MUST TO KNOW IN BACTERIOLOGY Flashcards
Germicidal
Soap
Catalase (+)
L. monocytogenes
3 C’s: Chicken, Coleslaw, Cheese
L. monocytogenes
Chlamydia
When delayed:
Freezing:
4’C
-20’C
Phenotypic
BAP
Gram (+) colonies: Dry, white, sometimes gray
Gram (-) colonies: Gray and moist
BAP
Destroyed by chlorine
M. gordonae
Utilizes 1N HCl
DNase test
For Gram (+)
For nonfermentative
For Enterobacteriaceae
LOA test
Inactivates HBV (10mins) and HIV (2mins)
Na hypochlorite
Inoculating needles:
(?) = F(+) on oxidase test
Not longer than (?)
Nichrome
5cm
Wire loop:
(?) diameter
(?) urine
2mm
0.001mL
Significant for UTI
50k CFU/mL
Pregnant
↑ C. albicans
↑ Lactobacillus
Carrier state
Lawn a culture
Toxic to Neisseria
Good for virus
Cotton swab
Removes the toxin inoculated by cotton
Charcoal
Basis of identifying organisms
Gram stain and colonies
Phenotype
Not Gram stained
Stool
Most definitive method of identification
PCR
Basis of serotyping
Somatic antigen
Father of microbiology
Microscopist
1st to describe bacteria
Anton van Leeuwenhoek
Germ theory: relationship of organisms to human disease
Robert Koch
Father of Modern Microbiology
Louis Pasteur
1st to use dyes for stain
Ehrlich
Bacteria Ave. size:
Reproduction:
0.4-2μm Binary fission (two-fold increase)
Cell wall:
(murein)
: wall less G(+)
: wall less G(-)
Peptidoglycan
Protoplast
Spheroplast
Thick peptidoglycan
Gram (+)
Teichoic acid
Gram (+)
Thin peptidoglycan
Gram (-)
LPS (Lipid A – exotoxin)jsjsb
Gram (-)
Somatic antigen
Gram (-)
Site for energy synthesis (ATP)
Plasma membrane
Osmotic/permeability barrier
Plasma membrane
Nucleoid
Chromosome:
Plasmid:
dsDNA
Extrachromosomal DNA
Carries the antibiotic-resistance gene
Plasmid
Drug-resistance
Chromosome and plasmid-mediated
Food reserves
Metachromatic granules
Ribosomes
Prokaryotic:
Eukaryotic:
70S
80S
Bacterial adherence
Gene transfer
Common pili
Sex pili
By Gram (-) bacteria
ESBL
Endospores
Calcium dipicolinate
Bacillus, Clostridium
Flagella
: one only
: one at both ends
: tuft at one end
: all around bacteria (most common)
Monotrichous
Amphitrichous
Lophotrichous
Peritrichous
Aerotolerant anaerobes
Lactobacillus
Inorganic compound as source of carbon (CO2)
Autotrophs/Lithotrophs
Organic compound as source of carbone (Glucose)
Pathogenic bacteria
Heterotrophs/
Organotrophs
Temperature requirements
Psychrophilic:
Mesophilic:
Thermophilic:
0-20’C (ref)
20-40’C (pathogenic)
40-60’C
pH requirement
Acidophilic:
Neutrophilic:
Basophilic:
Lactobacillus acidophilus (Doderlein bacillus)
pH 7.2-7.6 (optimal) – pathogenic
Vibrio (Halophilic)
Moisture:
Salt concentration:
Humidophilic
Halophilic
Halophilic:
Enterococcus and V. parahaemolyticus
Respiration (Aerobic)
Glucose → CO2 + H2O
Kreb’s cycle
Electron transport chain
Oxidation (Aerobic)
Glucose → Acid
Fermentation (Anaerobic)
Glucose → Acid/Alcohol
Embden-Meyerhoff pathway (glycolysis)
Adjustment
Lag phase
↑ in growth rate (cell division)
Log/Exponential phase
Susceptible to antimicrobial agents
Log/Exponential phase
No net growth
Death = Live cells
Depletion of nutrients
Accumulation of toxic wastes
Sporulation
Stationary/plateau phase
↑ Death rate
Death/Decline phase
Bacteria stain more by
basic stains
Capsule stain
India ink
Borris method
Nigrosin method
Not Gram stained
Chlamydia and Rickettsia = intracellular
Mycoplasma and Ureaplasma = no cell wall
Spirochetes
Gram Stain (Hucker’s
modification
Crystal violet =
Gram’s iodine =
Acetone-alcohol or 95% ethanol =
Safranin O =
1min
1min
30secs-1min
30 secs
Over-decolorization
Old dying
Acidic iodine
Penicillin: omits iodine
Gram (+) becomes (-)
Under-decolorization
Thick smear
Gram (-) becomes (+)
Acid Fast staining methods
Smear = 2 x 3cm
Pappenheim’s
M. smegmatis vs. M. tuberculosis
Baumgarten’s
M. leprae vs. M. tuberculosis
Fite Faraco
M. leprae
Fite Faraco
Counterstain:
Hematoxylin
Acid fast organisms
Mycobacterium
Nocardia = Mod. AFS (1% H2SO4 as decolorizer)
Cryptosporidium
Legionella micdadei
Rhodococcus equi
Best AFS
C-A-M
Ziehl-Neelsen (Hot method)
= 1’ stain
-Start timing: Vapor (10mins)
-Heat = Mordant
- Carbolfuchsin
= Decolorizer
-HCl + 95% etOH
-Until no more stain (Max: 3mins)
- 3% Acid alcohol
= counterstain
-30secs to 1min
- Methylene blue
Ziehl-Neelsen (Hot method) Results:
AFO = Red
NAFO = Blue
Not used
Kinyoun (Cold method)
Kinyoun (Cold method) Results:
AFO = Red
NAFO = Green
Kinyoun (Cold method)
= 1’ stain
-Phenol, Tergitol = Mordant
= Decolorizer
= Counterstain
- Carbolfuchsin
- 3% Acid alcohol
- Malachite Green
Most sensitive
Auramine-Rhodamine (Fluorochrome)
Auramine-Rhodamine (Fluorochrome)
= 1’stain
= Decolorizer
= Counterstain
- Auramine-rhodamine
- 0.5% Acid alcohol
- 0.5% KMnO4
Auramine-Rhodamine (Fluorochrome) Results:
AFO = Yellow fluorescence
NAFO = No fluorescence
Read 300 fields
AFB
Special stains
Capsule =
Spore =
Negative stain
Dorner, Wirtz, Conklin
Metachromatic granules
- Albert’s
-Loeffler’s Alkaline Methylene Blue (LAMB)
Flagella =
Nucleic acid =
Polar bodies (ex: Y. pestis) =
Rickettsia =
Spirochetes =
Leifson
Feulgen
Wayson
Gimenez
Levaditi
For study of living unstained organisms
Phase contrast microscope
For viruses
Electron microscope
Light source: Electrons
100,000x magnification
Electron microscope
Electron microscope Stains:
-Negative stain
-PTA
-Heavy metals (Gold, Silver)
DNA, RNA, chromosomes
Transmission EM
Surface structures (cell wall, capsule)
Scanning EM
For tissue culture
Inverted Microscope
Dual light source
Interference microscope
Non staining method
String’s test (3% KOH)
Pure culture:
Streak plate =
Pour plate =
[?] medium
Animal inoculation =
overlap method
Water and milk bacteriology
Selective
for virus, Chlamydia, Rickettsia
2 or more organisms
Mixed culture
Stored at refrigeratior or freezer (long term)
Stock culture
Liquid:
Semi-solid:
Solid:
Biphasic:
Broth
0.5-1% agar
2-3% agar
Both liquid and solid
Biphasic:
Ex.
