NEISSERIA ➢ Difference between N. meningitidis and N.

lactamica = LACTOSE
• Genus NEISSERIA is characterized to be coffee bean/kidney- FERMENTATION
shaped ➢ Which Neisseria spp is ONPG positive?
• EXCEPT FOR N. elongata (described as a rod) A. N. lactamica
• obligate aerobes AND capnophilic - 5-10% CO2 B. ONPG is a test for B-galactosidase
o USE CANDLE JAR (3-5% CO2) C. For the utilization of lactose
o but this cannot be used to support growth for
ANAEROBES since 15% O2 is also present DEFINITIVE ID: CHO UTILIZATION
MOTILITY: NONMOTILE PRESUMPTIVE ID: SUPEROXOL TEST
CATALASE: POSITIVE (except N. elongata) - Rgt: 30% H2O2 [modified catalase test]
OXIDASE: POSITIVE - Key ID of Neisseria - Positive; bubbling
Rgt - tetramethyl-p-phenylenediamine dihydrochloride - Species: N. gonorrhoeae
o PREPARE FRESH DAILY
➢ GN diplococci was isolated on BAP AT 25 DEGREES CELSIUS.
NOTE: This does not have DMSO Positive for utilization of Glucose and Maltose
positive result for oxidase; "dark purple"

PATHOGENS [2]
- Will only grow at 37 C

1. N. gonorrhoeae
- Gonorrhea/clap
- Disseminated inf
o Gonococcal arthritis (from joint fluid)
o Opthalmia Neonatorium
Opthalmia Neonatorium A. Do not answer N meningitidis.
• Acquired during BIRTH from infected mothers. B. N. meningitidis = BAP & CAP at 37C
• PREVENTION: Crede’s prophylaxis C. At RT, only NONPATHOGENIC Neisseria will grow
o 1% AgNO3

2. N. meningitidis MORAXELLA
- Meningitis, meningococcemia
- DIC GRAM: NEGATIVE DIPLOCOCCI
- Waterhouse-Friderichsen Syndrome MOTILITY: NONMOTILE
o Bleeding, hemorrhages in ADRENAL GLAND CATALASE: POSITIVE
OXIDASE; POSITIVE
NOTE: DO NOT REPORT AS N. GONORRHOEAE FOUND since there are DNASE: POSITIVE
many similar bacteria MEDIA: BAP
- Instead report as GN Diplococci and ESTIMATE WBC CHO UTILIZATION: NONFERMENTER
found. COLONIES: Wagon Wheel [wavelike periphery], and Hockey Puck [can be
moved/swept around the plate INTACT]
DEFINITIVE ID:
- Culture (Enriched CA) ➢ Hockey puck AND Wagon Wheel colonies
- Utilization of CHO A. M. catarrhalis
o CTA: Cytine Trypticase Agar

➢ NYC agar – will also support growth of genital MYCOPLASMAS

(Mycoplasma hominis, Ureaplasma urealyticum)
RECALL:
➢ Glistening, dry, claylike colonies
A. N. elongata

ACINETOBACTER

GRAM: NEGATIVE COCCOBACILLI

MOTILITY: NONMOTILE
CATALASE: POSITIVE
OXIDASE: NEGATIVE
MEDIA: MAC
 ➢ What is the test that will differentiate Acinetobacter from
Neisseria?
A. Acinetobacter is OXIDASE NEGATIVE

KINGELLA

GRAM: NEGATIVE BACILLI

CATALASE: NEGATIVE
OXIDASE: POSITIVE

EIKENELLA
➢ There are antisera directed towards the O Ag, but did not yield
GRAM: NEGATIVE BACILLI
reactions, what possible remedy can be done?
CATALASE: NEGATIVE
A. Boiling/Heating. The O Ag might be enveloped by the
OXIDASE: POSITIVE
capsule.
• Eikenella corrodens: CORRODING BACTERIUM
• Characterized by PITS or CORRODES the surface of the agar
TESTS TO ID ENTERICS:
SMELL: Bleach odor
- Aside from Gram staining
NOTE:
1. CULTURE
- CAPNOPHILIC (5-10% CO2) 2. TSI
- Part of HACEK 3. LIA = ONLY PPM ARE Pos for lysine deamination

ENTEROBACTERIACEAE

GRAM: NEGATIVE, NON-SPORE FORMING

FACULTATIVE ANAEROBES
➢ What are the genera that are NONMOTILE under 37 C
A. Klebsiella
B. Shigella
C. Yersinia – Most are motile only at RT

CATALASE: POSITIVE
OXIDASE: NEGATIVE (EXCEPT PLESIOMONAS)
NITROSOREDUCTASE: POSITIVE (except Erwinia and Pantoea
agglomerans)
- Significance: reduction of nitrate to nitrite
o UTI ➢ What is the meaning of IMVIC?
CHO UTILIZATION: Glucose A. Indole, Methyl red, Voges-Prokaeur, Citrate
- May be differentiated through lactate fermentation. 4. INDOLE
Detects: Tryptophanase
• Grow well on MAC, some as LF [pink colonies] and some as NLF Medium: tryptone broth, SIM
[colorless colonies]. Indicator: Kovac’s or Ehrlich’s rgt
Pos result: red ring
GAS PRODUCTION: AEROGENIC (Except SHIGELLA – VERY INERT)
- Variable for gas prod = Salmonella, Proteus, Providencia

