EP066 MM2 Instruction Leaflet 130318

For Detection
For andand
ForDetection
Detection Enumeration
andEnumeration of Contaminating
Enumeration Microbes
ofofContaminating
ContaminatingMicrobes
Microbes
in Fuels, Lubricants
ininFuels,
Fuels, andand
Lubricants
Lubricants Water
andWater
Water
Instructions for Use

Instructions
Instructionsfor
forUse
Use

Important NoticeNotice
Important
Important
Important Notice
Notice
Please
Please read the
Please read
Please
read thethe
read
the Instructions
Instructions for
Instructions for Use
Instructions
for Use
Use carefully carefully
before
for Use
carefully before
testing
carefully testing
samples.
before
before
testing samples.
testing
samples.
samples.
The
The test kit
The testtest
should
The
test kit
kit should
only
kit be
should
shouldonlyonly
used
only bepart
as
be used
be
used asan
ofas
used part
part of an
part
of an
asinvestigatory
of an investigatory
process into
investigatory
investigatory process
process
process intointo
contamination
into contamination
contamination
contamination in petroleum
in petroleum
in petroleum
inproducts
petroleum products
and
products andand
associated
products and associated
water.
associated water.
The
associatedwater. TheThe
water.
The
testtest
test kit must
test bekit used
kit must
must be be
kit must
be used
strictly
used in strictly
accordance
used
strictly in accordance
strictly
in accordance
with thesewith
in accordance with these
instructions
with
thesetheseinstructions
and/or
instructions and/or
Standard
instructions
and/or and/or Standard
Test
Standard
Standard TestTest
Method
Test Method
IP613
Method
Method IP613
or ASTM
IP613 or ASTM
D7978
IP613or ASTM D7978
or D7978
or ASTM other D7978 or other
other
instructions
or instructions
or other
instructions
instructions
authorised authorised
byauthorised
ECHA by
authorised by ECHA
Microbiology
ECHA
by ECHA Microbiology
Microbiology
Microbiology Ltd.Ltd.
Ltd. The results
Ltd. The
only
The results
The relate
results only
results
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toonly
the
relate to the
relate
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toofthe
sample
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portion oftested
of sample
sample
of sampletested
and not
tested andandnotnot
necessarily
testedand not necessarily
tonecessarily to other
other petroleum
necessarily to other
to other petroleum
product
petroleum
petroleum product
product
product
in the
in the system.
in the system.
inAlthough
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Although
Although guidance
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ECHAECHAMicrobiology
Ltd does not
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Microbiology Ltd Ltd
Ltd does
accept
does notany
does
not accept
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accept
accept any
liability
any forliability
any any
liability for any
decision
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for any
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decision
or assessment
or assessment
taken
taken or made
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of the
results
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obtained.
obtained.
ECHA, Sig Test(s), Sig Sulphide and MicrobMonitor are registered trademarks of ECHA Microbiology Ltd. in the UK, and are registered and
unregistered trademarks in other select regions globally.
ECHA Microbiology
ECHAECHA Ltd, UnitsLtd,
Microbiology
Microbiology 22 Ltd,
&Units
23 Units
Willowbrook
22 &2223 23 Technology
& Willowbrook
Willowbrook Park,
Technology Llandogo
Technology Park, Road,
Park,
LlandogoSt Road,
Mellons,
Llandogo St Cardiff,
Road, Mellons, UK,
St Mellons, CF3 0EF
Cardiff,
Cardiff,
UK, UK,
CF3CF3
0EF 0EF
Tel: + 44 (0)
Tel:2920
+ 44+365930
Tel: (0)
44 2920
(0) 2920
365930 Fax: +44 (0)
365930 2920
Fax: +44361195
Fax: +44
(0) 2920
(0) 2920
361195 [email protected]
361195 [email protected]
[email protected]
ECHA Microbiology Ltd, Units 22 & 23 Willowbrook Technology Park, Llandogo Road, St Mellons, Cardiff, UK, CF3 0EF
Tel: + 44 (0) 2920 365930 Fax: +44 (0) 2920 361195 [email protected]
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WHAT IS THE MICROBMONITOR2
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contamination in aviation, marine and ground fuels and is NATO Codified (6640-99-834-3573).
STORAGE
MicrobMonitor2
STORAGEprovides AND
AND SHELF a total
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LIFE
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box. Store tests between 2 to 22°C, in the dark. The expiry date applies only toto product
product
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ALSO
ALSO AVAILABLE
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The number of colonies formed is a direct estimate of the number of viable microbial particles (CFU) present in the sample. The
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MICROBMONITOR 2 2® by reference to the Test Results Chart (page 9). The volume tested can
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is considered in relation can
to the volume of sample added to the test andprocedure:
expressed as CFU/litre of fuel, or CFU/ml of oil or
The
The following
following items
items can assist
assist in
in performing the MicrobMonitor2
performing the MicrobMonitor2 test
test procedure:
water associated with fuel.
x
x MicrobMonitor
MicrobMonitor Sampling KitKit (ECHA16/SB)
(ECHA16/SB) -- A A sterile, polypropylene
polypropylene bottle for
for sampling
sampling fuel
fuel systems and
and which
MicrobMonitor2 is availableSampling
in packs of 5 and 50 tests, eachsterile,
44 containing everything bottle
that is required to performsystems
the test: which
enables
enables easy
easy separation
separation of of water
water phase
phase from
from fuel
fuel phase;
phase; includes
includes alcohol
alcohol wipes
wipes for
for decontaminating
decontaminating sample
sample
points.
points.
x MicrobMonitor2 test bottles
x
x Sterile syringes*
x Incubator
Incubator (ECHA14/IN)
(ECHA14/IN) -- enables
enables optimal,
optimal, consistent
consistent incubation
incubation temperature
temperature ofof MicrobMonitor2
MicrobMonitor2 tests;
tests; holds
holds 12
12
x Sterile loop dispensers*
MicrobMonitor2
MicrobMonitor2 test
test bottles;
bottles; available
available battery
battery or
or mains
mains (110/230
(110/230 Volt)
Volt) operated.
operated.
x Bottle labels
x
x
x MicrobMonitor
MicrobMonitor
This Bumpers
Bumpers (ECHA16/TB)
instruction leaflet. (ECHA16/TB) -- soft,
soft, rubber-like,
rubber-like, hemispheres
hemispheres which
which can
can be
be stuck
stuck to
to a
a table
table top
top and
and enable
enable
easy breaking
easy breaking and
and shaking
shaking of MicrobMonitor2 gel.
of MicrobMonitor2 gel.
50 pack is available with or without sterile syringes and loop dispensers.
* The 44
x
x Chlorine
Chlorine Release
Release tablets
tablets (ECHA21/CP)
(ECHA21/CP) -- to disinfect MicrobMonitor2
to disinfect MicrobMonitor2 test
test bottles
bottles showing
showing microbial
microbial growth
growth for
for easy
easy
MicrobMonitor2 has been developed and patented by ECHA Microbiology Ltd. and it is marketed worldwide by
disposal.
disposal.
ECHA and its distributors.
x
x Sterile
Sterile swabs
swabs (ECHA15/SW)
(ECHA15/SW) -- swabs
swabs for
for sampling
sampling surface
surface contamination
contamination (e.g.
(e.g. tank
tank surfaces,
surfaces, filter
filter cartridges
cartridges etc.).
etc.).
WHAT
x IS THE MICROBMONITOR
Long, 22 TEST USED FOR?enable water to be removed from the bottom of samples.
