BACTERIOLOGY

GRAM-NEGATIVE BACTERIA
Bacteria Morphology Catalase Oxidas Ferments Medium
e
Neisseriaceae Cocci & rods; + + Modified
Aerobic Thayer-
Martin
Neisseriaceae Kidney shape + + Glucose Capnophilic
gonorrhea in pairs Inside PMNs (neutrophil)
Gray-opaque colonies
Neisseriaceae + + Glucose,
meningitides maltose
Enterobacteriacea Bacilli + - Glucose Grow well in MacConkey
e /coccobacilli;
Aerobic
E. coli EMB = green metallic sheen
Salmonella Motile
Shigella NM

GRAM-POSITIVE BACTERIA
Bacteria Morphology Catalas BAP Coagulas Other
e e
Staphylococcus Cocci; Non- +
(General) motile;
Aerobic
Staphylococcus Cocci; NM; + beta + MSA = White-golden (cream)
aureus Aerobic Golden
yellow
color
Staphylococcus Cocci; NM; + non - Gray - white
epidermis Aerobic Novobiocin sensitive
Staphylococcus Cocci; NM; + non _ White - yellow
saphrophyticus Aerobic
Streptococcus Cocci; NM; -
(General) Anaerobic
Streptococcus Cocci; NM; - beta PYR = (+) Bacitracin Sensitive
pyogenes Anaerobic
(Group A)
Streptococcus Cocci; NM; - beta CAMP = Bacitracin Resistant
agalactiae Anaerobic (+) Normal flora in Female genitals
(Group B)
Streptococcus Cocci; NM; - beta
equismilis & Anaerobic
Streptococcus
equi (Group C)
Enterococcal: E. Cocci; NM; - PYR = (+)
faecalis (Group Anaerobic
D)
Non- Cocci; NM; -
enterococcal: S. Anaerobic
bovis (Group D)
Viridans: S. diplococci; - alpha Optochin Capnophilic
pneumoniae NM; = (+)
Anaerobic

MODULE 9: ENTEROBACTERICEAE
K. pneumoniae Friedlander’s bacillus
Yersinia pestis Plague: Bubonic, Septicemic, Pneumoniae
Yersinia enterocolitica Cefsulodin-Irgasan-Novobiocin (CIN) medium
Appear as bull eye
MODULE 10: NON-ENTERIC
Vibrio cholerae Comma-shaped, monotrichous flagella
Medium: TCBS, yellow colonies
String test, Cholera red test etc
Vibrio parahemolyticus Kanagawa phenomenon-pathogenicity
test
Campylobacter Motile; microaerophilic: “wings of seagull”
Warthin-Starry- stain, Cary-Blair, Butzler and Skirrow medium
Helicobacter pylori Rapid prod of urease
Medium: Skirrow’s medium
MODULE 11: NON-FERMENTIVE GRAM NEGATIVE BACILLI
Oxidative-Fermentive Medium (Hugh and Leifson)
Pseudomonas aeruginosa (+) Oxidase and Catalase; aerobic, M
Blue-green pigment: pyoverdine and pyocyanin
Burkholderia mallei NM; Aerobic; Rods
Glander’s disease – among animals like horse, goats and sheeps
Farcy – form of Glander’s disease
Burkholderia pseudomallei M; Aerobic; Rods
Meliodosis or Whitmore’s Disease – glader-like disease in humans and sea
animals
Medium: Ashdown
Burkholderia Most common with patients that has cystic fibrosis
Acinetobacter “Mima”
Oxidase Test:
Neisseria (+)
Acinetobacter (-)
MODULE 11: AEROBIC GRAM POSITIVE BACILLI
Bacillus anthracis The only NM in Bacillus spp.; Aerobic
Disjointed bamboo, medussa head colonies
3 Clinical Types:
Cutaneous – eschar formation
Pulmonary – Woolsorter’s disease
Intestinal – violent enteritis

Ascoli Test
Bacillus cereus “fried rice” Motile, Beta hemolytic
Associated with food poisoning
Emetic and Diarrheal Form
Bacillus subtilis Found in soil
Corynebacterium diphtheriae Kleb’s Loeffler’s bacillus
Chinese character arrangement
Intermedius, Mitis, Gravis
Pseudomembrane formation
In vivo test: Animal inoculation test
Aerobic Actinomycetes Nocardia, Nocardiolopsis, Streptomyces, Actinomadura, Rhodococcus, and
Dermatophilus
MODULE 12: SMALL PLEOMORPHIC GRAM NEGATIVE BACILLI
Grow slow in BAP and CAP and do not grow in MacConkey
Brucella / Bang’s bacillus Normal flora in animals
“Sandy appearance”
Malta & undulant fever
Bordetella Bordet-gengou medium: mercury droplets or pearl appearance
Causes Pertussis
Francisella tularensis Tularemia
Oculoglandular disease – conjunctiva of the eye
Medium: Blood-cysteine-glucose agar with Thiamine
Legionella pneuphilia “American Legion Convention in Philadelphia”
Pontiac Ferver; Legionnare’s disease
Medium: Buffered Charcoal yeast extract (BCYE)
Haemophillus Pfeiffer’s bacillus
“blood loving”
X and V Factor
Haemophillus ducreyi Chanchroid bacilli
Venereal disease
Requires X but not V factor
Haemophillus aegypticus Koch-Week’s bacillus
Sharing contaminated handkerchiefs and towels
Cause of Brazilian purpuric fever
Haemophillus parainfluenza Swine ‘hog’ influenza
V factor only
Pasteurella multocida Caused by dog and cat bite
HACEK Haemophillus, Aggregatibacter, Cardiobacterium, Eikenella, Kingella
Group of fastidious Gram-negative bacteria
Indigenous oral microbiota
MODULE 13: MYCOBACTERIUM GROUP
Gram +, Enhanced under increased CO2; NM; Aerobic
X,Y,V and L arrangement

