Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

GRAM + BACILLI - SPORE FORMERS

• Virulence factor
BACILLUS CLOSTRIDIUM
a. D-glutamate capsule (not polysaccharide)
Aerobic Anaerobic
b. Exotoxin with 3 components: edema factor, lethal factor, protective antigen
▪
Function of exotoxin: to mediate cellular destruction
GENUS BACILLUS
• Selective medium: PLET (Polymyxin Lysozyme EDTA Thallous acetate)
BACILLUS ANTHRACIS
BACILLUS CEREUS & BACILLUS SUBTILIS
• Characteristics that will differentiate B. anthracis from other Bacillus spp: Non-motile
and Gamma hemolytic • Both are motile and beta hemolytic
• Forms the so called “disjointed bamboo fishing rod appearance”
BACILLUS CEREUS BACILLUS SUBTILIS
o Typically has square ends
• Fried rice bacillus • Hay bacillus
• Colonies:
• Virulence factor: exotoxin/enterotoxin • Classified under BSL 1
o Colonies with tenacious (sticky) consistency cholera like toxin • Opportunistic pathogen
▪ When lifted using a loop it will stand like a beaten egg white • Causes food poisoning • Causes eye infection in heroin addicts
o Colonies may show swirling projections giving it a Medusa head or lion head • Produces 2 types of toxin Blood bank contaminant @ RT
appearance • Best specimen for testing: suspected food • Penicillin resistant
o Forms the so-called string of pearl pattern • (+) gelatin hydrolysis • Selective media: Mannitol egg yolk
▪ Due to susceptibility to penicillin • (+) growth on PEA polymyxin B agar
o Colonies with comet tail appearance
o Colonies with cut-glass appearance or frosted glass appearance
GENUS CLOSTRIDIUM
• Forms inverted fir tree/pine tree appearance in gelatin media (semi solid)
• Tests • Strict anaerobes but some are aerotolerant
o Lecithinase (+) • Catalase (-)
▪ Media to detect lecithinase production: Egg yolk agar (EYA)
▪ (+) result: opaque zone around colonies Histotoxic Neurotoxic Enteric
C. Gas C. tetani Tetanus C. Pseudomembranous
o Gelatin hydrolysis (-)
perfringens gangrene/ C. Food borne difficile colitis
o No growth in PEA agar myonecrosis botulinum botulism
o Ascoli test – serologic test for the diagnosis of anthrax Infant botulism Antibiotic
• Causes anthrax Enteritis SID (sudden associated diarrhea
necroticans infant death
Cutaneous Pulmonary / Intestinal anthrax Injectional syndrome)
woolsorter’s disease anthrax
• (+) Black eschar • Ragpicker’s disease • Ingestion of • Associated with VIRULENCE FACTORS
• Least severe • Hide porter’s dse improperly cooked use of drugs of C. perfringens Alpha toxin & enterotoxin
• Most common • Inhalation of spores infected meat abuse like C. tetani Tetanospasmin – blocks the release of neurotransmitters causing spastic
• Direct contact while handling wool • Most severe heroine paralysis
• Least common C. botulinum Botulinum toxin
• Ingestion C. difficile Toxin A – enterotoxin and Toxin B - cytotoxin

