MICROBIOLOGY LECTURE 10 - Enterobacteriaceae, Shigella,

Salmonella Microscopic Morphology

Notes from Lecture 1. small (0.5 by 3.0 um), gram negative non-sporeforming
USTMED ’07 Sec C AsM rods
2. motile by means of peritrichous flagella; non-motile –
ENTEROBACTERIACEAE Shigella and Klebsiella
Enterobacteriaceae General Characteristics 3. well-defined capsule
1. ubiquitous organisms that are found in the soil, water, • Klebsiella
decaying matter and the large intestines of animal and • Enterobacter
insects • Serratia
2. natural habitat in humans – gastrointestinal tract referred to
• Some Escherichia strains
as “enteric bacilli” or “enterics”
3. constitutes only a minor fraction of the total microbial flora - slime layer – loose ill defined coating
of the gastrointestinal tract 4. fimbriae or pili – responsible for attachment of the
4. includes some of the most important cause of gastrointestinal bacterial cells to other bacteria, host cells and
disease bacteriophage
a. salmonellae – typhoid fever 5. cell wall consists of:
20% of the cell wall; responsible for
b. shigella – bacillary dysentery a. murein cellular rigidity
5. some are members of the normal commensal flora that can b. lipoprotein
cause opportunistic infections c. phospholipids
a. Escherichia coli d. protein
b. Klebsiella pneumonia e. lipopolysaccharide – contains the specific
c. Proteus mirabilis polysaccharide side chain
6. Responsible for the majority of nosocomial (hospital- i. determines the antigenicity of the
acquired) infections various species
7. causes disease ii. portion of the cell responsible for
a. in the gastrointestinal tract the endotoxic activity
i. Shigella
b. Outside the gastrointestinal tract - 80% of the cell wall is joined to the lipid of the
i. Klebsiella lipoprotein to form a lipid bilayer
ii. Enterobacter
iii. Serratia Colonial Morphology
iv. Proteus 1. Similar colonial morphology in blood agar plate
c. Both inside and outside the gastrointestinal tract a. Moist, smooth, gray colonies
i. Escherichia coli b. Some strains are beta hemolytic
ii. Salmonella 2. Eschericia coli – convex, cicular, smooth with distinct
iii. Yersinia edges
8. Infection may originate from the: 3. Enterobacter – similar but somewhat more mucoid
a. Animal reservoir – most Salmonella infections 4. Klebsiella – large and very mucoid and tend to coalesce
b. Human carrier – Shigella and Salmonella typhi with prolonged incubation
c. Endogenous spread of organisms in susceptible 5. Salmonella and Shigella – similar to Escherichia coli but
patient – Escherichia coli do not ferment lactose
9. Possesses complex antigenic structure 6. Pigmented genera
10. Produces a variety of toxins and other virulence factors • Serratia – reddish orange
• Edwardsiella – yellowish white
Classification of Enterobacteriaceae is based on:
1. DNA homology Enterobacteriaceae on 5% sheep
2. Biochemical properties blood agar. Characteristic colonial
morphology on 5% sheep blood
3. Serologic reactions agar showing large, dull, grey,
4. Susceptibility to genus specific and species specific nonhemolytic colonies. Hemolysis
bacteriophages is variable and not characteristic
5. Antibiotic susceptibility pattern of any one genus.

Edwards and Ewing Classification of Enterobacteriaceae

Tribe Genus Species
I. Escherichiae I. Escherichia Eschericia coli

II. Shigella Shigella dysenteriae

Shigella flexneri
Shigella boydii
Shigella sonnei
II. Edwardsiellae I. Edwardsiella Edwardsiella tarda
III. Salmonellae I. Salmonella Salmonella choleraesuis
Salmonella typhi
Salmonella enteritidis

II. Arizona Arizona hinshawii

Citrobacter freundii

III. Citrobacter Citrobacter diversus

IV. Klebsiella I. Klebsiella Klebsiella pneumoniae
Klebsiella ozaenae
Klebsiella rhinoscleromatis

II. Enterobacter Enterobacter cloacae

Enterobacter aerogenes
Enterobacter hafnia
Enterobacter agglomerans

III. Seratia Serratia marcescens

Serratia liquefasciens
Serratia rubidaea
V. Proteae I. Proteus Proteus vulgaris
Proteus mirabilis
Proteus morganii
Proteus rettgeri

