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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
Campylobacteriaceae
Pseudomonaceae

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
1)
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
severe
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
stable
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
Disease:
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

MOT
= 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
electrolytes
= 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
seafood's
Morphology:
Morphology:
= 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)
MOT
Disease: = fecal–oral route (contaminated food & H2O)
Enterocolitis
= 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.
disease
370C
= symptoms includes: acute crampy abdominal
2) Histologic Examination
pain,
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


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

= gram negative, non-fermentative, bacilli that


inhabit
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
O2
= considered to be the 3rd most frequently
10% CO2 85% isolated agent of hospital-acquired pneumonia
N2
Rx.: Aminoglycosides, Tetracycline & Morphology:
Chloramphenicol
= gram negative rod, motile – single polar
flagellum
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
Enterobacter
Determinants of Pathogenicity:
Proteus
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
colonies
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
H2O,
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,
glutaraldehyde
= bacteriostatic to quaternary ammonium
compound
= chlorination of water impt. in controlling
dissemination
of intestinal pathogen like agent of TF A) presence of underlying disease of
= relatively sensitive to drying the
patient
Antigenic Structure: B) emergence of resistant organism
due to indiscriminate use of
= plays an important role in epidemiological antibiotics
classification of Enterobacteriaceae
O
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
A
= 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
causing
transduction of fluid into intestinal
lumen→diarrhea

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
(HUS)
4) Colonization factor
= cellular surface structure which plays a
role in the establishment of organism in the
host

Laboratory Diagnosis:

= specimen includes sputum, tissue, pus, body


fluids,
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
purposes)

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