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Gram Negative Rods = large, smooth, flat yellow colonies

with opaque center & transparent

Cytochrome oxidase periphery
TSI Acid/Acid (-) gas (-) H2S
(+) (-) IMViC (+ - + +)
Family: Vibronaceae Family: = Possess lysine and ornithine
Enterobacteriaceae decarboxylase

Biotypes V. cholerae
Family: Vibronaceae 1) El tor – most frequent causative agent of cholera
3 Genera: (Philippines)
A) Vibrio 2) Classical
B) Aeromonas
rarely cause disease in man V. cholerae
C) Plesiomonas V. cholerae
El Tor Classical
Genus Vibrio 1) Voges – Proskawer test +
Species: -
1) V. cholerae 2) Agglutination to chicken rbc. + -
2) V. parahaemolyticus 3) Sensitivity 50u Polymixin B R S
4) Choleraphage sensitivity R S
V. Cholerae
- most important human pathogen
Disease – Asiatic Cholera:
Morphology: = food and waterborne disease
= acquired fecal – oral route (contaminated
+ short gram negative rod (0.5u X 1.5u to 3.0u) food/H2O)
+ curved/comma – shaped (Kommabacillus) = man only host infection
+ on stained smear – appear lying parallel to one = incubation period 2-3 days
another®”fish in the stream“ arrangement ==== = serious disease charac. sudden onset of
+ motile ® single, thick polar flagellum voluminous, non–bloody watery diarrhea (rice
- water stool) containing
scintillating/darting flakes of mucus, non-odorous accompanied with
dehydration (hallmark of disease)
Biochemical/Cultural Charac.: = no fever, untreated cases 60% die
= high attack rate – children
= organism non – invasive (remains localized GIT)
= facultative anaerobe
= opt. temp. 18 – 370C
= grow best at alkaline ph Determinant of Pathogenicity:
(extremely basophilic up to pH 9) 1) Cholera enterotoxin/Choleragin (heat labile)
= Sensitive to acid ph → kill organism = resp. watery diarrhea
= Non – lactose fermenter but ferment sucrose and = activate adenylcyclase of intestinal
mannose mucosal cell
= Metabolism both respiratory and fermentative ↓
increase intracellular
cyclic adenosine
CM: monophosphate (CAMP)
MEA (Meat Extract Agar) ↓
hypersecretion of H2O
= Translucent colony with iridescent & Cl into the
green → red bronze color intestinal lumen
(viewed on ↓
oblique light) inhibition of
Selective CM: +
Na absorption
TCBS (Thiosulfate Citrate Bile Sucrose) ↓
diarrhea = virulence may be associated with prod. heat
hemolysin (kanagawa) that hemolyze
2) Invasiveness of organism human
+ ability to adhere or attached to microvilli red blood cell which is cytotoxic and
of brush border of epithelial cells. cardiotoxic
Serological types – based on antigenic differences Gastroenteritis/Foodpoisoning
1) Ogawa – A & B = self-limiting disease
2) Inaba – A & C = charac. nausea, vomiting, explosive,
3) Hikojima – A, B, C watery diarrhea (cholera – like)

(without blood and mucus)

= symptoms may persist for 10 days

= ingestion of raw or improperly cooked
Lab. Diagnosis: seafoods(shellfish)
Lab. Diagnosis:
1) Direct stool exam.– Alkaline Peptone broth 1) Rectal – Swab
incubated transport medium Cary – Blair/Amies
for 6 hours (darkfield microscopy → motility) ↓
2) Stool culture – MEA / TCBS incubated 2 – 3 days TCBS &
3) Direct Fluorescent Antibody Test Alkaline Peptone broth
Treatment: Treatment: Chloramphenicol, Kanamycin,
= Prompt replacement of fluid and Tetracycline
= Tetracycline/Furazolidone
(shorten the course of disease and
decrease) bacterial
secretion Differentiation:
Prevention: V. Cholera V. Parahaemolyticus

1) Maintenance of adequate sewage treatment 1. Growth 2% Na Cl - +

2) H2O purification 2. Sucrose fermentation + -
3) Prompt detection & treatment of cases 3. Kanagawa test - +
= vaccine no significant protection

V. Parahaemolyticus Genus Campylobacter

= marine/saltwater organism Campylobacter jejuni
= found estuaries throughout the world = worldwide
= major cause gastroenteritis involving = most important human pathogen
= gram (-) spirally curved rods (seagull-wing
= closely related V. Cholerae shaped)

Cultural/Biochemical charac.: = motile (single polar flagellum)

= extremely halophilic (salt-loving, req. 2% ↓
NaCl) = oxidase (+) darting motility
= non–lactose fermenter = oxidase and catalase (+)
CM – TCBS = asaccharolytic (non-carbohydrate fermenter)
= large, smooth, green colonies = microaerophilic (CO2 ¯O2)
(non–sucrose fermenter)
= grows best at 42 – 430C (thermophilic)
= susceptible to gastric acid
Determinants of Pathogenicity:
= H2S producer Morphology:
= fastidious
= curved or S – shaped gram negative organism
Culture Media: = motile (4 – 6 lopotrichous flagella)
= urease and oxidase (+)
Skirrous agar/Campy-BAP = microaerophile
incubated with 10% CO2 5%O2 85%N2 = killed at acid ph