II. Providencia Providencia stuartii

Providencia alcalifaciens
VI. Yersineae I. Yersinia Yersinia enterocolitica
Yersinia pseudotuberculosis
Yersinia pestis
VII. Erwiniae I. Erwinia
(plant pathogens)
 - citrate positive
Klebsiella pneumoniae on - differs from salmonella – (-) lysine decarboxylation
MacConkey agar. Rapid lactose - ferments lactose very slowly
fermenting colonies of Klebsiella
• Shigella
pneumoniae appears pink, large,
glistening, and mucoid. This strain is - nonmotile
probably encapsulated and therefore - nonlactose fermenter
appears mucoid. Although this - ferments other carbohydrates producing acid but
appearance is associated with not gas
Klebsiella pneumoniae. It is not - closely related to Escherichia coli
unique for that species. - share common antigen with one another and with
Pigmented Serratia sp. on
other enteric bacteria
MacConkey agar. These colonies
appear red and should not be • Salmonella
confused with the pink color due to - motile
lactose fermentation shown in 8.3. - ferments glucose and mannose without gas but do
Rare strains of Serratia spp. produce not ferment lactose or sucrose
pigment, which is seen on all solid
media including the blood agar - produces H2S
plate. Proteus species on 5% sheep
blood agar. Growth appears to
spread as a film on the plate from
the original colony or streak line,
Biochemical Characteristics often extending in waves. This
1. facultatively anaerobic or in low oxygen atmosphere characteristic of Proteus spp. is
ferments carbohydrates called swarming and suggests that
the microorganism is motile by
2. with sufficient oxygen – utilize the tricarboxylic acid means of flagella.
cycle and the electron transport system for energy
production
Antigenic Structure
Enterobacteriaceae Pseudomonadaceae 1. O-antigens – somatic – heat stable
All are facultative anaerobes Strict aerobes a. species specific polysaccharide which makes
All ferment glucose with acid or Does not ferment glucose up a part of the LPS component of the cell
acid and gas wall
Oxidase negative Oxidase positive b. endotoxic activity expressed by the LPS
Reduce nitrates to nitrites Does not reduce nitrates to complex reside in the lipid A molecule
nitrites 2. K or capsular antigen – heat labile
a. a polysaccharide component found in some
members of the enteric bacilli
b. called Vi antigen in Salmonella
• Escherichia c. basis of serologic typing of Klebsiella
1. positive indole d. cells possessing K antigen are more pathogenic than
2. positive lysine decarboxylation those that lack them
3. positive mannitol fermentation e. inhibits phagocytosis and the effects of serum
4. produce gas from glucose antibody
5. urine isolate – identified by: 3. H antigens or flagellar antigens – heat labile
a. hemolysis in blood agar a. protein found only in motile forms
b. typical colonial morphology in EMB – greenish b. motile bacteria may lose their capacity to produce
metallic sheen or iridescent sheen flagella but retain their O antigen specificity
c. positive spot indole test - Other antigens
d. positive β-glucoronidase using the substrate 4- 4. Enterobacterial common antigen or kunin antigen
methylumbelliferyl-β-glucoronide (MUG) a. haptenic substance occurring in two aggregative
e. negative oxidase test forms
o linked to the lipopolysaccharide and is
• Klebsiella-Enterobacter-Serratia Group immunogenic
1. Klebsiella o linked to a carrier molecule and
a. mucoid growth nonimmunogenic
b. large polysaccharide capsule b. LPS linked ECA rare but found in Escherichia coli
c. lack of motility 014
d. positive lysine decarboxylase and citrate c. Role in virulence and pathogenesis not established
2. Enterobacter d. Chemical nature undefined
a. positive tests for motility 5. Fimbrial antigens
b. positive citrate a. believed to be virulence factors because of their
c. positive ornithine decarboxylase adhesive properties
d. produce gas from glucose b. protein in nature and present in all enteric bacilli
e. enterobacter agglomerans – small capsule
3. Serratia
- produces DNase, lipase and gelatinase
- all are positive for Voges Proskauer test

• Proteus-Morganella-Providencia Group
- deaminate phenylalanine
- motile
- grow on KCN medium
- ferment xylose
1. Proteus
- swarming on solid media – actively motile by means
of peritrichous flagella
2. proteus species and Morganella morganii
– urease positive
3. Providencia
- urease negative
4. Proteus-Providencia
– ferment lactose very slowly or not at all
5. Proteus mirabilis
– more susceptible to antimicrobial drugs including
penicillin

