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Water surveillance
Dr. Mayuri Bhise
Assistant professor,Department of
Microbiology, AIIMS Rajkot
Contamination with enteric
pathogens
Indicator organism
 Coliforms ( other than
E.coli)
 Fecal (thermo tolerant)
E.coli
 Fecal streptococci
 Sulphite reducing
clostridia
 Pseudomonas
aeruginosa
 Bacteriophages specific
for E.coli
Collection and transport
of water sample
• Container- Screw capped
borosilicate glass or
plastic bottles (500 ml)
• Avoid contamination
• Volume- At least 150-200
ml of water
Microbiological testing methods for
water analysis
Test
Multiple-tube method
Membrane filtration
method
• F o r t h e e s t i m a t i o n o f
presumptive coliform count
• Extensively used for drinking
water analysis
• For highly turbid samples
 For testing dialysis water
 For testing clean water
 For testing large volume of water
 When bacterial count in water is
expected to be low
•Presence /absence
method
•Medium containing
sodium azide
50ml 10ml 1ml 0.1ml 0.01 0.001 0.000
ml ml 1ml
DS DS SS SS SS SS SS
Sample type
MacConkey purple broth (DS- double strength; SS-Single strength)
Unpolluted 1 5
water
Polluted - 5 5 5
water
Heavily - - 5 5 5 5 5
polluted
water
MULTIPLE TUBE METHOD
(Unpolluted)
McCrady statistical table
For Unpolluted
water
For polluted
water
Microbiological limits - drinking
water
Class Grading Coliform Count
/
100 mL (MPN)
E. coli count /
100 mL
Class-1 Excellent 0 0
Class-2 Satisfactory 1-3 0
Class-3 Suspicious 4-10 0
Class-4 Unsatisfactory >10 More than 0
Differential coliform count
(Eijkman test)
• To confirm that the coliform bacilli detected in
presumptive test are faecal E.coli
• Inoculate the positive tubes into brilliant green
bile broth and tryptone water. Incubate at 44°C
for 24 hours
• Positive for indole, acid, and gas production
confirms thermotolerant E. coli
Membrane filtration method
• Filtration of known volume
of water sample on
cellulose filter paper and
incubate it on suitable
culture media.
• Pore size = 0.2-0.45 μm
• Media : Membrane lauryl
sulphate broth
Count visually
identifiable colonies
× 100
× 100
THANK YOU

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Dr Mayuri -Water surveillance (Practical).pptx

  • 1. Water surveillance Dr. Mayuri Bhise Assistant professor,Department of Microbiology, AIIMS Rajkot
  • 2. Contamination with enteric pathogens Indicator organism  Coliforms ( other than E.coli)  Fecal (thermo tolerant) E.coli  Fecal streptococci  Sulphite reducing clostridia  Pseudomonas aeruginosa  Bacteriophages specific for E.coli Collection and transport of water sample • Container- Screw capped borosilicate glass or plastic bottles (500 ml) • Avoid contamination • Volume- At least 150-200 ml of water
  • 3. Microbiological testing methods for water analysis Test Multiple-tube method Membrane filtration method • F o r t h e e s t i m a t i o n o f presumptive coliform count • Extensively used for drinking water analysis • For highly turbid samples  For testing dialysis water  For testing clean water  For testing large volume of water  When bacterial count in water is expected to be low •Presence /absence method •Medium containing sodium azide
  • 4. 50ml 10ml 1ml 0.1ml 0.01 0.001 0.000 ml ml 1ml DS DS SS SS SS SS SS Sample type MacConkey purple broth (DS- double strength; SS-Single strength) Unpolluted 1 5 water Polluted - 5 5 5 water Heavily - - 5 5 5 5 5 polluted water MULTIPLE TUBE METHOD (Unpolluted)
  • 5. McCrady statistical table For Unpolluted water For polluted water
  • 6. Microbiological limits - drinking water Class Grading Coliform Count / 100 mL (MPN) E. coli count / 100 mL Class-1 Excellent 0 0 Class-2 Satisfactory 1-3 0 Class-3 Suspicious 4-10 0 Class-4 Unsatisfactory >10 More than 0
  • 7. Differential coliform count (Eijkman test) • To confirm that the coliform bacilli detected in presumptive test are faecal E.coli • Inoculate the positive tubes into brilliant green bile broth and tryptone water. Incubate at 44°C for 24 hours • Positive for indole, acid, and gas production confirms thermotolerant E. coli
  • 8. Membrane filtration method • Filtration of known volume of water sample on cellulose filter paper and incubate it on suitable culture media. • Pore size = 0.2-0.45 μm • Media : Membrane lauryl sulphate broth Count visually identifiable colonies