® ®
x Long, sterile
sterile bulb
bulb pipettes
pipettes (ECHA07/PP/EL23)
(ECHA07/PP/EL23) -- to
to enable water to be removed from the bottom of samples.
The MicrobMonitor2 is used to test for microbial contamination in fuel and oil products, and to monitor microbial growth in
storage tanks and supply facilities. MicrobMonitor2 can be used as a routine monitoring tool, for providing assurances
Other
Other Test Kits
Kits &
Testquality,
&forAncillary
Ancillary Items
Items
about product investigation of incidents and for confirming effectiveness of measures taken to prevent or
remediate microbial growth and contamination.
x
x SigTestsΠ:
SigTests®: Sig
SigTests®:
SigTestsΠ: Sig Sulphide
Sulphide (ECHA02/SS)
(ECHA02/SS) — — simple
simple test
test kit
kit for
for Sulphate
Sulphate Reducing
Reducing Bacteria
Bacteria (SRB)
(SRB) which
which can
can cause
cause serious
serious
corrosion
This instruction leaflet and
corrosion sulphide
provides
and spoilage
general
sulphide in
in fuel,
fuel, oil
instructions
spoilage forand
oil water
using
and water systems.
systems. Can
Can be
MicrobMonitor2. used
Further
be on-site
on-site or
used technicalor in the
the laboratory.
instruction
in on the use of
laboratory.
MicrobMonitor2 in specific applications is provided in the technical application leaflets listed below (free to download
from xwww.echamicrobiology.com).
x Biocide
Biocide Rapide
Rapide (ECHA01/BR) These—
(ECHA01/BR) — simple
leaflets rapid
simpleinclude test
test kit
rapid advice kit for assessing
assessing presence
on sampling,
for and
and approximate
sample preparation,
presence approximate concentration
interpretation of test of
concentration of
commonly used
results and appropriate
commonly used fuelwhen
actions
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and oil biocides.
oil biocides. is detected:
x x
x EP096Any
Howlevel
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level MicrobMonitor2
and bottom
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samplingTest Results(ECHA23)
devices —
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safeIATA from
Guidelines
sampling fromforfuel and
Aircraft
fuel oil
oil storage
andDrain Samples.tanks
storage tanks in
in
compliance with
compliance with industry
industry standards.
standards.
x EP119 Routine Monitoring of Aviation Fuels in Supply and Distribution Facilities, Airport Depots and Into-plane
x For
For our
x Operations full
ourwith MicrobMonitor2.
full range
range of
of test
test kits
kits and
and ancillaries
ancillaries see
see www.echamicrobiology.com.
www.echamicrobiology.com.
x Technical
Technical support
support and
and services
EP132 Routine Monitoring of Diesel Fuel Tanks and Distribution Systems with MicrobMonitor2.
services
x EP166
ECHA
ECHA Routine Monitoring
Microbiology
Microbiology Ltd. of Marine
Ltd. provides
provides a fullDiesel
a full on Ships
technical
technical andto
service
service toOffshore
support Installations
support our productwith
our product MicrobMonitor2
range.
range. ECHA
ECHA also .
also offers
offers laboratory
laboratory testing,
testing,
training courses,
training courses, consultancy
consultancy and
and on-site
on-site attendance
attendance for for audit
audit and
and investigation
investigation ofof microbial
contamination and and corrosion
corrosion
x associated with
with petroleum
EP157 Technical
associated Assistance
petroleum products
Reading
products and
and petroleum,
Results marine,
marine, aviation
of MicrobMonitor2.
petroleum, aviation and
and other
other industrial
industrial facilities.
facilities.
2
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PREPARATION
PREPARATION FOR
FOR TESTING
TESTING
Sampling
Sampling considerations
considerations
CFU/ litre or CFU/ ml
Sampling
Sampling should
should be
be conducted
conducted in in a
a clean
clean and
and consistent
consistent manner
manner which
which prevents
prevents introduction
introduction of of background
background contamination
Contamination in
contamination
into the
the sample.
sample. Once fuel ®
samples have been taken, any microbes present will tend to slowly die
die and
and it
it is
is important
important to
MICROBMONITOR
into Once fuel 2
samples have been taken, any microbes present will tend to slowly to
Microbial
test
test samples
samples as
as soon
soon as
as possible.
possible. IfIf the
the test
test is
is carried
carried out
out more
more than
than 24
24 h
h after
after sampling,
sampling, it
it is
is possible
possible that
that the
the results
results will
will not
not
sample
reflect the
reflect the viable
viable microbial
microbial content
content present
present atat the
the time
time of
of sampling.
sampling. To
To avoid
avoid this,
this, part
part or
or all
all of
of the MicrobMonitor2 test
the MicrobMonitor2 test
procedure
procedure can
can be
be conducted
conducted at at the
the sampling
sampling location.
location.
WHAT IS THE MICROBMONITOR®2 TEST?
Sample
Sample type
type and
and recommended
recommended test
test volumes
volumes
MicrobMonitor2 is a simple and easy to use test kit which enables the quantitative assessment of the viable microbial
MicrobMonitor2 can
MicrobMonitor2 can be be used
used toto test
test aa wide
wide variety
variety ofof petroleum
petroleum products
products andand associated
associated water.
water. TheThe volume
volume of of sample
sample
content in petroleum products including liquid fuels, oils and associated water. MicrobMonitor2 enables on-site or
which
which should be
shouldtesting
laboratory tested
be tested will depend
will depend
in accordance on the
onASTM
with the andtype of sample.
type IPofStandard Refer
sample. Methods to
Refer toASTMthe table
the table below
D7978below to determine
to determine
and IP613 the appropriate
the appropriate
- Determination of the viable
Day _______
volume
volume of
of sample
sample to
to test
test and
and which
which measuring
measuring device
device to to use.
use.
Counted
Colonies
aerobic microbial content of fuels and associated water - Thixotropic Gel Culture Method. MicrobMonitor2 is
microbial contamination in aircraft fuel tanks and is listed in the Aircraft Maintenance Manuals of Airbus, Boeing, BAE
Table
Table 11
Systems and other leading aircraft manufacturers. The test is used by a wide range of military services for monitoring
contamination in aviation, marine and ground fuels and is NATO Codified (6640-99-834-3573). Recommended
Recommended test test Measuring
Measuring
Sample
Sample
Sample
Sample volume
volume
volume
volume device
device
device
device
MicrobMonitor2 provides a total count of microbial colony forming units (CFU). CFUs are the conventional unit of measure
2 test results
Counted
Colonies
Day 4
Aviation kerosene
kerosene
for microbial
Aviation contamination used in a wide range of industries. MicrobMonitor20.5
kerosene
kerosene 0.5 ml
ml
detects
ml
ml the important bacteria, yeasts and
moulds which can contaminate petroleum products including Hormoconis resinae, Aspergillus, Candida and Pseudomonas
Other middle distillate
species.
Other middle distillate fuels
fuels (e.g.
(e.g. diesel,
diesel, marine
marine gas gas oil,
oil, heating
heating oil),
oil), biofuels
biofuels &&
0.25 ml
0.25 ml
ml
ml Syringe
Syringe
Syringe
Syringe
gasoline
gasoline
gasoline
gasoline
The MicrobMonitor2 test consists of a screw-capped bottle containing a thixotropic nutrient gel. A sample is added to the gel
usingphase
a sterile plastic loop ortank
syringe (provided). The bottle is shaken and the 0.1 gel
ml liquefies and the sample, and any
Counted
Water phase from aircraft fuel tank drains

drains ml
ml
Colonies
Water from aircraft fuel drains

drains 0.1 ml
Day 3
microorganisms in it are dispersed. The gel is re-set into a flat layer and the bottle is incubated in the dark. The gel
® ®
Heavy
Heavy andcomponents
and
contains residual fuels
residual fuels
fuels
fuels which sustain the growth of viable microorganisms and 0.01the
0.01 ml
ml
ml
ml sample itself contributes additional
MICROBMONITOR2
nutrients.
Lubricating, Viable
hydraulic microorganisms
and other oilsin the sample grow into visible spots known as
0.01 “colonies”, and a reactive compound
ml
Lubricating, hydraulic and other oils
oils ml Loop
Loop
changes the colour of these oils
colonies to red or purple so that they can be easily0.01 ml
ml
seen. Loop
Loop
Water phase from fuel storage
Water phase from fuel storage tanks tanks
tanks
tanks 0.01 ml
ml
0.01 ml
ml
counted
Colonies
The number of colonies formed is a direct estimate of the number of viable microbial particles (CFU) present in the sample. The
Day 2
actual number of colonies is counted or is estimated by reference to the Test Results Chart (page 9). The volume tested can
The
bevolume
The 0.01 ml of
volume of sample
sample
(using tested
loopcan
tested
a sterile can be
be changed
dispenser)changed in
in order
or between 0.1 to
order ml increase
to increase
and 0.5 ml or decrease
or(using
decrease the detection
thesyringe).
a sterile detectionThelevel
level of
of the
number oftest.
the test. However,
However,
colonies formed
testing
testing volumes
volumes in
is considered greater
greater
relation than
than those
to those recommended
recommended
the volume above
of sampleabove added may
may in some
in some
to the circumstances
circumstances
test and expressed leadlead to underestimation
to underestimation
as CFU/litre of
of microbial
of fuel, or CFU/ml microbial
of oil or
numbers.
numbers.
Determining whether to
to test fuel/oil
of 5 or
andwater phase
counted
Colonies
Determining whether
MicrobMonitor2 is availabletest fuel/oil
in packs or water phase
44 tests, each containing everything that is required to perform the test:
Day 1
Depending
Dependingx on
on the
the system
system sampled
MicrobMonitor2 sampled and
and the
test bottles the sampling
sampling location,
location, samples
samples may
may contain
contain fuel/oil
fuel/oil and/or
and/or free
free water
water phase.
phase.
MicrobMonitor2
MicrobMonitor2 can
can be
be used
used to
to test
test either
either fuel/oil
fuel/oil or
or associated
associated water.
water. Water
Water phase
phase will
will usually
usually contain
contain far
far more
more
x Sterile syringes*
microorganisms than than are
are present
present inin fuel
fuel or
or oil
oil phase
microorganisms phase andand therefore
therefore a a smaller
smaller volume
volume isis usually
usually tested.
tested. Because
Because free
free water
water
phase
phase may
xmay not
Sterile loop dispensers*
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always bebe recovered
recovered in in samples,
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ensure consistency
consistency when
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regular microbiological
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
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microbiological 0.5/0.25/0.1/0.01/other__
x itit Bottle
monitoring,
monitoring, is usually
is usually recommended that
labels
recommended that fuel
fuel or
or oil
oil phase
phase from
from above
above any
any free
free water
water is
is tested.
tested. However,
However, inin some
some cases
cases
it
it is
is recommended
x
recommended to
to additionally
additionally
This instruction leaflet. test
test water
water phase
phase (e.g.
(e.g. samples
samples from
from aircraft
aircraft fuel
fuel tanks,
tanks, and
and when
when investigating
investigating
suspected microbial
suspected microbial growth
growth in
in facilities).
facilities). Further
Further information
information is is provided
provided in
in the
the relevant
relevant technical
technical guidance
guidance documents
documents
listed
listed on
* Theon page
44 page 2.
pack is2.
available with or without sterile syringes and loop dispensers.
Sample & test details
To determine
determine ifif the
ToMicrobMonitor2 sample
thehas been
sample contains free
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contains andhold
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patented
hold up to the
to by light
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visually examine
Microbiology
and Ltd. it.
examine It
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access to to the
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provided, it
it may
may be be
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necessary to transfer
transfer some
some sample,
sample, after
after shaking,
shaking, to
to a
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smaller clean
clean container.
container.
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Preparing the
WHAT IS THE MICROBMONITOR2
the MICROBMONITOR 2® test
® ®
2 TESTbottles
test bottles
USED FOR?
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If The MicrobMonitor2
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the MicrobMonitor2 test bottles
bottles
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stored
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contamination in fuelthem
allow them to equilibrate
to products,
and oil to
to ambient
equilibrateand ambient
to monitortemperature
temperature before
before in
microbial growth
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)

Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Location
Location
Location
Location
Location
Incubation Temp. (°C)

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
they
they are used.
are used.
storage Remove
Remove
tanks and supplythe seal from
thefacilities. the cap
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the cap of thecan bottle
bottle before
be before using
used asusing each MicrobMonitor2
MicrobMonitor2
each monitoring
a routine test.
tool, test. Avoid prolonged
Avoid prolonged
for providing assurances
exposure
exposure of the MicrobMonitor2
of the
about product MicrobMonitor2 test
test bottles
quality, for investigation bottles to
to direct sunlight
direct and
of incidents for or
sunlight or other
other bright
confirming bright light
light during
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during
effectiveness preparation,
measures takenincubation
incubation and
and or
to prevent
examination
examination of
of tests.
tests. growth and contamination.
remediate microbial
Cleanliness during
This instruction
Cleanliness leaflettesting
during provides
testing general instructions for using MicrobMonitor2. Further technical instruction on the use of
from www.echamicrobiology.com).
MicrobMonitor2 These leaflets include advice on sampling, sample preparation, interpretation of test
MicrobMonitor2 tests
tests can
can be
be conducted
conducted on-site,
on-site, in
in an
an office
office or
or in
in a
a laboratory.
laboratory. However,
However, precautions
precautions should
should be
be taken
taken to to
results testing
ensure and appropriate actions when contamination isarea
detected:
ensure testing is conducted in a reasonably clean area to avoid contamination of the sample and the test by
is conducted in a reasonably clean to avoid contamination of the sample and the test by
microorganisms
microorganisms other
other than
than those
those inin the
the test
test sample.
sample. WashWash hands
hands before
before and
and after
after testing.
testing. During
During testing
testing wear
wear clean,
clean, nitrile,
nitrile,
x or
vinyl
vinyl or polythene
EP096 How
polythene gloves. Avoid
to Interpret
gloves. touching
touching areas
Avoid MicrobMonitor2areas of the
Test
of syringe
Results
the in and
syringe loop
loop dispenser
Accordance
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with IATA
dispenser come
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direct with
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contact the
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sample andand avoid
avoid touching
touching the
the inside
inside of the MicrobMonitor2
of the MicrobMonitor2 testtest bottle
bottle cap
cap and
and neck
neck when
when dispensing
dispensing sample.
sample.
Operations with MicrobMonitor2.
x Results that count

5HVXOWVWKDWFRXQW
EP132 Routine Monitoring of Diesel Fuel Tanks and Distribution Systems with MicrobMonitor2.
Results
Results
that that
count
count 3

3
11

5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW
x EP166 Routine Monitoring of Marine Diesel on Ships and Offshore Installations with MicrobMonitor2 .
x EP157 Technical Assistance Reading Results of MicrobMonitor2.

® ® ®
MICROBMONITOR2
MICROBMONITOR2 2 ®2®®
MICROBMONITOR
MICROBMONITOR2
MICROBMONITOR
TEST DISPOSAL
WHAT EPROCEDURE
Calculate
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MICROBMONITOR ® microbial CFU in the sample.
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MICROBMONITOR
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MicrobMonitor2 test
bottles
content in x
bottles showing microbial growth. Before disposal, disinfect all MicrobMonitor2 test bottles which show
showing
petroleum For microbial
products
tests of growth.
including
water phase Before
liquid
or oil, disposal,
fuels,
numbers oils
of disinfect
and all
associated
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water.
conventionally test bottles
MicrobMonitor2
expressed as which
enables
number growth
on-site
of
show CFU by
or ml.
per
growth by
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laboratory
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testing
of the
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accordance
opened bottles
with in
in a strong
ASTM
a strong
and disinfectant
IP Standard
disinfectant solution (e.g.
Methods
solution (e.g.
ASTM household
D7978
householdandbleach)
IP613
bleach) - overnight or by
Determination
overnight or by of
incineration
the viable
incineration or by
or by
Follow the appropriate steps below to add distillate fuel or biofuel (A.1), heavy/residual fuel or oil (A.2), water (A.3)
using
aerobic chlorine
a)or microbial release
content tablets
of to (available
fuels separately). - Thixotropic Gel Culture Method. MicrobMonitor2 is
using Results
chlorine
surfaces of test of
release
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tablets
(A.4) the and
phase sample:
(available associated
separately).
MicrobMonitor2 water
test bottle.
A.1 Decontaminated
microbial
If 0.5 contamination
Aviation
ml ofkerosene,
Decontaminated fuel has tests,
in unused
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other
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fuel or
or those
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multiply
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fuels
those isshowing
and
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in
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Aircraft
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no microbial growth
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Maintenance
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Manuals
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the number
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perlocal
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manufacturers.
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colonies after by
theaspecified
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are monitoring
less than 2000
contamination
A.1.1 microbial in aviation,
CFU to marine
per litre of fuel.and ground fuels and is NATO Codified (6640-99-834-3573).
Immediately prior testing, shake the sample for 30 seconds. If fuel samples contain
visible free water, the sample should be allowed to stand for 12 minutes to enable water to
settleSTORAGE
MicrobMonitor2
STORAGE
Iffrom
0.25 the
ml of AND
provides
AND
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sample SHELF
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SHELF total
be LIFE
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multiply thecolony
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colonies
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estimated by 4000 to unit giveofthe measure
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for microbial
microbial
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time for CFU
all fuel per litreused
samples in a
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If there
if free water is of
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industries.
observed. after the specified detects the important
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there areyeasts and 4000
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CFU contaminate
per litre of petroleum
fuel. products including Hormoconis resinae,
Aspergillus, Candida and Pseudomonas
The
The shelf
shelf life of MicrobMonitor2
life of MicrobMonitor2 is is 1
1 year
year from
from the the date
date of of production.
expiry date
date is is printed
printed on on the
the batch
batch label
label ofof each
each
species. expiry
Contamination in
MicrobMonitor2
MicrobMonitor2 box.
box. Store
Store MicrobMonitor2
tests between
between 2
2 to
to 22°C,
22°C, in
in the
the dark.
dark. The
The expiry date
date applies
applies only
only to
to product
product
If a different
stored as volume
as stipulated.
stipulated. If of fuel has
If refrigerated been
refrigerated (2 tested,
(2 -- 8°C)
8°C) the then a calculation
MicrobMonitor2 test
the MicrobMonitor2 can be
test will made
will usually as
usually still follows:
still be
be useable
useable for up
up to to aa year
year beyond
beyond the the
stored for
Microbial
The MicrobMonitor2 test consists of anot

screw-capped bottle containing
MicrobMonitor2 a thixotropic nutrient gel. Atemperatures
sample is added to the gel

sample
designated
designated expiry
expiry date.
date. Do
Do not store
store MicrobMonitor2 frozen.
frozen. Exposure
Exposure to
to freezing
freezing temperatures during
during transport
transport is
is
usingacceptable.
a sterileNumber
plastic
Avoid loop or syringe
of microbial
exposure of (provided).
CFU per litre = Number
MicrobMonitor2 The tests
bottle islight
shaken
oftocolonies and
counted
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or estimated use. xA and the
A1000
slightly pink sample, and anyin the
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microorganisms in Avoid
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The gel is re-set tests
into to
a
Volume light
flat during
layer
of fuel and storage
tested the and
bottle
(ml) use.
is slightly
incubated pink
in the discolouration
dark. The gel in the
MicrobMonitor2
MicrobMonitor2 gel
gel maymay develop
develop over
over time
time during
during storage
storage but
but this
this will
will not
not affect
affect the
the performance
performance of
of the
the test.
test.
contains components
the cap which sustain the growth of viable microorganisms and the sample
A.1.2 Remove of the MicrobMonitor2 test bottle taking care not to touch the inside itself contributes additional
nutrients.
b) Viable
Results microorganisms
and place it on a clean surface.of test of water in the
phase sample
or oil: grow into visible spots known as “colonies”, and a reactive compound
ALSO
ALSO AVAILABLE
If 0.1 mlAVAILABLE
of water hasFROM
FROM ECHA
ECHA multiply the number of colonies counted or estimated by 10 to give the number of
been tested,
The number
microbial of colonies
CFU performed ml of water. is a direct estimate
If there are no of the number
colonies afterof viable microbial
the specified particles (CFU)
incubation present
time there areinless
thethan
sample. The
10 microbial
For use with MICROBMONITOR 2® ®
2
Day _______
actual number ofofcolonies is counted or is estimated by reference to the Test Results Chart (page 9). The volume tested can
For use with MICROBMONITOR2
CFU per ml water. ®
Counted
Colonies
is considered
If 0.01 mlinofrelation
water or tooilthe has volume
been of sample
tested, added
multiply theto the test
number ofand expressed
colonies counted as CFU/litre
or estimated of fuel,
by 100or CFU/ml
to giveof theoilnumber
or
The
The following
following itemsitems can can assistassist in
in performing
performing the the MicrobMonitor2
MicrobMonitor2 test test procedure:
procedure:
waterofassociated
microbial with CFU fuel.
per ml of water or oil. If there are no colonies after the specified incubation time there are less than 100
microbial CFU per ml of water or oil.
x
x MicrobMonitor
MicrobMonitor Sampling
Sampling Kit (ECHA16/SB)
Kit5 and
(ECHA16/SB) -- AA sterile,
sterile, polypropylene
polypropylene bottle
bottle for
for sampling
sampling fuel
fuel systems
systems and which
MicrobMonitor2
A.1.3 Openenables theis peel
available
pack incontaining
packs of 44 tests,
a syringe at each
the containing
handle end everything
and remove thatthe is required
wipes forto perform the test:and which
enables easy
easy separation
separation of
of water
water phase
phase from
from fuel
fuel phase;
phase; includes
includes alcohol
alcohol wipes for decontaminating
decontaminating sample
sample
syringeIf ataking
different
care volume
not to of touchwater theorlower
oil hasbarrel
beenand tested,
nozzle. then a calculation can be made as follows:
points.
points.
Counted
Colonies
MICROBMONITOR®2 test results
x MicrobMonitor2 test bottles

Day 4
Number of microbial CFU per ml = Number of colonies counted or estimated

x
x Sterile syringes*
x Incubator
Incubator (ECHA14/IN)
(ECHA14/IN) -- enables enables optimal,
optimal,
Volume consistent
of waterincubation
or oil tested temperature
temperature
(ml) of MicrobMonitor2
of MicrobMonitor2 tests; tests; holds
holds 1212
x Sterile loop dispensers*
MicrobMonitor2
MicrobMonitor2 test
test bottles;
bottles; available
available batterybattery or or mains
mains (110/230
(110/230 Volt)Volt) operated.
operated.
xNote Bottle labels
x
x
x MicrobMonitor
MicrobMonitor
This instruction leaflet. Bumpers
Bumpers (ECHA16/TB)
(ECHA16/TB) -- soft, soft, rubber-like,
hemispheres which which cancan be be stuck
stuck to to a a table
table toptop and
and enable
enable
Counted
Colonies
easy breaking
If the number
easy breaking
of colonies andisshaking
and shaking
too numerous of MicrobMonitor2
of MicrobMonitor2
to count and gel.
gel.a sample volume of 0.5 ml or 0.25 ml of fuel or 0.1 ml or 0.01 ml
Day 3
of water
* The 44 or oil was tested,
withoutthe Test Results Chart can be used directly to determine the approximate number of
A.1.4 pack
Usingis the
available
syringe,with or
draw fuelsterile
fromsyringes and loop
approximately dispensers.
3 cm below the surface of the
x
microbial
x If there
sample.
CFU per
Chlorine
Chlorine
litre of tablets
Release
is lessRelease
fuel or per
than 6 tabletscm depth
ml of water- or
(ECHA21/CP)
(ECHA21/CP)
of fuel phase, to oil.
disinfect
- to disinfect
draw sample MicrobMonitor2
MicrobMonitor2 test
test bottles
from approximately bottles showing
showing microbial
microbial growth growth for
for easy
easy
MicrobMonitor2 disposal. has been developed and patented by ECHA Microbiology Ltd. and it is marketed worldwide by
halfway down disposal.
the fuel phase. Avoid the transfer of visible interfacial particulate, water
droplets or emulsion in theOF
INTERPRETATION aliquot
TESTtoRESULTS
be tested.
x
x Sterile
Sterile swabsswabs (ECHA15/SW)
(ECHA15/SW) -- swabs swabs for for sampling
sampling surface surface contamination
contamination (e.g. (e.g. tank
tank surfaces,
surfaces, filter filter cartridges
cartridges etc.).
counted
Colonies
etc.).
Day 2
There are no universally accepted limits or specifications for microbiological contamination in fuels or oils. The number of
WHAT x IS THELong, MICROBMONITOR
CFUsterile bulb pipettes 2 TEST
®
(ECHA07/PP/EL23)USED FOR? -- to
x
microbial Long, which
sterile define
bulb moderate
pipettes and heavy contamination
(ECHA07/PP/EL23) to enable
enable water levels
water to be
be removed
to will depend on
removed from
from the
the bottom
a number bottom of
of samples.
of factors including the
samples.
fuel type and intended use, the sampling location, whether the sample is taken from supply and distribution facilities or
The MicrobMonitor2
from point of end is used
use, tothetest for microbial
intended contamination
fuel storage time andinspecific fuel and oil products,
operational and to monitor
circumstances. microbial
Some guidancegrowth in are
limits
storage tanks and supply facilities. MicrobMonitor2 can to beprovide
used asearlya routine monitoring tool, for growthproviding assurances
A.1.5Other
shown
Other
Draw fuelTest
in Table Kits
Testquality,
Kits
into the&
2 Ancillary
below.
&syringe
Ancillary until itItems
These limits
Items are intended
is more than half full and then, with the syringe nozzle indication that microbial is occurring in the
counted
Colonies
about product
facility sampled. for
Higher investigation
levels of incidents and
of contamination willfuel for confirming
often bethe
required effectiveness the of measures taken toproblemsprevent oror any
end ofbefore onset of operational
Day 1
uppermost, press the plunger to expel air. Expel surplus until the plunger
remediate microbial
detrimental effect growth and
onSulphide
fuel contamination.
fitness for use. For further
alignsx
xwith the mark
SigTestsΠ:
SigTests®:
SigTestsΠ: forSig
the
Sig required
Sulphide volume
(ECHA02/SS)
(ECHA02/SS) of sample
—
— simple (seeinformation
simple table
test
test kit1).
kit for see the technical guidance
for Sulphate
Sulphate Reducing
Reducing Bacteria
Bacteria (SRB)
leaflets listed
(SRB) whichwhich can
on page 2.
can cause
cause serious
serious
This instruction corrosion
leaflet
corrosion and
and sulphide
provides
sulphide generalspoilage
spoilage in
in fuel,
instructions
fuel, oil
for
oil and
using
and water
water systems.
MicrobMonitor2.
systems. Can
Can be used
Further
be used on-site
technical
on-site or
or in
in the laboratory.
instruction
the on
laboratory. the use of
Table 2
from x Sample
www.echamicrobiology.com).
Biocide Moderate and approximate
Heavypreparation, Comment of test of
x Biocide Rapide Rapide (ECHA01/BR)
(ECHA01/BR) These— — simple
leaflets rapid
simpleinclude
rapid advice test
test kitkit for assessing
assessing presence
on sampling,
for sample
presence and approximate concentration
interpretation
concentration of
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
commonly used
results and appropriate
commonly used
actions fuelwhenand contamination
oil biocides.
biocides. is detected:
Fuel from aircraft fuel fuel tankand drain oil 4000 to 20,000 CFU / litre >20,000 CFU / litre IATA
Contamination
x x x EP096 Any
AnyHowlevel and
to Interpret
level andfuel MicrobMonitor2
bottom
bottom sampling
samplingTest devices
Results(ECHA23)
devices in Accordance — for
forwithsafe
safeIATAsampling
Guidelines from forfuel
Aircraftand
andDrain oil
oil storage
Samples.tanks
tanks in
Water from aircraft tank intodrain 1000 to(ECHA23)
10,000 CFU—/ ml sampling
>10,000 from
CFU fuel
/ ml Limits storage in
A.1.6 Insert the
compliance tip
compliance with of the syringe
with industry
industry standards. the
standards. neck of the MicrobMonitor2 test bottle and
dispense
x the representative
EP119
Bulk sample
RoutinebyMonitoring
fully depressing the from
of Aviation
fuel sample syringe plunger.
Fuels in Supply and Distribution Facilities, Airport Depots and Into-plane
x 4000see to 20,000 CFU / litre >20,000 CFU / ml
x For
Operations
supply & full
with
distributionMicrobMonitor2.
For our full range of test kits and ancillaries see www.echamicrobiology.com.
our range of test kits and ancillaries www.echamicrobiology.com.
e.g. Aviation fuel,
Fuel from bottom / drain of supply &
x Technical 10,000 to 100,000 CFU
of Diesel Fuel Tanks and Distribution Systems with MicrobMonitor2. / litre >100,000 CFU / litre diesel, marine gas
EP132
Technical
Routine
distribution support
support
tank Monitoringand
and services
/ filter services oil
Water phase from bottom / drain of
x ECHA EP166 Routine Monitoring of Marine 100,000 to 1,000,000 CFU / ml >1,000,000 CFU
MicrobMonitor2 / ml .
ECHA Microbiology
supply & distribution
Microbiology Ltd. provides
Ltd.fuel tank
provides /a fullDiesel
full
afilter on Ships
technical
technical service
service andto toOffshore
support our productwith
our product range.
range. ECHA
ECHA also also offers
offers laboratory
laboratory testing,
testing,
training
training courses,
courses, consultancy
consultancy and and on-site
on-site attendance
attendance for for audit
audit and
and investigation
investigation of of microbial
contamination and and corrosion
corrosion
x associated
Lubricating
EP157 with
Technical petroleum
/ hydraulic
Assistance oilproducts
inReading
circulation and petroleum,
Results 100 marine,
to 10,000
of MicrobMonitor2. CFU aviation
/ ml and and other industrial
>10,000 CFUfacilities.
/ ml
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)

Volume tested (ml)
Volume tested (ml)
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Location
Location
Location

Location
Location

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
associated with petroleum products and petroleum, marine, aviation other industrial facilities.
A.1.7 Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample
details and test date on the label provided and stick it on a narrow side of the bottle.
Lubricating / hydraulic oil sump 1000 to 10,000 CFU / ml >10,000 CFU / ml
Proceed to step B.

4 10

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D BLL
LEE
E
MICROBMONITOR®22® Test Results Chart
PREPARATION
PREPARATION FOR
A.2
FOR TESTING
Heavy and residual fuels and oils.
TESTING Number of colonies in
A.2.1 Immediately Microbial Content of
Appearance
Sampling after
priorincubation
to testing, shake the sample
considerations
for 30
test seconds.
bottle Volume tested
Sample
(counted or estimated)
A.2.2 Remove the cap of the MicrobMonitor2 test bottle and place it on a clean surface.
Sampling
Sampling should
should bebe conducted
conducted in in a
a clean
clean and
and consistent
consistent manner
manner which
which prevents
Contamination in
contamination
into the
into the sample.
sample. Once
Once fuel
fuel samples
samples have
have been
been taken,
taken, any
any microbes
microbes present
0.5 mlwill
present will tend
of fuel to slowly
(syringe)
tend to slowly die and
<2000
diethe it is
it CFU
andloop important
per litre to
is end
important to
A.2.3 Open the peel pack containing a loop dispenser and remove it taking care not to touch and the
Microbial
test
test samples
samples as
as soon
soon as
as possible.
possible. If
If the
the test
test is
is carried
carried out
out more
more than
than 24
24 h
h after
after sampling,
sampling, it
it is
is possible
possible that
that the
the results
results will
will not
not
sample
lower part of the handle.

reflect the
reflect the viable
viable microbial
microbial content
content present
present at at the
the time
time of
of sampling.
sampling. To
To avoid
avoid this,
this, part
part oror all
all of
of the MicrobMonitor2 test
the MicrobMonitor2 test
procedure can be 0.25 ml of fuel (syringe) <4000 CFU per litre
procedure
A.2.4 Immerse be conducted
can the conducted
loop end into
at
atthethe sampling
thefuel/oil
sampling location.
location.
to approximately 3 cm below the surface of the sample or, if there is less than 6
None
cm depth of fuel/oil phase, to halfway down the fuel/oil phase. Remove the loop from the sample and allow surplus fuel/oil
Sample
Sample type
type
to drain off; and
and
ensure recommended
recommended
a film test volumes
testwithin
of fuel/oil is trapped volumes
the loop. 0.1 ml of water (syringe) <10 CFU per ml
MicrobMonitor2
MicrobMonitor2 can be
can
A.2.5 Insert the loop be used
intoused to test
to
the neck test
ofaathe
wide
wide variety of
variety of petroleum
MicrobMonitor2petroleum products
products
test bottle and staband
and
0.01 associated
associated
it into
ml the gel
of water water.
water.
or and The volume
The
oil agitate volume of
it briefly of sample
to sample
transfer
which
which should be tested will depend on the type of sample.
should be tested will depend on the type of sample. Refer to the Refer to the table
table below to <100
determine CFU
the per ml
appropriate
below to determine the appropriate
the sample. (loop dispenser)
Day _______
volume
volume of
of sample
sample toto test
test and
and which
which measuring
measuring device
device toto use.
use.
Counted
Colonies
A.2.6 Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample details and test date on the label
provided
Table and stick it on a narrow side of the bottle. Proceed to step B.
Table 11
0.5 ml of fuel (syringe) 2 x 104 CFU per litre
Recommended
Recommended test
test Measuring
Measuring
Sample
Sample
A.3
Sample Water phase associated with fuel/oil. 10 Colonies
volume
volume
volume device
device
device
MICROBMONITOR2 ® test results