• Citrobacter Determinants of Pathogenicity

1. Endotoxin
a. LPS of the cell wall – endotoxin – associated with the
bacterial cell and is toxic to animals
b. Toxicity of LPS resides in the lipid A molecule
c. Effects when injected to animals
- fever
- fatal shock
- leukocyte alterations
- regression of tumors
- alterations in host response to infection
- Sanarelli-Shwartzman reaction
- Various metabolic changes
d. cellular targets are varied
e. exact mechanism of action – not clearly delineated
f. 30% of enteric bacteremia will develop endotoxic shock
chief defect – pooling of blood in the microcirculation –
causes cellular hypoxia and metabolic failure due to
inadequacy of blood in vital organs
2. Enterotoxins
a. toxins that usually affect the small intestine
causing a transduction of fluid into the intestinal
lumen and subsequent diarrhea
o salmonella
o shigella Laboratory Diagnosis
o strains of Escherichia coli, Klebsiella
pneumoniae Specimens – sputum, tissue, pus, body fluids, rectal swabs or feces
o citrobacter freundii
o Enterobacter 1. Culture
b. enterotoxin producing Escherichia coli – major a. cefsulodin-irgasan-novobiocin (CIN) – selective for
causes of travelers diarrhea and diarrhea in Yersinia
developing countries b. isolation and differentiation of lactose fermenters from
c. incidence of disease cause by enterotoxin nonlactose fermenters
producing Citrobacter, Klebsiella and Enterobacter - Eosin Methylene Blue agar
species – unknown - MacConkey agar
d. role of enterotoxins in salmonellosis and shigellosis c. Gram-negative broth (GN broth) – enrichment broth used
– unclear; tissue penetration important in to enhance isolation of enteric pathogens
pathology d. Isolation and differentiation of Salmonella and SHigella;
3. Shiga toxins and Shigalike toxins (verotoxins) inhibition of normal flora coliforms
a. Shiga toxin – interferes with protein synthesis of - Hektoen enteric agar
mammalian cells; role in shigellos is unclear
b. Certain E. coli strains – produce similar toxins called - Salmonella Shigella agar
verotoxins because of their action of Vero (African green - Xylose-lysine-desoxycholate (XLD)
monkey) tissue culture cells; important causes of e. enrichment broth used to enhance recovery of
hemolytic diarrhea and hemolytic uremic syndrome Salmonella and Shigella
4. Colonization factors - selenite broth
a. capsule of Klebsiella pneumoniae – to prevent - tetrathionate broth
phagocytosis
b. Vi antigen of Salmonella typhi – prevents intracellular 2. Biochemical Tests
destruction of the bacterial cell a. Triple sugar iron Agar – to distinguish morphologically
c. Fimbriae such as the CFA of human isolates – necessary similar bacteria of Enterobacteriacea all of which
for the attachment of the organism to target tissues ferment glucose to an acid end product
d. O antigen – may bind the organism to certain tissue b. IMVIC reactions – for the identification of lactose-
receptor sites fermenting members of Enterobacteriaceae.
Escherichia, Enterobacter and Klebsiella.
Clinical Infection i. Indol test
ii. Methyl red test
Types of infection iii. Voges Proskaeur test
1. leading cause of bacteremia and urinary tract infections iv. Citrate utilization test
2. can invade any body site and can cause wound c. Urease test – for the identification of:
infections, pneumonia, meningitis and various i. Rapid urease producers – Proteus and
gastrointestinal disorders Morganella
3. opportunistic infections occur outside of the intestine ii. Weak urease producers – Klebsiella and some
and require an alteration of the host by some enterobacter
mechanical, physiologic or infectious process before d. deaminase reactions – to identify organisms producing
they can cause disease the enzyme deaminase such as Proteus, Providencia and
4. true enteric pathogens – Salmonella, Shigella and Morganella
Yersinia e. decarboxylase reactions – to determine the production
5. potential danger – secondary bacteremia and endotoxic of decarboxylases by bacteria
shock f. motility – determines the motility of bacteria through
semisolid media
Sites of infections with members of the enterobacteriaceae i. Nonmotile – Shigella and Klebsiella
ii.Yersinia enterocolitica – nonmotile at 37oC but
motile at 22oC
g. ONPG reaction
i. Determines the presence of late or slow
lactose fermenting strains
o useful in detecting late-lactose
fermenting strains of Escherichia coli
o distinguishes Citrobacter species and
arizonae subspecies from similar
Salmonella subspecies
o useful in the speciation of shigella