H2S PRODUCTION: [TSI] SPACEd = Salmonella, Proteus, Arizonae,

Citrobacter, and Edwardsiella
[LIA] – SACED

ANTIGENIC STRUCTURE:
 5. MRVP TEST - Detects the production of the UREASE ENZYME
Medium: Christensen’s urea agar, Stuart’s urea broth
pH Indicator: Phenol red [ A= Y; K=R]
Positive; RED
Product: AMMONIA (Alkaline product)

• Rapid Urease producers: pos in 2-4 hours [PPM]

➢ What is the ONLY Providencia that is urease pos?

Glucose Fermentation pathways:
1. Mixed Acid Production = detected by MR TEST
LAFS = Lactic, Acetic, Formic, Succinic
2. Butylene Glycol pathway = detected by VP TEST
Reagent: KOH, alpha-napthol
Product: acetoin (acetylmethylcarbinol)
- Methyl red [A = red; K = yellow]
➢ At which pH will the methyl red indicator turn red/
A. pH < 4.5
➢ What test in Microbiology uses KOH and alpha-napthol?
A. VP TEST
NOTE: FOR ENTEROBACTERIACEAEA, THE MRVP RESULTS ARE
USUALLY OPPOSITE
A. P. rettgeri
• Slow Urease Producers: pos in 4 hours [CKEYS]
o Citrobacter, Klebsiella, Enterobacter, Yersinia, and
Serratia
➢ What is the pH indicator of TSI, MSA, and Urease medium?
A. Phenol red
9. ONPG
- Rapid test to detect BETA-GALACTOSIDASE
- Help ID LATE lactose fermenters
Substrate: ONPG (o-nitrophenyl-beta-galactopyranoside)
Positive: YELLOW
➢ The ONPG test will detect for what enzyme?
A. BETA-GALACTOSIDASE
➢ What is the VALLUE of the ONPG test?
A. Help differentiate the LATE lactose fermenters from
the NON FERMENTERS of lactose
➢ What is the pos result of ONPG test?
A. YELLOW

6. CITRATE UTILIZATION TEST 10. Decarboxylase test/decarboxylation test

- Detects if an organism can UTILIZE CITRATE as a sole Medium: MOLLER’S MEDIUM
source of CARBON - Uses 4 TUBES
o 1 lysine
o 1 ornithine
o 1 arginine
- Primary component of SCA (Na citrate) o 1 control – no AA inside
Positive: PURPLE
Dye: BROMCRESOL PURPLE

➢ How many tubes are needed for the DECARBOXYLATION

TEST?
Pos result: BLUE A. FOUR [lysine, ornithine, arginine, control]
7. MALONATE TEST
- Detects if an organism can UTILIZE MALONATE as a sole
source of CARBON

- Similar to Citrate utilization test, but the carbon source is

changed to malonate

➢ What is the value of the Malonate test?

A. To help distinguish Salmonella from Shigella
B. Salmonella = malonate pos = Blue
C. Shigella = malonate neg = Yellow/green

8. UREASE
 • SIMILAR TO VIBRIO TOXIN
11. PHENYLALANINE DEAMINASE TEST • Traveler’s diarrhea (for micro recall)
• If Para recall, etiologic agent is G.
lamblia
EIEC Watery • Invades intestinal epith
with Blood • SIMILAR TO ACTIVITY OF SHIGELLA
Medium: Phenylalanine Medium SPP.
Rgt: 10% FeCl3 (like in CM, the screening test for Aminoaciduria ) EPEC Watery w/ • No toxins produced, non-invasive
Positive: GREEN FLUID/SLANT Mucus, no • Nosocomial line
o PPM blood • Responsible for DIARRHEA
OUTBREAKS IN NURSERIES
➢ What is the reagent for the Phenylalanine test? EHEC, • Produces TOXIN SIMILAR TO THE
A. 10% Ferric Chloride VTEC, or SHIGA TOXIN
➢ From the members of Enterobacteriaceae, who are positive for STEC • Associated with HUS – form of
Phenylalanine deamination? hemorrhagic colitis associated with
[verotoxic improperly cooked meat
A. PPM
– • Associated with TTP
cytotoxic
12. GELATIN LIQUEFACTION TEST to Vero
- Determine if an organism can break down gelatin into AA cells]
Positive: Liquefaction
➢ This test will detect if an organism can break down GELATIN into [STEC –
Amino acids. SHIGA
A. Serratia, Proteus (one way to differentiate Proteus TOXIN]
from Providencia and Morganella. EAEC Watery • Adheres to HOST CELLS
diarrhea • MOT is not well understood

