Guideline – SOP
Microbial Analysis Report of Water
Date of Sampling :
Sampled By
:
Date of Completion :
Method Used
: Membrane filtration Method/ Pour plate
Incubator ID:Sample Location &Sample Point ID No.
………..…... …….….………
…….….…... …………..
Date of Testing
Analyzed By
…….….……
…….…….…
…………………
……………
:
:
……………...…
….………
Test
Media/ Incubatio Incubation Incubati Count
Count
Count
Count
Count
lot No.
n
Started on
Condition Date/sign Complet
ed
Date/sig
n
Total
R2A 30-35°c
Plate Plate Plate
Plate Plate Plate Plate
Plate
Plate
Plate
aerob ………..
for
I
II
I
II
I
II
I
II
I
II
ic
SCDA 5 days
=
=
=
=
=
=
=
=
=
=
micro ………..
Mean
Mean
Mean
Mean
Mean
bial
=
=
=
=
=
count
+ve
Contr
ol
-ve
Con
trol
+ve
Contr
ol
-ve
Con
trol
Test for specific Microorganism
Test
Media
Enric
hmen
t for
Patho
SCDM
………
Incubati Incubatio Incubati Observation
on
n Started on
Conditio Date/sig Complet
n
n
ed
Date/sig
n
30Growth
35°c
Observed/
18Not observed
24hrs
Observation
Growth
Observed/ Not
observed
1
Observation Observation
Observation
Growth
Observed/
Not
observed
Growth
Observed/
Not observed
Growth
Observed/ Not
observed
Guideline – SOP
Microbial Analysis Report of Water
gen
E.coli
MCB
42Growth
Growth
Growth
Growth
Growth
44°c
Observed/
Observed/ Not
Observed/
Observed/ Not
Observed/
……… 24Not observed
observed
Not
observed
Not observed
48hrs
observed
MCA
30Growth
Growth
Growth
Growth
Growth
35°c
Observed/
Observed/ Not
Observed/
Observed/ Not
Observed/
………
18Not observed
observed
Not
observed
Not observed
72hrs
observed
Identification Test: Aseptically prepare culture suspension from the suspected colonies which are appears on MacConkey agar in a
sterilized tube. Add 0.1 ml of the prepared suspension to the tube containing 10 ml of peptone water; incubate the tube at 30°C to 35°C for
18 - 24 hours. From incubated peptone water tube perform Indole test as follow:
Add 0.5 ml of Kovac’s reagent to peptone water tube; allow to stand for one minute. The presence of cherry red colour ring along the side of
test tube indicates the positive test (the presence of Escherichia coli). Presence of Escherichia coli shall be confirmed by Gram staining
(Gram-ve rods).
Remarks:-
Test
s.
aureus
Media
MSA
Incubati Incubati Incubati
on
on
on
Conditio Started Complet
n
Date/si ed
gn
Date/sig
n
3035°c
………
Observation
Observation
Growth
Observed/
Not observed
Growth
Observed/ Not
observed
18-
2
Observation Observation
Growth
Observed/
Not observed
Growth
Observed/
Not observed
Observation
Growth
Observed/
Not observed
+ve
Contr
ol
-ve
Con
trol
Guideline – SOP
Microbial Analysis Report of Water
72hrs
Identification Test: With the aid of an inoculating loop, individually transfer suspected colonies to separate tubes containing 0.5 ml of
mammalian plasma (preferably rabbit or horse). Incubate in a water-bath at 37°C for 3 to 24 hours, in parallel with positive control using
known strain of Staphylococcus aureus and negative control using Plasma alone. Examine after 3 hours and at suitable intervals thereafter
for the presence of coagulation
If coagulation in any degree is observed, the presence of Staphylococcus aureus is indicated
Remarks:Test
Incubati Incubati Incubati Observation
Observation
Observation Observation
Observation +ve
on
on
on
Contr
Conditio Started Complet
ol
n
Date/si ed
gn
Date/sig
n
p.
CA
30Growth
Growth
Growth
Growth
Growth
aerugin
35°c
Observed/
Observed/ Not
Observed/
Observed/ Not
Observed/
osa
………
18Not observed
observed
Not observed
observed
Not
72hrs
observed
Identification Test: If growth of suspected colonies occurs carry out Oxidase test.
Transfer suspected colonies using inoculating loop on to Oxidase disc; Standard result: Development of purple colour in 5 to 10 seconds
Remarks:Test
Salmo
nella
specie
s
Media
Media Incubation Incubat Incubati Observation
Condition
ion on
Started Complet
Date/si ed
gn
Date/sig
n
RVS
30-35°c
Growth
18Observed/
………
24hrs
Not observed
Observation
Observation
Growth
Observed/ Not
observed
Growth
Observed/ Not
observed
3
Observation
Growth
Observed/ Not
observed
Observation
Growth
Observed/
Not
observed
+ve
Con
trol
-ve
Con
trol
-ve
Con
trol
Guideline – SOP
Microbial Analysis Report of Water
XLDA
30-35°c
1848hrs
Growth
Observed/
Not observed
Growth
Observed/ Not
observed
Growth
Observed/ Not
observed
Growth
Observed/ Not
observed
Growth
Observed/
……...
Not
observed
Identification: If colonies found transfer a few of suspect colonies by first Sub culturing the slope of slant of TSI agar and then stabbing with
inoculating straight wire well in the butt; Media lot No. ____________ , Incubate at 300C to 350C for 24 to 48 hrs; Incubator ID
No._______________.
Remarks:Test
Media Incubation Incubati Incuba Observation
Observation
Observation Observation
Observation
+ve -ve
Condition
on
tion
Con Co
Started Compl
trol
ntr
Date/si eted
ol
gn
Date/s
ign
GN
30-35°c
Growth
Growth
Growth
Growth
Growth
Broth
24Observed/ Not
Observed/
Observed/ Not Observed/ Not
Observed/
48hrs
observed
Not
observed
observed
Not observed
Shigella
……
observed
…
XLDA 30-35°c
Growth
Growth
Growth
Growth
Growth
24Observed/ Not
Observed/
Observed/ Not Observed/ Not
Observed/
……...
48hrs
observed
Not
observed
observed
Not observed
observed
Identification Test: If growth of suspected colonies occurs Subculture the growth on Modified SS agar Media lot No. ____________. If
the characteristic growth than confirms the presence of Shigella in the sample, sample fails the test for absence of Shigella.
Remarks:Conclusion of analysis
Complies/
does not
complies
Complies/
does not
complies
4
Complies/
does not
complies
Complies/ does
not complies
Complies/
does not
complies