3. Serologic grouping of the O, H and K antigens to

characterize certain enteric isolates
a. Salmonella and Shigella – for complete identification
b. Other organisms – for epidemiologic purposes
Rapid, presumptive identification of gram negative bacteria

Lactose fermented rapidly

Escherichia coli
- metallic sheen on differential media; motile, flat,
nonviscous colonies

Enterobacter aerogenes
- raised colonies, no metallic sheen; often motile; more
viscous growth

Klebsiella pneumniae
- very viscous, mucoid growth; nonmotile

Lactose fermented slowly

Edwardshiella
Serratia
Citrobacter
SHIGELLA
Arizona
Providencia
1. Gram negative nonsporeformers
Erwinia
2. none are motile, hence they do not contain flagellar
Lactose not fermented
antigens
3. some resemble anaerogenic type of coliform bacilli and
Shigella species
typhoid bacilli in that they ferment carbohydrates with
- nonmotile; no gas from dextrose
the production of acid without gas
Salmonella species
- motile; acid and usually gas from dextrose 4. Differ from one another biochemically and
Proteus species immunologically
- swarming on agar, urea rapidly hydrolyzed (smell of
ammonia)
5. Facultative anaerobes; optimum temperature for growth
is 37oC
6. Grow upon the ordinary nutrient (beef extract medium)
7. relatively resistant to the bacteriostatic action dyes
8. can grow in EMB, MacConkey, SS agar, deoxycholate agar
9. Only Shigella sonnei is a slow lactose fermenter

Properties that distinguish Shigella from most Salmonellae

1. lack of motility
2. failure to produce gas during fermentation except for
some biotypes of Shigella flexneri 6 and rare strains of
Shigella boydii 13
3. lack of lysine decarboxylase
4. possess specific polysaccharide O antigens but no H
antigens
5. much less invasion rarely causing bacteremia
6. much narrower distribution in nature inhabiting only the
intestinal tracts of primates

Classification is based on fermentation of mannitol

Treatment
1. factors in the difficult treatment of enterobacterial
infections
• underlying disease of patient
• emergence of resistant strains
2. The appropriate antibiotic must be chosen by:
• careful evaluation of the isolate’s susceptibility
pattern
• condition of the host
• site of infection
3. Treatment of shock – centers on the cardiovascular system
• restoration of the intravascular volume
• digitalization
• administration of isoproterenol
• steroids, pressor amines and norepinephrine
I. Nonfermenting group
– serogroup A – Shigella dysenteriae
II. Mannitol fermenters
– serogroup B – Shigella flexneri
- serogroup C – Shigella boydii
- serogroup D – Shigella sonnei
• Group A – Shigella dysenteriae
1. Shigella dysenteriae type I – Shiga bacillus
o produces an endotoxic LPS which is
responsible for O antigenicity of the
bacillus
o unique in its production of an exotoxin, a
neurotoxin, which affects the central
nervous system
o Similar to diphtheria toxin in potency and
mechanism of action
o Produces an enterotoxin which closely
resembles cholera and coliform
enterotoxins in its mechanism of action
2. Shigella dysenteriae type 2 – Schmitz bacillus
o encountered in institutional and other
outbreaks of dysentery
• Group B – Shigella flexneri – Flexners bacillus, Hiss and
Russell’s bacillus
1. Worldwide in distribution
2. most commonly found of the dysentery bacilli
making up more than half of isolates
• Group C – Shigella boydii – Newcastle Manchester
bacillus
- similar in pathogenicity to Shigella flexneri but
differs immunologically
• Group D – Shigella sonnei – Duval’s bacillus
1. most common species in the US – 72% of isolates
2. slow lactose fermenter, producing acid in one week
to 10 days