↓ Habitat:
colorless convex colonies = gastric mucosa where it invade tissues and
watery and spreading causes gastritis and ulcers
= grows at ph 6 –7
Determinants of Pathogenicity:
1) heat – labile enterotoxin Disease:
Anthral Gastritis/Chronic Gastritis
= activate adenylcyclase enzyme →
Duodenal ulcers (no direct evidence that H. pylori)
increase CAMP → split of Cl → causes duodenal ulcers but there is strong
prevent absorption of Na+ → evidence between the presence of H. pylori and
massive diarrhea duodenal ulcer
(bloody mucoid stool)
Disease: = fecal–oral route (contaminated food & H2O)
= self – limiting disease lasting less than 7 days Lab. Diagnosis:
= one of the most common cause of infectious 1) Microaerophilic culture of biopsy material incub.
= symptoms includes: acute crampy abdominal
2) Histologic Examination
3) Urease test
vomiting, bloody diarrhea with pus, malaise
and fever 4) Culture – Skirrous agar (selective CM) 370C
= incubation period 1 – 7 days ↓

Reservoir and MOT: small, circular, translucent colonies after 7 days

= organism exist as normal GI tract flora of many
wild and domestic animals (dogs) Treatment: Bismuth + Metronidazole +
= human infection is acquired through ingestion of
raw or undercooked meat Genus Pseudomonas
Specie: Pseudomonas aeruginosa

= gram negative, non-fermentative, bacilli that

Lab. Diagnosis: soil and water
1) Gram staining (demonst. gull-wing shaped = important opportunistic pathogen among
of organism) individual
with impaired host defense
2) Darkfield microscopy for motility = human infection severe and difficult to treat
3) Microaerophilic culture on special media = major agent of nosocomial infection
= exhibit broad resistant to many antimicrobials
(Skirrous) incubated at 420C with 5%
= biochemically inactive
= considered to be the 3rd most frequently
10% CO2 85% isolated agent of hospital-acquired pneumonia
Rx.: Aminoglycosides, Tetracycline & Morphology:
= gram negative rod, motile – single polar
Genus Helicobacter = piliated and slime layer present
Biochemical/Cultural Charac.:
Species: Helicobater pylori (Campylobacter pylori)
= strictly aerobic
= opt. temp. for growth 350C
= beta–hemolytic BAP
= Pseudomonas agar- special media which enhance
pigment production: Members of the Family:
colonies appears turquioise
blue due to (pyocyanin) pigment which is
water Opportunistic members
soluble and fluorescent (pyoverdin) pigment Escherichia
which is greenish color Klebsiella lactose fermenters-pink colonies
Determinants of Pathogenicity:
1. Colonization to appropriate site by the pathogen Morganella
= Pili (enable org. to attach to host cell) Citrobacter
= Slime layer/glycocalex enhances Serratia non-lactose fermenter-colorless
adherence to tissue colonies
2. Hemolysin (phospholipase and Glycolipids) Providencia
= contribute to invasiveness of organism Edwardsiella
3. Protease enzyme = resp. hemorrhagic skin
lesions Intestinal pathogens
4. Enterotoxins = resp. diarrhea Salmonella
5. Elastase = impt. in corneal infection Shigella non-lactose fermenter-colorless
Clinical infections: Yersenia

= occurs in individual with altered host defenses

= Meningitis, burned-wound infection, GUT,
malignant Gen. Morphology:
disease, etc.
= small, gram negative non-sporeforming rod
= most are motile (peritrichous flagella) except
Lab. Diagnosis: Shigella
1. Isolation & bacteriological demonstration of and Klebsiella which are non-motile
organism = all are noncapsulated, except Klebsiella
2. Culture = some are provided with fimbriae/pili
Rx: Aminoglycosides (Amikacin, Gentamycin,
Tobramycin) Biochemical/Cultural Charac.:
Prevention: = facultative anaerobe
= grows readily on ordinary media
1. Proper isolation procedure = ferment CHO under anaerobic or low O2
2. Hyperimmune gamma – globulin atmosphere
= decrease mortality rate of infection = ferment glucose with the production of
3. Vaccine – Pseudogen (heptovalent vaccine) acid or both acid and gas
= reduce nitrate to nitrite
= oxidase (-)
Family Enterobacteriaceae = culture medium for isolation:
Differential: EMB, Mc Conkey
= composed large groups of organism found in soil, Highly selective: Hektoen Enteric agar, SSA, XLA
decaying matter and large intestine of
humans, Resistance:
animals and insects
= habitat → Gastrointestinal tract of humans = destroyed easily by heat and low concentration
“Enteric bacilli” of
common germicide and disinfectants
= bactericidal to phenols, formaldehyde,
= bacteriostatic to quaternary ammonium
= chlorination of water impt. in controlling
of intestinal pathogen like agent of TF A) presence of underlying disease of
= relatively sensitive to drying the
Antigenic Structure: B) emergence of resistant organism
due to indiscriminate use of
= plays an important role in epidemiological antibiotics
classification of Enterobacteriaceae
H major antigen used in serologic typing
K of the organism

Determinants of Pathogenicity:
1) Endotoxin
= LPS of the cell wall®toxicity resides in lipid
= heat - stable
= effect of endotoxin causes:
Fever, Fatal shock, Leucocytic alteration,
Disseminated intravascular coagulation
(DIC), Shwartzman reaction
2) Enterotoxin
= heat-labile usually affect small intestine
transduction of fluid into intestinal

3) Shiga toxin and Verotoxins

= interferes with protein synthesis of
mammalian cells
= unclear role in Shigellosis
VTEC toxin - important cause of hemolytic
diarrhea and hemolytic uremic syndrome
4) Colonization factor
= cellular surface structure which plays a
role in the establishment of organism in the

Laboratory Diagnosis:

= specimen includes sputum, tissue, pus, body

rectal swab/feces
A) Culture (commonly used media)
Differential (EMB, Mc Conkey)
Selective (Hektoen agar, SS agar)
B) Biochemical test
TSI, IMViC, LIA, Carbohydrate fermentation
C) Serologic (used primarily for epidemiologic

= remains a major therapeutic problem
= several factors contribute to the difficulty
treating these infection
= one of the most important factor is: