Gram Rods

Gram Negative Rods
Cytochrome oxidase
(+) (-)
Family: Vibronaceae Family: Enterobacteriaceae
Campylobacteriaceae
Pseudomonaceae
Gram Negative Rods
Cytochrome Oxidase Positive

Family: Vibronaceae
3 Genera:
A) Vibrio
B) Aeromonas rarely cause disease in man
C) Plesiomonas
Genus Vibrio
Gen. Characteristics:
- gram negative, comma – shaped bacilli
- non-sporing, non-capsulated
- metabolism both respiratory and fermentative
- oxidase (+)
- motile – single polar flagellum
Species:
1) V. cholerae
2) V. parahaemolyticus
*Vibrio cholerae
(Komma bacillus)
- most important human pathogen
Morphology:
- short gram negative rod (0.5u X 1.5u to 3.0u)
- curved/comma shaped
- on stained smear – appear characteristically lying
parallel to one another (”fish in the stream“
arrangement)
- motile  single, thick polar flagellum

scintillating/darting
Vibrio cholerae
(Komma bacillus)
Categories :
1st group - ( V.cholerae O1 ) classic epidemic cholera
- caused by organisms that agglutinate in

antisera
- directed against the O1 antigen &

produce disease primarily by means
of a specific enterotoxin*
Vibrio cholerae
(Komma bacillus)
Categories :
2nd group - (atypical or nontoxigenic V.cholerae O1)

- biochemically similar to the 01
organisms & type with the O1
antisera
* - do not produce enterotoxin
3rd group – ( non-O1 V. cholerae ) does not

agglutinate in O1 antisera
- referred to as nonagglutinating vibrios
( NAG ) or noncholera vibrios (NCV)*
Biochemical / Cultural Characteristics:
- facultative anaerobe
- optimum growth temp. 18 – 370C
- metabolism is both respiratory & fermentative
- grow best at ph 7.0
- can tolerate alkaline ph of 9.0 (extremely basophilic)
- extremely sensitive to acid ph*  kill organism
- susceptible to heat, drying and to common
chemical disinfectant
- grow best on simple ordinary culture media
- possess lysine and ornithine decarboxylase
- reduce nitrate to nitrite
- urease (-), H2S (-)
- non-lactose fermenter but ferment sucrose,

maltose and glucose.
CM:
*MEA (Meat Extract Agar)
- translucent colony with iridescent green 
red brown color (viewed on oblique light)*
Selective - Differential CM: (vibrio colonies from fecal

matter)
- TTGA ( Tellurite Taurocholate Gelatin Agar )
- TCBS (Thiosulfate Citrate Bile Sucrose)
large, smooth, flat yellow colonies with
opaque center & transparent periphery
MacConkey’s Agar – most strains grow luxuriantly and on

short incubation produce a colorless colonies,
prolonged incubation result in the production of pink
colored colonies.
* Antigenic structure:
- “O” or somatic antigen
- major Ag for serologic typing of V. cholera
- part of the lipopolysaccharide (LPS) component of
the cell wall*
- “H” or flagellar antigen commonly shared by all vibrios

*2 Biotypes El tor V.cholerae
*1) Voges – Proskawer + -

2) Agglutination to chicken rbc. + -
3) Sensitivity to polymixin B R S
4) Choleraphage sensitivity R S
- both biotypes are responsible for classic epidemic cholera

and are divided into 3 antigenic subtypes:
1. Ogawa A, B
2. Inaba A, C
3. Hikojima A, B, C
Determinants of Pathogenicity:
* 1) Cholera enterotoxin/Choleragen (heat labile)
- major pathogenic factors that act on the cells of the

small intestine
- composed of two major subunits:
subunit A
subunit B
Determinant of Pathogenicity:
* - subunit A – responsible for the biologic activity of the

organism
stimulate adenylcyclase activity of the epithelial cell,

converting ATP to cAMP

over production of intracellular cAMP

hypersecretion of H2O & Cl

inhibition of Na+ absorption

watery diarrhea _dehydration leading to
hemoconcentration, anuria and hypovolemic shock
due to massive loss of water and electrolyte
*subunit B - responsible for cellular binding of the
toxin to the intestinal epithelial cell
*2) Invasiveness of organism
- ability to adhere/penetrate to the intestinal mucosa

and attached to microvilli of brush border of
epithelial cells.
Disease – Cholera:
- food and waterborne disease
- *human carrier serves as source of new cases of

cholera
- *acquired fecal – oral route (contaminated food/H2O)
- man only host infection
- incubation period 2-3 days
- serious disease charac. sudden onset of voluminous,

non–bloody, non-odorous watery diarrhea
(rice water stool) containing flakes of mucous
(hallmark of disease)
* Disease – Cholera:
- fluid loss severe 15 – 20 L / day
- patient severely dehydrated:

eyes – sunken, lips – dry and cracking,
skin – loss turgor, hands – washerwoman
- no fever, untreated cases 60% die
- high attack rate – children
- organism non – invasive (remains localized GIT)

Lab. Diagnosis:
1) Direct microscopy of stool specimen
- observe for comma-shaped bacilli

- inoculation into Alkaline Peptone broth incubated
6 hours exam. darkfield microscopy motility
2) Stool culture – MEA / TCBS incubated 2 – 3 days
3) Direct Fluorescent Antibody Test

Treatment:
- Prompt replacement of fluid and electrolytes
- Tetracycline (DOC) & Doxycycline
Prevention:
1) Maintenance of adequate sewage treatment

2) H2O purification
3) Prompt detection & treatment of cases
cholera vaccine (no significant protection)
V. Parahaemolyticus
- marine/saltwater organism
- found in estuaries throughout the world
- major cause of gastroenteritis involving seafoods
Morphology:
- closely related to V. Cholerae

V. Parahaemolyticus
Cultural/Biochemical characteristics:
- extremely halophilic
- oxidase (+)
- non–lactose and non-sucrose fermenter
* CM – TCBS = large, smooth, green colonies

(non–sucrose fermenter)
- hemolysis on Wagatsuma agar is used to identify
pathogenic strains ( Kanagawa phenomenon )
- most strain prod. diarrhea due to heat – stable

hemolysin (Kanagawa hemolysin) which is cytotoxic
and cardiotoxic
- can lyze human rbc.
- Kanagawa Hemolysis Test

Disease: Gastroenteritis/Foodpoisoning
- self-limiting disease acquired through ingestion of

raw or undercooked seafood (shellfish, oysters)
- incubation period 12-24 hours
Clinical manifestations :
- nausea, vomiting, explosive watery diarrhea(without
blood and mucus)
- headache
- fever (not common)
- clinical manifestations may last for 3-5 days

Lab. Diagnosis:
1) Rectal Swab
transport medium Cary – Blair/Amies

TCBS & Alkaline Peptone broth
Treatment: Chloramphenicol, Kanamycin, Tetracycline

Differentiation: V. Cholera V. Parahaemolyticus
1. Growth 2% Na Cl - +
2. Sucrose fermentation + -
3. Kanagawa test - +
Genus Campylobacter
Species:
Campylobacter jejuni
Campylobacter Coli
- worldwide
- closely related to V. cholerae morphologically and

physiologically
- most important human pathogen associated with

disease production
Genus Campylobacter
Morphology:
- gram (-) spirally curved or seagull-wing shaped rod
- motile (single polar flagellum) darting motility
- oxidase and catalase (+), H2S producer
- asaccharolytic (non-carbohydrate fermenter)
- microaerophilic, fastidious
- grows best at 42 – 430C
- susceptible to gastric acid

1) heat - labile enterotoxin
- acts in the same manner as choleragen

toxin
- activate adenylcyclase enzyme 

increase cAMP  split of Cl  prevent
absorption of Na+ massive diarrhea
(bloody mucoid stool)
Culture Media:
- slow growing and fastigious
Skirrous agar (Vancomycin, Polymixin, Trimethoprin)
Campy-BAP agar – incubated w/ 10% CO2

 5%O2 85%N2
colorless convex colonies, watery and spreading
Disease: Enterocolitis
- one of the most common cause of diarrhea among

pediatric patients in U.S. (infectious type)
- self–limiting disease lasts less than 7 days
- causes gastroenteritis producing inflammation

of the intestine with ulceration of the GIT mucosa
Reservoir and MOT:
- organism exist as normal GI tract flora of many wild

and domestic animals (dogs)
- human infection is acquired through ingestion of raw

or undercooked meat
- secrete an exotoxin that is antigenically similar to

cholera toxin
- incubation period last 1– 7 days
- symptoms includes: acute crampy abdominal pain,

vomiting, fever, malaise & diarrhea (bloody with pus)
Lab. Diagnosis:
1) Gram staining (demonst. seagull-wing shaped

organism)
2) Darkfield microscopy for motility
3) Culture on special media

Skirrous or Campy-BAP incubated
microaerophilically at 420C for 72 hours
Treatment : Erythromycin, Tetracycline & Chloramphenicol

Ciprofloxacin - alternate drug
Prevention : no vaccine available

Genus Helicobacter
Species: H. pylori (Campylobacter pylori)

- isolated from biopsied gastric tissue of stomach
among patients with ulcer
- associated with gastritis, gastric and duodenal ulcer
and gastric carcinoma
- does not invade gastric tissue but is found in
association with gastric mucous secreting cells
- now classified by WHO as type 1 carcinogen
Morphology:
- curved or spiral – shaped gram negative

organism that have strong urease activity
- motile (4 – 6 lopotrichous flagella)
- urease and oxidase (+)
- microaerophile
- killed at acid ph
Habitat:
- human gastric mucosa in the antrum and

fundus of the stomach
- exact mode of transmission unknown but

oral-oral, fecal oral are possible route
of transmission
1.Ability to adhere to gastric epithelium by the
production of urease and mucinase
Urease – to neutralize stomach acid on its migration

to the lining epithelium of the stomach
Mucinase – enhance penetration to the mucous
layer of the GIT
2. Production of cytotoxin – induce intracellular vacuolation
of cultured cells
- frequently assoc. with peptic ulcer and gastritis
Disease: Anthral Gastritis/Chronic gastritis
- acquired by fecal–oral route (contaminated food & H2O)
- characterized by epigastric pain, anorexia, nausea

and vomiting
Lab. Diagnosis:
1) Microaerophilic culture of biopsy material in Skirrous

agar medium incubated for 7 days at 370C
presence of small, circular, translucent colonies
2) Histologic Examination (biopsied gastric tissue)

Disease: Anthral Gastritis/Chronic gastritis
Lab. Diagnosis:
3) Direct detection of organism from biopsied specimen

using Warthin - Starry stain
4) Urease breath test – radioactive urease swallowed
5) Serological - EIA
Treatment: Bismuth + Metronidazole + Tetracycline

Genus: Pseudomonas
- gram negative, non-fermentative bacilli that inhabit

soil, water, sewage and air*
- most specie do not infect human
- * some are important opportunistic pathogen

(infect individual with impaired host defense)
- human infection is severe and difficult to treat

Specie: Pseudomonas aeruginosa
Morphology:
- slender gram negative rod, arranged singly or in

short chain
- actively motile with single polar flagellum*
- non-sporing and non-capsulated
- *piliated and most strain have mucoid slime layer

Specie: Pseudomonas aeruginosa
- most frequently isolated human pathogen
- major agent of nosocomial infection
- leading cause of death among patient with cystic

fibrosis, neoplastic diseases and severe burns
- exhibit broad resistant to many antimicrobials
- biochemically inactive
*Biochemical/Cultural Characteristics:
- strictly obligate aerobic
- optimum temp. for growth 370C, opt. pH of 7.2 – 7.4
- can grow on ordinary culture medium which helps

in the identification
*Blood agar - beta–hemolytic

Biochemical/Cultural Charac.:
*Pseudomonas agar- special medium which enhance

pigment production
pyocyanin – bluish-green diffusible phenazine

pigment which is soluble in H2O and
chloroform
pyoverdin - greenish-yellow fluorescent pigment

soluble in H2O not in chloroform
Biochemical / Cultural Characteristics:
Mac Conkey’s agar – colorless non-lactose fermenting

colonies
Nutrient agar - large, smooth, convex, grayish-blue

colonies that emits fruity aromatic
odor
- resistant to chemical disinfectant and

destroyed by boiling
1. Colonization to appropriate site utilizing pili and slime
2. Hemolysin (phospholipase and glycolipids)

- contribute to invasiveness of organism
3. Production of extracellular enzyme (Protease and Lipases)

which help in the formation of hemorrhagic skin lesions
4. Production of exotoxin- which inhibits protein synthesis
5. Elastase-digest elastin of the arterial wall causing tissue

necrosis
Clinical infections:
- occurs in individual with altered host defenses

(immunocompromised individual)
- meningitis, extensive burn, urinary tract infection

following catheterization, eye, ear, wound infection
Lab. Diagnosis:
1. Microscopic examination – demonstrate characteristic

morphologic appearance of the organism
2. Culture
3. Antibiotic susceptibility test (obligatory because of

multiple antibiotic resistant of the organism)
Treatment : Aminoglycosides (Amikacin, Gentamycin,

Tobramycin)
Prevention:
1. Isolation of patient
2. Hyperimmune gamma – globulin

- decrease mortality rate of infection
3. Vaccine – Pseudogen (heptovalent vaccine)

Family Enterobacteriaceae
Genera : Escherichia Klebsiella

Shigella Serratia
Salmonella Proteus
Enterobacter
- also referred to as coliforms ( gm neg.rods & enteric

bacteria/ bacilli )
- composed of large groups of closely related bacterial

species that inhabit the large intestine of man,
animals, soil, water and decaying organic matter
- includes some of the most important intestinal pathogen

affecting human (Salmonella and Shigella)
- most species are opportunistic organisms that can infect

any body sites if their habitat is changed
- responsible for majority of nosocomial infections seen today

Gen. Morphology, Cultural /Biochemical Characteristics :
- small, gram negative non-sporing rod
- most are motile (peritrichous flagella) except Shigella

and Klebsiella which are non-motile
- all are noncapsulated, except Klebsiella

(thick polysaccharide capsule)
- some provided with fimbriae/pili (for attachment)

- biochemically diverse
- all ferment glucose with the production of acid or both

acid and gas
- reduce nitrate to nitrite
- oxidase (-) catalase (+) except Plesiomonas

- lactose fermentation is useful in differentiating pathogens

from non pathogenic members of the family
- grow on peptone or MEA without addition of NaCl or other

supplements
- grow well on MacConkey agar

Ultrastructure: Cell Wall
- composed of Murein, Lipoprotein, Phospholipids,

Lipopolysaccharides (LPS)
- Lipopolysaccharides determines antigenicity of species

and responsible for endotoxin activity
- 20% is made up of murein-lipoprotein layer - responsible

for cellular rigidity
- 80% forms the lipid bilayer

Classification of gram negative bacilli based upon
lactose fermentation:
Rapid lactose fermenter
- ferment lactose producing pink colonies on

Mac Conkey’s agar
E. coli, Klebsiella, Enterobacter

Classification of gram negative bacilli based upon
lactose fermentation:
Slow lactose fermenter
Serratia, Citrobacter, Arizona
Non-lactose fermenter
- do not ferment lactose, produce colorless colonies

on Mac Conkey’s agar
Salmonella, Shigella, Proteus
Antigenic Structure:
- plays an important role in the epidemiology and

classification of members of the family
Enterobacteriaceae
- major antigenic component used in serological

typing of the organism: O, H, K antigens
- enterics share a common antigen (ECA) present

in outer surface of bacterial cell  useful
for taxonomic and epidemiologic study
Flagellar (H) antigens
- protein antigen found in the flagella
- denatured with acid or alcohol
- removed by heat at 1000C for 20 minutes
- preserved by treating with formalin
- anti H AB forms light fluffy clumps

Somatic (O) antigens
- found at the most external part of the outer membrane

of the cell wall (LPS)
- consist of repeating units of oligosaccharide
- diversity allows serologic subgrouping within genera
- single organism may carry several O Ag
- resistant to heat and alcohol
- usually detected by bacterial agglutination
- anti O AB are produced as a closely packed granular
clumps
Capsular (K) antigen
- found external to O antigen

- may interfere with agglutination by O antisera
- polysaccharide antigen
- destroyed by boiling for 2 hours
- may be associated with virulence
- may cross react with other genera
- Vi antigen plays a role by preventing intracellular
destruction of the org. and is the virulence Ag
of S. typhi
1) Endotoxin (heat stable)
- LPS of the cell walltoxicity resides in lipid A layer
- effect of endotoxin causes:
1.fever
2. shock
3. hypotension
4. hemorrhage
5. Disseminated intravascular coagulation (DIC)
6. Shwartzman reaction – necrosis around
infection site on the skin
2) Enterotoxin (heat labile)
- affects the small intestine causing transduction

of fluid into intestinal lumen resulting to diarrhea
- activate adenyl cyclase  increase excretion of

Cl & H2O  prevent reabsorption of sodium
 massive diarrhea
3) Shiga toxin and Verotoxins
- interferes with protein synthesis of mammalian cells
- Shiga toxin - 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 includes:
capsule, slime layer, fimbriae/pili
- capsules of Klebsiella prevents phagocytosis
- Vi Ag of Salmonella prevents intracellular destruction/

phagocytosis
Laboratory Diagnosis:
- specimen includes sputum, tissue, pus, body fluids,
rectal swab/feces
A.) Microscopic demonstration of organism
B.) Culture - commonly used media for isolation
a) differential (EMB, Mac Conkey agar, DCA)
b) selective media (Brilliant green agar, BSA)

- Hektoen Enteric agar
- Xylose Lysine Desoxycholate agar
C) Enrichment Media
(Selinite F broth, Mueller Tetrathionate broth)
- useful for culture of stool specimen
D) Screening Media (TSI)

- contain 3 sugars at different concentration
(Glucose, Lactose, Sucrose)
E) Media for biochemical & carbohydrate fermentation test

- final identification of many species based on
antigenic structure (serotyping)
IMVIC, SIM, LIA
G) Serologic (used primarily for epidemiologic purposes)

Treatment:
- remains a major therapeutic problem
- several factors contribute to the difficulty of treating

these infection :
1. ) presence of underlying disease of the patient

2. ) emergence of resistant organism due to
indiscriminate use of antibiotics
Enteropathogenic /
True intestinal pathogen
Enterobacteriaceae
- are clinically divided into:
I –Enteropathogenic /True intestinal pathogen
- those that induce GIT diseases

Salmonella, Shigella, Yersinia, Enteropathogenic E. coli
II – Coliform / Opportunistic group
- organism capable of infecting any body sites

Enterobacter, Klebsiella, Providencia, E. coli,
Proteus, Serratia, Citrobacter, Arizona
Genus Escherichiae
Specie: Escherichia coli
- may be non-pathogenic, opportunistic or true

intestinal pathogen
- capable of causing primary intestinal disease as
well as extra intestinal infection
- predominant facultative anaerobe found in large
intestine of human
- use as microbial indicator for fecal contamination
of water
Morph., Biochemical & Cultural Characteristics:
- short plump, motile rod (peritrichous flagella)
- piliated
- most members are encapsulated
- rapid lactose fermenter
- majority of isolates are non-pigmented
- lysine decarboxylase (+)
- use acetate as a carbon source
- hydrolysis of tryptophan to indole

Morph., Biochemical & Cultural Characteristics:
- non-fastidious, grows well on commonly used ordinary

culture media (NA, BA, Mac Conkey’s agar)
Nutrient agar - smooth, translucent colonies

Blood agar - beta-hemolytic
Mac Conkey’s - pink colonies due to lactose
fermentation
Selective differential EMB  smooth, circular,
convex colonies with a greenish black
metallic sheen
- Serotyping is based on
O Ag
H Ag
K Ag
1) Surface factors/ antigens – (KI/capsular antigen)
a. Polysialic acid capsule ( K1 )

- found in 80% of E. coli causing neonatal meningitis and
sepsis
- enable the organism to resist killing/phagocytosis
- plays an important role in the dissemination of organism
from primary infection site
2) Enterotoxins (target organ  small intestine)

2) Heat – stable enterotoxin ( ST )
- activates guanylate cyclase enzyme of

intestinal mucosa leading to the
formation of cGMP in the epithelial cell

causing inhibition NA+ and Cl absorption by the
brush border of the intestine producing diarrhea
3) Verotoxins ( Shigalike toxin)
- produced by certain strain of E. coli causing

irreversible cytotoxic effect on Vero tissue
cell culture
- cytotoxic toxin associated with 3 syndromes:
1. Diarrhea
2. Hemorrhagic colitis
3. Hemolytic uremic syndrome (HUS)
Clinical Infections: E. coli
- most frequently isolated pathogen causing:

1. GIT disorders
2. Urinary tract infection (most common)
3. Neonatal meningitis
4. Gram negative sepsis
5. Infantile and traveler’s diarrhea
6. Peritonitis 2O ruptured appendicitis
Strains of E. coli producing gastroenteritis/diarrhea
particularly in infants and children.
A)ETEC (Enterotoxigenic E. coli)
- major cause of “travelers” & infantile diarrhea

in developing countries
*- toxin prod. LT/ST - binds to CAMP

*- induce secretion of H2O and electrolyte
of intestinal lumen

loss of fluid

dehydration

death
B) EIEC (Enteroinvasive E. coli)
- causes dysentery – like syndrome in all age groups

- charac. fever, abdominal cramp, blood & pus stool
- ability to invade intestinal wall  inflammation
site of entry
C)*EPEC (Enteropathogenic E. coli)
- major cause of chronic diarrhea

- do not produce LT/ST toxin
- do not invade the intestinal wall
- causes diarrhea in neonates (infantile)
D) EHEC (Enterohemorrhagic E. coli)
- verotoxin producing strain of E. coli

- causes hemorrhagic colitis with bloody stoo
E ) EAEC ( Enteroaggregative E.coli )
- causes acute & chronic diarrhea ( > 14 days induration)

- associated with persistent diarrhea in patients with HIV
* Laboratory Diagnosis:
1. Microscopy (gram stain) – observe gram (-) bacilli

morphology
2. Culture (BA, Mac Conkey’s agar, EMB)
3. Animal Pathogenicity test (Sereny test)

- instillation of bacterial susp. eyes of guinea pig
(+) conjunctivitis
4. Biochemical test IMViC (++ - - )

TSI A/A (+) gas (-) H2S
5. Antibiotic sensitivity test

*Treatment :
- fluid and electrolyte replacement
- sensitive to most antimicrobials
Prophylaxis : Trimethoprim – Sulfamethoxazole

*Genus Shigella
Species Mannitol
fermentation
1. S. dysenteriae -
- causes the most
severe disease
2. S. flexneri +
3. S. boydii +
4. S. sonnei +
General Characteristics :
- are the major causes of bacillary dysentery
- characterized by severe abdominal cramps
- frequent passage of low volume bloody,mucoid stools
- mostly seen in pediatric age of 1-10 yrs old
- characterized by 4 major serogroups :

serogroup A - S. dysenteriae
B - S. flexneri
C - S. boydii
D – S.sonnei
Biochemical Properties & Characteristics:
- slender,gram (-) non-motile, non-capsulated & non-

sporeforming
- non – lactose fermenter
- does not produce lysine decarboxylase
- does not produce H2S except S. flexneri
- do not produce gas from CHO fermentation
- less resistant than Salmonella to physical and
chemical agent
- do not utilize acetate as a carbon source
Resistance to Physical & Chemical Agents
- high concentration of acid is detrimental*

- requires well buffered media for transport of
specimen & growth of the organisms
- can tolerate low temp if adequate moisture is
present
- can survive for more than 6 mos. in H20 @ room
temperature
- high concentration bile inhibit growth of some strain
- grows in ordinary culture medium
Enrichment Medium – Selinite F broth

Selective Differential – Salmonella-Shigella agar
*Antigenic Structure:
- All species have O antigen
A – S. dysenteriae (12 serologic types)

B – S. flexneri (6 serologic types)
C – S. boydii (18 serologic types)
D – S. sonnei ( 1 serologic types)
- only few have K antigen which can easily be removed

by boiling the cell suspension before typing the
organism
- all Shigella have no H antigens
* Determinants of Pathogenicity:
1)* Endotoxin (smooth LPS antigen)

- enhances ability to survive through host defenses
- thermo stable lipopolysaccharide found in the cell wal
2) Invasiveness
- invade and proliferate within epithelial cells of
intestinal mucosa intracellular multiplication
 inflammation  cell death ulceration impaired
colonic fluid absorption discharge of blood, mucus &
pus
3)* Shiga toxin
- has multiplicity of effects, acts as:
A) neurotoxic
- produce damage of the endothelial of the small
blood vessels of the central nervous system
resulting in neurological complication such as
convulsion
B) Cytotoxic
- interferes with protein synthesis causing cellular
death resulting to bloody diarrhea with
mucus and pus
C) Enterotoxic
- acts as enterotoxin in the intestinal mucosa
blocking the absorption of electrolyte from
intestinal lumen causing watery diarrhea
Disease: Bacillary Dysentery/Shigellosis
- produced by all species and is acquired by fecal–oral

route (4 F’ S – finger, flies, food, fomites)
- incubation period 1- 4 days
- characterized by sudden onset frequent, painful

passage of low volume, bloody, mucoid
stools
- fever, chill, convulsion present
- organism is non-invasive
Disease: Bacillary Dysentery/Shigellosis
1. Microscopic detection of pus cells, rbc and

non-motile bacilli by wet mount
2. Rectal swab/Sigmoidoscopic exam (ulcer)
3. Culture – colonies grown are confirmed by:

a.) Biochemical test
b.) Slide agglutination test
Treatment:
- Fluid and electrolyte replacement

- Ampicillin (drug of choice)
Prevention:
1) Persons with disease should be isolated until
stool culture is negative
2) Carriers should not be allowed to handle/
prepare foods
3) Proper sewage disposal
4) H2O chlorination is important
Genus Salmonella
- biochemically more complex, serologically more diverse
- infect animal besides human
- capable of invading extra-intestinal tissues

Genus Salmonella
Species:
S. typhi – most important specie

S. paratyphi A, B, C
S. cholerae-sius
S. enteritidis – S. typhimurium
Morphological , Cultural & Biochemical Characteristics:
- short, gram (-) non-sporeforming, non-capsulated
- motile  peritrichous flagella
- non–lactose fermenter
- ferment glucose and mannose without production of

gas
(anaerogenic-ferment without gas)
Morphological, Cultural &Biochemical Characteristics :
- produce H2S except S. paratyphi A & S. cholerae-suis
- grow well on ordinary media (BA, MC, NA)

Selective Differential–SS agar, Wilson-Blair med.
- killed at 600C in 15 minutes
- survive in iced water and snow for weeks to months

Morphological, Cultural & Biochemical Characteristics :
- can tolerate large concentration of bile
- oxidase –
- IMViC - + - +
O & H - are major antigens for Salmonella serotyping
O Ag - (similar to the O antigen of other member)

H Ag - diphasic and not shared, can exist in 2 phases :
- Specific – shared by few organisms and react

only with homologous antisera
- Non-specific – shared by many organisms and

react with heterologous antisera
Vi Ag (Capsular antigen)
- plays minor role in classif. but with important

pathogenic significance for detecting typhoid carrier
- impart virulence by preventing intracellular

destruction of organism
- not found in other species of Salmonella

1. Surface antigen
- resp. attachment of organism  host receptor cell
and survive intracellularly
Vi Ag - when present, organism is more virulent than

those lacking it
- found in capsule and prevent phagocytosis or

intracellular destruction of organism
2. Invasiveness
- Salmonella can penetrate epithelial lining of small

intestine  subepithelial tissue and lamina
propia by a process similar to phagocytosis
3. Endotoxin
- resp. for fever which directly / indirectly is the result
of the release of endogenous pyrogens from
leukocyte during bacteremic stage of disease
4. Enterotoxin
- similar to E. coli which is heat stable and labile
- associated with penetration  mucosa to induce
diarrhea
Disease : Salmonellosis
Clinical Syndrome:
1.) Enteric fever (Typhoid Fever & Paratyphoid fever)

S. typhi ,S. paratyphi A, B, C
- uniquely adopted to human

- 3% of patients become chronic carrier
- majority of chronic carrier are females with
gallbladder diseases
- organism resides in the gallbladder or biliary tree
*Pathogenesis:
- food and waterborne disease
- man is the only host for infection
- sources of infection:
1. infected human carrier

2. contaminated food / H2O
Pathogenesis:
- after ingestion  organism penetrate the submucosa

of the small intestine infect the mesenteric
lymph nodes  bloodstream causing primary
bacteremia  disseminate to the internal organ
(liver, gallbladder, spleen, lungs, kidney and
bone marrow )
 undergo multiplication  reenter the
bloodstream causing secondary bacteremia *
Clinical Manifestations:
1st week of infection:

- includes : lethargy
fever
malaise
myalgia
constipation
2nd week of infection:

- organism reenter to bloodstreamcauses prolonged bacteremia
 liver  biliary tree (bile duct)
- patient is severely ill, high grade fever 104OF and delirious
- abdominal tenderness with the presence of rose spots *
Clinical Manifestations:
- bacteremia common
- diarrhea may or may not be present
- organism reinfect the GIT & gall bladder causing necrosis
of Payer’s patches
3rd week of infection:
- pat. exhausted , but still febrile

- shows improvement if no complication develop
- death rate 2-10% relapse rate 20%
Collection of specimen (blood, feces and urine)
1.) Blood Culture

- blood (best specimen to detect presence of org.)
- during 1st two weeks of illness 80% of
Ty patients give + result
- OX-Bile medium incubated overnight
2.) Stool culture - McConkey agar

3) Serological: Widal test (tube agglutination test for

antibody titer determination)
- not very reliable and gives a positive result

during the 2nd week of illness that is 7-
10 days after start of infection
- measures specific antibody against O and H Ag
- ideally should be performed on 2 serum sample

to demonstrate 4-fold increase in titer
Interpretation:
Active infection - high or rising titer of anti O Ab (1:160)
Post infect./vaccination - high titer of H Ab.
Anti Vi Ab - carrier person

Typhi Dot
- detect specific IgG and IgM in patient’s serum
Treatment:
Chloramphenicol - drug of choice
Chronic carrier - (Ampicillin + Cholecystectony)
Ciprofloxacin - for resistant cases - 85% cure rate

Prevention:
1. Chlorination of H2O
2. Proper sanitary control of food and eating places
3. Detection and Rx of carriers
4. Efficient waste disposal
5. Vaccine – most promising

oral vaccine T21 A (attenuated typhoid bacilli)
Complications :
1. Intestinal perforation
2. Intestinal hemorrhage
3. Abscess formation
4. Pneumonia
5. Cholecystitis
Clinical Syndrome :
2) Gastroenteritis (S. enteritidis)

- self-limiting disease2-5 days (extreme
casesweeks)
- incubation period 18 – 24 hours after

ingestion of organism  invades intestinal
mucosa and underlying tissues
 localized inflammatory reaction with PMN
S/Sx - diarrhea, fever, abdominal pain
- organism is isolated from feces

- chronic carrier rare
- bacteremia uncommon
Clinical Syndrome :
3) Septicemia (S. cholerae-suis)
- early invasion of organism to the blood stream

follows infection by oral route
- no intestinal involvement
- focal lesions may develop in any organ or

tissue producing:
Osteomyelitis
Pneumonia,
Pulmonary abscess
Meningitis
Endocarditis
Genus Yersenia
Species: Yersenia enterocolitica
- gram (-) coccobacilli

- motile (peritrichous flagella)
- non-lactose fermenter
- urease (+)
- TSI A/A H2S (-) gas (+)
- ferment glucose with gas
- rapid sucrose fermenter
- produce ornithine decarboxylase
Disease: Gastroenteritis
- associated with gastroenteritis in human
- self – limiting
- commonly affect young children
- acquired by fecal-oral route or through ingestion

of contaminated food / H2O
Disease: Gastroenteritis
- primary lesion results from invasion of organism

in wall of small intestine  ulceration
- Symptoms:
abdominal pain (mimic appendicitis)
bloody diarrhea,
fever,
headache
malaise
- extra-intestinal lesion and septicemia may occur
Laboratory Diagnosis :
Culture (Blood agar/Nutrient agar)
- cold enrichment medium

specimen includes feces, blood, aspirate
from mesenteric lymph nodes
Treatment:
Ampicillin, Tetracycline
Supportive (maintenance of fluid and electrolyte)
Coliform /
Opportunistic organisms
Genus Citrobacter
- isolated from environmental sources, feces of

human and animals
- opportunistic (can infect any organ)
*Species:
C. freundii
C. diversus
C. amalonaticus
*Biochemical Characteristics :
- C. freundii prod. H2S, others do not
- grow well on any enteric media
Genus Klebsiellae
Capsule Pigment Motility

Production
Klebsiella + - -
Enterobacter - - +
Serratia - + +
Genus Klebsiella
K. pneumoniae
(Friedlanders bacillus)
Morphologic, Cultural, & Biochemical Characteristics :
- gram (-), non-motile, non-sporeforming bacilli
- capsulated  polysaccharide
- aerobic and facultative anaerobe

Genus Klebsiella
K. pneumoniae
(Friedlanders bacillus)
- can grow on ordinary media
NA- large mucoid, grayish white colonies
MacConkey- red, mucoid gelatinous colonies
- H2S (-), Urease (+) IMViC - - + +

Antigenic structure:
- possess O and K antigen
*K antigen - polysaccharide capsular Ag
- most useful in serologic typing
- all species share the same capsular antigen
1. Capsule  resist phagocytosis (encapsulated organism)

most virulent
2. Elaboration of endotoxin (similar LT & ST E. coli)*

Clinical Infection:
1. Primary community acquired pneumonia

- thick, non-putrid, bloody sputum
2. Urinary tract infection
3. Meningitis
4. Nosocomial infection
5. Wound infection
Lab. Diagnosis:
- microscopic examination (Gram staining)
demonstration morphological characteristic
- India Ink staining  Capsule
- culture medium: Mac Conkey

red, large, mucoid gelatinous colonies
(String test)
- animal inoculation  death 12 – 18 hours
*Treatment: Cephalosporin, Cefuroxime, Cefotaxime)

Genus Enterobacter
8 species associated with human disease
E. cloacae E. sakasakii
E. aerogenes E. taylorae
E. agglomerans E. hafniae
E. gergoviae E. hormaechii
Genus Enterobacter
Morphology:
- short gram (-) rods
- motile ( peritrichous flagella )
- some strains are encapsulated

Biochemical & Cultural Characteristics :
- grow readily on media used for isolation of enteric
- can ferment all CHO with production of acid and gas
- all specie decarboxylate ornithine, except

E. agglomerans
- IMVC - - + +
- TSI A/A gas (+) H2S (-)

*Antigenic structure
- all species possess O & H antigen
* Clinical infection:
2. Meningitis
3. Bacteremia
Genus Serratia
S. marcescens
- gram (-) motile rod
- slow lactose fermenter
- produce extracellular enzymes: Dnase, gelatinase, lipase
- resistant to colistin and cephalosporin
- H2S (-)
Genus Serratia
- produce magenta–red/salmon pigmented colonies when

incubated at room temperature
- most commonly associated with:
1) nosocomial infection
2) urinary tract infection
3) pneumonia
4) wound infection, septicemia
Treatment: Amikacin, Chloramphenicol, Gentamycin, Ciprofloxacin

Tribe Proteeae
3 Genera :
1) Proteus
2) Providencia
3) Morganella
- all members can deaminate phenylalanine

diaminase
- majority are isolated from urine

Genus Proteus
Species of medical importance :
1) Proteus mirabilis
2) Proteus vulgaris
- both actively motile producing swarming
growth on blood agar medium
- H2S +
Genus Proteus
- non – lactose fermenter
- urease producer (break down urea  NH3 & CO2)
- can grow in alkaline ph
- phenylalanine diaminase (+)
- IMViC + + - +/-
- TSI = Alkaline / acid H2S + gas +

Differentiation P. mirabilis P. vulgaris
1. Indole test - +
2. Ornithine
Decarboxylase + -
- possess O, H & K Ag
- *can cross react with rickettsial organism using

Proteus vulgaris strain 0X-19, OXK, OX- 2*
(Weil – Felix test)
Clinical Infection:
P. mirabilis – account for majority of Proteus

infection in human
1) community acquired UTI
2) wound infection
3) pneumonia
4) septicemia
5) bacteremia
Treatment: Ampicillin
Cephalosporin (sensitive)
- all Proteus are resistant to Tetracycline

Genus Morganella
Species: M. morganii (formerly P. morganii)
- infection similar to other members of Tribe Proteeae
Genus Providencia
P. rettgeri (previously classified as Genus Proteus)
P. alcalifacience
Genus Morganella
P. stuartii
P. rustiganii
- both genus are closely related to Proteus group

- Urease - (used to diff. Providencia from
Proteus)
- Indole +
- H2S –
- both species are resistant to most antimicrobial

agent
Clinical Disease:
1. Nosocomial infection
3. Respiratory tract infection
4. Wound infection
P. stuartii P. alcalifacience
1. Prod. of gas from glucose + -
2. Utilization of inositol - +
Gram Negative Coccobacilli Organism
[intermediate between spherical and rods]
- composed of several genera showing considerable

variations but with common characteristics:
1. small, pleomorphic, gram negative rods
2. non-motile and non-sporeformer

Gram Negative Coccobacilli Organism
[intermediate between spherical and rods]
3. fastidious and requires specific accessory

substances for growth such as:
hemoglobin
serum
acetic fluid
amino acid
enzymes
4. difficult to stain - visualization by Polychrome

staining:
1. Giemsa
2. Wayson
Genera:
1. Haemophilus
2. Brucella
3. Francisella
4. Bordetella
5. Calymatobacterium
6. Pasteurella
7. Gardnerella
8. Bartonella
9. Legionella
Genus Haemophilus
- comprises a group of small gram negative, pleomorphic

rods with fastidious growth requirement
- name of genus derived from need of accessory growth

substances X and V factor  BLOOD
- both are produced by heating blood for 15 min. at 80OC

Genus Haemophilus
X factor - protoporphyrin lX (precursor of hematin) heat stable
V factor - NAD (nicotinamide adenine dinucleotide) –

heat labile (coenzyme for pyridine-linked
dehydrogenase)
Species :
H. influenzae
H. aegyptus
H. ducreyi
Haemophilus influenzae
[Pfeiffers bacillus]
- smallest known pathogenic bacilli
- isolated in the sputum of patient with influenza
- most invasive organism
Morphology:
- small, pleomorphic, gram negative coccobacillus
- size 0.2-0.3 x0.5-0.8u