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

CLOSTRIDIUM PERFIRNGENS CLOSTRIDIUM BOTULINUM

• a.k.a. Frankel’s Bacillus; C. welchii • A.k.a Canned good Bacillus; Von Ermengen’s Bacillus
• Microscopically forms box-car morphology • Causes Botulism -fatal type of food poisoning
• On BAP: Double/Target hemolysis • On BAP: Alpha hemolytic
o Colony surrounded by: • Produces Botulinum toxin that when release will cause flaccid paralysis
▪ Inner: Beta • Infant Botulism may develop as a result of spore ingestion via breast feeding, baby
▪ Outer: Alpha will manifest FLOPPY BABY SYNDROME
• (+) Lecithinase test and Nagler’s test using EYA • Spores are sub terminally located
• (+) Stormy fermentation of milk – Litmus milk • (+) Lipase
• Reverse CAMP Test (BAP)
o Known organism: S. agalactiae Motility Lecithinase Lipase Lactose Glucose
o Unknown organism: C. perfringens C. - + - + +
perfringens
o (+) result: enhanced hemolysis as shown by arrow head zone of beta
C. botulinum + - + - +
hemolysis
C. tetani + - - - -
D. difficile + - - - +
CLOSTRIDIUM TETANI *(+) Swarming: C. tetani and C. septicum
• Lollipop Bacillus, Tack head Bacillus, tennis racket bacillus, Drumstick bacillus
• Can produce TETANOSPASMIN GRAM + BACILLI-NON-SPORE FORMERS
• Diagnosis is often made by: CORYNEBACTERIUM DIPHTHERIAE
a. Clinical symptoms
• AKA Kleb Loeffler’s Bacillus
▪ Risus sardonicus – sardonic smile / distorted grin (lock jaw)
• Non-motile & highly pleomorphic
▪ Triggered by the release of tetanospasmin
• Morphologic arrangement
b. Observation of terminal swollen spores
o Club shaped appearance - one end typically swollen
o Palaside arrangement – side by side arrangement
CLOSTRIDIUM DIFFICILE
o Picket fence arrangement
• Causes Pseudomembranous colitis or _______________________________________ o X, y, v, l formation – Chinese letter arrangement
• Major cause of diarrhea in hospitals • On BAP: Beta hemolytic
• Normal flora of the colon • MEDIA for isolation
• Can produce Toxins A & B a. Leoffler’s serum & PAI’s coagulated egg
• Media to isolate: CCFA- Cycloserine cefoxitin fructose agar ▪ will stimulate granule formation & pleomorphism
o Horse stable or barn yard odor b. CTBA -cystine tellurite blood agar
• (+) growth on BAP it develops colonies that FLUORESCE CHARTREUSE ▪ Primary isolation media
• Diagnosis is usually thru detection of cytotoxin through immunoassay (EIA) ▪ Gray to black colonies
• Freshly passed stool – liquid/unformed stools for culture & toxin assay c. Tinsdale medium
• Formed stool/rectal swab – carrier state
▪ Black colonies with brown halo

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

• Toxigenic C. diphtheriae causes DIPTHERIA – low grade fever, mild sore throat & o Decontamination must be carried out to remove normal flora and other
body malaise. contaminating organisms
• Virulence Factor: Diptheria toxin o Digestion must be carried out to dissolve disulfide bond in mucus which could
• Characteristic symptom: Pseudomembrane – BULL’S NECK APPEARANCE trap MTB
o Initial site of infection: Epithelial cells of tonsils • Agents:
• (+) growth on media will require detection of toxin production o NALC (N-acetyl-L cysteine) and NaOH: Gold standard
• Appears beaded when stained with methylene blue because of Babes Ernst o 4% NaOH
metachromatic granules o Zephiran (benzalkonium chloride) and Trisodium phosphate / Z-TSP
• Colony types: Gravis, mitis, belfanti, and intermedius – based on the phenotypic o 1% Cetylpyridium chloride: prolong sputum shelf life
characteristics of size, texture, color, hemolysis, and presence of metachromatic o Oxalic acid 5%: remove gram (-) organisms like pseudomonas and proteus
granules
o Largest: Gravis PROCESSING OF SPECIMENS FOR MYCOBACTERIUM DETECTION
o Smallest: Mitis
Do SCREENING
TOXIGENECITY TEST Sterile (CSF) Centrifuge
Smear prep TEST
Modified Elek’s Animal Inoculation / Guinea Pig Lethal Test
• In vitro toxigenicity test • Suspension of isolated strain of C. and culture
Stain the smear
• On agar media, place a filter paper diphtheriae (0.3 ml) is injected
Non-sterile Digestion and w/ acid fast
strip with diphtheria anti-toxin, subcutaneously into thigh of two
(Sputum) Decontamination staining
streak + control at right angle to the guinea pigs, one protected
strip of anti-toxin, streak negative intramuscularly with 500 units of
control in the same manner. diphtheria anti-toxin 18-24 hours
Unknown culture suspected of C. before the test.
diphtheriae is streaked parallel to + • (+) result: The unprotected animal CULTURE MEDIA
and - control. Incubate at 35 degC dies within 2-3 days with evidence NON-SELECTIVE AGAR BASED MEDIA SELECTIVE LIQUID
for 24 – 48 hrs. of hemorrhage in the adrenal Egg based media with Clear media, easy
• (+) result: Lines of precipitation with glands These are non-
malachite green, inhibits examination of
arc of identity With antibiotics conventional
contaminating organism; colonies growth in 10-
media
growth in 6-10 weeks 12 days
Growth is
ACID FAST BACILLI – GENUS MYCOBACTERIUM Common Disadvantage: slow growth
detected earlier
• Lowenstein Jensen Media:
• Aerobic, non-motile
most common Gruft Modified
• Slow growers and requires whole egg for growth
• Petragnani- more Middlebrook 7H10 Lowenstein
• Difficult to stain but once stained difficult to decolorize due hydroxy methoxy acid / Bactec 12B,
inhibitory: best for non- with dextrose Jensen
mycolic acid Bactec 13A
sterile specimen Middlebrook 7H11 Selective
Septi-chek
• American Thoracic with casein Middlebrook
Middlebrook
SPECIMEN PROCESSING Society /ATS: less hydrolysate 7H11
7H9
inhibitory; best for sterile Mitchison 7H11 Mitchison
DIGESTION & DECONTAMINATION specimens Selective 7H11
• Dorset Egg Media
• Purpose:

