Professional Documents
Culture Documents
Bacteriology
Bacteriology
Soap Germicidal
L. monocytogenes Catalase (+)
3 C’s: Chicken, Coleslaw, Cheese
Chlamydia When delayed: 4’C
Freezing: -20’C
BAP Phenotypic
Gram (+) colonies: Dry, white, sometimes gray
Gram (-) colonies: Gray and moist
M. gordonae Destroyed by chlorine
DNase test Utilizes 1N HCl
LOA test For Gram (+)
For nonfermentative
For Enterobacteriaceae
Na hypochlorite Inactivates HBV (10mins) and HIV (2mins)
Inoculating needles Nichrome = F(+) on oxidase test
Not longer than 5cm
Wire loop 2mm diameter
0.001mL urine
50k CFU/mL Significant for UTI
Pregnant C. albicans
Lactobacillus
Cotton swab Carrier state
Lawn a culture
Toxic to Neisseria
Good for virus
Charcoal Removes the toxin inoculated by cotton
Phenotype Basis of identifying organisms
Gram stain and colonies
Stool Not Gram stained
PCR Most definitive method of identification
Somatic antigen Basis of serotyping
Anton van Father of microbiology
Leeuwenhoek Microscopist
1st to describe bacteria
Robert Koch Germ theory: relationship of organisms to human disease
Louis Pasteur Father of Modern Microbiology
Ehrlich 1st to use dyes for stain
Bacteria Ave. size: 0.4-2μm
Reproduction: Binary fission (two-fold increase)
Cell wall Peptidoglycan (murein)
Protoplast: wall less G(+)
Spheroplast: wall less G(-)
Gram (+) Thick peptidoglycan
Teichoic acid
Gram (-) Thin peptidoglycan
LPS (Lipid A – exotoxin)
lec.mt 04 |Page | 55
Somatic antigen
Plasma membrane Site for energy synthesis (ATP)
Osmotic/permeability barrier
Nucleoid Chromosome: dsDNA
Plasmid: Extrachromosomal DNA
Plasmid Carries the antibiotic-resistance gene
Drug-resistance Chromosome and plasmid-mediated
Metachromatic Food reserves
granules
Ribosomes Prokaryotic: 70S
Eukaryotic: 80S
Common pili Bacterial adherence
Sex pili Gene transfer
ESBL By Gram (-) bacteria
Endospores Calcium dipicolinate
Bacillus, Clostridium
Flagella Monotrichous: one only
Amphitrichous: one at both ends
Lophotrichous: tuft at one end
Peritrichous: all around bacteria (most common)
Lactobacillus Aerotolerant anaerobes
Autotrophs/Lithotrop Inorganic compound as source of carbon (CO2)
hs
Heterotrophs/ Organic compound as source of carbone (Glucose)
Organotrophs Pathogenic bacteria
Temperature Psychrophilic: 0-20’C (ref)
requirements Mesophilic: 20-40’C (pathogenic)
Thermophilic: 40-60’C
pH requirement Acidophilic: Lactobacillus acidophilus (Doderlein bacillus)
Neutrophilic: pH 7.2-7.6 (optimal) – pathogenic
Basophilic: Vibrio (Halophilic)
Moisture Humidophilic
Salt concentration Halophilic
Enterococcus and V. parahaemolyticus
Respiration (Aerobic) Glucose CO2 + H2O
Kreb’s cycle
Electron transport chain
Oxidation (Aerobic) Glucose Acid
Fermentation Glucose Acid/Alcohol
(Anaerobic) Embden-Meyerhoff pathway (glycolysis)
Lag phase Adjustment
Log/Exponential phase in growth rate (cell division)
Susceptible to antimicrobial agents
Stationary/plateau No net growth
phase Death = Live cells
Depletion of nutrients
Accumulation of toxic wastes
lec.mt 04 |Page | 56
Sporulation
Death/Decline phase Death rate
Staining 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 Crystal violet = 1min
modification Gram’s iodine = 1min
Acetone-alcohol or 95% ethanol = 30secs-1min
Safranin O = 30 secs