Cultural characteristics :
- aerobe and facultative anaerobes
- req. 5-10% CO2 for growth
- optimum temp.370C ph 7.4 –7.8
- req. X and V factor for growth
- produce indole
- fastidious grows on blood containing enriched media
1. Chocolate agar
- small, colorless, transparent “dew-drop” colonies
with mousy/bleach-like odor
2. Levinthal-Fildes agar
- colorless, mucoid, glistening iridescent colonies
- useful in differentiating capsulated from non-
capsulated strain
3. Blood agar cross-streaked with staphylococcus

- large glistening colonies growing near
staphylococcus which provides the V factor
(Satellite colonies/Satellite phenomenon/Satillitism)
Antigenic Structure
1) capsular polysaccharide antigens

- major antigenic determinant of encapsulated
strain
- basis for serotyping organism
2) somatic antigens
- cell wall Ag found in the outer and inner
membrane containing protein and
lipopoliopsaccharide.
1. Polysaccharide capsule (phosphoribosylribitol phosphate-
PRP)
- most important virulence factor
- plays critical role in the pathogenesis of invasive disease
caused by H. influenzae type B
-antiphagocytic
2. Endotoxin
- contribute to the virulence / invasiveness
3. IgA Proteases
- help in the attachment and resistance of organism to
phagocytosis
Epidemiology:
- worldwide occurrence
- 60-90% of healthy young children harbors non-

capsulated strain
- strain in the nasopharynx (symptom free)
- 50% of children harbor the encapsulated strain
- 25% harbors the type B encapsulated H. influenzae

Pathogenesis:
Portal of entry  URT (Aerosol / Inhalation)

Source of Infection
1. Clinically Active cases
2. Convalescent patient
3. Carriers
Nasopharynx  Sinuses
(colonize) Middle ear
Bronchi
 
Bloodstream Tracheobronchial tree
Site of Predilection :
1. CNS 2. URT
Diseases Produced:
1.Bacterial meningitis
- most serious and most common infection in infants

3 months - 6 years old
- capsulated type b
2. Acute Bacterial Epiglotitis

- occurs older children
- characterized by presence of micro-abscess in the infected
epiglottis with edema and swelling  complete
airway obstruction  respiratory distress
Diseases Produced:
3. Cellulitis
- common site  cheek
4. URT infection (Otitis media, Sinusitis)
5. Chronic Pulmonary Disease

- common among adult especially in patients with
COPD
1. Bacteriological (Presumptive) - gram staining of material:

- CSF, middle ear aspirate, sputum
2. Culture
- specimen  Chocolate agar incubated aerobically
5-10% CO2
3. Quellung test
4. Serological – Antigen detection
A) CIE – detect capsular Ag. (specific test)
B) Latex Particle Agglutination Test
C) Elisa
Treatment: 3rd generation Cephalosporin- drug of choice
Ceftriaxone, Cefotaxime, Cefixime
Alternate drugs:
Ampicillin, Chloramphenicol
Prevention:
- Hib vaccine containing type B polysaccharide given to
children 24 months and above
- Rifampicin – recommended as prophylaxis for children

who are in close contact with infected patient
Haemophilus aegyptus
(Kock – Weeks Bacillus)
- closely related to H. influenzae with regards to morphology

- differentiated from H. influenzae by:
Indole Test Fermentation of Xylose
H. influenzae + -
H. aegyptus - +
- Requires X and V factor

- Do not require CO2 for growth
- Chocolate Agar  greenish sheen colony
Disease: Pink Eye
- communicable form of acute conjunctivitis

characterized by redness and swelling
of conjunctiva accompanied with mucopurulent
discharge
- common among children
- acquired through contaminated hands, towels and

other infected materials
- incidence higher during hot season

Disease: Pink Eye
Lab. Diagnosis:
1. Bacteriological
2. Culture
Treatment:
1.Topical Sulfonamides (Gantrisin)

2.Tetracycline ophthalmic
Haemophilus ducreyi
(Ducreyi bacillus)
Pathogenicity: Chancroid/Soft chancre
- a sexually transmitted disease
- incubation period 2-14 day
- presence of single or multiple circumscribed,

non-indurated, painful genital ulcer accompanied with
marked swelling, tenderness and enlargement of
regional lymph nodes.
Haemophilus ducreyi
(Ducreyi bacillus)
Pathogenicity: Chancroid/Soft chancre
- a sexually transmitted disease

- incubation period 2-14 day
- presence of single or multiple circumscribed,non-indurated,
painful genital ulcer accompanied with
marked swelling, tenderness and enlargement
regional lymph nodes.
Lab. Diagnosis:
1. Bacteriological
- gram stained smear from ulcer exudate or bubo
aspirate  “school of red fish” (classic microscopic
diagnostic appearance)
2. Culture
3. Hypersensitivity test: Ducreyi test
- intradermal injection H. ducreyi suspension
(+) = redness and induration
(-) = absence of above reaction
Treatment: Sulfonamide / Tetracycline

Genus Brucella
- essentially obligate parasite of animals and human

- normal flora urinary tract & genitalia dogs, cow, goat, pig
- inactive metabolically
- man (accidental host)
Species most often found causing
human zoonotic infection
B. abortus - cattle
B. melitensis - goat/sheep
B. suis - swine
= killed by pasteurization
Genus Brucella
Morphology:
- gram negative, non-motile, non-capsulated,

non-sporeforming organism
- arranged singly/cluster
- facultative intracellular parasite
- moderately sensitive to heat and acid
- killed by pasteurization
Cultural characteristics:
- strict aerobe
- catalase and oxidase (+)
- growth enhance by erythritol in placenta of animal

(not in human)
- fastidious, requires serum/blood for 10 isolation
- CM - Serum Glucose Agar – translucent slightly

opalescent smooth, convex moist colonies w/out
hemolysis
Disease: Brucellosis (Undulant / Malta fever)
- disease common among slaughterhouse worker,

veterinarians and farmers
- acquired through:
1. Direct contact w/ infected animals or their products

2. Ingestion of infected meat
3. Drinking unpasteurized milk
4. Occupational exposure
Disease: Brucellosis (Undulant / Malta fever)
- incubation period 1-6 weeks
- characterized by gradual onset of intermittent fever with

diurnal variation, weakness, chill, muscle pain and
profuse sweating
- mental changes common
- granulomatous nodule develop all over the body
- acute bacteremia followed by chronic stage may last

for many years
- liver, spleen, bone marrow are also involved

- blood, CSF, urine or bone marrow (specimen)
1. Culture – done during acute phase of the disease
2. Hypersensitivity test - Brucellergin test (skin test)
3. Serological - Agglutination test

Elisa
C-F test
Treatment: Combination of Streptomycin and Tetracycline

B. abortus B.melitensis B.suis
1. Need of CO2 for growth + - -

2. Ability to produce H2S + - +
3. Growth on dye media:

Basic fuchsin + + -
Crystal violet + - +
Thionine - + +
Genus Francisella
Specie: F. tularensis
- small, non-motile, gram (-) pleomorphic rod with a

characterized by a safety–pin appearance by bipolar
staining
(Polychrome stain)
- encapsulated  organ of virulence
- found intracellularly
Genus Francisella
Specie: F. tularensis
- strictly aerobic
- catalase (-)
- H2S (+)
- fastidious requires cysteine for primary isolation
CM – Glucose Cysteine Blood agar

small drop-like colonies with greenish discoloration
around without hemolysis
Disease: Tularemia (Deerfly fever/Rabbit fever)
- zoonotic
- primarily a disease of persons who have the most

contact with wild animals and more exposed to
insect vectors
- animals are primary reservoir host

(rodents, rabbit, wild and domestic animal)
Disease: Tularemia (Deerfly fever/Rabbit fever)
Acquired through:
1. Handling carcasses / skin of infected animals
2. Ingestion infected meat
3. Drinking infected milk / H2O
4. Inhalation
5. Bites of insect vectors

(tick, lice, mosquito, deerflies)
- organism is extremely invasive, can penetrate intact skin
- intracellular parasite,able to survive in the RES
- clinical manifestation includes chill, fever, back pain,
sweating and prostration, non-productive cough,
nausea,
vomiting and abdominal pain
- granulomatous lesions may develop in various organ
Capsule (major determinants of pathogenicity)

- loss of capsule  loss of virulence
4. Clinical forms of infection affecting humans (accdg. to site of entry)
1. Oculoglandular
primary site conjunctiva granulomatous conjunctivitis
MOT – direct inoculation / bite of arthropods

- characterized by enlargement periauricular LN
2. Glandular / Pneumonic
- primary site -- lungs

- associated with severe systemic manifestation with
generalized lymphadenopathy without ulceration.
- due to inhalation of infected dust
4. Clinical forms of infection affecting human (accdg. to site of
entry)
3. Ulcerogladular
- most common
- focal ulcer develop at site of bite accompanied with
swelling, tenderness & enlarged lymph nodes
- fingers, lower extremities most common site
4. Typhoidal
- GIT disorders due to ingestion contaminated-food / H2O

- characterized by nausea, vomiting & abdominal pain
1. Bacteriological – Fluorescent antibody staining to

demonstrate organism from infected tissue
2. Culture - Blood Cystiene Glucose agar
3. Serology – most important and more commonly used

lab. diagnosis (detect rise of AB titer)
Elisa
Direct-Indirect Fluorescent Antibody test
4. Foshay test
5. Hypersensitivity test – Tularemia skin test

Treatment: Streptomycin
Gentamycin
Prevention:
1. Avoidance of contact with animals

2. Protection from bites of arthropods
3. Avoid eating food / drink contaminated H2O
4. Person at high risk (laboratory personnel)

Immunization - vaccine made of live attenuated strain of
F. tularensis
Genus Bordetella
- highly communicable obligatory parasites of man and

animals
- resides in the mucous membrane of the respiratory tract
- colonized in the ciliated epithelial cell mediated by fimbriae

Genus Bordetella
Species:
B. pertussis - (Bordet Gengou bacillus)
B. parapertussis
B. bronchiseptica
Morphologic & Cultural characteristics :
- small, gram (-) coccobacillus, non-motile, except

B. bronchiseptica
- capsulated (organ of virulence)
- poorly stain with gram staining

- visualization  tolouidine blue stain

safety–pin appearance by bipolar staining
Morphologic & Cultural characteristics :
- strict aerobe
- fastidious – blood & nicotinic acid required for growth
A) Bordet – Gengou medium

(Potato-glycerol-blood agar)
B) Charcoal Oxoid medium
- smooth, convex, pinpoint glistening

colonies with pearl-like luster resembling
mercury droplet appearance
Disease produced : Pertussis (Whooping Cough)
- highly communicable childhood disease

- charac. by paroxysm of coughing with a whoop
Mode of transmission:
1. Aerosol
2. Contaminated material
3. Direct contact
Pathogenesis:
- after inhalation  colonize in the ciliated epithelial

 cell of the respiratory tract
multiply
initial manifestation
conjunctivitis
rhinitis
cough
sneezing
produce toxin - systemic manifestation
- incubation period 5-20 days (1 – 3 weeks)

- no bacteremia, no tissue invasion
3 Stages of Infection:
1. Catarrhal / Prodromal stage
- begins 10 days after exposure and last from 1-3 weeks
- characterized by mild symptoms of uncomplicated URT

infection - coughing, sneezing, rhinitis (common cold)
- most contagious stage
- organism easily isolated from URT at this stage

3 Stages of Infection:
2. Spasmodic / Paroxysmal stage

- followed 1 week later by a dry non-productive cough
which becomes paroxysmal
- paroxysm is characterized by a series of short cough

producing copious mucus followed by a whoop
which is a characteristic sound produced by an
inspiratory gasp of air
- most severe stage
3. Convalescent - cough persist for months with gradual

decrease of s/sx of disease
- plays an important role in the systemic manifestation of

disease
1. Pertussis exotoxin
- respiratory signs and symptoms of the disease
A) Histamin - Sensitizing Factor (HSF)

B) Lymphocytosis - Promoting Factor (LPF)
C) Islet - Activating Protein (IAP)
2. Fimbiae hemagglutinins (F-HA)
- fimbrial antigen derived from surface of organism

- responsible for the adherence of organism in the
respiratory epithelium
3. Tracheal cytotoxin
- role in pathogenesis is unknown
4. Endotoxin (lipopolysaccharide)
- heat stable endotoxin – found cell wall
1. Bacteriological – throat swab / washing
2. Culture (Cough Plate method)

- should be done during catarrhal stage
3. Fluorescent – antibody staining

- direct smear from nasopahrynx
- specific test
4. Serological – Elisa
- Slide agglutination test
Treatment:
1. Erythromycin – drug of choice
Tetracycline
Chloramphenicol ALTERNATE
Trimethoprim
2. Supportive measures
A) Careful suction–to remove tenacious secretion

B) Hydration
C) Electrolyte balance
Prevention:
1) Routine immunization should be given to infants

with killed suspension of B. pertussis
(whole cell inactivated vaccine) - given to

children in 3 doses together with Diphtheria
and Tetanus toxoid (DPT)
2) Erythromycin – given to children under 4 years with
history of exposure to infected person
3) Hyperimmune gamma–globulin for exposed infants

under 2 years old
Immunity:
- permanent immunity acquired after attack

Genus Calymmatobacterium
Specie: Calymmatobacterium granulomatis
Morphology:
- pleomorphic, non-motile, heavily capsulated organism
which exhibit single or bipolar condensation of
chromatin  safely-pin appearance
- replicate intracellularly within mononuclear cells and are

liberated when cell rupture
Disease - Granuloma inguinale / Donovanosis
- classified as a minor venereal transmitted disease
- characterized by presence of beefy red painless

ulcerating lesion of the skin and mucous
membrane of genitalia and inguinal area that
bleeds easily on contact
- may progress very slowly  heal with scarring 

genital elephantiasis
- direct exam. of smear / biopsy material stain with

Wright / Giemsa
- demonstrate pathognomonic cell  Donovan bodies

(large, histiocytic, endothelial cells containing
numerous encapsulated, red – oval bacilli )
Treatment : Tetracycline
Ampicillin
Trimethoprim - Sulfamethoxazole
Genus Pasteurella
Specie: Pasteurella multocida
- forms part of the normal oral flora of cat, dog and

other domestic and wild animals
- only specie associated with human infection
- worldwide distribution
- habitat  respiratory and GIT of animals

Genus Pasteurella
Morphology:
- small gram (-) coccobacillus with a safety pin

appearance by bipolar staining
- occur singly, pairs and occasionally short chains
- non-motile and capsulated (hyaluronic acid)


organ of virulence
- catalase and oxidase (+)
- metabolism fermentative
- optimum temperature 370C
- grows readily on any CM. contg. blood or hematin
- BA - small translucent colonies with brownish
discoloration which often have a characteristic
musty odor, non – hemolytic
Epidemiology:
- most common organism isolated in human wounds

inflected by bite of cat and dog
- survives poorly in soil and water
- tonsils of dog - common site of colonization of

organism
Epidemiology:
- transmitted by direct contact- usually a bite
- human infection may be due to:
1) localized wound infection / cellulitis from

bites or scratches of dogs and cats
2) superinfection of a chronically diseased lung
3) septicemia
1.capsular Ag.
- 4 serotypes- A, B, D, E
- A and D – most frequent type assoc. with
human infection
2. somatic Ag.
1. capsule
2. somatic Ag.
3. endotoxin
4. neuraminidase
5. hyaluronidase
Culture
Specimen: early AM sputum, bronchial washing,
nasal swab, pus from animal bite,
blood, CSF
Treatment: PCN – drug of choice.