Counted
Colonies
A.3.1 If akerosene
test of water phase is required, allow the sample
If possible, count to the
stand until free water0.5
exact hasml
mlsettled to the bottom of the sample. It
Day 4
Aviation
Aviation kerosene
kerosene 0.5 ml
is recommended the water phase test is conducted numberafter any fuel/oil
of colonies andphase test. The test procedure is essentially the same as
described
Other above
middle but an
distillate appropriate
fuels technique
(e.g. diesel,
diesel, for
gasseparating/removing
calculate
marine the
oil, actual
heatingnumber 0.1
oil), biofuels
biofuelsml&from
water of water (syringe)needs to100
the sample be CFU per ml
used.
Other middle distillate fuels (e.g. marine gas oil, heating oil), & 0.25 ml
ml Syringe
Syringe
gasoline
gasoline of CFU present in the sample 0.25 ml Syringe
gasoline
A.3.2 As appropriate for the volume of sample being tested (see table 1)0.01 use ml
theofloop dispenser
water or oil or syringe to measure the
required volume ofaircraft
water. fuel tank drains 1000 CFU per ml
(loop0.1dispenser)
Counted
Water phase from drains ml

ml
Colonies
Water phase from aircraft fuel tank drains 0.1 ml

Day 3
Heavy
Heavy and
a) To test
and residual
0.1residual fuels
ml of water
fuelsusing the syringe;
fuels 0.01 ml
0.01 ml
ml
Lubricating,
Lubricating, hydraulic
hydraulic and other oils oils
oils 100 Colonies 0.01 mlml
ml the MicrobMonitor2 Loop
Loop
The water phase can andbe drawnother directly from the bottom of the sample 0.5 andmladded 0.01
of fuel to (syringe) Loop
c. 10 testper
5 CFU bottle.
litre To
enable
Water easy
phase access
from to
fuel the
storage water it
tanks
tanks may be necessary to first decant off some fuel from
0.01 ml
ml the sample and/or transfer the water
Water If possible, count the exact
phase phase from fuelsmall,
to a separate storage tanks
sterile container using a syringe or a long sterile pipette 0.01 ml (supplied separately). Mix gently by
counted
Colonies
number of colonies and

Day 2
swirling but avoid mixing the fuel and water layers together. 0.25 ml of fuel (syringe) c. 105 CFU per litre
The calculate the actual number
The volume
volume of of sample
sample testedtested can can be be changed
changed in
in
of CFU present
order
order to
to increase
increase
inmay
the in sample
or
or decrease
decrease the
the detection
detection level
level of
of the
the test.
test. However,
However,
testing
testing volumes greater than those recommended above may in some circumstances lead to underestimation of
Open volumes
the peel greater
pack than
containing those a recommended
syringe at the above
handle end and some removecircumstances
the syringe lead
taking to underestimation
care not to touch microbial
of the
microbiallower
barrel and nozzle. Insert the barrel of the syringe into the sample so that0.1
numbers.
numbers. the mlnozzle
of water is in(syringe)
the water phase. c. 103Draw CFU perwater ml into
or
the syringe, remove from the sample and then, with the syringe nozzle uppermost, expel air. Expel surplus water until the end
Determining whether with to thetest fuel/oil
mark. or water phase
counted
Colonies
Determining
of the plunger aligns whether to test
0.1 ml fuel/oil or water
Insert the tip phase
of the syringe into 0.01 themlneck of the
of water orMicrobMonitor2
oil test bottle and
Day 1
Estimate 100 colonies if 104 CFU

c.bottle pertighten.
ml
dispense the sample by fully depressing the syringe plunger. Replace the cap (loop the MicrobMonitor2
ondispenser) and
result is similar to picture
Record
Depending
Depending on the sample
on the details
the system and
system sampled test
sampled and date
and theon the label
the sampling provided
sampling location, and
location, samples stick
samples may it on a narrow
may contain side
contain fuel/oil of the
fuel/oil and/or bottle.
and/or free Proceed
free water
water phase. to step
phase.
B.
MicrobMonitor2 can be used to test either fuel/oil or associated water. Water phase will usually contain far more
microorganisms than
microorganisms than are are present
present in in fuel
fuel or or oil
oil phase
phase and and therefore
smaller volume
volume is is usually
usually tested.
tested. Because
Because free free water water
b) To test 0.01not
phase ml of water using the loop dispenser;
phase may may not always always be be recovered
ensure consistency 0.5 mlwhen of fuelconducting
(syringe) regular
c. 106microbiological
CFU per litre
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
consistency when conducting regular microbiological 0.5/0.25/0.1/0.01/other__

monitoring,
monitoring, it it is
is usually
usually recommended
recommended that that fuelfuel or or oil
oil phase
phase fromfrom above
above any any freefree water
water is is tested.
tested. However,
However, in in some
some cases cases
itA is
it issyringe can be used
recommended
recommended to to
to draw watertest
additionally
additionally fromwater
test the bottom
water phase of
phase the samples
(e.g.
(e.g. sample and
samples fromthen
from a drop
aircraft
aircraft fuel
fuel of tanks,
water placed
tanks, and
and when
wheninto the loop and
investigating
investigating
added to the
suspected
suspected MicrobMonitor2
microbial
microbial growth in
growth intest bottle. Alternatively,
facilities).
facilities). Further information
Further transfer is
information the
is water 0.25
provided
provided phase mlto
in the
in the afuel
separate
relevant
of
relevant small, sterile
technical
(syringe)
technical c. 10container
guidance
guidance per using
6 CFUdocuments
documents litre a
syringe
listed on or a
page
listed on page 2. long 2. sterile pipette (supplied separately).
Estimate Avoid
1000 transfer
colonies if of fuel. Invert the container with the separated water
three times to mix. result is similar to picture