Gram (+) becomes (-) Over-decolorization
Old dying
Acidic iodine
Penicillin: omits iodine
Gram (-) becomes (+) Under-decolorization
Thick smear
Acid Fast staining Smear = 2 x 3cm
methods
Pappenheim’s M. smegmatis vs. M. tuberculosis
Baumgarten’s M. leprae vs. M. tuberculosis
Fite Faraco M. leprae
Counterstain: Hematoxylin
Acid fast organisms Mycobacterium
Nocardia = Mod. AFS (1% H2SO4 as decolorizer)
Cryptosporidium
Legionella micdadei
Rhodococcus equi
Ziehl-Neelsen (Hot Best AFS
method) C-A-M
1. Carbolfuchsin = 1’ stain
-Start timing: Vapor (10mins)
-Heat = Mordant
2. 3% Acid alcohol = Decolorizer
-HCl + 95% etOH
-Until no more stain (Max: 3mins)
3. Methylene blue = counterstain
-30secs to 1min
Results:
AFO = Red
NAFO = Blue
Kinyoun (Cold method) Not used
C-A-M
1. Carbolfuchsin = 1’ stain
-Phenol, Tergitol = Mordant
2. 3% Acid alcohol = Decolorizer
lec.mt 04 |Page | 57
3. Malachite Green = Counterstain
Results:
AFO = Red
NAFO = Green
Auramine-Rhodamine Most sensitive
(Fluorochrome) 1. Auramine-rhodamine = 1’stain
2. 0.5% Acid alcohol = Decolorizer
3. 0.5% KMnO4 = Counterstain
Results:
AFO = Yellow fluorescence
NAFO = No fluorescence
AFB Read 300 fields
Special stains Capsule = Negative stain
Spore = Dorner, Wirtz, Conklin
Metachromatic granules
- Albert’s
-Loeffler’s Alkaline Methylene Blue (LAMB)
Flagella = Leifson
Nucleic acid = Feulgen
Polar bodies (ex: Y. pestis) = Wayson
Rickettsia = Gimenez
Spirochetes = Levaditi
Phase contrast For study of living unstained organisms
microscope
Electron microscope For viruses
Light source: Electrons
100,000x magnification
Stains:
-Negative stain
-PTA
-Heavy metals (Gold, Silver)
Transmission EM DNA, RNA, chromosomes
Scanning EM Surface structures (cell wall, capsule)
Inverted Microscope For tissue culture
Interference Dual light source
microscope
Non staining method String’s test (3% KOH)
Pure culture Streak plate = overlap method
Pour plate = Water and milk bacteriology
Selective medium
Animal inoculation = for virus, Chlamydia, Rickettsia
Mixed culture 2 or more organisms
Stock culture Stored at refrigeratior or freezer (long term)
Liquid Broth
Semi-solid 0.5-1% agar
Solid 2-3% agar
Biphasic Both liquid and solid
Ex. Castañeda = Brucella
lec.mt 04 |Page | 58
General purpose media Nonfastidious organisms
1. Sheep BAP = Hemolysis
2. Horse BAP = Haemophilus
-Heat-stable, provides X-factor
3. Nutrient agar
Enriched media Solid
Fastidous organisms
1. CAP = Heat-labile, provides X & V factor
Enrichment media Liquid
1. Selenite F
2. Alkaline peptone water
3. Thioglycollate broth
Differential media 1. BAP = hemolysis
2. MAC
3. EMB
4. XLD
5. HEA
Selective media Inhibitory media
1. TCBS
2. SSA
3. TMA
4. CBAP
lec.mt 04 |Page | 59
Calcium alginate swab Toxic to virus
Good for Neisseria
Needle aspiration Anaerobic and aerobic cultures
Catheterization Needle and syringe for collection
Intubation Gastritis
Vomitus
Gastric washing (aerobic culture only)
Delay in processing Refrigerate except:
1. CSF = Room temp. or 35’C
2. Blood
3. Swab of N. gonorrhoeae (sensitive to cold)
4. Urine = Boric acid
5. Rectal swab = Cary-Blair
Transport medium 1. Cary Blair = for stool pathogen
2. Stuart’s
3. Amies = Respiratory specimen
4. Transgrow = Neisseria
5. JEMBEC = Neisseria
6. Todd-Hewitt = Vaginal carriage (S. agalactiae)