➢ The E coli strain that produces a toxin similar to SHIGA TOXIN

A. EHEC/VTEC/STEC

➢ E coli strain associated with IMPROPERLY COOKED hamburger

patties.
A. EHEC/VTEC
➢ E coli serotype most commonly associated with HUS
A. E coli O157:H7
B. Before sending and unknown isolate pf E coli to a
➢ In general, E coli is positive for MUG test EXCEPT what strain? reference laboratory, it should be tested for the
A. E coli O157:H7 [associated with HUS] fermentation of the CHO Sorbitol – Sorbitol
B. MUG Test = Check for beta-d glucuronidase Substituted Mac
(electric blue fluorescence) C. Original MAC contains Lactose.
D. ONLY E coli serotype that is NON FERMENTER OF
SORBITOL = leading to colorless colonies

• All Klebsiella are Indole negative EXCEPT K. oxytoca = INDOLE

POS
• Similar to IMVIC of ENTEROBACTER
o It is differentiated through MOTILITY. Enterobacter is
MOTILE

TYPE STOOL NOTES

ETEC Watery • Grows only in BAP
• Produces toxins that mediate secretion
of water (lower electrolytes)
 • Same IMVIC with KLEBSIELLA. Differentiate through MOTILITY

➢ This is the most predominant isolate of Enterobacter spp.

A. E. cloacae
➢ This isolate of Enterobacter produces a yellow pigment that is
enhanced by incubation at RT/25 C
A. E. sakazakii (Chronobacter)

LDC = Lysine decarboxylase -> produces CADAVERINE

ODC = Ornithine decarboxylase -> produces PUTRESCINE
ADH = Arginine dihydrolase

Bacteria IMVIC LOA

K. pneumoniae - - ++ +--
K. oxytoca +-++ +--
E. aerogenes --++ LOLO =
++-
E. cloacae --++ OA SYA =
-++

➢ What is the product of Ornithine DC?

o Putrescine

• S. typhi – MOST COMMON CAUSE of typhoid fever

➢ During the second week what is the specimen where we can

collect the causative agent of Typoid fever?
GELATINASE: POSITIVE
A. Urine/stool
DNASE: POSITIVE
1ST WEEK BLOOD B
LIPASE: POSITIVE
2ND WEEK URINE/STOOL U
VP: POSITIVE (together with Kleb and Enterobacter)
3RD WEEK SERO TEST S
• S. MARCESCENS and S. RUBIDEA produce a RED PIGMENT = if both are present in choices, PRIORITIZE
PRODIGIOSIN – color is limited to the colonies only and will STOOL and then urine.
not diffuse to the media Blood > Stool > Urine
• S. ODORIFERA – produces a potato-like oder
• VP positive together with Enterobacter and Klebsiella. Producer of
LARGE AMOUNTS OF GAS for TSI as well NOTE:
1. Salmonella can also be recovered from BILE
2. SSA: Salmonella grows as COLORLESS COLONIES WITH A
HAFNIA ALVEI BLACK CENTER
• Formerly known as ENTEROBACTER 3. BSA; Bismuth sulfite agar
• Differentiated through LACTOSE FERMENTATION - SELECTIVE for S. typhi
H. alvei Enterobacter sp. COLONIES: BLACK COLONIES WITH METALLIC SHEEN
• LATE LF • RAPID LF
• CITRATE NEGATIVE • CITRATE POSITIVE 4. WIDAL’S TEST – Classic Tube Dilution Agglutination test for
Typhoid fever
Ab titers Interpretation Notes
O Ag ≥ 160 ACTIVE INF Ab from O Ag are
IgM
 H Ag ≥ 160 PAST INF or IgG A. Shigella
IMMUNIZTION B. Klebsiella
High titer of Ab to CARRIER STATE C. Yersinia [motile at 22 C/RT]
Vi Ag D. Yersinia pestis = NONMOTILE AT RT and at 37 C

5. Shewanella ➢ The bacteria associated with plague or BLACK DEATH

• Like salmonella, NLF, H2S+ A. Yersinia pestis
• Differentiated from salmonella through HEA B. Characteristic inclusion: BIPOLAR BODIES
o SALMONELLA = SUCROSE FERM NEG = GREEN [Aggregate on both poles] = WAYSON STAIN
o SHEWANELLA = SUCROSE FERM POS = YELLOW C. SAFETY PIN APPEARANCE on Wayson stain

➢ Name of the cook who was a carrier of Typhoid. ➢ Bacteria associated with MESENTERIC LYMPHADENITIS
o Mary Mallon = Typhoid Mary A. Y. enterocolitica
B. Culture medium; CIN [Cefsulodin Irgasan Novobiocin]
C. Y. enterocolitica - RED BULL’S EYE COLONIES

NOTE: DO NOT CONFUSE RED BULLS EYE COLONIES (Y. enterocolitica)

with FISH EYE (Enterobacter)

• Known for Bacillary Dysentery

• VERY INERT
• ALL Shigella spp are mostly CATALSE POS and MANNITOL
FERM POS EXCEPT for S. dysenteriae.