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

• Wallenstein medium: may SALT TOLERANCE TEST • For identification of rapid growers & M. triviale
increase chances of 5% NaCl
recovering M. avium T2H/ TCH susceptibility • M. tuberculosis: Resistant
*If there is (+) growth in any of these media, next step is to do series of biochemical tests to Thiopene-2-carboxylic • M. bovis: Sensitive
identify species hydrazide

BIOCHEMICAL TESTS
DIRECT SPUTUM SMEAR MICROSCOPY (DSSM)
BIOCHEMICAL TEST REQUIRED REAGENT (+) RESULT • Smear size: 2x3 cm
MEDIA • Dry prior to heat fixation – to prevent aerosol
NIACIN TEST Lowenstein Strip impregnated with YELLOW • Examine 300 fields before reporting negative result
Jensen Media CYANOGEN BROMIDE (-) no color change
NITRATE REDUCTION Sodium After incubation we Initial (+) RED
Nitrate Broth add: (-) no color change SIGNIFICANT SPECIES
n-n -dimethyl-l-
naphthylamine After adding zinc M. TUBERCULOSIS
sulfanilic acid powder:
(+) no color change • a.k.a Koch’s Bacillus or Tubercle Bacillus
To confirm a (-) result (-) RED • Virulence Factors:
we add: a. CORD FACTOR
zinc dust/zinc powder b. SULFATIDES
(to detect unreduced
• With MUCH granules
nitrate)
• Produces cauli flower like growth at 35 – 37 deg C
IRON UPTAKE TEST Lowenstein 20% Ferric ammonium (+) Rust brown color
Jensen Media citrate • Obligate aerobe requires CO2 for growth
ARYLSULFATASE TEST Phenolphthale 2N Sodium carbonate (+) Pink • Causes Pulmonary TB
Detection of rapid in media o BCG vaccine/Bacillus Calmette Guerin - to prevent occurrence of TB; from M.
growers bovis
• To detect Exposure to M. tuberculosis:
(+) M. fortuitum-
a. Skin Testing – MANTOUX TEST – involves intradermal injection of PPD on the
chelonae
skin
PYRAZINAMIDASE TEST Pyrazinamida 1% ferrous ammonium Pink color band
se agar sulfate ▪ PPD (Purified protein derivative) – protein extracted and purified
HEAT STABLE CATALASE Tween 80 Heating of colonies at Vigorous bubbling from the cell wall of M. tuberculosis
TEST Media 68 deg C prior to the ▪ Von Pirquet – (+) redness on site
To detect heat labile addition of reagent is b. Gene expert
catalase Mycobacterium required (20 mins)
like M. tuberculosis *30% H2O2
▪ Rapid and sensitive method
TWEEN 80 HYDROLYSIS • Useful in the identification of M. kansasii – rapid tween (+) c. Culture
TEST • M. tuberculosis (+) in 10-20 days ▪ Gold standard for detection
• (+) result: Pink color • DOTS: Direct Observation Treatment Strategy
• Indicator: Neutral red
TELLURITE REDUCTION • For identification of M. avium
TEST • (+) result: black metallic tellurium