Pathogenesis
 1. natural habitat of dysentery bacilli – large intestine of all the consequences of such a mistake
humans – BACILLARY DYSENTERY
2. infection limited to gastrointestinal tract
3. blood stream invasion is rare Epidemiology
4. highly communicable 1. occurs in children from 6 months to 10 years of age
5. infective dose – less that 10 microorganisms 2. endemic disease in adults is frequently due to contact
with infected children
Bacilli penetrate epithelial cells of the colonic mucosa 3. epidemic outbreaks of disease are associated with day
care centers, nurseries and custodial institutions
4. transmitted by the fecal oral route, primarily by
Multiplication of bacilli contaminated hands and less commonly in water or food
– food, fingers, flies, feces
5. spreads rapidly in communities where sanitary standards
Microabscesses develop in the lamina propria and the level of personal hygiene are low

Laboratory Diagnosis
Necrosis of mucosa due to action of enterotoxin Specimens
1. fresh stool, mucus flecks, rectal swab – culture
2. serum – serology
ulcerations
1. Culture
a. EMB or MacConkey – colorless colonies
b. Salmonella-Shigella agar – colorless colonies without
Formation of diphtheritic or pseudomembrane consisting
black centers
of fibrin, WBC, debris, tissues and bacteria
c. Hektoen enteric agar – green colonies without black
centers
Granulation and scar formation
2. Biochemical tests
Cllinical Manifestations
1. incubation period – 1 to 4 days a. TSIA – acid butt, alkaline slant, no gas, no H2S
2. begins with fever, abdominal cramping and pain, and b. MR positive
diarrhea c. Citrate – negative
3. Two stages of the infection d. ODC – negative
a. First stage – involves a watery diarrhea, which e. ADH – negative
may last up to 3 days f. Deaminase (phenylalanine) – negative
b. Dysenteric phase – characterized by frequent g. Urease – negative
stools, with red blood cells, white blood cells, h. Carbohydrate fermentation
and mucus present o sucrose – negative
o the bacteria have invaded the epithelial o salicin – negative
lining of the intestine, causing severe o adonitol – negative
inflammation o dulcitol – negative
o D-mannose – positive
DIFFERENTIAL DIAGNOSIS BETWEEN AMEBIC DYSENTERY AND TISSUE-
INVASIVE BACTERIAL GASTRO-ENTERITIS 3. Slide agglutination by specific Shigella antisera
Amebic Dysentery Tissue-invasive bacterial Reaction Dysenteriae Flexneri Boydii Sonnei
Colitis (Shegella dysentery)
Fermentation
Incubation period Variable (usually 2- Short (24-72 hours)
of:
4 weeks or longer)
Lactose - - - +
Onset Insidious Acute
Mannitol - + + -
Fever Usually absent, Common, moderately
ODC - - - +
unless complicated high
ONPG - - - +
Toxemia Usually absent Often present
Dehydration Unusual usual
Treatment
Hepatomegaly Common (if uncommon 1. supportive
complicated or a. prompt replacement of fluids and electrolytes
disseminated) b. opiates should be avoided
Colonic mucosal Gross findings: Diffuse erythema with 2. specific
changes segmented: ulcers loss of vascular pattern; a. potent specific antitoxin against Shigella
with undermined mucopurulence, mild dysenteriae
borders and friability, occasional b. antimicrobials
normal intervening ophthoid ulcers i. Ampicillin or the analogue
mucosa; lack of amoxicillin
inflammation ii. Chloramphenicol
iii. Tetracycline
Microscopic iv. Norfloxacin or Ciprofloxacin
findings – Edema, capillary v. Trimethoprim sulfamethoxazole
trophozoites in congestion, focal
flask-shaped hemorrhages, cyst, Prevention and Control
ulcers; wet mount hyperplasia, goblet 1. mass chemoprophylaxis
shows motile depletion, mononuclear 2. eliminate organism from reservoir hosts
amoeba containing polymorphonuclear a. sanitary control of water, food and milk;
RBC leukocyte infiltrate; loss sewage disposal and fly control
of epithelial cells with b. isolation of patients and disinfection of
microulcerations excreta
Stool appearance Bloody mucoid Bloody mucoid less c. detection of subclinical cases particularly in
semiformed formed food handlers
Fecal leucocytes Uncommon or few Usually abundant with
(methylene blue macrophages SALMONELLAE
staining)
Red cells Numerous and in Numerous, discrete
1. Gram negative, nonencapsulated, nonsporulating rods
clumps
2. All species are actively motile by means of peritrichous
E. histolytica Hematophagous absent flagella except Salmonella pullorum and Salmonella
trophozoites gallinarum.
present 3. stain readily with the usual dyes such as methylene blue
Indirect Positive in invasive negative and carbol fuchsin
hemogglutination amebiasis (85-95%
test (IHA) of cases) 4. have simple nutritional requirements growing readily on
* Confusion in differentiating amebic cysts from fecal leucocytes and amebic cysts from the usual nutrient media
fecal leucocytes and amebic trophozoites from large motile macrophages may occur with
 5. optimum temperature is 37 C but growth occurs at a
reasonable rate at room temperature
6. facultative anaerobes growing equally well under either
aerobic or anaerobic conditions
7. characterized biochemically by failure to ferment
lactose or salicin and inability to liquefy gelatin or
produce indole
8. ferments sugar with gas but there are anaerogenic
strains of Salmonella enteritidis, Salmonella
typhimurium and Salmonella paratyphi C
Enteric Fevers
Kauffman White Classification scheme – based on agglutination
1. Paratyphoid fever
tests with absorbed antisera, permitting identification of different
a. relatively mild course with sudden chills but
O and H antigens in an unknown organism.
otherwise similar to a mild typhoid fever
b. most often milk borne and transmitted by
Salmonella Nomenclature
carriers
c. predominant in younger age groups
Salmonella subgroup Former Genus Subspecies
d. causative agents
1 Salmonella Choleraesuis
 Salmonella paratyphi A – sewage
2 Salmonella Salamae
contaminated food
3a Arizona Arizonae
 Salmonella paratyphi B (Salmonella
3b Arizona Diarizonae
schottmulleri) – often in carriers
4 Salmonella Houtenae
 Salmonella paratyphi C (salmonella
5 Salmonella Bongor hirschfeldii) – tends to produce
6 Salmonella idnica endocarditis
2. Typhoid fever
Antigenic Structure a. Salmonella typhi – a strict parasite of man
1. somatic O antigen – LPS b. Not only an intestinal infection but a general
a. associated with the cell substance invasion particularly the lymphatic system
b. antibodies are mostly IgM
2. flagellar or H antigen
a. associated with the flagella
b. antibodies are mostly IgG
3. Vi antigen
a. surface antigen of the K class
b. occurs in Salmonella typhi, Salmonella paratyphi A
and B, and some strains of Citrobacter
c. blocks agglutination by antiserum against the O
antigen
d. virulence antigen – presence associated with
virulence and antibody; it is protective
4. Additional K antigen – R antigen