Castañeda = Brucella
Nonfastidious organisms
General purpose media
General purpose media:
- Sheep BAP = Hemolysis
- Horse BAP = Haemophilus
-Heat-stable, provides X-factor - Nutrient agar
Fastidous organisms
Enriched media Solid
= Heat-labile, provides X & V factor
- CAP
Enrichment media Liquid:
- Selenite F
- Alkaline peptone water
- Thioglycollate broth
Differential media
- BAP = hemolysis
- MAC
- EMB
- XLD
- HEA
Selective media
- TCBS
- SSA
- TMA
- CBAP
Inhibitory media
Selective media
Inhibitory agents
Antibiotics
Dyes, bile salts = inhibit Gram (+)
Alcohol (PEA) = inhibit Gram (-)
Gram (+) bacteria
Gram (+) bacteria
Gram (-) cocci
PEA
Columbia CNA
Gonococci Agar (GCA)
Gentamicin BAP
S. pneumoniae
Bacitracin CAP
H. influenzae
Cystine Tellurite Blood Agar
C. diphtheriae
Cystine Blood Glucose Agar
F. tularensis
Cystine Trypticase Agar Confirm:
Neisseria
Charcoal Cephalexin Blood agar
B. pertussis
Bordet-Gengou Agar (Potato Blood Glycerol Agar)
B. pertussis
L. pneumophila
BCYE
Cl. trachomatis
McCoy
Brucella
TSB
Sterile specimen
Nonsterile specimen
(-) normal flora
(+) normal flora
Toxic to virus
Good for Neisseria
Calcium alginate swab
Anaerobic and aerobic cultures
Needle aspiration
Needle and syringe for collection
Catheterization
Intubation
Gastritis
Vomitus
Gastric washing (aerobic culture only)
Delay in processing
Refrigerate except:
- CSF = Room temp. or 35’C
- Blood
- Swab of N. gonorrhoeae (sensitive to cold)
- Urine = Boric acid
- Rectal swab = Cary-Blair
Transport medium
- Cary Blair = for stool pathogen
- Stuart’s
- Amies = Respiratory specimen
- Transgrow = Neisseria
- JEMBEC = Neisseria
- Todd-Hewitt = Vaginal carriage (S. agalactiae)
HEPA filter: filters air
Negative pressure
Biologic safety cabinet
Environment and MT protected
BSC Class I
Air velocity = 75 linear ft/min
BSC Class I
Exhaust air thru HEPA filter
BSC Class I
Product contaminant
BSC Class I
Vertical laminar airflow
BSC Class II
MT, environment and product are protected
BSC Class II
Air velocity = 75-100 linear ft/min
BSC Class II
Recommended for hospitals
BSC Class II
Supply and exhaust air thru HEPA filter
BSC Class III
Maximum protection
BSC Class III
Contains HEPA filter
N95 Mask
For Mycobacterium
N95 Mask
No direct exam in Microbiology
Stool
Resistant gene
Mobile or jumping
Transposons
F. nucleatum
Capnocytophaga
Fusiform
No risk
Moderate risk
High risk
BSL I
BSL II
BSL III , BSL IV
Treatment available
BSL III
BSL III Inhalation of aerosols, Ex. Mycobacteria (BSC Class II)
No treatment available
BSL IV
Inhalation of aerosols, Ex. Small pox
BSL IV
BHIB + 0.25% SPS
Blood culture bottle
Dilution = 1:10 (1mL blood, 9mL broth)
Blood culture bottle
Anti-complementary, anticoagulant, antiphagocytic
Blood culture bottle
Neutralizes aminoglycosides
Blood culture bottle
Disadvantages of SPS Inhibits:
-Neisseria
-G. vaginalis
-S. moniliformis
-P. anaerobius
Counteract SPS to allow the growth of organisms
1% gelatin
Indications of growth (Blood culture)
Hemolysis
Turbidity
Pellicle
Subculture (Blood culture)
BAP
MAC = no CO2
CAP
If blood culture = negative
= Bacteremia (Typhoid)
= Brucellosis, SBE
7 days
21 days
Urine culture
Specimen:
Quantitative:
= significant for UTI
= not significant (contaminants)
Catheterized, Midstream, Suprapubic
BAP, MAC
->100,000 CFU/mL (or >50,000 CFU/mL)
-<10,000 CFU/mL
CSF culture
DO NOT refrigerate
Agents:
Media:
C. neoformans:
Neisseria, Haemophilus (Meningitis)
BAP, MAC, CAP, BHI
-India ink method
-Latex agglutination
Wound specimen
Gram stain
Media:
BAP, MAC, Thioglycollate broth
Stool specimen
Media:
Oxidase test
Biochemical tests
Serologic typing
MAC, BAP+Ampicillin, CBAP, SSA, Selenite F, TCBS, APW, HEA
Respiratory specimen
Sputum, NPS
TB = 3 sputum specimen
Media:
BAP, MAC, GBAP, BCAP, Amies, Gram stain and Acid fast stain
Throat swab
Sore throat
2 specimen
Media:
BAP, MTM, Thioglycollate broth
Vaginal, Urethral swab
Media:
Gram stain
CAP, MTM
TB culture
= Gold standard
= digestion, lyse the mucus
= decontamination
= Pseudomonas
Centrifuge (4’C) for [?]
Media:
Incubate at [?] —-(NG)—-> Report as (-)
If (+), after [?]: growth is seen
NALC-NaOH
-NALC
-NaOH
6% Oxalic acid
15 mins at 3000g
LJ, Middlebrook 7H11, 7H10 (AST)
37’C for 8 weeks
2-3 weeks
Genetic Pro
DNA test
Result → 2 hrs
GenPro
Moist heat sterilization
- Autoclave (sporicidal)
- Inspissation (sporicidal)
- Tyndallization (sporicidal)
- Boiling (Nonsporicidal, disinfectant)
- Pasteurization (Nonsporicidal, disinfectant)
Dy heat sterilization
- Hot air oven (Sporicidal)
- Incineration (Sporicidal)
- Cremation (Sporicidal)
- Flaming (Sporicidal)
- Gas: Ethylene oxide (sporicidal)
-121’C at 15 lbs/psi for 15 mins
- Autoclave (sporicidal)
-Culture media, bandages, gauze
- Autoclave (sporicidal)
-QC: B. stearothermophilus
- Autoclave (sporicidal)
-75-80’C for 2 hrs on 3 days
- Inspissation (sporicidal)
-Disinfect and solidify protein containing medium (LJ, Loeffler’s)
- Inspissation (sporicidal)
-Water is heated from below and slanting surface gets heated
- Inspissation (sporicidal)
-100’C for 30mins on 3 days
- Tyndallization (sporicidal)
-100’C for 30mins
- Boiling (Nonsporicidal, disinfectant)
-Kills vegetative cells only
- Boiling (Nonsporicidal, disinfectant)
-Milk
-63’C for 30mins
-72’C for 15secs
-Phosphatase: to determine if successful. (+): Not pasteurized
- Pasteurization (Nonsporicidal, disinfectant)
-170-180’C for 2 hrs
- Hot air oven (Sporicidal)
-Glasswares, cottonswabs, metallic instruments, oils, powders
- Hot air oven (Sporicidal)
-QC: B. subtilis
- Hot air oven (Sporicidal)
-Waste disposal
- Incineration (Sporicidal)
-Not recommended
- Incineration (Sporicidal)
-Prevents communicable disease
- Cremation (Sporicidal)
-Needles
- Flaming (Sporicidal)
-Heat-labile machine instruments
- Gas: Ethylene oxide (sporicidal)
-Preservation
- Cold temperature/Freezing (Bacteriostatic)
-Freeze drying
- Lyophilization (Powderized)
-Best to preserve culture
- Lyophilization (Powderized)
-Preservation
- Osmotic pressure (Bacteriostatic)
= removal of water
- Dessication
= produce pyrimidine dimer to DNA → mutation
- UV light
-Reduces airborne infection
- UV light
-For disposable materials (gloves, syringe)
- Ionizing radiation
-Air: HEPA filter
- Filtration
-H2O: cellulose membrane/ membrane filter
- Filtration