➢ TAKE NOTE OF THE REPRESENTATIVES FROM EACH

GROUP
• NONLACTOSE FERMENTERS
• MOTILITY: MOTILE
• MR: POSITIVE
• LYSINE DEAMINATION: POSITIVE
• DNASE: POSITIVE (Proteus)
• OXIDASE: NEGATIVE

➢ What is the only UREASE POSITIVE member of

PROVIDENCIA
A. P. rettgeri
• Biochemically resembles Salmonella
o Differentiated through ONPG, LDC, and KCN ➢ Bacteria associate with the formation of renal calculi
A. Proteus spp.
NOTE: C.freundii = H2S POS B. STAGHORN CALCULI – composed of struvite =
C. diversus = H2S NEG mineral name of Triple phosphate (Magnesium
Ammonium Phosphate)
➢ The source of Ag for the Weil-felix rxn (Dx rickettsial inf)
A. Proteus spp. [P. vulgaris = OX2 & OX -19; P. mirabilis
= OX-K (KINGSBURY)]
B. It SHARES A COMMON POLYSACCHARIDE WITH
RICKETTSIA

➢ What are the Proteus Ag’s used for the Weil-felix rxn?
A. P. vulgaris = OX2 & OX -19
B. P. mirabilis = OX-K (KINGSBURY)]

NOTE: Proteus and Serratia are the ONLY Enterics with DNASE
- They are differentiated by:
PAD H2S DNASE
➢ Genera that are NONMOTILE at 37 C Proteus POSITIVE POSITIVE POSITIVE
Serratia NEGATIVE NEGATIVE POSITIVE
 • P. vulgaris vs P. mirabilis
o vulgaris = Indole POSITIVE
o mirabilis = Indole NEGATIVE

IMVIC: + + - - [similar to E coli]

• To differentiate, E. tarda is NLF AND H2S POS
• Thus, this also resembles SAMONELLA ➢ What is the organism that exhibits SERRATE, CONFLUENT
GROWTH?
Edwardsiella resembles; A. Pseudomonas aeruginosa
1. E coli = through IMVIC – differentiated through Lact ferm and
H2S prod ➢ What organisms grows in CETRIMIDE medium
2. Salmonella = Through Lact ferm and H2S prod – differentiated A. Pseudomonas aeruginosa
through
CHARACTERISTIC ODORS:

• ESBL STRAINS ARE RESISTANT TO;

o CAP
o Cephalosporins
o Aztreonam
o Penicillin

OTHER GN BACILLI

CHARACTERISTIC PIGMENTS:
Campylobacter
P. aeruginosa

OXIDASE: POSITIVE
• RGT: Tetramethyl-p-phenylenediamine dihydrochloride
• Result: Dark purple
PIGMENTS: Pyocyanin, Pyoverdin (Fluorescein)
• These pigments are DIFFUSABLE to the agar media, UNLIKE
SERRATIA MARCESCENS where the pigment is limited to the
colonies alone [PRODIGIOSIN]
COMMON INFECTIONS:
1. Blue-green pus in would & burn px
2. UTI
3. Nosocomial pneumonia
4. Septicemia in immunocompromised px and infants
5. Septic arthritis in IV drug abusers
6. Swimmer’s ears
7. Destructive eye inf (keratitis, corneal ulcers) in contact lens
wearers
8. Chronic lung inf in px with cystic fibrosis
9. Hot tub/ whirlpool dermatitis
• In spas
10. Skin lesions of P. aeruginosa = ECTHYMA GANGRENOSUM

NOTE:
Swimmer’s ears = P. aeruginosa
Swimming pool granuloma = Mycobacterium marinum
Swimmer’s itch = BLOOD FLUKES (in parasitology)
Contact lens care/cleaning solution = A PA
• Acanthamoeba
• Pseudomonas aeruginosa ➢ How do you differentiate B. gladioli from S. maltophilia?
o OXIDATION OF MALTOSE AND LYSINE DC
➢ RESPIRATORY INFECTIONS IN PATIENTS WITH CYSTIC o DO NOT ANSWER OXIDASE TEST = GLADIOLI IS
FIBROSIS OXIDASE NEG
A. P. aeruginosa
▪ OXIDASE POSITIVE
▪ GROWTH AT 42 C
B. Burkholderia cepacia
▪ OXIDASE POSITIVE
▪ Wont exhibit growth at 42 C
▪ Belonged to genus Pseudomonas before
C. Stenotrophomonas maltophilia
▪ OXIDASE NEGATIVE
▪ CHARACTERISTIC PIGMENT;
LAVENDER GREEN COLONIES MEDIA:
▪ ODOR: Ammonia odor APW • Alkaline peptone water (pH 8.4) may be used as
an enrichment broth for obtaining growth of Vibrios
• Causative agent of GLANDERS DSS from stool
BURKHOLDERIA MALLEI in horses, mules, and donkeys; NOT TCBS • SELECTIVE & DIFFERENTIAL medium
PART OF HUMAN FLORA
• Causative agent of MELIOIDOSIS INIHIBITORS:
BURKHOLDERIA (Glanders-like dss of ANIMAL and • 1% NaCl, Bile salts = inhibit the growth of GP
PSEUDOMALLEI MAN) organisms,
• In several forms, including the • sucrose for the differentiation of the various Vibrio
[Whitmore’s Bacillus] formation of skin abscesses, sepsis, spp.
[VIETNAMESE TIME and septic shock, abscess formation in
BOMB DSS] several internal organs, and acute pH INDICATORS:
pulmonary dss. • Bromothymol blue [A=YELLOW; K=BLUE]
• Environment (soil, water, plants); • thymol blue
survives well in HOSPITAL
BURKHOLDERIA • High pH of the medium inhibits the growth of other
ENVIRONMENT; not part of human
CEPACIA intestinal flora
flora; may colonize RESPIRATORY
TRACT OF PX with Cystic Fibrosis
RECALL CHO UTILIZATION: SUCROSE (V. cholerae is SF, it will form YELLOW
• Environmental pathogen of plants COLONIES in TCBS; V. parahaemolyticus is non-SF, it will form GREEN
• Occasionally found in respiratory tract COLONIES)
of px with Cystic Fibrosis but NOT
BURKHOLDERIA PART OF NORMAL FLORA STRING TEST:
GLADIOLI • Included with the OXIDASE • Value of the test: Used to differentiate Vibrio spp. from
NEGATIVE organisms because Aeromonas spp AND Plesiomonas spp..
oxidase reactions are frequently weak. • Organisms are EMULSIFIED in 0.5% NA DEOXYCHOLATE,
which LYSES VIBRIO CELLS, but not those of Aeromonas