Biologic safety HEPA filter: filters air
cabinet Negative pressure
BSC Class I Environment and MT protected
Air velocity = 75 linear ft/min
Exhaust air thru HEPA filter
Product contaminant
BSC Class II Vertical laminar airflow
MT, environment and product are protected
Air velocity = 75-100 linear ft/min
Recommended for hospitals
BSC Class III Supply and exhaust air thru HEPA filter
Maximum protection
N95 Mask Contains HEPA filter
For Mycobacterium
Stool No direct exam in Microbiology
Transposons Resistant gene
Mobile or jumping
Fusiform F. nucleatum
Capnocytophaga
BSL I No risk
BSL II Moderate risk
BSL III High risk
Treatment available
Inhalation of aerosols
Ex. Mycobacteria (BSC Class II)
BSL IV High risk
No treatment available
Inhalation of aerosols
lec.mt 04 |Page | 60
Ex. Small pox
Blood culture bottle BHIB + 0.25% SPS
Dilution = 1:10 (1mL blood, 9mL broth)
Anti-complementary, anticoagulant, antiphagocytic
Neutralizes aminoglycosides
Disadvantages of SPS Inhibits:
-Neisseria
-G. vaginalis
-S. moniliformis
-P. anaerobius
1% gelatin Counteract SPS to allow the growth of organisms
Indications of growth Hemolysis
(Blood culture) Turbidity
Pellicle
Subculture (Blood BAP
culture) MAC = no CO2
CAP
If blood culture = 7 days = Bacteremia (Typhoid)
negative 21 days = Brucellosis, SBE
Urine culture Specimen: Catheterized, Midstream, Suprapubic
Quantitative: BAP, MAC
->100,000 CFU/mL (or >50,000 CFU/mL) = significant for UTI
-<10,000 CFU/mL = not significant (contaminants)
CSF culture DO NOT refrigerate
Agents: Neisseria, Haemophilus (Meningitis)
Media: BAP, MAC, CAP, BHI
C. neoformans:
-India ink method
-Latex agglutination
Wound specimen Gram stain
Media: BAP, MAC, Thioglycollate broth
Stool specimen Media: MAC, BAP+Ampicillin, CBAP, SSA, Selenite F, TCBS,
APW, HEA
Oxidase test
Biochemical tests
Serologic typing
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: CAP, MTM
Gram stain
TB culture NALC-NaOH = Gold standard
-NALC = digestion, lyse the mucus
-NaOH = decontamination
lec.mt 04 |Page | 61
6% Oxalic acid = Pseudomonas
Centrifuge (4’C) for 15 mins at 3000g
Media: LJ, Middlebrook 7H11, 7H10 (AST)
Incubate at 37’C for 8 weeks ----(NG)----> Report as (-)
If (+), after 2-3 weeks: growth is seen
GenPro Genetic Pro
DNA test
Result 2 hrs
Moist heat 1. Autoclave (sporicidal)
sterilization -121’C at 15 lbs/psi for 15 mins
-Culture media, bandages, gauze
-QC: B. stearothermophilus
2. Inspissation (sporicidal)
-75-80’C for 2 hrs on 3 days
-Disinfect and solidify protein containing medium (LJ,
Loeffler’s)
-Water is heated from below and slanting surface gets heated
3. Tyndallization (sporicidal)
-100’C for 30mins on 3 days
4. Boiling (Nonsporicidal, disinfectant)
-100’C for 30mins
-Kills vegetative cells only
5. Pasteurization (Nonsporicidal, disinfectant)
-Milk
-63’C for 30mins
-72’C for 15secs
-Phosphatase: to determine if pasteurization is successful. (+):
Not pasteurized
Dy heat sterilization 1. Hot air oven (Sporicidal)
-170-180’C for 2 hrs
-Glasswares, cottonswabs, metallic instruments, oils, powders
-QC: B. subtilis
2. Incineration (Sporicidal)
-Waste disposal
-Not recommended
3. Cremation (Sporicidal)
-Prevents communicable disease
4. Flaming (Sporicidal)
-Needles
5. Gas: Ethylene oxide (sporicidal)