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

TYPES OF TB NON-TUBERCULOUS MYCOBACTERIA (RUNYON’s CLASSIFICATION)

Primary Secondary MDR (Multi drug XDR (Extensively
resistant) drug resistant) Photochromogens Scotochromogens Non-photochromogens Rapid growers
Resulting from initial Type that may Shows resistance to Shows resistance to M. kansasii M. szulgai M. avium complex M. abscessus
exposure; primary develop in those who RIPES (primary TB both primary and M. marinum M. scrofulaceum M. xenopi M. fortuitum
complex in children had TB in the past drugs) secondary drugs M. asiaticum M. gordonae M. ulcerans M. chelonae
Primary TB drugs: Rifampin, Isoniazid, Secondary line TB drugs: Ofloxacin, M. interjectum M. genavense
Pyrazinamide, Ethambutol, Streptomycin kanamycin, ciprofloxacin M. haemophilum

Other TB: FAMILY ENTEROBACTERIACEAE

• Pott’s disease – TB of the spine
• Military TB - disseminated form of tuberculosis GENERAL CHARACTERISTICS
• Gram (-) bacilli or coccobacilli
M. LEPRAE • Facultative anaerobe
• a.k.a HANSEN’s Bacillus • Non-spore former
• Forms the so-called Cigar packet arrangement • Some are with pili or fimbrae; grows well on MAC
• Non culturable on artificial media, but can be maintained in the lab using MICE FOOT • Oxidase (-)
PADS OR FOOT PADS OF ARMADILLO • Motile except: Klebsiella, Shigella, Y. pestis (at 37 deg C)
• Causes Leprosy • All Ferments glucose and often with gas production aerogenic except: Shigella
o Chronic disease affecting the skin, mucous membrane, peripheral nerves (loss • Catalase (+) except S. dysenteriae type 1
of sensation in severe cases) • Cytochrome oxidase (-) except: Plesiomonas
• Based on Clinical Infections produced there are 2 types of
Tuberculoid type/paucibacillary Lepromatous type/multibacillary a. Primary Intestinal Pathogens
• Milder and less sensitive • More severe ▪ Never normal flora
• Less contagious • Can become malignant
• Localized form • Disseminated form
▪ Salmonella, Shigella, Y. enterolitica
• Less/rare organism b. Opportunistic pathogens
▪ GIT flora that can produce infections outside the GIT

• Mode of transmission: direct contact Antigenic determinants (for characterization of E. coli, Klebsiella, Shigella, Salmonella)
o It has tropism for peripheral nerves O or somatic antigen K or envelope antigen H or flagellar antigen
• Basis of diagnosis: clinical symptoms – Leonine facies • For E. coli tehre are 164 • Consist of capsular • Protein in nature
types of O antigen and polysaccharide • Heat labile
• Lepromin Test: skin test to determine the type of leprosy a person has
specific types maybe • Heat labile • Unique to those motile
o Intradermal injection of inactivated M. leprae
associated with a • With K antigens: Klebsiella, only
• WADE FITE FARACO – to detect AFB in paraffinized tissues particular disease E. coli, Salmonella
o Instead of methylene blue, we use hematoxylin as counterstain • Serotype 0111: diarrhea in • Vi antigen = S. typhi
infants
• Serotype 0157: associated
with verotoxin production

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

o K. pneumoniae: pink-purple mucoid colonies

Use colonies for smear
prep
Do a series of
o E. coli: pink to purple colonies w/ green metallic sheen
Inoculate on Do gram staining to
STOOL plated media check if it is really gram
biochemical tests to o Enterobacter: pink-purple colonies with dark center giving it a fish eye
(-) bacilli identify
(MAC/EMB) genera/species appearance
Use colonies to do
cytochrome oxidase test