Ziel-Neelsen Stain
Auramine-Rhodamine Stain

Exudative (Acute inflammation) & Granulomatous (Tubercle Formation)

Lesion
Extrapulmonary tuberculosis Pott’s disease
(Military TB) Lupus vulgaris
Scrotula

ON THE SPOT-EARLY MORNING

Lowenstein Jensen Culture – gold standard

Tuberculin Skin Test (TST)
Nucleic Acid Amplification Tests (NAATs)
Niacin Accumulation Test
Nitrate Reduction
Mycobacterium leprae Hansen’s bacillus
Lepromin Test
 (+) Tuberculoid (-) Lepromatous

DIFFERENT TEST

Procedure Result
Gram Stain Crystal Violet Gram + = blue color
Lugol’s Iodine Gram - = red color
Alcohol
Safranin
Acid Fast Stain Carbol Fuchsin AFB = pink or red (will retain the color
Heat of Carbol Fuchsin)
Methylene Blue / Malachite Green Non AFB – blue or green color
CARBOHYDRATE FERMENTATION TEST
Triple Sugar Iron (TSI)  To test the ability of org in fermenting  K = Red Color = Alkaline Prod.
sugars and producing hydrogen sulfide  A = Yellow Color = Acid Prod
 Identification of Gram – enteric  H2S = Black precipitate = Hydrogen
pathogens Sulfide Prod.
 G = Bubbles = Gas Prod.

Note:
o Ferrous sulfate & sodium
thiosulfate = for H2S prod.
o Fermenters attack glucose first
then lactose or sucrose

IMVIC Test  To identify Gram – bacteria and E. coli + + - -

differentiating members of K. pneumonia - - + +
Enterobacteriaceae

Indole: Indole
To detect the ability of the bacteria to (+) red ethereal/ring layer
produce a product, Indole in utilizing (-) no color change
tryptophanase.

Methyl Red: Methyl Red:

To detect the ability of bacteria to produce (+) red color
mixed acids from glucose metabolism (-) yellow color
through Mixed Acid Fermentation Pathway

Vogues-Proskauer: Vogues-Proskauer:
To detect the ability of bacteria to (+) pink – red color in top
metabolize pyruvate to acetoin. (-) no color change
Using a-naphthol & KOH

Citrate: Citrate:
Utilization of sodium citrate as sole source of (+) blue color
carbon. (-) green color
Medium: Simmon’s Citrate Medium
Oxidase Test To detect the bacteria that possess (+) deep purple color
cytochrome oxidase enzyme that will react (-) green color
 with the reagent
Reagent: tetra-p-phenylene-diamine or
paraaminodimethylaniline
Urease Test To detect the ability of the bacteria to (+) pink – red color
produce urea from enzyme, urease. (-) NCC or yellow
Urea = NH3 + CO2
Medium: Christensen Urea Medium
Phenylalanine deaminase To detect if org has deaminase that will (+) green color complex
Test deaminate phenylalanine to phenylpyruvic (-) no green color complex
acid after adding 10% FeCl, ferric chloride
Lysine Iron Agar (LIA) To detect if the org can deaminase or Deaminase
decarboxylase lysine and formation of H2S (+) Red
(-) remains purple
Deaminase (Slant) (Aerobic)
Decarboxylase (Butt) (Anaerobic) Decarboxylase
(+) Violet
(-) Yellow
Catalase Test To detect org that has catalase enzyme or (+) Bubbles; Gas
obligate aerobe org.
To differentiate staph and strep
Coagulase Test To identify staphylococcus which produces Slide Coagulase
coagulase (+) clumping/agglutination
Slide Coagulase – bound coagulase/clump
factor Tube Coagulase
Tube Coagulase – free coagulase (+) clot formation
Mannitol Salt Agar (MSA) To detect S. aureus (+) Yellow (Acid prod)
(-) Not S.aureus
CAMP Test To identify Strep Group B (+) formation of distinct arrow head at
intersection
PYR Test To detect pyrolidonyl arylamidase from (+) Deep cherry red color
group A and enterococci
Optochin Test To determine the effect of optochin towards
the org.
To differentiate alpha hemolytic strep.
Pneumonia from alpha strep. viridans

CULTURE MEDIUMS

Purpose Result
MacConkey  To grow Gram –  lactose fermenters – pink
 Ferments: Lactose colonies from
 non lactose fermenters – white
colonies
Salmonella – Shigella Agar (SSA)  To grow Gram –  Shigella = colorless
 Ferments: Lactose  Salmonella = small colonies;
black center (H2S production)
Bismuth Sulfite Agar (BSA)  Ferments: Glucose Salmonella = black colonies (H2S
prod.)
Eosin Methylene Blue (EMB) Ferments: Lactose E. coli = green metallic sheen
Thiosulfate Citrate Bile Salt Sucrose Ferments: Sucrose V. cholerae = small; yellow color
(TCBS) colonies
V. parahemolyticus = blue-green
colonies
Xylose – Lysine – Desoxycholate Ferments: Xylose,Lactose and
(XLD) Sucrose
Mueller-Hinton Agar Specific for Antibiotic Sensitivity
testing