VIBRIO = POSITIVE (LYSIS)

AEROMONAS AND PLESIOMONAS = NEGATIVE
 • Divided into three major subgroups: V. cholerae O1, V. cholerae
O139, and V. cholerae non-O1
• Somatic Ag O1 and O139 are associated with the V. cholerae cell
envelope are POSITIVE MARKERS for strains capable of
epidemic and pandemic spread of dss
• CHOLERAGEN TOXIN - Cholera toxin causes mucosal cells to
HYPERSECRETE WATER and ELECTROLYTES into the lumen
of the GI tracts OXIDASE: POSITIVE
• RICE WATER STOOLS composed of fluids and mucous flecks CATALASE: POSITIVE
are the HALLMARK OF CHOLERA TOXIN ACTIVITY INCUBATION: 5-10% CO2 (Capnophilic, HACEK)
• Fecal-oral route, by ingestion of contaminated washing, • Blood-loving bacilli
swimming, cooking, or drinking water; also by ingestion of • GN, non-spore forming.
contaminated shellfish or other seafood
➢ What are the spore forming bacilli?
Vibrio = Rice water stool – Salmonella = Pea soup stool o Clostridium, Bacillus
watery, gray, w/ mucin
1. X factor/ hemin
• From degradation of hemoglobin
SEROGROUPS:
• HEAT STABLE
1. Inaba – PHILIPPINES
2. Ogawa – INDIA
2. V Factor/NAD/Coenzyme I
3. Hikojima - JAPAN
• Produced by some bacteria (Staph aureus) and yeasts.
BIOTYPES (BIOVARS): o H. influenzae will produce SATELLITE COLONIES
around S. aureus.
CLASSICAL EL TOR
[pandemics of the past] [recent pandemics] • HEAT LABILE
RBC HEMOLYSIS NEG POS
VP NEG POS ➢ D-ALA TEST meaning
POLYMYXIN B Susceptible Resistant o aminolevulinic acid test
AGGLUTINATION NEG POS o a PRECURSOR OF HEME
WITH CHICKEN
RBCs ➢ Clinical significance for the D-ALA test
o For members of genus Haemophilus
• CLIN SIG: wound and ear infections
associated with marine environment. ➢ The D-ALA test detects requirements for which factors?
• Halophilic o X FACTOR (ONLY)
V. ALGINOLYTICUS
• SUCROSE FERMENTATION: o D-ALA is a precursor of HEME, so it detects HEMIN
POSITIVE
• TCBS: Yellow colonies o Organisms that DO NOT require the X- factor are D-ALA
• CLIN SIG: Gastroenteritis, usually POSITIVE = since they have enzymes that will convert
associated with contaminated seafood. porphobilinogen and porphyrin to HEMIN
• Halophilic o Those organisms that require the X-factor are D-ALA NEGATIVE
V. PARAHEMOLYTICUS
• SUCROSE FERMENTATION:
POSITIVE
• TCBS: Green colonies
• CLIN SIG: Gastroenteritis and ear
infections associated with marine
environment.
V. MIMICUS • Non-halophilic
• SUCROSE FERMENTATION:
NEGATIVE
• TCBS: Green colonies
• CLIN SIG: Septicemia and wound NOTE:
infections involving MARINE • ALL Haemophilus spp. with “PARA” ONLY REQUIRE FACTOR V
ENVIRONMENT (wounds from (NAD/coenzyme 1)
V. VULNIFICUS
shells/shellfish) • ONLY H. haemolyticus and H. parahaemolyticus are beta-
• Halophilic hemolytic
• TCBS: Variable • ALL organisms requiring X factor are D-ALA NEGATIVE =
influenzae, haemolyticus, aegypticus, ducreyi
• Hemophilus spp NEED HORSE BLOOD for BAP
o To demonstrate beta-hem (hemaolyticus and
parahemolyticus)