BIOCHEMICAL TESTS
CULTURE MEDIA CARBOHYDRATE FERMENTATION TEST
CULTURE CLASSIFICAT FERMENTAB INDICATO FERMEN • To detect acid production
INHIBITOR NON-FERMENTER
MEDIA ION LE CHO R TER
EMB/ Eosin Y • Media: TSI/Triple sugar Iron dispensed as Butt & Slant
Eosin Y &
EOSIN and
Methylene • Composition: lactose, sucrose, glucose
METHYLENE Methylene
Blue Pink -
BLUE Blue Lactose Colorless
purple Fermentable CHO pH indicator H2S indicator
MAC Crystal violet
Neutral
CONKEY
Selective
and
red 10 parts lactose Phenol red Sodium thiosulfate
AGAR Bile salts
& 10 parts sucrose Ferric ammonium sulfate
XLD
Xylose
Differential Xylose 1 part glucose
Media Bile salts Lactose Phenol red yellow Red/colorless
Lysine
Sucrose
deoxycholate
• Possible results:
Yellow –
HEKTOEN Salicin Green/colorless
Bromthym w/ or o In an acid pH: Yellow (A)
ENTERIC Bile Salts Lactose W or w/o black
ol blue w/o o In an alkaline pH: Red (K)
AGAR Sucrose w/black:
black
Colorless with Black o H2S production: Blackening
center: Salmonella o Gas production: Splitting of media; pulling away of media; cracks on media
SSA
Neutral
Salmonella Bile Salts Lactose Red
red Colorless without black Color: Slant / Report Interpretation Classification of Inoculated
Shigella Agar Selective
center: Shigella Butt results as organism
media
Red / Red K/K No sugar was fermented NON-FERMENTER
BSA Red/Yellow K/A Only 1 sugar was fermented NON-LACTOSE
Brilliant Bismuth
Bismuth
green
Glucose
sulfite
Salmonella = black (glucose) FERMENTER
sulfate agar
Yellow/ Yellow A/A 2-3 sugars were fermented LACTOSE FERMENTER

Other media:
• GN broth: Gram (-) broth enrichment media Lactose Fermenters Late Lactose Fermenters Non-Lactose Fermenter
• Selenite broth: Enrichment broth for salmonella and shigella Enterobacter Citrobacter Proteus
• Tetrathionate broth: Enrichment broth for salmonella Escherichia Serratia Providencia
• CIN (Cefsulodin Irgasan Novobiocin): for yersinia Klebsiella S. arizonae Morganella
• BGA (brilliant green agar) for other Salmonella except S. typhi S. sonnei Edwardsiela
Y. enterocolitica All Salmonella except S.
o Inhibitory agent: Brilliant green
Arizona
o pH indicator: phenol red All Shigella except S. sonnei
o Salmonella colonies growing in BGA may appear like snow flakes All Yersinia except Y.
• On EMB/ MAC: enterocolitica

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

IMVIC TEST M. morganii + + - - P. mirabilis - + - +

P. rettgeri
VOGUES Y. pestis - + - - + + - +
P. stuartii
INDOLE TEST METHYL RED TEST PROSKAUER CITRATE UTILIZATION K. pneumoniae
TEST - - + + Enterobacter spp. - - + +
K. ozaenae
• Based on the ability • Detect acid • Detects acetoin • Based on the ability of
of the organism to production or acetylmethyl the organism to utilize
produce indole from when glucose is carbinol citrate as a sole source ONPG TEST
tryptophan metabolized of carbon
• Detects • O-nitrophenyl-beta-d-galactopyranoside
Tryptophanase • To detect late lactose fermenters
MRVP BROTH MRVP BROTH • To differentiate Citrobacter which are ONPG (+) from Salmonella which are ONPG (-)
SIM MEDIA OR OR SIMMON CITRATE
• Salmonella are ONPG (-) except S. arizonae
TRYPTOPHAN BROTH CLARK LUBBS CLARK LUBBS AGAR
BROTH BROTH • May be carried out using ONPG broth/disk (+) result: Yellow
5% ALPHA
NAPHTOL ONPG POSITIVE ONPG NEGATIVE
No reagent will be Escherichia coli Proteus spp
KOVAC'S &
METHYL RED added after incubation Klebsiella spp Salmonella spp.
OR 40% KOH
INDICATOR since the media has Y. enterocolitica Shigella spp.
EHRLICH'S REAGENT (Barritt’s method)
incorporated indicator Enterobacter spp Providencia
Incubation: 24 hrs which is Bromthymol Citrobacter spp Y. pestis
Incubation: 18-24 hrs Colbelntz: 40%
blue S. sonnei Morganella
KOH wirh
creatine H. alvel Y. pseudotuberculosis
(+) Distinct red S. arizonae
(+) Pink to red
color pH less than
(+) red ring (-) No color color (+) Blue
4.5
development (-) No color (-) Green
(-) No color UREASE TEST
change
change
• Useful in the Identification of PPM -Proteus-Providencia-Morganella
• Based on the ability of the organism to convert urea to ammonia thru the action of
• KOVAC’S – paradimethyl aminobenzaldehyde the enzyme urease
• MRVP reactions are opposite • PPM are regarded as the rapid urease producers
• MR tubes must be incubated for 48 hrs before reading the result • Slow Urease (+)
o RESULTS read before 48 hrs = false positive o (+) after 4 hours are the following (KEY-SC) = Klebsiella; Enterobacter;
Yersinia; Serratia; Citrobacter
Sp. I M Vi C Sp. I M Vi C • Media is Christensen Urea Agar /Urea broth
E. coli + + - - C. freundii - + - + • Indicator Phenol red
S. marcescens
Y. enterocolitica + + - - - V + + • (+) result: pink; red
H. alvei
• (-) yellow
P. vulgaris + + - - S. sonnei - + - -
E. tarda + + - - S. typhi - + - - • (+) result after 4 hrs – orange
S. dysenteriae - + - - K. oxytoca + - + +

