassezia furfur
- erythrasma (chronic infection)
Infection of keratinized layer
Hidradentis suppurativa
- obstructed apocrine gland (sweat gland)
- S. aureus, anaerobic strep, Bacteroides
Cellulitis
- usually in extremities, usually staph or strep
- anaerobic and aerobic mixed
- E. coli, Enterics, S. aureus, Strep, Anaerobes
Necrotizing fasciitis pathogens
- Group A Strep, S. aureus, Bacteroides, Clostridium
- Gas gangrene: C. perfringens
Myositis pathogens
- extensive muscle involvement
- S. aureus, Clostridium, Group A and B strep
- Virbio vulnificus, Aeromonas, other wound-associated bacteria
Wound infection pathogens
- human bite wounds: mouth flora, strep, S. aureus, Eikenella, Anaerobes
- animal bite wounds: pasteurella, strep, staph, anaerobes
- burns: Pseudomonas, S. aureus, Enterobacter, E. coli
Lymphadenitis pathogens
M. scrofulaceum, Group A strep, S. aureus, Y. pestis
Skin infections associated with Bacteremia
- Petechiae with meningococcemia
- Cutaneous ulcers and Vibrio vulnificus sepsis
- S. aureus, P. aeruginosa, M. leprae, T. pallidum, Rickettsia
CSF functions
- cushions brain
- reduces effective weight by factor of 30
- supplies metabolites
- removes wastes
Routes of CSF Infection
- Hematogenous spread (most common)
- Direct spread from infected site (close to or contiguous with CNS - sinusitis, etc.)
- Anatomic defects in CNS (surgery, trauma, abnormalities)
- Direct intraneural (along nerves to brain): least common
CSF Infections
- Meningitis: infection within subarachnoid space or leptomeninges
- Encephalitis: inflammation of brain parenchyma
- Meningoencephalitis: meningitis and encephalitis
Acute Bacterial Meningitis Pathogens
S. pneumoniae, N. meningitidis, H. influenzae, S. agalactiae, E. coli, Elizabethkingia meningosepticum, Listeria monocytogenes, GNR’s
Newborn meningitis
GNR’s, Group B Strep, Listeria monocytogenes
Infant meningitis
Group B Strep, H. influenzae, S. pneumoniae, N. meningitidis, E. coli
Kids/Young Adult meningitis
S. pneumoniae, N. meningitidis, Group B strep, S. aureus, Enterobacter
Adult meningitis
S. pneumoniae, N. meningitidis, Listeria, GNR’s
CSF Shunt Infections
- 2/3 of infections are Staph
- rest are gram negatives (E. coli, Kleb, Proteus, Propionibacterium)
- Candida
Meningoencephalitis pathogens
- most commonly caused by viruses
- involvement of cerebral cortex
Brain abscess pathogens
- commonly bacterial
- Streptococcus
CSF Fluid Collection and Processing
- 3-4 tubes collected
- first tube is most contaminated
- tubes 2 and 3 go for cell count and differential
- other tubes go to micro and chemistry
- CSF must be hand-delivered immediately
- NEVER refrigerate CSF
Pleural fluid (location and organisms)
- between lungs and chest wall
- organisms isolated here are pneumonia associated
- S. pneumoniae, S. aureus, H. influenzae, Enterobacteriaceae, Pseudomonas, TB
Transudate vs. Exudate
- Transudate: fluid passed through membrane or extracted from tissues (renal, hepatic or cardiac disease)
- Exudate: fluid from blood vessels in adjacent spaces; associated with infection (contains WBC’s); bloody fluid associated with malignancy
Peritoneal fluid
- between abdominal wall and organs
- small amount of fluid is normal
- ascites: increased fluid in cavity due to infection or inflammation containing WBC’s and proteins
Peritoneal fluid organisms
- gain entry via bowel perforation, surgery, trauma
- kids: S. pneumoniae, Group A strep, Enterics, Staph
- adults: E. coli, S. pneumoniae, Group A strep, N. gonorrhoeae, Chlaymdia (sexually active women)
Intra-abdominal abscess pathogens
- Anaerobes (B. frag, Clostridium)
- Enterobacteriaceae
- Strep, Enterococcus
Peritoneal dialysis fluid pathogens
- S. epidermidis and S. aureus (most common)
- Strep, GNR’s, etc.