Host factors that contribute to the resistance to Salmonella

infection
1. Gastric acidity
2. normal intestinal microbial flora
3. local intestinal immunity

Three Clinical Entities

1. Salmonella gastroenteritis or Salmonella food poisoning
2. enteric fevers
3. septicemia

Salmonella Gastroenteritis or Salmonella food poisoning

1. Most common manifestation of Salmonella infection Clinical Manifestations
2. 8-48 hours after the ingestion ---------à nausea, 1. First week
headache, vomiting and profuse diarrhea with few a. gradual onset with increasing remittent fever
leukocytes in the stools but rarely blood b. dull, continuous headache
3. Low grade fever c. anorexia, malaise
4. Recovery within 2-3 days d. nonproductive cough
5. Blood stream invasion quite rare e. epistaxis
6. an infection not an intoxication because no toxin is f. vague abdominal discomfort
involved g. constipation
7. Large numbers of bacilli cause irritation of mucosa 2. Second week
8. Causative agents a. temperature sustained at 40oC
a. Salmonella typhimurium b. appears acutely ill and weak
b. Salmonella enteritidis(Gartners baciluus) c. dull, lethargic, expressionless face
c. Salmonella choleraesuis d. mental state normal to mental confusion to
delirium
e. abdominal discomfort and distention
f. diarrhea more common with blood
- Physical Findings
1) spleen enlarged and soft
2) maculopapular lesions on anterior chest and
upper abdomen
3) rose spots 2-3 mm. in diameter, blanch on
pressure lasting 2-4 days
4) slow pulse rate or bradycardia inspite of high
temperature
3. 3rd and 4th week – recovery when fever subsides; death
when untreated
4. Complications
a. intestinal hemorrhage
b. perforation and peritonitis
c. splenic rupture
 5. Typhoid carriers black sheen or with dotted black or greenish
a. more in females than males between 50-59 years gray
b. maybe convalescent carriers or chronic carriers
c. bacilli in the gall bladder or in the urinary bladder 2. Biochemical identification
d. best treated with massive ampicillin and a. TSI: KI
cholecystectomy b. H2S: positive
c. Motility: positive
Septicemia
d. MR: positive
1. may occur as a sequelae of enteric fever or rarely from
e. VP: negative
gastroenteritis or may have no intestinal focus
f. Indole: negative
2. characterized by rapid rise with spiking temperature
g. LDC: positive
3. Signs and symptoms refer to the area involved
h. Urease: negative
a. pneumonia
i. Deaminase (phenylalanine): negative
b. meningitis
j. Fermentation of:
c. conjunctivitis
o sucrose – negative
d. sinusitis
o myoinositol - positive
e. suppurative arthritis
f. pyelonephritis o sucrose – positive