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

DEAMINASE TEST • DECARBOXYLASE TEST:

o (+) Purple
• Useful in the identification of PPM, these are the only deaminase (+) enterics o (-)Yellow
• Uses lysine, tryptophan or phenylalanine • DEAMINATION:
• PAD + overnight culture + 10% Ferric chloride o (+) Burgundy/red
• (+) result with Phenylalanine agar: Green o (-) Purple
• (+) result with tryptophan agar: Brown
LIA – INTERPRETATION OF RESULTS
Color of Slant/Butt Report it as Interpretation Possible Organism
DECARBOXYLASE TEST
Purple/purple K/K (-) deamination Salmonella
• Based on the ability of the organism to remove carboxyl group from specific amino (+) decarboxylation Others:
acid thru the release of the enzyme decarboxylase • E. coli
• Klebsiella
• There are 3 amino acids that can be used:
• E. aeorgenes
• Serratia
AMINO ACID CARBOXYL
• E. gergoviae
LYSINE CADAVERINE
• Hafnia
ORNITHINE PUTRESCINE
• E. tarda
ARGININE CITRULLINE Purple/Yellow K/A (-) deamination and Shigella
decarboxylation Others:
• E. cloacae
MOELLER’S BROTH LYSINE IRON AGAR • Citrobacter
• BROMCRESOL PURPLE + AMINO • Media which can be used to detect • Yersinia
ACID + 1%GLUCOSE deamination and decarboxylation Red/Yellow R/A (+) deamination PPM:
• Tubes are overlaid with Mineral Oil • It is a media dispensed as slant & (-) decarboxylation Proteus
• (+) result: Purple butt Providencia
• (-) result: Yellow • SLANT is observed for Deaminase Morganella
• USE OF MINERAL OIL: oxygen test
barrier • BUTT is observed for Decarboxylase
test

MUST KNOW IN BACTERIOLOGY

COLUMN I
DOME shaped colonies after 24 hrs and
After 48 hrs: nail head colonies appearing, crater like colonies
or colonies with checker appearance
Colonies appearing like a coin with a raised rim
COLUMN II
Photochromogens _________________________________________
_________________________________________
Scotochromogens _________________________________________
_________________________________________