Prevention:
After animal bite:
- wound should be cleaned with soap and H2O

and debrided
- avoid suturing
- PCN / Tetracycline should be given at the

time of the bite
- no vaccine available
Genus Gardnerella
Specie: Gardnerella vaginalis
General characteristics:
- pleomorphic, gram negative coccobacilli with

pointed ends (club-shaped form with
metachromatic granule)
- non-motile, non-capsulated
- fastidious in nutritional requirement

Genus Gardnerella
Clinical infection: nonspecific vaginitis (Bacterial Vaginosis)
- present in vagina 40% healthy women
- 95% women with vaginitis
- sexually transmitted disease
- characterized by copious malodorous

vaginal discharge with fishy-like odor
- vaginal pH greater than 4.5
- no symptoms in male
Lab. Diagnosis:
Specimen - Cervical
Urethral swab
Vaginal
1. microscopic exam. ( wet mount )
- demonstrate presence of characteristic
“Clue Cells”

(mass of bacteria on the surface of the)
vaginal epithelial cell
2. culture medium - Human Blood Tween agar

- convex, opaque gray colonies with
beta - hemolysis
3. tissue culture
Treatment: Metronidazole
Genus Bartonella
Specie: Bartonella bacilliformis

- only specie pathogenic to human
Morphology:
- pleomorphic, motile (unipolar flagellum), gram negative
coccobacilli
- stain weakly with aniline dye
- appears bright redpurple with Wright/Giemsa stain

Genus Bartonella
- obligate aerobe
- growth temp. requires 30oC
- Culture media :
1. Cell free medium containing fresh serum and
hemoglobin of rabbit / horses
2. Yolk sac chick embryo
3. Tissue culture
Disease: Bartonellosis / Carrion’s disease
- exclusively human disease
- transmitted by bites of sandfly (Genus phlebotomus)
- bite usually occurs during nighttime when female

flies takes their blood meal
2 Clinical forms of infection:
1) Oroya fever (initial stage)

- serious form with systemic manifestation
- highly fatal hemolytic disease characterized by

a) intermittent fever
b) severe bone & muscles pain
c) anemia (due to blood destruction)
- incubation period 2-5 weeks

2) Verruga Peruana (eruptive stage)
- chronic and non-fatal

- characterized by the presence of bright red
angiomatous wart-like skin lesions
which maybe localized or generalized
- can occur to patient who recover from Oroya fever or
persons w/ no prior clinical evidence of
Bartonellosis
1. demonstration & Isolation of organism in peripheral

blood smear stained with Geimsa stain
2. blood culture
3. skin biopsy (Verruga Peruana)
4. serological – CF test
Treatment:
Chloramphenicol (drug of choice)
Penicillin, Tetracycline, Streptomycin (alternate drugs)
Prevention:
- no vaccine available
- anti-vector measures must be instituted
a) use of insecticide DDT or insect repellent

b) use mosquito net (personal protection)
Genus Legionella
Specie: Legionella pneumophila
Morphology:
- gram negative motile rod

- size 0.3-0.9 x 2-3 u
- difficult to stain (stained poorly with gram stain)
- special stain to visualize organism from tissue
1. Dieterle Silver impregnation stain
2. Direct fluorescent antibody

Cultural & Biochemical Characteristics :
- facultative intracellular parasite

- do not grow on routine laboratory media
- fastidious and requires enriched media containing
cysteine and iron for growth (Charcoal Yeast
Extract )
- oxidase and catalase (+)
- produce catalase, gelatinase, B- lactamase enzymes
Clinical Infection:
2 clinical forms :
1) Acute Atypical Pneumonia / Legionnaires disease

- characterized by severe pneumonia with abrupt onset
of fever, chill, headache and non-productive
cough
- usually accompanied with extra pulmonary
manifestation
2) Pontiac Fever
- characterized by mild influenza–like manifestation
w/out pneumonia
- self–limiting
- incubation period1-2 days
Clinical Infection:
2) Pontiac Fever
- characterized by mild influenza–like manifestation

w/out pneumonia
- self–limiting

Mode of transmission : airborne/ inhalation of
contaminated H2O from environmental source
- primarily a water – borne organism

- no documented person–person transmission
- common sources of infection:
1. exposure to air conditioning system, cooling towers

or evaporative condenser
2. potable water supply from hotels, hospitals and

factories
3. sprinkling system and from showers

1. Direct demonstration of organism from clinical

materials (sputum, lung / trachea
aspirate )
- Dieterle silver impregnation FAT
2. Culture – Charcoal Yeast extract Mueller-

Hinton
3. Serology – detection of specific AB pat. serum
Indirect Immunoflourescence, ELISA, RIA

Treatment:
Erythromycin (may be combined with Rifampicin)
or Ciprofloxacin
Prevention: none – no vaccine available
- depends upon maintenance of hot water and air

conditioning system particularly in large
buildings, offices, hospitals and hotels

Gram Rods

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Gram Negative Rods

Cytochrome Oxidase Positive

- most important human pathogen

1st group - ( V.cholerae O1 ) classic epidemic cholera

- caused by organisms that agglutinate in

- directed against the O1 antigen &

2nd group - (atypical or nontoxigenic V.cholerae O1)

3rd group – ( non-O1 V. cholerae ) does not

- possess lysine and ornithine decarboxylase

- reduce nitrate to nitrite

- urease (-), H2S (-)

- non-lactose fermenter but ferment sucrose,

Selective - Differential CM: (vibrio colonies from fecal

MacConkey’s Agar – most strains grow luxuriantly and on

- “H” or flagellar antigen commonly shared by all vibrios

*1) Voges – Proskawer + -

- both biotypes are responsible for classic epidemic cholera

* 1) Cholera enterotoxin/Choleragen (heat labile)

- major pathogenic factors that act on the cells of the

- composed of two major subunits:

* - subunit A – responsible for the biologic activity of the

stimulate adenylcyclase activity of the epithelial cell,

*2) Invasiveness of organism

- ability to adhere/penetrate to the intestinal mucosa

- food and waterborne disease

- *human carrier serves as source of new cases of

- *acquired fecal – oral route (contaminated food/H2O)

- man only host infection

- incubation period 2-3 days

- serious disease charac. sudden onset of voluminous,

- fluid loss severe 15 – 20 L / day

- patient severely dehydrated:

- no fever, untreated cases 60% die

- high attack rate – children

- organism non – invasive (remains localized GIT)

1) Direct microscopy of stool specimen

- observe for comma-shaped bacilli

2) Stool culture – MEA / TCBS incubated 2 – 3 days

3) Direct Fluorescent Antibody Test

- Prompt replacement of fluid and electrolytes

- Tetracycline (DOC) & Doxycycline

1) Maintenance of adequate sewage treatment

- found in estuaries throughout the world

- major cause of gastroenteritis involving seafoods

- closely related to V. Cholerae

- non–lactose and non-sucrose fermenter

* CM – TCBS = large, smooth, green colonies

- most strain prod. diarrhea due to heat – stable

- can lyze human rbc.

- Kanagawa Hemolysis Test

- self-limiting disease acquired through ingestion of

- incubation period 12-24 hours

- fever (not common)

- clinical manifestations may last for 3-5 days

Treatment: Chloramphenicol, Kanamycin, Tetracycline

- closely related to V. cholerae morphologically and

- most important human pathogen associated with

- gram (-) spirally curved or seagull-wing shaped rod

- motile (single polar flagellum) darting motility

- oxidase and catalase (+), H2S producer

- asaccharolytic (non-carbohydrate fermenter)

- grows best at 42 – 430C

- susceptible to gastric acid