3. Serology
a. slide agglutination with specific antisera – clumping
b. Widal Test – tube dilution agglutination
o Patients serum + known antigen
o Serum obtained twice 7-10 days apart
o Significant reaction: rising titer or high titer
(1:160) 2nd-3rd week

Interpretation:
o if predominantly O – active or acute infection
o if predominantly H – past infection or
vaccination
o If predominantly V – carrier state
What makes Salmonellae pathogenic?
c. Typhidot – the 60 minute test
1. rarely, formation of exotoxins. But all contain endotoxins
o Dot EIA for the rapid detection of specific
2. ECA or kunin antigen – inhibits phagocytosis and decreases
IgM and IgG antibodies
their susceptibility to bactericidal action of serum
3. fimbriae – mediate attachment of bacteria to intestinal
mucosa, but have no pathogenic function extraintestinally
4. tendency to plasmid-mediated multiple drug resistance
5. tendency to a carrier state especially in highly nutritive Result Clinical Interpretation
environment of gall bladder 1. IgM positive only Acute typhoid fever
6. Past infection and vaccination confer only partial or 2. IgM and IgG positive Acute typhoid fever (in the middle stage
temporary active immunity of infection)
7. Enterotoxin similar to Escherichia is produced by Salmonella 3. IgG positive Previous infection (not due to typhoid);
typhimurium, causative agent of food poisoning. or relapse or reinfection
4. IgG and IgG Probably not typhoid
Epidemiology
Sources of infection – food and drink contaminated with
salmonellae Treatment
a. water 1. Enterocolits
b. milk and other dairy products (ice cream, cheese, a. fluid and electrolyte replacement
custard) b. antibiotic treatment not needed
c. shellfish o may prolong excretion of the organisms
d. dried or frozen eggs o increase the frequency of carrier state
e. meats and meat products
o select mutants resistant to the antibiotic
f. recreational drugs
2. Enteric fevers and bacteremias
g. animal dyes
a. antimicrobial treatment
h. household pets
o ceftriaxone or ciproflaxin – treatment of
Laboratory Diagnosis choice
Specimens o chloramphenicol
o trimethoprim-sulfamethoxazole
• Blood – culture + 1st week o ampicillin
• Bone marrow – culture b. carriers
ampicillin alone
• Urine – culture + after 2nd week
o
o cholecystectomy combined with drug
• Stool – culture treatment
o Typhoid fever + 2nd week or 3rd week
Prevention and Control
o Enterocolitis + 1st week
1. Through cooking of infected poultry, meats and eggs
• Serology 2. carriers should not be allowed to work as food handlers
3. sanitary measures to prevent contamination of food and
1. Culture water
a. enrichment cultures – selenite F broth or 4. immunization
tetrathionate broth a. two injections of acetone killed bacterials
b. differential media suspensions of Salmonella typhi followed by booster
o MacConkey – colorless colonies, injection
nonlactose fermenter b. oral administration of a live avirulent mutant strain
c. selective of Salmonella tyhi
o Salmonella Shigella agar – colorless colonies,
sometimes with black colonies due to H2S
production (black center with clear periphery)
-fin-
o Hektoen enteric agar – green with black
centers (lactose negative; H2S positive)
o Bismuth sulfite agar – rapid detection of lapit na quiz 3…aral kayo ng mabuit!..
Salmonella typhi producing jet black colonies,
thanks to marc ostrea and coycoy
congrats to Terps esp to ate osang! Galing niyo!

Hi c3!!!

audsmartinez@gmail.com
ustmedc3@yahoogroups.com