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY
Lancet shaped diplococci
Virulence factor __________________________
Causes:
________________________________________
Otitis media
Lobar pneumonia -rusty colored sputum
Hockey Puck colonies – colonies remaining intact when pushed
across the plate using loop
Oxidase (+) because of indophenol oxidase
A cause of ____________________________
Butyrate Disk Test _____________________
(+) result _____________________________
Erysipelothrix rhusiopathiae Bottle brush like/test tube brush like or pipe cleaner pattern of
growth
An occupational hazard
H2S _______ on TSI
Causes: erysipeloid or ____________________
Abiotrophia Nutritionally Variant / NVS
Granulicatella Thiol Requiring Strep, Pyyridoxal requiring
Satelliting strep
Needs _________________________________
Colonies appearing like Bird’s nest on Corn meal agar
Water droplet colonies, TB in cattles intestinal TB in man
Niacin and Nitrate Reduction _______________
M. kansasii Cross barred bacillus, yellow Bacillus
# 2 NTM to cause TB in those with AIDS
“ ____________________________”
Non-photochromogen _________________________________________
_________________________________________
Rapid Growers _________________________________________
_________________________________________
Genus Vibrio Oxidase _________ except V. metschnikovii
Facultative anaerobe
Halophilic Except:
_________________________________________
All are glucose (+)
all are lactose (-) except: _________________
DIAGNOSTICS:
Specimens for diagnosis: _______________
Transport media : _____________________
Enrichment media _____________________
Selective-differential media ______________
CHO: _______________________________
Indicators ____________________________
Sucrose fermenter _____________________
Non-sucrose fermenter __________________
Vibrio cholerae Causes Cholera/ asiatic cholera or epidemic cholera – a
diarrheal disease characterized by the production of
_________________________________
Virulence Factor _________________________
Motility __________________________________
String test (+) ____________________________
Helicobacter pylori Found in the human gastric mucosa
Causes:
________________________________________
________________________________________
Virulence Factor: _________________________
Diagnosis:
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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

Runyoun’s Classification Photochromogens 1) Endoscopy

NTM / MOTT
Genus Proteus
Genus Providencia
Genus Morganella
Klebsiella-Enterobacter-
E. coli are
Serratia marcescens
Scotochromogens 2) Copro Stool antigen test
Non-photochromogens 3) Urea Breath test
Rapid growers 4) Culture
5) Tissue biopsy – Warthin Starry Stain
Swarming on BAP, Genus Shigella S. dysenteriae produces endotoxin and neurotoxin
Colonies with burnt chocolate and burnt gun powder odor Causes Bacillary
dysentery / Shigellosis S. sonnei ___________________________
Source of antigens in Weil Felix test Spread via fecal oral route
P. vulgaris ______________________
P. mirabilis _____________________
Rapid Urease ____ & Deaminase _________ Resembles E. coli, invasive, non-motile, H2S negative,
Decarboxylase negative
Resembles Citrobacter, motile, H2S positive, decarboxylase
positive
Produces red pigment ____________________________ E. coli # 1 cause of __________________
Produces gelatinase, lipase, Dnase Primary marker of fecal contamination
DNase test _____________________ IMViC___________________________________

Yersinia pestis Safety pin appearance/bipolar staining E. coli serotype that causes SEVERE BLOODY DIARRHEA due
Use of ____________________ stain to production of very powerful neurotoxin VERO toxin,
Causes Plague transmitted thru flea bites Hemolytic Uremic syndrome

On Broth- Stallactite growth /Flocculent growth Specific example is

On BAP -Hammered copper colonies E. coli serotype 1057:H7

Yersinia enterocolitica
Yersinia pseudotuberculosis
Genus Yersinia
Causes enterocolitis which may mimic symptoms
appendicitis
________________________________________
Can tolerate cold enrichment
(+) Bull’s eye colonies on CIN
________________________________________
On TSI ___________________________________
Causes mesenteric lymphadenitis
Triple decarboxylase (+)
Motile at RT but non-Motile at 37 degC except
_____________________________________
of E. coli serotype 0157:H7
Cronobacter sakazakii
Lactose __________ but Sorbitol ____________
Media: MACSOR / SMAC
MUG TEST ______________________
MUG TEST – detects enzyme beta -d- glucuronidase
Reagent:
4 methyl-umbelliferryl beta d-glucoronide
(+) result _______________________________
Causes Necrotizing colitis due to powdered infant milk formula
Produces ____________ at RT
Salmonella species considered as human pathogen
Test that will differentiate K. pneumoniae and K. oxytoca