➢ Factor Requirement of H. aprophilus

o NONE
 ➢ The use of sheep blood will not demonstrate beta-hemolytic
Hemophilus. What should be our source of blood?
o Horse blood

➢ Respiratory spx, Beta-hemolytic organism requires X and V

factor.
o H. haemolyticus – BETA HEMOLYTIC = MISTAKEN
AS S. PYOGENES, THE PREDOMINANT
PATHOGEN of resp tract.
o H. influenzae is - GAMMA HEMOLYTIC
GROWTH REQ: X and V factors
COMMON NAME: Koch-Week’s bacillus
CLIN SIG: Pink eye conjunctivitis

➢ How do we differentiate H. haemolyticus from S. pyogenes? NOTE: KOCH’S BACILLUS IS NOT THE SAME AS KOCK-WEEK’S
o Gram staining BACILLUS

➢ CSF sample, nonhemolytic, requiring X and V factors Koch’s bacillus = MTB

o H. influenzae Koch-Week’s bacillus = H. aegypticus

GROWTH REQ: X factor ONLY

CLIN SIG: Chancroid/Soft chancre = venereal dss characterized by painful
ulcers in the genitalia
DIRECT EXAMINATION:
• COMMON NAME: Pfeiffer’s bacillus • Short bacilli in a SCHOOL OF FISH ARRANGEMENT.
• RAIL ROAD TRACK PATTERN
➢ What is the most common serotype of H. influenzae associated
with infection?
o H. influenzae Serotype b ➢ The SMALLEST PATHOGENIC BACILLI
o H. ducreyi
• NOT ALL STRAINS are encapsulated.
o The encapsulated strains are ➢ LARGEST PATHOGENIC BACILLI
PATHOGENIC/VIRULENT o Bacillus anthracis
o Nonencapsulated strains are NON-PATHOGENIC
• Associated with respiratory conditions including EPIGLOTTITIS. ➢ SMALLEST FREE-LIVING ORGANISM/BACTERIA
o MYCOPLASMA SPP.
GROWTH REQ: X and V factors
Soft chancre/CHANCROID = H. ducreyi [PAINFULLLL]
STAPHYLOCOCCAL STREAK • DUCRYYYYYYY KAY SAKIT
• BAP streaked with a known beta-hemolytic organism (S. aureus)
Hard chancre = T. pallidum [PAINLESS]
• The UNKNOWN ORGANISM is streaked PERPENDICULARLY to
S. aureus. • Hard, gahi na, tough love mao manhid
• H. influenzae requires X and V factors. When blood is hemolyzed,
IT WILL RELEASE X FACTOR and S. AUREUS CAN PRODUCE
V FACTOR CAMPYLOBACTER, ARCOBACTER, HELICOBACTER
• This leads to SATELLITE/DEW DROP COLONIES around S. - Related organisms
aureus. This phenomenon is known as SATELLITISM. GN, Small, Curved, Motile Bacilli

APPEARANCE: S-shaped; similar to Seagull’s wings

HIPPURATE HYDROLYSIS (hippuricase): POSTIVE
O2 CO2 AVAILABILITY: Microaerophilic = 5% O2, 10% CO2, 85% N2
 ➢ How to differentiate Campylobacter from Helicobacter HELICOBACTER INF USALLY ARE ID BY NONCULTURE METHODS
Campylobacter Helicobacter
OPTIMAL 42-43 C 35-37 C 1. Presumptively ID in a GASTRIC BIOPSY SPX by testing for the
TEMP presence of a rapid urease rxn
FOR [PA. C = 2. Urease activity can also be detected be detected by the UREA
GROWTH Pseudomonas BREATH TEST which is reportedly sensitive and specific and is
aeruginosa & recommended for monitoring therapy.
Campylobacter] • In this test, the px is given 13 C and 14 C- labeled urea orally.
MOTILITY Darting • Motile 4-6 POLAR Urea degraded by the urease activity of H. pylori in the
• Motile by means of FLAGELLA stomach releases 13 CO2 or 14 CO2 which is ABSORBED
a SINGLE POLAR into the bloodstream and detected in the exhaled breath by a
FLAGELLUM SCINTILLATION COUNTER
MEDIA Campy’s BAP Campy’s BAP 3. FECAL AG DETECTION
Skirrow’s medium Skirrow’s medium 4. MICROSCOPIC EXAMINATION of stained gastric tissue
5. DNA AMPLIFICATION TESTS (PCR)

➢ Which group of tests best differentiates H. pylori from C. jejuni?

