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Examination of Faecal[1]

The document outlines the examination of fecal specimens, detailing possible pathogenic bacteria, normal gut flora, specimen collection, transport, and laboratory examination procedures. It specifies criteria for specimen rejection and describes various culture media used for identifying specific pathogens. Additionally, it includes guidelines for biochemical tests and antimicrobial susceptibility testing.

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2 views20 pages

Examination of Faecal[1]

The document outlines the examination of fecal specimens, detailing possible pathogenic bacteria, normal gut flora, specimen collection, transport, and laboratory examination procedures. It specifies criteria for specimen rejection and describes various culture media used for identifying specific pathogens. Additionally, it includes guidelines for biochemical tests and antimicrobial susceptibility testing.

Uploaded by

m7hghjfc
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Examination of faecal

specimens
By: Mazen Mohammed Qaid
Possible Pathogenic Bacteria
• Gram positive • Gram negative
1. Shigella species
1. Clostridium perfringens
types A and C 2. Salmonella serovars
3. Campylobacter species
2. Clostridium difficile 4. Yersinia enterocolitica
3. Bacillus cereus (toxin) 5. Escherichia coli (ETEC, EIEC,
EPEC, EHEC, EAEC)
4. Staphylococcus aureus
(toxin) 6. Vibrio cholerae 01, 0139
7. Other Vibrio species
8. Aeromonas species

Also Mycobacterium tuberculosis


Commensals
• The normal microbial flora of GIT include:–
• Coliform bacilli and species of Proteus
• Pseudomonas,
• Clostridium,
• Bacteroides,
• Enterococcus,
• Lactobacilli, and
• Mycoplasma.
• Also Candida species and a variety of protozoa and
viruses.
Types of specimen

• Stool

• Rectal swab
Collection and transport of faeces
• Collected during the acute stage of diarrhoea prior
to initiation antibiotics.
• Stool collected in clean, dry, disinfectant-free
wide-necked, leak-proof container.
• Deliver to the laboratory within 1 hour.
• If a delay longer than 1 hour, collect the specimen
in Cary-Blair medium.
Salmonella serovars, Shigella,Vibrio and Yersinia species survive well in
Cary-Blair medium for up to 48 hours
Campylobacter for upto 6 hours.
Continue,
When cholera is suspected:
• Transfer about 1 ml of specimen into 10 ml of
sterile alkaline peptone water and label.
• The specimen should reach the Microbiology
Laboratory within 8 hours of collection.
Criteria of specimen rejection

• Specimen contaminated with urine, residual


soap, or disinfectants.
• Leaking transport containers;
• diapers; dry specimens;
• Specimens submitted in fixative or additives.
Laboratory examination

Day 1
1. Describe the appearance of the specimen:-
•Color •Consistency •Presence
of blood, mucus or pus
2. Culture the specimen
When the specimen is formed or semi-formed, make a thick suspension of
it in about 1 ml of sterile peptone water and culturing on:
 Selenite
F broth (After Overnight incubation do subculture onto fresh XLD or SSA)

XLD or SSA
ADDITIONAL
• APW and TCBS agar when cholera is suspected
1. Inoculate several loopfuls of specimen in APW
incubate at 35–37 °C for 5–8 hours.
2. Subculture several loopfuls of the APW (taken from the surface) on (TCBS) agar.
Incubate at 35–37 °C overnight.
• Sorbitol MacConkey agar, when an outbreak of E. coli 0157
is suspected
• Inoculate a loopful of specimen on sorbitol MacConkey agar.
Incubate 35–37 °C overnight.
• MacConkey agar
• Examine the specimen microscopically
-Saline and eosin preparations
Day 2

1. Examine and report the cultures.(see next slide)


2. Perform gram stain.
3. perform Biochemical tests.(Catalase,
Coagulase,Oxidase,Urease,KIA, SIM,Indole test,...)
4. Antimicrobial susceptibility test(Using Muller
Hinton agar)
On XLD agar culture
Look for colonies that could be Shigella or Salmonella.
-Salmonella produce red colonies with black centers.
-Shigella produce red colonies without black centers.
-Proteus produce red colonies with black centers.
-Coliform produce yellow colonies.
On MacConkey agar,
-Shigella, and salmonellae and other NLF ,
produce colorless colonies.
-E. coli and other LF produce pink colonies.
TCBS agar culture
-V. cholerae is sucrose fermenting produces yellow colonies
with a yellow color in the medium.
• Note: With prolonged incubation (48 h or more) the colonies may
become green.
-Vibrio parahaemolyticus is non-sucrose fermenting and therefore
produces green-blue colonies.
Day 3

1. See the biochemical tests results and record the


name of bacteria

2. Measure the zone of inhibition of antibiotics


and record the report
• Cary-Blair medium • Selenite F broth
• Transport medium • An enrichment medium
• These media are used • Allows the growth of specific
when specimens cannot bacterial species, however,
enrichment media are
be cultured soon after supplemented with a reagent
collection. Transport that allows, rather than
media prevent drying of inhibits, the growth of a
the sample and inhibit particular species.
the growth of • Example: Selenite F broth
undesirable bacteria. (selenite is an inhibitor for
• Example: Cary-Blair coliforms and beneficial for
agar, Amies Medium. the recovery of Salmonella
species), Thayer Martin agar.

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