Filter heat-labile filter
Seitz filter
Made up of cellulose nitrate, cellulose diacetate, polycarbonate or polyester
Membrane filter
New: cellulose diacetate w/ a pore diameter of 0.015 to 12 microns
Membrane filter
Best filter used
Membrane filter
Spillage disinfectant
Sodium hypochlorite (Clorox)
Sporicidal
Iodine/Iodophor
Formaldehyde
Iodine + Detergent =
Iodine alone =
Betadine (Best antiseptic)
toxic to skin
Nonsporicidal
70% ethyl alcohol
Cleansing of wound
H2O2
Crede’s prophylaxis (New: Erythromycin eye droplets)
1% AgNO3
Prevents ophthalmia neonatorum
1% AgNO3
Sterilant
Glutaraldehyde
Standard disinfectant
Phenol (Carbolic acid)
Multipurpose
Lysol (Cresol)
Inhibit Gram (+)
Dyes
For decontaminating sputum
Zephiran (Benzalkonium chloride)
Instrument caused
Iatrogenic
Antagonistic
Synergistic
1 antibiotic > 2 antibiotics
2 antibiotics > 1 antibiotic
Extensively Drug Resistant Tuberculosis
Quinolone resistant
No treatment at all
XDR-TB
QC for beta-lactamase
H. influenzae
Extended spectrum beta-lactamase
Produced by Gram (-) = E. coli, Klebsiella
Plasmid mediated
Test: Beta-lactamase = Keyhole effect (overlapping zones)
-Clavulanic acid and cephalosporin
ESBL
Chromosome mediated
Produced by Gram (+) and (-) bacteria
Test: Beta-lactamase = D zone
-(+) to MRSA
-Imipenem and cefotixin
Amp C
Cell wall inhibitors
Broad spectrum:
Penicillin
Cephalosporin
Vancomycin = Tx: MRSA
Bacitracin
Cycloserine
Carbapenems/Imipenem
Penicilinase-resistant: Methicillin, Cloxacillin, Nafcillin
Cell membrane inhibitors
Colistin = against Gram (-)
Polymixin = against Gram (-)
Amphotericin B = drug of choice for systemic fungi
Nystatin = antifungal
Ribosome (Protein) inhibitors
Aminoglycosides (30S)
Tetracycline (30S)
Chloramphenicol (50S)
Erythromycin/Macrolide (50S)
Clindamycin (50S)
-False-resistant = P. aeruginosa (Mg2+ and Ca2+)
-Discovered by Bernardo Aguilar
-For penicillin allergic patients
Aminoglycosides (30S)
Erythromycin/Macrolide (50S)
Nucleic acid (DNA) inhibitors
Mitomycin
Quinolones
Metronidazole (Flagyl)
Trimethoprim-Sulfamethoxazole (SXT/Bactrim)
= inh. folate synth., synergistic
Trimethoprim-Sulfamethoxazole (SXT/Bactrim)
Anti-TB
Pyrazinamide
Rifampin
Isoniazid
Streptomycin
Ethambutol
Reference method (AST)
Det. MIC/MBC
Micro/Macrobroth dilution
Many organisms vs. single drug
Pure culture vs. many drugs
Agar dilution
Disk diffusion
Agar gradient diffusion
Antibiotic strip diffusion MIC test
MIC = Ellipse zone at intersection
E test (Epsilometer)
Kirby-Bauer Disk Diffusion
Std. Inoculum:
Medium:
pH:
Depth:
Condition:
Temp:
Incub. time:
Std:
Antibiotic disc:
1.5 x 10^8
MHA
7.2-7.4
4mm
Aerobic, No CO2
35-37’C (MRSA: 35’C)
16-18 hrs
0.5 McFarland (1% H2SO4 + 1.175% BaCl2)
6mm (refrigerated/frozen)
For bacterial count
Petroff-Hausser counting chamber
Distance of antibiotic disc to each other
Time for the medium to absorb the bacteria after inoculation
Inoculation of discs → Incubation
15mm
15mins
w/in 15mins
False resistant
Heavy inoculums
Thick medium
Delay in disc application
↑ Ca2+ and Mg2+ = Aminoglycoside (vs. P. aeruginosa)
↑ Thymine-Thymidine = SXT (vs. Enterococcus)
↑ pH = tetracycline
↓ pH = aminoglycoside, erythromycin
Expired discs
False sensitive
Light inoculums
Thin medium
If double zone of inhibition
If there are colonies inside the zone of inhibition
Measure the outer zone
Ignore swarming
Gram stain the colonies
AST media
1. MHA =
2. MHA + 2% NaCl =
3. MHA + 5% Sheep blood =
4. Haemophilus test medium:
5. GC agar =
6. Middlebrook 7H10 =
std. media
MRSA
S. pneumoniae (w/ CO2)
MHA + Yeast extract + Hemin + NAD + CO2
Neisseria (w/ CO2)
Mycobacteria (w/ CO2)
Specific
Regular basis
Checking media and reagents w/ specific organisms to check expected results
Set by CLSI (formerly NCCLS)
QC
General
Snap shot
Total process whereby the quality of lab. reports can be guaranteed
QA
Daily QC
Oxidase
Catalase
Incubator
Gram stain
Refrigerator/Freezer
Water bath
Each use (QC)
GasPak Jar
ONPG
Weekly QC
Antibiotic (Newly opened: 30 days QC weekly)
Autoclave
Biochemical tests
Semi-annually
Safety hood
ATCC (American Type Culture Collection)
For AST
Stock culture:
Working culture:
-20 or -70’C
2-8’C
ATCC-1234
Beta-lactamase producers:
-S. aureus
-N. gonorrhoeae
-H. influenzae
-Enterococcus
-E. coli
-P. aeruginosa
Catalase test
Rgt:
(+):
F (+):
3% H2O2
Gas bubbles
BAP
Coagulase test
Rgt:
(+) Clot formation after
F (+):
F (-):
= detects clumping factor/bound coagulase
= detects free/unbound coagulase
Rabbit EDTA plasma
4hrs
Citrate
Reading result after 6 hrs (Staphylokinase)
- Slide test (Screening)
- Tube test (Confirmatory)
Mannitol fermentation
Medium:
Indicator:
(+):
(-):
MSA (7.5% NaCl)
Phenol Red
Yellow
Red
DNase test
1. [?] (pink zone)/ [?] (clear zone)
2. [?]: no pptn. after adding 1N HCl when DNase (+) = pink
Toluidine blue / Methyl green
HCl precipitation
Novobiocin test
Amt.:
(R):
(S):
5μg
<16mm
> 16mm
Modified oxidase test
Rgt:
(+):
tetramethyl-p-phenylenediamine dihydrochloride in dimethylsulfoxide
Purple
Pinhead colonies
Staphylococcus
Mod. oxidase (-)
Staphylococcus
Stomatococcus
Lysostaphin and Furazolidone (S)
Staphylococcus
Ferments sugar
Staphylococcus
Mod. oxidase (+)
Micrococcus
Lysostaphin and Furazolidone (R)
Micrococcus
Stomatococcus
Oxidizes sugar
Micrococcus
S. aureus
Virulence factors:
Identification:
Infections:
-Protein A (cell wall)
-Leukocidin (Panton-Valentine)
-Exfoliatin (SSS/Ritter’s disease)
-TSST-1 (Tampons)
-Staphyloxanthin (Lipochrome): Yellow-orange colony
-(+) Phosphatase, ONPG, Arginine, NO3, VP, Gelatin
-(-) PYR
-Carbuncles, furuncles, folliculitis, cellulitis, impetigo, bacteremia, endocarditis, osteomyelitis
Slide coagulase (+)
PYR (+)
Slide coagulase (+)
VP (-)
S. lugdunensis
S. intermedius
Staphylococcus
: carrier of S. aureus
Culture:
Nasal swab
-Vogel-Johnson: Black colonies
-Chapman: Black colonies
-Tellurite Glycine: Black colonies
-P agar
-PEA: selective
-Columbia CNA: selective
1 skin flora
Blood culture contaminant
S. epidermidis
Biofilm/slime production: Prosthetic heart valve → Endocarditis, bacteremia
UTI: catheterized
S. epidermidis
UTI: sexually active women
S. saprophyticus
Pinpoint colonies
Capnophilic: 5-10% CO2
Streptococcus
Streptococcus
Culture:
SBA: Medium of choice
PEA: Selective medium
Smith and Brown’s classification