-Heat-labile machine instruments
lec.mt 04 |Page | 63
-(+) to MRSA
-Imipenem and cefotixin
Cell wall inhibitors Penicillin
Cephalosporin
Vancomycin = Tx: MRSA
Broad spectrum:
Bacitracin
Cycloserine
Carbapenems/Imipenem
Penicilinase-resistant: Methicillin, Cloxacillin, Nafcillin
Cell membrane Colistin = against Gram (-)
inhibitors Polymixin = against Gram (-)
Amphotericin B = drug of choice for systemic fungi
Nystatin = antifungal
Ribosome (Protein) Aminoglycosides (30S)
inhibitors -False-resistant = P. aeruginosa (Mg2+ and Ca2+)
Tetracycline (30S)
Chloramphenicol (50S)
Erythromycin/Macrolide (50S)
-Discovered by Bernardo Aguilar
-For penicillin allergic patients
Clindamycin (50S)
Nucleic acid (DNA) Mitomycin
inhibitors Quinolones
Metronidazole (Flagyl)
Trimethoprim-Sulfamethoxazole (SXT/Bactrim) = inh. folate
synth., synergistic
Anti-TB Pyrazinamide
Rifampin
Isoniazid
Streptomycin
Ethambutol
Micro/Macrobroth Reference method (AST)
dilution Det. MIC/MBC
Agar dilution Many organisms vs. single drug
Disk diffusion Pure culture vs. many drugs
E test (Epsilometer) Agar gradient diffusion
Antibiotic strip diffusion MIC test
MIC = Ellipse zone at intersection
Kirby-Bauer Disk Std. Inoculum: 1.5 x 108
Diffusion Medium: MHA
pH: 7.2-7.4
Depth: 4mm
Condition: Aerobic, No CO2
Temp: 35-37’C (MRSA: 35’C)
Incub. time: 16-18 hrs
Std: 0.5 McFarland (1% H2SO4 + 1.175% BaCl2)
Antibiotic disc: 6mm (refrigerated/frozen)
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Petroff-Hausser For bacterial count
counting chamber
15mm Distance of antibiotic disc to each other
15mins Time for the medium to absorb the bacteria after inoculation
w/in 15mins Inoculation of discs Incubation
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 Measure the outer zone
inhibition Ignore swarming
If there are colonies Gram stain the colonies
inside the zone of
inhibition
AST media 1. MHA = std. media
2. MHA + 2% NaCl = MRSA
3. MHA + 5% Sheep blood = S. pneumoniae (w/ CO2)
4. Haemophilus test medium:
-MHA + Yeast extract + Hemin + NAD + CO2
5. GC agar = Neisseria (w/ CO2)
6. Middlebrook 7H10 = Mycobacteria (w/ CO2)
QC Specific
Regular basis
Checking media and reagents w/ specific organisms to check
expected results
Set by CLSI (formerly NCCLS)
QA General
Snap shot
Total process whereby the quality of lab. reports can be
guaranteed
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
lec.mt 04 |Page | 65
ATCC (American Type For AST
Culture Collection) Stock culture: -20 or -70’C
Working culture: 2-8’C
ATCC-1234 Beta-lactamase producers:
-S. aureus
-N. gonorrhoeae
-H. influenzae
-Enterococcus
-E. coli
-P. aeruginosa
Catalase test Rgt: 3% H2O2
(+) Gas bubbles
F (+): BAP
b. M. chelonei
-(-) NO3 reduction
-(-) 5% NaCl
-(-) Iron uptake
2. M. smegmatis
3. M. phlei (“Hay bacillus”)
Growth: 10-21 days Groups I, II, III (Runyon’s Classification)
Growth: 3-7 days Group IV
(Rapid growers)
lec.mt 04 |Page | 76
Molar tooth colony, A. israelii
sulfur granules
Breadcrumb colony Fusobacterium
Gram (+) Anaerobic Actinomyces
Bacilli Bifidobacterium
Eubacterium
Propionebacterium
Lactobacillus
Mobiluncus
Gram (-) Anaerobic Bacteroides
Bacilli Porphyromonas
Prevotella
Fusobacterium
Gram (+) Anaerobic Peptostreptococcus
Cocci Peptococcus
Gram (-) Anaerobic Veillonella