Pericardial fluid
- between epicardium and pericardium
- normally clear fluid
- organisms: Viruses (usually), parasites, bacteria, fungi, non-infectious fluid buildup
Synovial fluid
- joint fluid
- secondary to hematogenous spread of bacteria
- bone infection, PROSTHETIC DEVICES
Synovial fluid pathogens
- S. aureus (70%)
- N. gonorrhoeae in young adults
- H. influenzae in kids < 2
- Group A and B strep, S. pneumoniae, B. fragilis, Fusobacterium
Pathogens of prosthetic devices
- gain entry during surgery
- S. epidermidis, CoNS, Corynebacterium, Propionibacterium, S. aureus
Osteomyelitis
- # 1 cause is S. aureus secondary to bacteremia
- Salmonella, Haemophilus
- hematogenous spread: S. AUREUS, Salmonella, Haemophilus, Pseudomonas, Enterics
- Bites: Eikenella, Pasteurella
VP + Enterobacteriaceae
Klebsiella, Enterobacter, Hafnia, Serratia
H2S + Enterobacteriaceae
Salmonella, Proteus, Edwardsiella, Citrobacter
PAD + Enterobacteriaceae
Proteus, Providencia, Morganella
Primary Genital Pathogens
- Chlamydia trachomatis
- Neisseria gonorrhoeae
- Treponema pallidum
- Human papilloma virus
- Gardnerella vaginalis
- Trichomonas vaginalis
- Haemophilus ducreyi
- Klebseilla granulomatis
- Mycoplasma hominis
- Ureaplasma urealyticum
- cervicitis with mucus
- PID leading to infertility
- Preterm births
Chlamydica trachomatis
- may be #1 STI
- penile malignancy, cervical carcinoma
- genital warts
Human Papilloma virus (HPV)
- bacterial vaginosis, premature labor, low birth rates
- CLUE CELLS (epi’s surrounded by bacteria)
- KOH + (Whiff test)
Gardnerella vaginalis
- questionable role in genital infection
- PID, pyelonephritis
- postpartum fever with morbidity and still births
Mycoplasma hominis
- acute non-gonococcal urethritis
- acute urethral syndrome in females with reproductive failure
Ureaplasma urealyticum
Cervicitis
- inflammation of cervix
- increased number of PMN’s
- N. gonorrhoeae, C. trachomatis
Anorectal lesions
- HPV, HSV, viruses, parasites, N. gonorrhoeae, C. trachomatis
- itching, discharge, anal pain
- proctitis: N. gonorrhoeae and C. trachomatis
- due to HIV in ICP’s
Pelvic Inflammatory Disease (PID)
- cervical microorganisms travel to endometrium, fallopian tubes and other pelvic structures
- N. gonorrhoeae and C. trachomatis
- associated with IUD’s (Actinomyces)***
Chorioamnionitis
- infection of uterus and contents during pregnancy
- anaerobes, genital mycoplasmas, Group B strep, E. coli
Gonococci media
Modified Thayer Martin, New York City agar, JEMBEC
Herpes Simplex Virus (HSV)
- genital herpes
- extensive PAINFUL vesicles
- culture and look for cytopathogenic effect
- DFA, EIA, PCR
Gram positive cell wall
- thick peptidoglycan layer
- penicillin and vancomycin effective
Gram negative cell wall
- thin peptidoglycan layer surrounded by membrane
- substances cannot diffuse into cell wall (Vanco’s too large)
- substances must be actively transported into cell
Antibiotic target sites
- cell wall integrity
- cell membrane structure and functions
- inhibition of protein synthesis
- inhibition of nucleic acid mechanisms
- inhibition of essential metabolites
Intrinsic resistance
- natural resistance
- all gram negatives resistant to vancomycin
- anaerobes resistant to aminoglycosides
Acquired resistance
- low affinity target sites (drugs bind poorly)
- reduced permeability
- efflux pumps
- bypass mechanisms
- enzymatic inactivation
Resistance expression
- constitutive: constantly expressing resistance
- inducible: only when exposed to agent
- constitutive-inducible: constantly expressed at low levels
- homogeneous: entire population is resistant
- heterogeneous: some of population is resistant (i.e. MRSA)
Full susceptibility testing required for
- Staphylococci
- S. pneumoniae
- Viridans strep (if from normally sterile site)
- Enterococci
- Enterobacteriaceae
- Pseudomonas aeruginosa
Beta lactamase testing required for
- H. influenzae
- N. gonorrhoeae
- Moraxella catarrhalis
- Anaerobes
Penicillins mechanism of action
Inhibits cell wall synthesis by binding PBP
Penicillin (resistance mechanisms)
- Beta lactamase
- low-affinity PBP (MRSA mecA gene)
- ESBL’s (Extended Spectrum Beta Lactamases)
Glycopeptides: drugs and mechanism of action
- Vancomycin
- Inhibits cell wall synthesis
- gram negatives intrinsically resistant
- treats MRSA, C. difficle
Aminoglycosides: drugs and mechanism of action
- Gentamycin, Tobramycin, Amikacin
- protein synthesis inhibitor acts on 30S ribosome component
- has toxic side effects (only for serious GN infections)