Hemolysis:
1. Alpha =
2. Beta =
3. Gamma =
4. Alpha prime =
incomplete (green)
complete (clear)
no zone
alpha (around colonies) + beta (around alpha)
Universally susceptible to antibiotics
S. pyogenes
(Group A)
(Beta-hemolytic)
S. pyogenes
(Group A)
(Beta-hemolytic)
Virulence factors:
Diseases:
-SLO = O2-labile, subsurface hemolysis, immunogenic
-SLS = O2-stable, surface hemolysis, non-immunogenic
-Erythrogenic toxin (Scarlet fever)
-Pharyngitis, AGN, RHF, erysipelas, impetigo
-Scarlet fever:
a. Dick’s test (red): Skin test
b. Schultz-Charlton (rash fade/blanching): Immunity test
Vaginal and URT flora
#1 neonatal meningitis
S. agalactiae
(Group B)
(Beta-hemolytic)
Group C
(Beta-hemolytic)
S. equisimilis
S. equi
S. zoopedemicus
S. dysagalactiae
Group F
(Beta-hemolytic)
S. anginosus
Group D Enterococcus
(Alpha, beta or gamma- hemolytic)
E. faecalis
E. faecium
E. durans
E. avium
Cause UTI
Group D Enterococcus
(Alpha, beta or gamma- hemolytic)
Group D non-Enterococcus
(Alpha, beta or gamma-
hemolytic)
Drug-resistant: VRE
Group D Enterococcus
(Alpha, beta or gamma- hemolytic)
Group D non-Enterococcus
(Alpha, beta or gamma-hemolytic)
S. bovis
S. equinus
Lancet-shaped, diplococci
S. pneumoniae
(Alpha-hemolytic)
Colonies: Mexican hat/ Dome-shaped
S. pneumoniae
(Alpha-hemolytic)
Encapsulated
S. pneumoniae
(Alpha-hemolytic)
1 Adult bacterial meningitis
S. pneumoniae
(Alpha-hemolytic)
Most common cause of Otitis media
S. pneumoniae
(Alpha-hemolytic)
Lobar pneumonia: Rusty sputum
S. pneumoniae
(Alpha-hemolytic)
S. pneumoniae
(Alpha-hemolytic)
Lab. Diagnosis:
- Neufeld Quellung (pptn. test, capsular swelling)
- Bile solubility
-BAP: 10% Na desoxycholate
-Tube: 2% Na desoxycholate - Francis test: skin test
- Mouse virulence test: (+) death
Viridans Streptococci
S. mitis (mitior)
S. salivarius
S. uberis
S. constellatus
S. intermedius
S. mutans = dental plaques/caries
S. sanguis = SBE
Nutritionally Variant
Abiotrophia
Granulicatella
Require Vit. B6 (pyridoxine)
(+) Staph. Streak test
Nutritionally Variant
Vancomycin Resistant
Leuconostoc = LAP (-)
Pediococcus = LAP (+)
Aerobic
Gram (-) diplococci
Oxidase (Taxo N): Presumptive test (+)
CTA: Confirmatory test
Capnophilic: 5-10% CO2
Neisseria
Pili: Adherence
N. gonorrhoeae
N. gonorrhoeae
Diseases:
Lab. Diagnosis:
-Culture:
-Gonorrhea (“Clap”)
-Ophthalmia neonatorum (Tx: Erythromycin eye drops)
-Salphingitis
-Epididymitis
a. Sterile:
= CAP: (+) Growth
= BAP: (-) Growth (Fastidious)
b. Nonsterile:
= GC agar: AST media
= TMA (Vancomycin-Colistin-Nystatin)
= MTM (V-C-N-Trimethoprim lactate)
= MLA (V-C-Anisomycin-T)
= NYCA (V-C-Amphotericin B-T)
Carrier: Nasopharynx
N. meningitidis
N. meningitidis
Virulence factors:
Diseases:
Lab. Diagnosis:
-Culture:
-Capsule
-Endotoxin
-Pili
-IgA protease
-Meningitis
-Meningococcemia
-Waterhouse-Friderichsen syndrome (Adrenal gland hemorrhage)
-DIC
a. BAP = (+) Growth
b. CAP = (+) Growth
-Serotypes: A, B, C, Y, W135 (Capsular Antigens)
Commensal of URT
M. catarrhalis
(+) NO3 → NO2
(+) Butyrate disk
(+) Tributyrin hydrolysis
(G) Nutrient Agar
M. catarrhalis
Colony: Hockey Puck
M. catarrhalis
3rd cause of Otitis media
M. catarrhalis
Breadcrumb/wrinkled colony
N. sicca
(+) ONPG
N. lactamica
Superoxol catalase test:
30% H2O2
(+) N. gonorrhoeae
Beta-lactamase test
1. Chromogenic cephalosporin test or Nitrocefin/Cefinase disk test
-(+):
2. Acidimetric
-Phenol red → (+):
3. Acidimetric
-I2 → (+):
Pink/red color
Colorless
Yellow
AFB (Mycolic acid)
Much granules = metachromatic
Aerobic nonsporeformer
Mycobacteria
Mycobacteria 3 Groups:
1. M. tuberculosis complex = cause TB
a. M. tuberculosis =
b. M. bovis =
c. M. africanum =
2. MOTT (New: NTM)
3. M. leprae
pulmonary TB
intestinal TB, bovine TB (BCG)
pulmonary TB in Africa
“Koch bacillus”
Capnophilic
Virulence: Cord Factor and Sulfatides
M. tuberculosis
M. tuberculosis Lab Dx:
1. GS = qualify specimen
> 10 epithelial cells = [?]
< 10 epithelial cells (>25 pus cells) = [?]
2. Decontamination-Digestion = Best
-[?]: remove metallic compound
3. AFS
4. Culture
saliva
sputum
Na citrate
Bone TB
Pott’s disease
Agar based media
Duboi’s oleic acid albumin medium
Mitchison’s medium
Middlebrook 7H10 and 7H11 (AST)
Egg based media (Sterilization: Inspissation)
Petragnani medium
Löwenstein Jensen (Slant) medium
American Thoracic Society medium
Dorset egg medium
Liquid media
BACTEC 12B
Septi-Chek
Middlebrook 7H9
Heat stable catalase test
Temp:
Medium:
Rgt:
(+)
(-)
68’C
Tween 80
30% H2O2
M. kansasii
M. tuberculosis
Tween 80 hydrolysis test
Tween 80:
(+) [?] and
Differentiates [?] (+) from (-) [?]
Polyoxyethylene sorbitan mono-oleate
M. kansasii
M. gordonae
M. scrofulaceum
Arylsulfatase test
For:
(+)
rapid growers
M. fortuitum-chelonae
TCH Susceptibility test
TCH:
(S):
(R):
Thiopene-2-carboxylic acid hydrazide
M. bovis
M. tuberculosis
Old Skin Test (TB)
= intracutaneous (most sensitive)
= scratch on skin
= piece of cloth w/ PPD skin
4. Moro percutaneous test
= multiple puncture technique
- Mantoux
- Von Pirquet
- Vollmer’s patch test
- Tuberculin time test
PPD
0.1 mL (5 TU)
(+):
Doubtful:
Induration (≥10mm)
5-<10mm/(-): <5mm
Photochromogens
(Group I)
- M. marinum (“Of the sea”)
- M. asiaticum
- M. simiae
- M. kansasii (“Yellow bacillus”)
-(+) pyrazinamidase
-(+) urease
- M. marinum (“Of the sea”)
-1st: Macaca rhesus monkey
- M. simiae
-(+) Heat-stable catalase
-(+) Tween 80 hydrolysis
-(+) NO3 reduction
-Pulmonary infection
-#2 isolate
- M. kansasii (“Yellow bacillus”)
Scotochromogens
(Group II)
- M. scrofulaceum
- M. szulgai
- M. gordonae (“Tap water bacillus”)
- M. flavescens
- M. xenopi
- M. thermoresistible
-(+) Urease
-Scrofula: cervical lymphadenitis
- M. scrofulaceum
-Photochromogen at 25’C
-Scotochromogen at 35-37’C
- M. szulgai
-Hot and cold H2O taps
-Cornmeal agar: “Bird’s nest”
-Optimal growth at 42’C
-(G) 37’C
-(NG) 25’C
-(-) Tellurite reduction
-(-) Catalase
- M. xenopi
-(G) 52’C
- M. thermoresistible
Colony: Cream/buff
Non-photochromogens
(Group III)
Non-photochromogens
(Group III)
- M. terrae-triviale
- M. avium-intracellulare (“Battey bacillus”)
- M. gastri
- M. haemophilum = require hemin (CAP)
- M. ulcerans (Buruli)
- M. malmoense
- M. tuberculosis
-(+) Catalase
-(NG) NaCl
-(G) NaCl
- M. terrae-triviale
a. M. terrae (“Raddish bacillus”)
b. M. triviale