Cocci Megasphera
Acidaaminococcus
A. bovis Lumpy jaw
P. acnes Acne vulgaris
(+) Catalase, Indole
L. acidophilus Tomato juice agar
(-) Catalase
Mobiluncus Vaginitis (G. vaginalis)
(-) Catalase
Anaerobic infections Foul odor
B. fragilis Needs 20% bile
F. necrophorum Vincent’s angina (B. vincenti)
(F. moriferum)
P. anaerobius SPS sensitive (Neut: 1% gelatin)
(-) Indole, Catalase
P. asaccharolyticus (-) Catalase
P. niger Staphylococcus-like
Black
H2S indicators TSI = Ferrous sulfate
LIA = Ferric ammonium citrate
SIM = Lead acetate
Phenol red Acid = Yellow
Alkaline = Red
Methyl red Acid = Red
Alkaline = Yellow
Neutral red Acid = Pink/red
Alkaline/neutral = Colorless
Bromthymol blue Acid = Yellow
Neutral = Green
Alkaline = Blue
Enterobacteriaceae All ferment glucose
lec.mt 04 |Page | 77
All are motile except:
-Shigella
-Klebsiella
-Y. pestis
-S. gallinarum
-S. pullorum
All are catalase (+)
All reduce NO3 NO2
All are oxidase (-) except P. shigelloides (+)
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
lec.mt 04 |Page | 78
Y. enterocolitica
Aerogenic Klebsiella
E. coli
Salmonella (except S. typhi, S. gallinarum)
Proteus
Arizona
Citrobacter
Enterobacter
Enteric Media
Inhibitory CHO Indicator LF NLF
EMB Eosin Y Lactose Eosin Y Red purple Colorless
Methylene blue Methylene blue (E. coli:
GMS)
MAC Bile salts Lactose Neutral red Red/pink Colorless
Crystal violet
SSA Bile salts Lactose Neutral red Red Colorless
DCA Bile salts Lactose Neutral red Red/pink Colorless
TCB Bile salts Sucrose Bromthymol blue Yellow Green
S
HEA Bile salts Salicin Bromthymol blue Yellow Green
Lactose
Sucrose
XLD Bile salts Xylose Phenol Red Yellow Red
Lactose
Sucrose
BSA Brilliant green Glucose Bismuth sulfite S. typhi = black colonies
(Old: Wilson-Blair)
Lysine-Ornithine-Arginine Reactions
“(LO)9(OA)3(L)4(O)6(N)6” - Enterobacteriaceae
L O A L O A
Escherichia coli + + - Shigella sonnei - + -
Enterobacter aerogenes + + - Proteus mirabilis - + -
Enterobacter gergoviae + + - Morganella morganii - + -
Edwardsiella tarda + + - Citrobacter - + -
Arizona + + - Yersinia enterocolitica - + -
Serratia + + - Salmonella paratyphi - + -
Hafnia + + - Providencia - - -
Salmonella enteritidis + + - Proteus vulgaris - - -
Salmonella choleraesuis Klebsiella
+ + - - - -
rhinoscleromatis
Enterobacter cloacae - + + Pantoea agglomerans - - -
Enterobacter sakazakii Yersinia
- + + - - -
pseudotuberculosis
Enterobacter taylorae - + + Yersinia pestis - - -
Salmonella typhi + - - Plesiomonas shigelloides + + +
Klebsiella pneumoniae + - - Vibrio + + -
Klebsiella oxytoca + - - Aeromonas hydrophila + - +
lec.mt 04 |Page | 79
Klebsiella ozaenae + - - Pseudomonas aeruginosa - - +
E. coli “Colon Bacillus”
#1 UTI
#2 Neonatal meningitis (K1 Ag)
MUG (+) except E. coli O157:H7
-MUG (+): Electric blue fluorescence
Lab Diagnosis:
-1st week: Blood
-2nd week: Urine and stool (carrier)
S. paratyphi C (S. Septicemia
choleraesuis)
S. enteritidis “Gardner’s Bacillus”
Shigella Biochemically inert
SSA: colorless
Related to E. coli:
-E. coli: Acetate (+)
-Shigella: Acetate (-)
Intestinal pathogen: Dysentery
Lab. Diagnosis:
-BEST: Culture of fresh stool w/ mucous flecks
-Rectal swab of ulcer
Shigella Subgroups (O A = S. dysenteriae (“Shiga Bacillus”)
Ag) B = S. flexneri (“Strong Bacillus”)