➢ ID the organism: Skirrow’s media at 42 C o Urease, nitrate, Hippurate hydrolysis
o Campylobacter

➢ ID the organism: Skirrow’s media at 37 C

o Helicobacter

Human pathogens
• Associated with GASTRITIS and DIARRHEA
1. C. coli
2. C. jejuni o
o Include REACTIVE ARTHRITIS
o GUILLAIN-BARRE SYNDROME ➢ Organisms that exhibit HIPPURATE HYDROLYSIS
o ONLY Campylobacter that is HIPPURATE o S. agalactiae
HYDROLYSIS POSITIVE o L. monocytogenes
o G. vaginalis
➢ C. jejuni is associate with Guillain-Barre Syndrome and is o C. jejuni
Hippurate Hydrolysis POSITIVE

Animal pathogens
1. C. fetus
• Associated with ABORTIONS in animals

➢ How to differentiate animal pathogens from human pathogens?

o Animal Campylobacter spp show NO GROWTH at 42
C GRAM NEGATIVE NON-SPOREFORMING BACILLI
MOTILITY: NONMOTILE (except B. brociseptica)
CLIN SIG: Whooping cough (B. pertussis)
• 3 stages of pertussis (CPC)
1. CATARRHAL – gen flulike symptoms
2. PAROXYSMAL – repetitive coughing episodes
3. CONVALESCENT – recovery phase

IDEAL SPX: NPS

APPEARANCE: S-shaped; similar to Seagull’s wings (like Campylobacter)
O2 CO2 AVAILABILITY: Microaerophilic = 5% O2, 10% CO2, 85% N2
CLIN SIG: PEPTIC ULCER
KEY ENZYME: UREASE
NOTE: To differentiate between Helicobacter and Campylobacter spp, kindly
refer to the table above
➢ Helicobacter pylori can survive the acidity of stomach acid
through the KEY ENZYME {{UREASE}}
- Urease converts UREA to AMMONIA, thus neutralizing the
acidic pH
MEDIA:
• Bordet-Gengou medium (potato-blood-glycerol)
ENRICHED MEDIA ONLY
• Regan-Lowe (horse blood, charcoal, amphotericin, cephalexin) –
PREFERRED since it is SELECTIVE w/ antibiotics
COLONIES: Mercury droplet colonies
B. parapertussis – pertussis-like syndrome [MILD FORM]
B. bronchiseptica – inhabits respiratory tract of canines (KENNEL’S
COUGH),
* infrequently responsible for chronic respiratory tract infections in humans
[MOTILE BORDETELLA]
 UREASE MOTILITY NITRATE OXIDASE COMMERCIAL BREAK!!!!
B. PERTUSSIS - - - + Grade A Milk
B. • Obtained ONLY from healthy cows
+ - - -
PARAPERTUSSIS • Bacterial content
B. RAW: below 75,000 bacteria/mL
+ + + +
BRONCHISEPTICA • PASTEURIZED: below 15, 000 bacteria/mL

• B. pertussis = only UREASE NEG ➢ What is the ESTIMATE OF THE TOTAL NUMBER of viable
• B. bronchiseptica = ONLY MOTILE AND NITRATE TEST aerobic bacteria present in raw milk
POSITVIE o STANDARD PLATE COUNT
• B. parapertussis = only OXIDASE NEGATIVE

• Naturally found in both NATURAL and ARTIFICIAL water

sources; bacteria have bee found in ponds, creeks, and streams,
wet soil, WATER-COOLING TOWERS for Air Conditioning and
Heating system, showerheads, and plumbing systems
• Not usually detectable directly in clinical material by Gram stain;
the use of 0.1% FUCHSIN substituted for SAFRANIN in the Gram
stain procedure may enhance the visibility of the organisms

SOURCE: H2O sources

GRAM NEGATIVE NON-SPOREFORMING BACILLI 1. Natural: ponds, creeks, and streams
MOTILITY: NONMOTILE 2. Artificial: water cooling and heating systems
CLIN SIG: Brucellosis, undulant fever (up and down/on and off temperature),
MALTA FEVER ➢ When gram staining Legionella, what gram stain will you use?
DIRECT EXAMINATION: Small nonmotile aerobic coccobacilli or short rods o 0.1% FUCHSIN
MOT: Infection through [1] ingestion of contaminated animal products (milk MEDIA: BCYE medium
and dairy products) [2] meats and milk, [3] handlers/farmers can directly IDEAL SPX: Sputum, Bronchial wash
acquire inf through direct animal contact
IDEAL SPX: Blood or Bone marrow L. PNEUMOPHILA Legionnaire’s dss, Pontiac fever [mild form of
MEDIA: Castañeda bottles (biphasic medium) – with agar and broth LEGIONNAIRE’S DSS]
component • History: the American legions had a
meeting and ALL the inhabitants acquired
pneumonia
NOTE: Brucella is a SLOW GROWER. Maintain culture for 3-4 weeks. Only
L. MICDADEI Pittsburgh pneumonia
report as negative with 3-4 weeks of No Growth
L. BOZEMANNI Wiga’s agent of pneumonia
• If blood culture is mentioned:
1. Routine: retained for 7 DAYS
2. SUSP BRUCELLOSIS: retained for 3-4 weeks