-(+) Tellurite reduction
-(+) Urease
- M. avium-intracellulare (“Battey bacillus”)
- M. gastri
Rapid Growers
(Group IV)
- M. fortuitum-chelonae
- M. smegmatis
- M. phlei (“Hay bacillus”)
-(+) Arylsulfatase
-(G) MAC
- M. fortuitum-chelonae
-(+) NO3 reduction
-(+) 5% NaCl
-(+) Iron uptake
a. M. fortuitum
-(-) NO3 reduction
-(-) 5% NaCl
-(-) Iron uptake
b. M. chelonei
Growth: 10-21 days
Growth: 3-7 days
Groups I, II, III (Runyon’s Classification)
Group IV (Rapid growers)
Principle: RIA
14C Palmitic acid + organisms ———-> 14CO2
(+): >10 growth index
BACTEC 460 Middlebrook 7H12
Fluorometric based
↑ O2 consumption = ↑ Fluorescence
Mycobacteria Growth Indicator test (MGIT)
Rgt: p-nitroacetylamino-beta-hydroxypropiophenone (NAP)
(NG) = (+) M. tuberculosis
BACTEC 12B + NAP (Growth inhibition test)
Cigarette-packet/picket-fence
M. leprae
Not cultivable in agar
M. leprae
Can hydrolyze DOPA
M. leprae
Tropism to peripheral nerves
M. leprae
M. leprae Tx:
Dapsone
(-) Lepromine
↑ ↑ ↑ AFB
(+) Lepromine
↓ ↓ ↓ AFB
Lepromatous
Tuberculoid
Lab. Diagnosis (Leprosy)
Specimen:
Culture:
Stain:
Ear lobe, nasal scraping
Foot pads of Armadillo
Fite Faraco
Disseminated infection in AIDS
BACTEC (+)
M. genavensi
Crohn’s disease
M. paratuberculosis
Pleomorphic (rod-cocci)
Rhodococcus equi
(+) pink colonies
Rhodococcus equi
Partially acid fast (Mod. AFS: H2SO4)
Nocardia
(+) Urease
Nocardia
Cause pneumonia
Nocardia
= casein hydrolysis (-)
= casein hydrolysis (+)
N. asteroides
N. brasiliensis
Pleomorphic
Corynebacteria
Club shape: X, Y, V, L (Chinese characters)
Corynebacteria
Palisade appearance: diphtheroids
Corynebacteria
Nonmotile, nonsporeforming, nonencapsulated
Corynebacteria
(+) catalase and oxidase
Corynebacteria
“Klebs-Loeffler” bacillus
C. diphtheriae
C. diphtheriae
Virulence:
Diphtheria:
Lab. Diagnosis:
Exotoxin (A and B) – heat labile
pseudomembrane of the pharynx
-Elek test: definitive
-Roemer test: incubates the bacteria in pig
-Schick’s test: skin test (Ab’s)
-(+) DNase
-(-) Urease
-Ferments dextrose
Biotypes of C. diphtheriae
= gray, large, beta-hemolytic, starch/glycogen fermentation (+)
= black, medium-size, beta-hemolytic, starch/glycogen fermentation (-)
= black, small, nonhemolytic
- Gravis
- Mitis
- Intermedius
“Hoffman’s bacillus”
C. pseudodiphtheriticum
Ferments glucose, maltose and sucrose
C. xerosis
Agent of erythrasma
C. minutissimum
Coral red fluorescence on Wood’s lamp (porphyrin)
C. minutissimum
Lysogeny (no lysis)
Transduction
Aerobic, catalase (+)
Zoonotic
B. anthracis
B. anthracis
Virulence:
Diseases:
Lab. Diagnosis:
-Exotoxin: edema and lethal
-Capsule: D-glutamate
-Malignant pustule (Black eschar)
-Woolsorter’s disease (Respiratory)
-Gastroenteritis (Bloody diarrhea)
-PLET: sensitive medium
-Colony: medusa-head, inverted pine tree growth
-(+) catalase
-String of pearls on BAP + 0.05 U of penicillin
-Gamma phage susceptible
-Ascoli test
“Fried rice bacillus”
(-) capsule
B. cereus
B. cereus
Exotoxin (cholera-like):
↑ cAMP = ↑ H2O, Na+, K+ = dehydration
Common lab. contaminant
B. subtilis
Causes eye infection in heroin addicts
B. subtilis
Anaerobic, catalase (-)
Clostridium
Clostridium Saccharolytic except:
C. tetani, C. septicum
- Neurotoxic:
- Histotoxic:
- Enteric:
C. tetani (spastic paralysis), C. botulinum (flaccid paralysis)
C. perfringens, C. septicum
C. difficile
(Old: C. welchii)
Encapsulated, nonmotile
C. perfringens
Double zone of hemolysis:
-Inner (complete): due to theta toxin
-Outer (incomplete hemolysis): due to alpha toxin
C. perfringens
C. perfringens
Lab. Diagnosis:
-Chopped meat:
-Nagler test:
= Medium:
= (+) Opalescence on agar w/o antitoxin
= (-) No opalescence on agar w/ antitoxin
growth + gas (anaerobic growth)
lecithinase test (alpha toxin)
McClung or Neomycin egg yolk ;
“Canned-good bacillus”
C. botulinum
C. botulinum
Virulence:
Diseases:
-Heat labile toxin
-Botulinum toxin
-Not cultured
-Wound botulism
-Infant botulism
“Tack head bacillus”
C. tetani
Tennis head, drumstick
C. tetani
Oval, terminal spore
C. tetani
Swarms on BAP
C. tetani
-Not cultured
C. tetani
: block release of acetylcholine (muscle) → flaccid paralysis
: Most potent toxin
: grow in gut and honeybee (SIDS)
-Heat labile toxin
-Botulinum toxin
Infant botulism
C. tetani
Virulence:
Disease:
Exotoxin
-Tetanolysin
-Tetanospasmin
-Lockjaw
-Risus sardonicus: Sardonic smile
-Opisthotonus
: binds to ganglioside receptors and inhibit neurons in CNS → spastic paralysis
: Sardonic smile
-Tetanospasmin
-Risus sardonicus
Normal flora of the colon
C. difficile
Clindamycin-associated pseudomembranous colitis
C. difficile
C. difficile Lab. Diagnosis:
-Cytotoxin assay
-Culture: CCFA (Yellow colonies w/ horse manure odor)
Anaerobic bacteriology
Collection:
Reduced media:
Needle aspiration
-Anaerobic BAP
-Schaedler medium
-Bacteroides bile medium
-Laked Kanamycin-Vancomycin BAP
-Anaerobic PEA
-Egg yolk agar
-Chopped meat
-Peptone yeast extract glucose
-Thioglycollate broth
-Lombard Dowell Agar
Methods to promote anaerobiosis
- GasPak Jar or Mcintosh Fildesh, Brewer, Torbal or any anaerobic jar
- PRAS
- GasPak Jar or Mcintosh Fildesh, Brewer, Torbal or any anaerobic jar
-CO2, N2, no O2
-H + O2 = H2O
-Candle jar = not to be used
- PRAS = roll tube of Hungate
Anaerobic indicators:
a. Resazurin = pink
b. Methylene blue = colorless
Brick red fluorescence
Prevotella
Porphyromonas
Red fluorescence
Veillonella
Pitting of agar
B. ureolyticus
Swarming
C. tetani
C. septicum
Molar tooth colony, sulfur granules
A. israelii
Breadcrumb colony
Fusobacterium
Gram (+) Anaerobic Bacilli
Actinomyces
Bifidobacterium
Eubacterium
Propionebacterium
Lactobacillus
Mobiluncus
Gram (-) Anaerobic Bacilli
Bacteroides
Porphyromonas
Prevotella
Fusobacterium
Gram (+) Anaerobic Cocci
Peptostreptococcus
Peptococcus
Gram (-) Anaerobic Cocci
Veillonella
Megasphera
Acidaaminococcus
Lumpy jaw
A. bovis
Acne vulgaris
P. acnes
(+) Catalase, Indole
P. acnes
Tomato juice agar
L. acidophilus
(-) Catalase
L. acidophilus
Mobiluncus
P. asaccharolyticus
Vaginitis (G. vaginalis)
Mobiluncus
Foul odor
Anaerobic infections
Needs 20% bile
B. fragilis
Vincent’s angina (B. vincenti)
F. necrophorum (F. moriferum)
SPS sensitive (Neut: 1% gelatin)
P. anaerobius
(-) Indole, Catalase
P. anaerobius
Staphylococcus-like
Black
P. niger
H2S indicators
TSI =
LIA =
SIM =