C = S. boydii
D = S. sonnei (Cross reacts w Plesiomonas)
Serratia marcescens Red pigment (Prodigiosin)
(+) DNase, Gelatinase, LPS
Red milk
Resistant to many antibiotics
Serratia odorifera Rancid, potato-like odor
Y. pestis “Plague Bacillus”
(+) Stalactite on broth medium
V and W antigens
MOT: Rat flea bite
Infections:
-Bubonic plague: buboes, lymphadenopathy
-Pneumonic plague
-Septicemic plague
-Black death
Y. enterocolitica Motile at RT’
Appendicitis-like infection
(+) ONPG
Y. pseudotuberculosis Motile at RT’
lec.mt 04 |Page | 81
Mesenteric lymphadenitis, septicemia
Vibrio “Comma Bacillus”
Comma-shaped
Darting motility
Oxidase (+) except V. mitschnikovii (-)
Halophilic except:
-V. cholerae
-V. mimicus
V. cholerae (+) Indole
(+) String test (0.5% Na desoxycholate)
Cholera: Ricewater stool
“Fish in the stream” appearance
Classical Biotype El Tor Biotype
Polymyxin Susceptibility S R
Lysis by bacteriophage + -
Chicken RBC
- +
agglutination
Classical Biotype El Tor Biotype
Hemolysis of sRBC - +
VP test - +
Hikojima
Ogawa Serotype Inaba Serotype
Serotype
Anti-Ogawa + - +
Anti-Inaba - + +
Lab. Diagnosis (V. Culture:
cholerae) -TCBS: Sucrose fermenter
-TTGA: Tellurite Taurocholate Gelatin Agar
-APW: 6-8 hrs Subculture (TCBS)
-Cary Blair: Transport medium (stool)
-Rectal swab: Carrier
O129 Susceptibility test = (S)
V. parahaemolyticus Halophilic (8% NaCl)
(+) Indole
#1 Gastroenteritis in Japan (seafood)
Nonsucrose fermenter (TCBS)
Kanagawa (+) = Beta hemolysis on Wagatsuma agar
V. vulnificus Stool pathogen
(+) Blood culture
(+) Lactose fermentation, ONPG
Nonsucrose fermenter (TCBS)
V. alginolyticus Sucrose fermenter (TCBS)
Aeromonas Motile (monotrichous)
(+) Oxidase
Beta-hemolytic
(R) O129
(+) DNase
(+) Bile esculin hydrolysis
lec.mt 04 |Page | 82
Plesiomonas (+) Oxidase
Non-hemolytic
(V) O129
(-) DNase
(-) Bile esculin hydrolysis
Campylobacter (G) 42’C
Curved, S-shaped (Wings of Seagull)
(+) Catalase
(+) Oxidase
Darting motility
C. jejuni: (+) Hippurate
H. pylori (+) Catalase
(+) Oxidase
(+) Urease
Test: Urease breath test
OF test Fermentative (close) = O/F = +/+ = Y/Y
Oxidative (open) = O/F = +/- = Y/G
Nonutilizer = O/F = -/- = G/G
Medium: Hugh and Leifson (1% glucose, 1% agar)
Indicator: Bromthymol blue
P. aeruginosa Old: P. pyocyanea
Pyocyanus: Blue pus agent
(+) Oxidase
Motile (monotrichous)
Capsule: Slimy layer
Grapelike odor (aminoacetaphenone)
(G) 42’C
Cetrimide medium: selective
Pigments:
-Pyocyanin: Best
-Pyoverdin/Fluorescein: (+) P. putida/P. fluorescens
Infections:
-#2 Burns (#1: S. aureus)
-Wound: Ecthyma gangrenosum
-Swimmer’s ear (Otitis externa)
-Dermatitis: whirlpool baths
-Contact lens infection
-#1 Cystic fibrosis
B. cepacia (+) Oxidase
Motile (lophotrichous)
#2 Cystic fibrosis
(+) Lactose, ONPG
lec.mt 04 |Page | 92
Streptococcus lactis Acid forming, causes normal souring of milk
Coliform bacteria Gas-forming, hydrolyze the milk protein casein
Bacillus subtilis Hay bacteria, proteolytic action on coagulated milk
Clostridium butyricum Stormy fermentation of milk
Clostridium perfringens
Pseudomonas syncyanea Blue milk
Serratia marcescens Red milk
Pseudomonas aeruginosa Blue green milk
Flavobacterium Yellow milk
synxanthum
Alcaligenes viscosus Slimy or ropy milk (capsule formation)
lec.mt 04 |Page | 93