ANIMAL CO2 H2S THIONINE BASIC • The organisms (NORMAL FLORA) is carried in the oral cavity
SOURCE REQ FUCHSIN and respiratory and GI tracts of CATS and DOGS
B. Cattle + + Inhibited Grow • Humans may acquire the inf following contact with domestic
ABORTUS CAPNOPHILIC
(5-10% CO2) animals that harbor the bacterium; most frequently the route of inf
BANG’S is from the BITE or SCRATCH of an infected animal, usually a cat
BACILLUS • Wound inf are the MOST COMMON type of Pasteurella inf
B. Sheep, - - Grow Grow • Pasteurella multocida (agent of pasteurellosis)
MELITENSIS goat
o
B. SUIS Pigs - - Grow Inhibited
B. CANIS dogs - - Grow Inhibited • Agent of SHIPPING FEVER in cattle, a hemorrhagic septicemia
• Take note of B. ABORTUS AND B. MELITENSIS
In MAN Pasteurellosis Wound inf from bites and scratches of
CATS AND DOGS
INHIBITOR DYES USED: Thionine and Basic Fuchsin In CATTLE Shipping Hemorrhagic Septicemia
Fever
➢ Capnophilic Brucella, Pos H2S, will not grow in the presence of NOTE: Do not interchange!!!
THIONINE, will grow in the presence of Basic Fuchsin
o B. ABORTUS 1. Dog/Cat Bite or Scratch – P. multocida

➢ Brucella spp. that DOES NOT REQUIRE CO2, H2S NEG, GROW 2. Cat Scratch Dss (CSD) – Bartonella henselae, Afipia felis
in the presence of THIONINE and BASIC FUCHSIN.
o B. melitensis
 • F. TULARENSIS – Tularemia – dss of rodents (rabbits) • CALYMMATOBACTERIUM GRANULOMATIS
transmissible to man • Based on DNA-DNA hybridization, it has been recently proposed
GROWTH REQ: CYSTINE that the name of this organism be changed to KLEBSIELLA
MEDIA: Blood-glucose-cystine medium GRANULOMATIS
• TULAREMIA is a dss of rodents, primarily RABBITS
o Transmissible to man!!! [ZOONOSIS THROUGH CLIN SIG: Granuloma inguinale / DONOVANOSIS
DIRECT CONTACT WITH BLOOD OR ANMAL BITE
OR SCRATCH]
MOT:
1. Animal bite/scratch • Normally found the UPPER RESPIRATORY TRACT and is
2. Infected blood isolated as a RARE CAUSE OF ENDOCARDITIS.
3. Vectors = TICKS • PART OF HACEK [ Hemophilus, Actinobacillus, Cardiobacterium,
Eikenella, Kingella]

REVIEW!
1. CIN – Y.enterocolitica
2. Bordet Genou – Bordetella
3. Blood-glucose cystine – FRANCISELLA
4. Biphasic medium – Brucella
5. BCYE medium – Legionella

PIGMENT: Violacein – purple pigment

ODOR: Ammonium cyanide odor/ Almond like odor
CLIN SIG: Bacterial vaginosis
• Characterized by FOUL SMELLING vaginal discharge
• FORMERLY KNOWN as Haemophilus vaginalis, PREVIOUS NAME: Chryseobacterium meningosepticum, Flavobacterium
Corynebacterium vaginalis meningosepticum
DIAGNOSIS:
1. Cytology – CLUE CELLS COLONY: Yellow colonies on BAP
• Epith cells studded with bacteria; dirty cytoplasm with coccobacilli • Exposure of px to contaminated MEDICAL DEVICES OR
2. WHIFF/SNIFF TEST SOLUTIONS, but source is not always known
• Needs vaginal discharge and Reagent (10% KOH) • OCCASIONALLY may be transmitted from BIRTH CANAL to
NEONATE
• Pos result: fishy aminelike odor
3. CULTURE: HBT MEDIUM (Human blood Bilayer TWIN MEDIUM)

• Associated with GRANULOMATOUS DSS in ANIMALS and

have been associated with soft tissue inf in humans following
• NORMAL FLORA of rodents and rats
animal bites
• GN bacillus that requires media containing blood, serum, or
• Staining morphology gives the overall appearance of the DOTS
ascites fluid as well as incubation under CO2 for isolation from
AND DASHES OF MORSE CODE
clinical spx
CLIN SIG:
1. RAT BITE Fever: (SM/SM) -> Streptobacillus moniliformis (GN
bacilli), Spirillum minor/minus (Spiral organism associated with
➢ CAUSES GRANULOMATOUS DSS IN ANIMALS
RAT BITE FEVER
o Actinobacillus
2. HAVERHILL Fever - from INGESTION of CONTAMINATED
MILK

Corynebacterium diptheriae – CHINESE CHARACTER ARRANGEMENT

Actinobacillus spp. – gives the overall appearance of the DOTS AND
CHARACTERISTIC MOTILITY: GLIDING DASHES OF MORSE CODE
• GN, fusiform-shaped bacilli with ONE ROUNDED END and ONE
TAPERED END and OCCASIONAL filamentous forms
• Capnophilic, fusiform or filamentous bacilli

REVIEW!

1. Tumbling motility – Listeria (GP bacilli)

2. Darting – Campylobacter (GN Bacilli)
3. Gliding – Capnocytophaga