Ferrous sulfate
Ferric ammonium citrate
Lead acetate
Phenol red
Acid =
Alkaline =
Yellow
Red
Methyl red
Acid =
Alkaline =
Red
Yellow
Neutral red
Acid =
Alkaline/neutral =
Pink/red
Colorless
Bromthymol blue
Acid =
Neutral =
Alkaline =
Yellow
Green
Blue
All ferment glucose
All are catalase (+)
All reduce NO3 → NO2
Enterobacteriaceae
Enterobacteriaceae
All are motile except:
All are oxidase (-) except:
-Shigella
-Klebsiella
-Y. pestis
-S. gallinarum
-S. pullorum
P. shigelloides (+)
Enterobacteriaceae Antigens:
-K = Capsular (for serotyping)
-O = Somatic (cell wall, heat-stable)
-H = Flagellar (heat-labile)
VP (+) “KESH”
Klebsiella
Enterobacter
Serratia
Hafnia
H2S (+) “SPACEd”
Salmonella
Proteus
Arizona
Citrobacter
Edwardsiella
Lactose Fermenters “ACH” (Slow) “EKE” (Rapid)
Arizona
Citrobacter
Hafnia
E. coli
Klebsiella
Enterobacter
—————————————————–
Y. enterocolitica = (+) ONPG, TSI: A/A
Deaminase (+) “PMP”
Proteus
Morganella
Providencia
Urease (+)
“PMP” (except P. alcalifaciens)
“K.pn/ox”
“E”
“Y.ps/en”
Proteus
Morganella
Providencia
K. pneumoniae
K. oxytoca
E. gergoviae
Y. pseudotuberculosis
Y. enterocolitica
Aerogenic
Klebsiella
E. coli
Salmonella (except S. typhi, S. gallinarum)
Proteus
Arizona
Citrobacter
Enterobacter
“Colon Bacillus”
E. coli
1 UTI
#2 Neonatal meningitis (K1 Ag)
E. coli
MUG (+) except:
MUG (+):
E. coli O157:H7
Electric blue fluorescence
Traveler’s diarrhea; Turista
ETEC
Montezuma revenge
ETEC
Belly Delhi
ETEC
Cholera like, watery diarrhea (SL, LT)
ETEC
Infantile diarrhea
EPEC
Pili: pathogenicity island
EPEC
Dysentery (Shigella)-like diarrhea
EIEC
Invasin: Pathogenicity
EIEC
Stool: bloody w/ mucus
EIEC
Sereny test
EIEC
HUS
EHEC/VTEC
Hemorrhagic colitis
EHEC/VTEC
Verotoxin: Shigella-like toxin
EHEC/VTEC
EHEC/VTEC
Sorbitol MAC (-)
E. coli O157:H7
Acute and chronic diarrhea
EAEC
Aggregative adhesion fimbriae
EAEC
Other Escherichia spp
= yellow colonies
= yellow colonies
-Nonmotile, (-) H2S, NLF, PG (-), Vi (-)
E. hermanni
E. vulneris
Alkalescens dispar (Anaerogenic E. coli/E. coli inactive)
UTI, wound, septicemia
= yellow pigment
= yellow pigment
Enterobacter
E. sakazakii
E. (Pantoea) agglomerans
“Friedlander’s bacillus”
(+)Capsule, ESBL
K. pneumoniae
K. pneumoniae Diseases:
-Pneumonia: “Currant jelly-like sputum”
-Wound infections
-Meningitis
-UTI
Biochemically inert
K. rhinoscleromatis
PMP Group (Proteeae) All are indole (+) except
P. mirabilis (-)
Swarm on BAP but not on MAC
Proteus
2 UTI
Proteus
Renal stone association
Proteus
Odor: Burnt gun powder/Burnt chocolate
Proteus
: Rapid urease producer
P. mirabilis
Indole (-)
Salmonella
BSA & SSA: Black colonies
Salmonella
Gold standard: Culture
Salmonella
Kaufmann White schema
-Vi Ag = heat-labile, masks the O Ag
-O Ag
-H ag
Salmonella
Salmonella Tx:
Chloramphenicol
S. typhi
Diseases:
Complication:
Lab Diagnosis:
Typhoid fever
Meningitis
Osteomyelitis
Psychosis
-1st week: Blood
-2nd week: Urine and stool (carrier)
Septicemia
S. paratyphi C (S. choleraesuis)
“Gardner’s Bacillus”
S. enteritidis
Biochemically inert
Shigella
SSA: colorless
Shigella
Shigella Related to E. coli:
-E. coli:
-Shigella:
Acetate (+)
Acetate (-)
Intestinal pathogen: Dysentery
Shigella
Shigella Lab. Diagnosis:
-BEST: Culture of fresh stool w/ mucous flecks
-Rectal swab of ulcer
Shigella Subgroups (O Ag)
A =
B =
C =
D =
S. dysenteriae (“Shiga Bacillus”)
S. flexneri (“Strong Bacillus”)
S. boydii
S. sonnei (Cross reacts w Plesiomonas)
Red pigment (Prodigiosin)
Red milk
Serratia marcescens
(+) DNase, Gelatinase, LPS
Serratia marcescens
Resistant to many antibiotics
Serratia marcescens
Rancid, potato-like odor
Serratia odorifera
“Plague Bacillus”
Y. pestis
(+) Stalactite on broth medium
Y. pestis
V and W antigens
Y. pestis
MOT: Rat flea bite
Y. pestis
Y. pestis Infections:
-Bubonic plague: buboes, lymphadenopathy
-Pneumonic plague
-Septicemic plague
-Black death
Motile at RT’
Y. enterocolitica
Appendicitis-like infection
Y. enterocolitica
(+) ONPG
Y. enterocolitica
Motile at RT’
Y. pseudotuberculosis
Mesenteric lymphadenitis, septicemia
Y. pseudotuberculosis
“Comma Bacillus”
Comma-shaped
Vibrio
Darting motility
Vibrio
Vibrio Oxidase (+) except
V. mitschnikovii (-)
Vibrio Halophilic except:
-V. cholerae
-V. mimicus
(+) Indole
V. parahaemolyticus
Cholera: Ricewater stool
V. cholerae
“Fish in the stream” appearance
V. cholerae
V. cholerae Lab. Diagnosis
Culture:
-: Sucrose fermenter
-: Tellurite Taurocholate Gelatin Agar
-: 6-8 hrs → Subculture (TCBS)
-: Transport medium (stool)
-: Carrier
= (S)
TCBS
TTGA
APW
Cary Blair
Rectal swab
O129 Susceptibility test
Halophilic (8% NaCl)
V. parahaemolyticus
(+) String test (0.5% Na desoxycholate)
V. cholerae
1 Gastroenteritis in Japan (seafood)
V. parahaemolyticus
Nonsucrose fermenter (TCBS)
V. parahaemolyticus
V. vulnificus
Kanagawa (+) = Beta hemolysis on Wagatsuma agar
V. parahaemolyticus
Stool pathogen
V. vulnificus
(+) Blood culture
V. vulnificus
(+) Lactose fermentation, ONPG
V. vulnificus
Sucrose fermenter (TCBS)
V. alginolyticus
Sucrose fermenter (TCBS)
V. alginolyticus
Motile (monotrichous)
Aeromonas
(+) Oxidase
Aeromonas
Plesiomonas
Campylobacter
H. pylori
P. aeruginosa
B. cepacia
Beta-hemolytic
Aeromonas
(R) O129
Aeromonas
(+) DNase
Aeromonas
(+) Bile esculin hydrolysis
Aeromonas
Non-hemolytic
Plesiomonas
(V) O129
Plesiomonas
(-) DNase
Plesiomonas
(-) Bile esculin hydrolysis
Plesiomonas
(G) 42’C
Campylobacter
Curved, S-shaped (Wings of Seagull)
Campylobacter
(+) Catalase
Campylobacter
H. pylori
Darting motility
Campylobacter
(+) Hippurate
C. jejuni
(+) Urease
H. pylori
H. pylori Test:
Urease breath test
OF test
Fermentative (close) =
Oxidative (open) =
Nonutilizer =
Medium:
Indicator:
O/F = +/+ = Y/Y
O/F = +/- = Y/G
O/F = -/- = G/G
Hugh and Leifson (1% glucose, 1% agar)
Bromthymol blue
P. aeruginosa Old:
P. pyocyanea
Pyocyanus: Blue pus agent
P. aeruginosa
Motile (monotrichous)
P. aeruginosa
Capsule: Slimy layer
P. aeruginosa
Grapelike odor (aminoacetaphenone)
P. aeruginosa
(G) 42’C
P. aeruginosa
B. pseudomallei
Cetrimide medium: selective
P. aeruginosa
P. aeruginosa Pigments:
-Pyocyanin: Best
-Pyoverdin/Fluorescein: (+) P. putida/P. fluorescens
P. aeruginosa Infections:
-#2 Burns (#1: S. aureus)
-Wound: Ecthyma gangrenosum
-Swimmer’s ear (Otitis externa)
-Dermatitis: whirlpool baths
-Contact lens infection