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY
K. pneumonia
OF media of Hugh and Leifson
OF MEDIA -CDC method
Pseudomonas aeruginosa
Genus Haemophilus
X factor / Hemin
V factor or co-enzyme I
Causes community acquired pneumonia E. coli serotypes ETEC – causes severe watery diarrhea that is cholera like
a.k.a Friedlander’s Bacillus (Traveller’s diarrhea, Turista, Montezuma’s revenge)
Colonies ____________________ EIEC – causes bloody diarrhea, shigella like
Neufeld Quellung __________________ EPEC- causes diarrhea in infants
Wings of seagulls, Satellitism – luxuriant growth near the source of incorporated
microaerophilic & capnophilic V factor
CHO ____________________ H. influenzae a.k.a Pfeiffer’s bacillus
Indicator _________________________ causes:
(+) result _________________________ meningitis in children less than 5 y/o
(-) result _________________________ major virulence factor ____________________
with 6 serotypes,
Interpretation most significant is serotype ____________
(+) open and closed tubes _________________ School of fish appearance, finger print appearance, rail road
(+) open and (-) closed tube _________________ track appearance
(-) p[en and closed tubes ___________________ Glander/s Bacillus
pH indicator Detected in PAP’s-stained smear, Whiff test/sniff test (+)
(+) ________________ Clue cells in PAP’s smear
(-) _________________
#1 non- fermenter and ICU isolate Streptobacillus Puff balls/fluff balls growth on broth
Causes: moniliformis String of beads
Ecthyma gangrenosum Aggregatibacter
Swimmer’s ear infection/diver’s ear infection actinomyctemecomitans
Jaccuzi / hot tub syndrome Gram positive cocci, cause of SBE
________________________________________ Gram negative coccobacilli which may cause SBE
Media for initial isolation ____________________
Acetamide utilization (+)
Colonies with corn tortilla like/corn taco odor, overripe grapes
Pyocyanin _______________________________ Vietnam Time bomb, Mellioidosis
Pyoverdin _______________________________ Colonies with earthy odor on Ashdown media
Mostly normal flora Requiring cysteine, cysteine and thiosulfate for growth
Requires ______________ and __________ blood Media: BCGA blood cystine glucose agar
(-) growth on ________________________ Water rat trapper disease, Market men dse
Rabbit fever, lemming’s disease
Human Bite wound/Clenched fist wounds
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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

Requires only X factor Rat Bite fever

H. haemolyticus, H. aegypticus, H. influenza Animal Bite wound
Needs H. parainfluenzae, H. parahaemolyticus, H. paraphrophilus Legionella pneumophila Isolated from aircondition & cooling towers
______________________ May cause a form of Pneumonia
for growth Media: __________________________________

Leptospirosis detection
T. pallidum subspecies
T. cuniculi
T. vincentii
Tightly coiled spirochetes Pits and corrodes the agar, colonies with bleach like odor
Media: _________________________________
Gold standard Sero test
_________________________________________
Specimen on ist week: blood and CSF Colonies appearing like dew drops
2nd week __________________________-
Loosely coiled spirochetes Colonies with musty odor/smells like mushroom
Blood spirochete
Arthropod borne
Can be visualized using Brightfield microscope
Caused by Borrelia burgdorferi Colonies with MOLAR TOOTH appearance
Vector: Ticks (Ixodes spp), deer ticks
Black legged ticks
Causes Louse Borne relapsing fever Atypical pneumonia
Pediculus spp.
Primary syphilitic lesions Psittacosis/ornithosis/parrot fever
Secondary syphilitic lesions Ehrlichia and Anaplasma
infects
Tertiary syphilitic lesions Bartonella infects
Non-culturable in vitro but can be maintained alive using Rochalimaea quintana
testicular chancre of rabbits
PERTENUE causes Frambesia or ____________ Colonies appearing like bread crumbs
CARATEUM causes _______________________ Colonies with speckled appearance
ENDEMICUM causes Bejel / endemicum May appear like toothpick because of tapered ends
syphilis
MOT ____________________________________
Causes venereal spirochetosis TRIC/ Lymphogranuloma venereum
Causes Trench mouth / Vincent stomatitis
Rapid test for Brucella Cat scratch disease
Significant titer ____________________________
Brucella species which is NOT an agent of Bioterrorism Q fever
Potato Blood glycerol agar, Regan Lowe Mycoplasma/ Mollicutes Fried egg appearing colonies

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 Gram + Bacilli - Spore Formers and Non-Spore Formers • Mycobacterium • Family Enterobacteriaceae • MUST KNOW IN BACTERIOLOGY

Cell wall less

R. rickettsi Rickettsia infects
O. tsutsugamushi Brick red fluorescence under UV
R. prowazeki With vector: Resistant to kanamycin-vancomycin and colistin in Potency disk
W/o vector: Brill Zinsser disease test
Lumpy Jaw Colonies appearing like mercury droplets

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