Tetracyclines: drugs and mechanism of action
- tetracycline, doxycycline
- protein synthesis inhibitor acts on 30S ribosome component
- broad spectrum
- leads to discolored teeth, yeast infection
Chloramphenicol mechanism of action
- protein synthesis inhibitor acts on 50S subunit
- can cause aplastic anemia
Macrolides: drugs and mechanism of action
- erythromycin, clindamycin
- protein synthesis inhibitor acts on 50S subunit
- D zone test for inducible resistance
Quinolones: drugs and mechanism of action
- Ciprofloxacin, Norfloxacin
- inhibits DNA synthesis
- broad spectrum for P. aeruginosa and aerobes
Sulfonamides: drugs and mechanism of action
- Sulfamethoxazole
- inhibits folic acid synthesis (DNA synthesis)
Mycobacterial Chemotherapy: drugs and mechanism of action
- Isoniazid
- inhibits synthesis of mycolic acid
Susceptibility testing
- disk diffusion or dilution
- no susceptibility on organisms from normal site
- gram-positive battery, gram negative battery, sometimes UTI battery
lowest concentration of antibiotic that visibly inhibits growth of an organism
Minimum inhibitory concentration (MIC)
lowest concentration that results in the death or killing of > 99.9% of test organism
Minimum bactericidal concentration (MBC)
An interpretive category that indicates an organism that is inhibited based on the amount of drug safely achieved in patient
Susceptible or Sensitive
represents an organism that may require a higher dose of antibiotic for a longer period of time to be inhibited
Intermediate
indicates an organism that is not inhibited by the recommended dose or achievable level
Resistant
Preparation of Inoculum
Mcfarland standards used
Standardized methods
- Micro dilution method (automated)
- Disk diffusion method (antibiotic disks, Kirby Bauer)
- Broth dilution method (antibiotic in broth)
- Agar dilution method (antibiotic in agar)
- Special methods (beta lactamase)
Breakpoint MIC panels
- point where effective tissue concentration can be achieved for bacterial inhibition
- results based on two wells (both turbid = R, one = I, none = S)
Meuller-Hinton Agar (QC and troubleshooting) – Kirby Bauer method
- QC weekly with E. coli, S. aureus, P. aeruginosa
- too thin agar = increased zones for disks
- too thick agar = decreased zones for disks
- decreased pH of agar = decreased activity
- increased cation concentration = decreased aminoglycoside and tetracycline activity
Disk Diffusion Test Interpretaion
- Susceptible: large zone and low MIC
- Resistant: small zone and high MIC
Broth Dilution Test
tests for bactericidal activity
E Test
- used for testing uncommon drugs for an organism or new drugs
- used for S. pneumo and fastidious organisms
Normal eye flora
S. epidermidis, Lactobacillus
Blepharitis
- infection of the eyelid margins
- redness, burning, irritation, itching
- Chlamydia trachomatis (leading cause of blindness)
- S. aureus, S. epidermidis, viruses
Conjunctivitis
- most common eye infection
- inflammation, swelling
- H. aegyptius, Chlamydia trachomatis, gonococcal, Moraxella, M. TB, F. tularensis, T. pallidum, Y. enterocolitica, viruses
- adults: S. pneumoniae, S. aureus, S. epidermidis
- kids: H. influenzae, S. pneumoniae, S. aureus
Keratitis
- inflammation of the cornea
- S. aureus, S. pneumoniae, P. aeruginosa, Acinetobacter, T. pallidum, fungi (Fusarium, Acanthamoeba)
Keratoconjunctivitis
N. gonorrhoeae, Chlamydia trachomatis
Endophthalmitis
- infection of ocular cavities
- S. aureus, S. epidermidis
- C. albicans
Dacryocystitis
- inflammation of lacrimal sac
- S. pneumoniae, S. aureus, S. pyogenes, H. influenzae
Dacryoadenitis
- acute inflammation of the lacrimal gland
- S. pneumoniae, S. aureus, S. pyogenes
Periocular infections
- acute infection of orbital contents
- S. aureus
- H. influenzae in kids
- Mucormycosis in diabetics
Eye Infection Specimen
- sterile swab from BOTH EYES
- gram stain
- inoculate plates
Normal ear flora
Pneumococci, Propionibacterium acnes, S. aureus, Enterobacteriaceae, P. aeruginosa, Candida
External Ear Infections
- S. aureus causes pustules
- Group A strep causes erysipelas
- swimmers ear caused by P. aeruginosa**
Middle Ear Infections
- kids: Pneumococci, H. influenzae, Group A Strep
- M. catarrhalis, viruses
Sinusitis
- maxillary sinusitis is most common
- H. influenzae, S. pneumoniae, Group A Strep, Moraxella
Neck infections
normally oral flora is responsible
Stomatitis
- inflammation of mucus membranes of oral cavity
- herpes simplex virus
Thrush
Candida
Parotitis
S. aureus
Dental infections
usually anaerobes