-#1 Cystic fibrosis
Motile (lophotrichous)
B. cepacia
B. pseudomallei
2 Cystic fibrosis
B. cepacia
(+) Lactose, ONPG
B. cepacia
“Whitmore’s Bacillus”
B. pseudomallei
Vietnamese timebomb
B. pseudomallei
Melioidosis/Glanders-like disease
B. pseudomallei
Ashdown medium: Wrinkled colony
B. pseudomallei
(+) Lactose
B. pseudomallei
Long incubation period: 10-20 years
B. pseudomallei
Nonmotile
B. mallei
Glander’s disease (Horses)
B. mallei
Brown (buff colored) wrinkled colony
P. stutzeri
(+) 6.5% NaCl
P. stutzeri
(+) NO2 → N2
P. stutzeri
(-) Lactose
P. stutzeri
Found in stagnant water
P. stutzeri
(-) Oxidase
(+) DNase
(+) Maltose and Glucose
Motile (lophotrichous)
S. maltophilia
Colony: Lavender green
S. maltophilia
Common w/ the use of catheter
S. maltophilia
Blue milk
P. syncyanea
TSI: K/K + H2S
(+) Oxidase
Shewanella putrefaciens
(-) Oxidase
(+) Catalase
Nonmotile
Causes UTI
Acinetobacter
MAC: purple colonies
Acinetobacter
Mistaken as Neisseria – (+) oxidase
Acinetobacter
Oxidizer
A. anitratus (A. baumanii)
Herella vaginocola
A. anitratus (A. baumanii)
Nonoxidizer
A. lwoffi
Mima polymorpha
A. lwoffi
(+) Oxidase
(+) Catalase
Alcaligenes faecalis
Motile (peritrichous)
Alcaligenes faecalis
Asaccharolytic
Alcaligenes faecalis
M. lacunata
“Fruity” odor
Alcaligenes faecalis
(+) Pitting of agar
(+) Oxidase
(+) Catalase
M. lacunata
(NG) MAC
M. lacunata
Infection: Blepharoconjunctivitis
M. lacunata
Mistaken as Neisseria
M. lacunata
Flavin = Yellow pigment
Flavobacterium (Chryseobacterium) meningosepticum
(+) Oxidase
(+) DNase
(+) Indole
(+) Gelatin hydrolysis
Nonmotile
Flavobacterium (Chryseobacterium) meningosepticum
Causes neonatal meningitis
Flavobacterium (Chryseobacterium) meningosepticum
Twitching motility
E. corrodens
MOT: Human bite
E. corrodens
“Clenched fist”
E. corrodens
Corrodes agar
E. corrodens
Odor: Bleach-like
E. corrodens
(-) Catalase
(+) Oxidase
E. corrodens
Cause SBE (HACEK)
Kingella spp
Pits agar
Kingella spp
Require X factor (hemin) and V factor (NAD)
Haemophilus
(+) Satellitism
(+) Oxidase
Haemophilus
HCAP: medium of choice (5% CO2)
Haemophilus
X factor
D-ALA → (+) Protoporphyrin (Red)
Porphyrin test
“Pfeiffer’s Bacillus”
H. influenzae
H. influenzae Serotype polysaccharide:
-Hib = Haemophilus type B: capsular antigen type B (polyribose ribitol PO4)
H. influenzae
Infections:
Lab. Diagnosis:
-Epiglotitis (Major)
-#3 Meningitis (#1: S. pneumoniae/#2: N. meningitidis)
-Otitis media, CF, conjunctivitis, pneumonia, URTI, sepsis
-Culture: Grayish, dew drop colony w/ Mousy odor
a. CAP
b. Levinthal
c. Fildes
“Chancroid Bacillus”
H. ducreyi
(G) CAP + Vancomycin (33’C)
H. ducreyi
Soft chancre (painful)
H. ducreyi
School of red fish
H. ducreyi
“Koch Week’s Bacillus”
H. aegyptius
Pink eye conjunctivitis
H. aegyptius
Brazilian purpuric fever
H. aegyptius
“Whooping cough Bacillus”
B. pertussis
Strictly aerobic
B. pertussis
Encapsulated, nonmotile
B. pertussis
Require cysteine and methionine for growth
B. pertussis
Toluidine blue: Bipolar granules
B. pertussis
B. pertussis Stages: Whooping cough
: mild
: successive cough
: recovery
- Catarrhal
- Paroxysmal
- Convalescence
B. pertussis Lab. Diagnosis:
-NPS: Carrier
-Culture media:
a. BG (PBGA) = mercury droplet colonies (pearl-like)
b. RL (Charcoal horse blood)
c. Jones Kendrich (Charcoal, yeast extract)
d. CCBA
e. Stainer and Scholte
f. Casamino broth
“Kennel cough Bacillus”
B. bronchiseptica
(-) Capsule
Brucella
Obligate aerobe
Nonmotile
Facultatively intracellular (phagocytes)
Brucella
Erythritol: enhances growth
Brucella
Zoonotic: found in animal placenta
Brucella
Brucella
Infections:
Lab. Diagnosis:
- Undulant fever
-Malta fever
-Aboriton in cattle
-Endocarditis
-Castañeda broth: Best medium
-TSB: Best medium
-W (Wisconsin) medium: selective
-(+) Rose Bengal & 2-ME agglutination
“Bang’s Bacillus”
B. abortus
(+) Capsule
F. tularensis
Nonmotile
Facultatively intracellular
Obligate aerobe
F. tularensis
Lab. acquired infection (BSL II)
F. tularensis
Tularemia:
-Lymphadenopathy
-Deerfly, lemming, rabbit, water rat trapper’s disease
F. tularensis Lab. Diagnosis:
1. GCBA:
2. PCA:
3. CHA:
4. Biochem:
Glucose cysteine blood agar
Peptone cysteine agar
Cysteine heart agar
-(+) Catalase
-(-) Oxidase
-(-) Urease
-(-) MAC
Multocida: Multiple killing
P. multocida
(+) Capsule
Nonmotile
(+) Oxidase
(+) Catalase
(+) Glucose
(+) Ornithine
(+) Indole
(+) Urease
(G) BAP
(NG) MAC
P. multocida
P. multocida Infections:
-Animal (cat) bite wound infection
-Shipping fever
(-) Catalase
Dx: Serology
Treponema
(+) Catalase
Dx: Culture
Leptospira
(-) Catalase
Dx: Giemsa (blood smear), Serology
Borrelia
Noncultivable on agar
T. pallidum
Obligate intracellular (Rabbit’s testicle)
T. pallidum
Syphilis
1’ =
2’ =
Latent:
3’:
Congenital syphilis:
Hard chancre (Painless)
Condylomata lata (HPV: Condylomata acuminata)
Asymptomatic, Dx: Serology
Gummas, neurosyphilis
Hutchinsonian triad, abortion, still birth
Neurosyphilis
Tabes dorsalis
Treatment for syphilis (Jarisch-Herxheimer reaction)
Penicillin
Yaws, Framboesia
Pinta
Bejel
Rabbit syphilis
T. pertenue
T. carateum
T. endemicum
T. cuniculi
Lab. Diagnosis (Treponemes)
1. Darkfield microscopy:
- Stain:
- Serology:
-Corkscrew motility
-1’ and 2’ syphilis
Levaditi silver impregnation
-Latent, 3’ syphilis
-Reagin test (VDRL, RPR, TRUST)
-Trep. Antibody test (FTA-ABS [confirm], MHA-TP, HATTS)
Obligate aerobes
L. interrogans icterohemorrhagiae
Spiral w/ hooked ends
L. interrogans icterohemorrhagiae
L. interrogans icterohemorrhagiae Weil’s disease (Zoonotic):
-1st week:
-2nd week:
blood, CSF
urine (chronic)
L. interrogans icterohemorrhagiae Animal serum:
30’C for 6 weeks
L. interrogans icterohemorrhagiae Culture media:
-Fletcher’s
-Noguchi
-EMJH
Blood spirochete
Borrelia
Borrelia
1. Relapsing fever:
Diagnosis:
2. Lyme disease:
-MOT:
a. 1’ stage:
b. 2’ stage:
c. 3’ stage:
Diagnosis:
-Culture on Barber Stoenner Kelly medium =
a. B. recurrentis: louse-borne
b. B. anserine: tick-borne
c. B. turicatae: tick-borne
d. B. parkeri: tick-borne
Wright’s/Giemsa (blood/BM)
B. burgdorferi
Tick bite (I. dammini)
Erythema chronicum migrans (Bull’s eye rash)
neurological (meningitis), cardiac
Arthritis (joint fluid)
Culture and Serology
33’C for 6 weeks
Star-like colony
A. actinomycetemcomitans
Dots and dashes of Morse code
A. actinomycetemcomitans
(+) Indole
C. hominis
Teardrop shape
C. hominis
Rosette formation
C. hominis
Asaccharolytic
E. corrodens
Obligate intracellular
Chlamydia (Old: Bedsonia)
Energy (ATP) parasites
Chlamydia (Old: Bedsonia)
Chlamydia (Old: Bedsonia) Inclusion body: diagnostic (stained by Giemsa), not Gram stained
= infectious
= reproductive
a. Elementary body
b. Reticulate body
Chlamydia (Old: Bedsonia) Immunofluorescence:
det. Chlamydia antigen (N. gonorrhoeae)
Contains glycogen —(I2)—> Brown
C. trachomatis
TRIC agent: Trachoma Inclusion Conjunctivitis
C. trachomatis
C. trachomatis Diseases:
-LGV (Frei’s test)
-PID (pelvic inflammatory disease)
-#1 STD in US
-#1 NGU
(S) Sulfonamide
C. trachomatis Lab. Diagnosis:
1. Inclusion body (Halberstadter Prowazeik)
-Stain (Glycogen):
-Elementary body:
2. Fitz-Hugh Curtis
3. McCoy (shell vial) =
4. Direct IF =
5. DNA amplification =
6. Transport =
Iodine (brown)/Giemsa (purple)
Round-shaped
Best medium (cell culture)
Ag
most sensitive
4’C/-70’C (long term)
Parrot fever/Psittacosis/Ornithosis
C. psittaci
Parrot fever/Psittacosis/Ornithosis
C. psittaci
Man: Pneumonia
C. psittaci
Inclusion body (Levinthal Cole Lillie)
Elementary body: Round-shaped
C. psittaci
(-) glycogen
(R) Sulfonamide
C. psittaci
TWAR agent
C. pneumoniae
Pneumonia: Human-to-human
C. pneumoniae
Growth on:
-Human lines
-Hep-2 cell
C. pneumoniae
Guillain-Barré syndrome: ascending paralysis
C. pneumoniae
Obligate intracellular (endothelial cells) except Coxiella (extracellular)
Rickettsia
Arthropod borne except Coxiella (inhalation)
Rickettsia
Arthropod borne except Coxiella (inhalation)
Rickettsia
Rickettsia Lab. Diagnosis:
: cross react w/ Proteus (det. Rickettsial Ab)
: Gimenez, Macchiavelo
: Embryonated egg, cell culture
- Weil-Felix test
- Special stain
- Culture
Intracellular (WBCs)
Intracellular (WBCs)
Ehrlichia
Morulae: diagnostic form
Ehrlichia
Tick-transmitted
Ehrlichia
Cause sennetsu fever
Ehrlichia
No cell wall (pleomorphic)
Mycoplasma
Fried egg/mulberry
Mycoplasma
Cell membrane: Sterol (bilipid layer)
Mycoplasma
Require sterol for growth
Mycoplasma
(R) Penicillin
Mycoplasma
Diene’s stain: stain for Mycoplasma colonies (blue)
Mycoplasma
Eaton agent
M. pneumoniae
PPLO: Pleuropneumonia-like organism
M. pneumoniae
PAP: Primary atypical pneumonia
M. pneumoniae
Walking pneumonia
M. pneumoniae
M. pneumoniae Lab. Diagnosis:
1. Media:
2. Confirm:
3. Screening:
4. Confirmatory:
PPLO agar
Hemadsorption test (not a serological test), RBCs → Colonies
Cold agglutination
IF
M. pneumoniae Tx:
Tetracyclin/Erythromycin
Large fried egg colony
M. hominis
Infections: Post abortal, post partum fever, PID
M. hominis
Media:
-A7/A8: selective
-NYCA
-SP4: arginine
M. hominis
T strain = tiny fried egg
U. urealyticum
(+) Urease = brown
U. urealyticum
Cause NGU
U. urealyticum
Media:
-A7/A8
-SP4: urea
-NYCA
U. urealyticum
Old: Haemophilus v., Corynebacterium v.
G. vaginalis
(-) Catalase
(-) Oxidase
G. vaginalis
(S) SPS (Counteracted by 1% gelatin)
G. vaginalis
Whiff’s test: 3% KOH Fishy amine-like odor
G. vaginalis
Media: HBTA (selective), V (vaginalis) agar, Columbia CNA
G. vaginalis
Gram (V)
G. vaginalis
(+) Capsule
C. granulomatis
Safety pin
C. granulomatis
MOT: Sexual contact
C. granulomatis
Causes granuloma inguinale (Donovanosis)
C. granulomatis
Giemsa stain: Donovan bodies (macrophage w/ Gram [-] bacilli)
C. granulomatis
Not cultured
C. granulomatis
String of beads
S. moniliformis
Broth: fluff balls
S. moniliformis
HI: fried egg
S. moniliformis
(w/ cell wall) —(Penicillin)—> L forms (no cell wall)
S. moniliformis
(S) SPS
S. moniliformis
S. moniliformis Infections:
-Rat bite fever
-Haverhill disease
Spiral organism
S. minus (S. minor)
Causes rat bite fever
S. minus (S. minor)
Cannot be cultivated
S. minus (S. minor)
Old: Achromobacterium
C. violaceum
Violacein: violet colored
C. violaceum
MAC: NLF
C. violaceum
NH4 cyanide
C. violaceum
Fusiform
Capnocytophaga
Gliding motility
Capnocytophaga
Periodontal disease (oral flora) = periodontitis
Capnocytophaga
Large spreading colonies
Capnocytophaga
(-) Oxidase
(-) Catalase
(-) Indole
(+) Esculin hydrolysis, NO3 reduction
Capnocytophaga
Infections:
Cat scratch disease
Bacillary angiomatosis
Peliosis
Hepatitis
B. henselae
(NG) MAC
B. henselae
Vector: Sandfly
B. bacilli formis
Infections:
-Carrion’s disease
-Verruga peruana: skin eruptions
-Oroya fever: anemia
B. bacilli formis
Aerobic
Facultatively intracellular
L. pneumophila
Require cysteine and iron for growth
L. pneumophila
(NG) MAC = no cysteine
L. pneumophila
Isolated from airconditioners and water-cooling towers
L. pneumophila
Infections:
-Broadstreet pneumonia
-Pontiac fever
L. pneumophila
(+) Catalase
(+) Oxidase
(+) DNase
(+) Beta-lactamase
L. pneumophila
L. pneumophila Lab. Diagnosis:
: Ag (IF: Ab)
: Blue green cut glass colony
: 10 mins
: 4’C or frozen
- DF Ab test
- BCYE
- Safranin
- Transport
Tumbling motility: head over heels/end-to-end motility
L. monocytogenes
Listeriolysin O
L. monocytogenes
Infections:
-Granulomatosis infantiseptica
-Meningitis
L. monocytogenes
Lab. Diagnosis: Anton’s test (Skin test)
L. monocytogenes
Growth: Test tube brush
E. rhusiopathiae
Erysipeloid: Butcher’s cut
E. rhusiopathiae
E. coli: Index of fecal contamination
Water Bacteriology
Det. coliforms
Water Bacteriology
Neutralizes chlorine
Sodium thiosulfate
Lactose broth/ + H2O ———-(35’C for 24 hrs)—-> (+) Gas
Lauryl tryptose broth (-) No gas after 48 hrs
- Presumptive test
EMB/Endo agar + Inoculum —–(24 hrs)———–> (+) Colony
- Confirmatory test
Lactose broth fermentation tube —(35’C for 24-48 hrs)—> (+) Acid + Gas
- Completed test
Most common method for H2 analysis
Estimation of coliform group density
# of positive coliforms from multiple decimal dilution
MPN (Most Probable Number)
Reference laboratories (Water Bacteriology)
EAMC
FDA
Acid forming, causes normal souring of milk
Streptococcus lactis
Gas-forming, hydrolyze the milk protein casein
Coliform bacteria
Hay bacteria, proteolytic action on coagulated milk
Bacillus subtilis
Stormy fermentation of milk
Clostridium butyricum
Clostridium perfringens
Blue milk
Pseudomonas syncyanea
Red milk
Serratia marcescens
Blue green milk
Pseudomonas aeruginosa
Yellow milk
Flavobacterium synxanthum
Slimy or ropy milk (capsule formation)
Alcaligenes viscosus