To 4 gently
To determine
determine if if the
the sample
sample contains
contains free free water
water hold hold it it up
up toto the
the light
light andand0.1 visually examine
ml of water
visually examine it.
it. It
(syringe) It may
may help c.10
help to
to CFU per
gently swirl
ml the
swirl the
Remove
sample the cap of the MicrobMonitor2 test bottle and place
sample to create a vortex. In order to enable access to the sample with the measuring devices provided, it
to create a vortex. In order to enable access to it
the on a
sample clean withsurface.
the Open
measuring the peel
devices pack containing
provided, it may
may a loopbe
be
dispenser
necessary to
necessary and remove
to transfer
transfer some it taking
some sample, care
sample, after not to
after shaking, touch
shaking, to to a the loop
a smaller end
smaller clean and
clean container. the
container. lower part of the handle. Immerse the loop end
into the separated water or fill the loop with a drop of water using a syringe. Allow surplus water to drain c.10 0.01 ml of water or oil away but per
ensure
5 CFU ml the
if a residue of fuel is present(loop dispenser)
impair the ability of the loop to fill with
circle of the loop remains filled with water;®
Preparing
Preparing the MICROBMONITOR 2 ®note,
test
2 MicrobMonitor2bottles this can
water. Insertthe the MICROBMONITOR
loop into the neck of the test bottles test bottle and stab it into the gel and agitate it briefly to transfer
the sample. Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample details and test date on the
If the MicrobMonitor2
the
Iflabel MicrobMonitor2
provided and stick
test
testitbottles
bottles
on a narrow
have
have been been stored refrigerated,
side ofstored refrigerated,
the bottle. Proceed
allow
allow them
them
to step
to equilibrate to ambient temperature before
B. to equilibrate to ambient temperature before
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)

Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Location
Location
Location
Location
Location

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
they are used. Remove the seal from

they are used. Remove the seal from the cap of the bottle before using the cap of the bottle before using each
each MicrobMonitor2
MicrobMonitor2 test.
c.107Avoid
test. CFU per
Avoid prolonged
litre or
prolonged
exposure of the MicrobMonitor2 test bottles to direct sunlight or other 0.5 ml
bright oflight
fuel (syringe)
during preparation, incubation and
exposure of the MicrobMonitor2 test bottles to direct sunlight or
Note The MicrobMonitor Sampling Kit (supplied separately) is a sample bottle which has a side siphon and water collectionother bright light during preparation, incubation
above and
examination
examination of tests.
chamber to of tests. easy separation of water from fuel samples.
enable
c.107 CFU per litre or
Estimate 10,000 or more 0.25 ml of fuel (syringe)
Cleanliness
Cleanliness during during testingtesting above
colonies if result similar to
picture
MicrobMonitor2
MicrobMonitor2 tests tests can
can be be conducted
conducted on-site, on-site, in in an
an office
office or or inin aa laboratory.
0.1 ml of However,
laboratory. water (syringe)
However, precautions
c.105 CFU
precautions should
should perbeml taken
be or above
taken to
to
ensure
ensure testing is conducted in a reasonably clean area
testing is conducted in a reasonably clean area to to avoid
avoid contamination
contamination of of thethe sample
sample and and the the test test by by
microorganisms
microorganisms other other than
than those
those in in the
the test
test sample.
sample. Wash Wash hands
hands beforebefore and and after testing. During testing wear
wear clean,
clean, nitrile,
0.01after ml oftesting.
water or During
oil testing nitrile,
vinyl
vinyl or or polythene
polythene gloves. gloves. AvoidAvoid touching
touching areas areas of of the
the syringe
syringe and and looploop dispenser
dispenser which
which come
come into
into direct
c.10 6 CFU
direct contact
per ml or
contact with
with the
above
the
(loop dispenser)
sample and
sample and avoid
avoid touching
touching the the inside
inside of of the MicrobMonitor2 test
the MicrobMonitor2 test bottle
bottle cap cap and and neckneck whenwhen dispensing
dispensing sample.
sample.
Results
Results that that
Results that count
5HVXOWVWKDWFRXQW
count
count
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW 3

5
9

MICROBMONITOR2
MICROBMONITOR2
® ®
2® ®2®®®
MICROBMONITOR2
MICROBMONITOR
MICROBMONITOR
MICROBMONITOR2
MICROBMONITOR2
A.4 DISPOSAL
A.4
Testing
Testing
surfaces.
surfaces.
WHATDISPOSAL
IS THE MICROBMONITOR 2 ® TEST?
A.4.1A.4.1MicrobMonitor2
MicrobMonitor2 can be can used
be used
to testtofor
testmicrobial
for microbialcontamination
contamination on surfaces,
on surfaces,
e.g. to
e.g.
test
todeposits
test deposits
on tankon surfaces
tank surfaces
or or
Wear gloves
MicrobMonitor2 is and avoid
a simple touching
and easy to the inside
theuse testof the
ofkit bottle
which or
enables captheandquantitative
wash hands after handling
assessment MicrobMonitor2
anyviable
of the microbial test
MicrobMonitor2
filters.Wear
filters. gloves
SterileSterile
swabsand avoid
swabs
(availabletouching
(available
separately) inside
separately)
should the
be bottle
should used or
be usedto capto and
removeremovewashsurface
hands
surface after handling
contamination
contaminationandanyand
transfer
transfer thisa totest
this to a
content bottles showing
in petroleum microbial
products growth.
including Before disposal,
liquid fuels, oils disinfect
and associatedall MicrobMonitor2 test bottles which
water. MicrobMonitor2 enables show growth
on-site or by
bottles
MicrobMonitor2 showing
MicrobMonitor2 test bottle.
test bottle. growth. Before disposal, disinfect all MicrobMonitor2 test bottles which show growth by
microbial
immersion
laboratory of
testing the
in opened
accordance bottles
with in a
ASTM strong
and disinfectant
IP Standard solution
Methods (e.g.
ASTM household
D7978 bleach)
and IP613 -
immersion of the opened bottles in a strong disinfectant solution (e.g. household bleach) overnight or by incineration or byovernight or
Determination by incineration
of the viable or by
aerobic usingmicrobial
chlorine release
contenttablets (available
of fuels separately).
and associated water - Thixotropic Gel Culture Method. MicrobMonitor2 is
A.4.2using
A.4.2
Open chlorine
Open
the peel release
the packtablets
peel pack (available
containing
containing
a swab separately).
a swab
at the athandle
the handleend and
end remove
and remove
the swab, taking
the swab, taking
care care
not to not
touch
to touch
the the
cotton cotton
end of end
theofswabthe swab
and theandhandle
the handlenear near
the cotton
the cotton end. end.
Decontaminated
Decontaminated tests, unused
in aircraft
tests, unusedfuel tests
tests or
tanks those showing
and isshowing
or those listed in nono microbial
themicrobial growth
Aircraft Maintenance can
growth can be be disposed
Manuals of as
of Airbus,
disposed normal
of as Boeing,
normal BAEwaste
waste in in
Systems accordance
and other with local
leading waste
aircraft regulations.
manufacturers. The test is used by a wide range of military services for monitoring
A.4.3accordance
A.4.3
Rub theRubsurface
the with local
to bewaste
surface be regulations.
totested tested
usingusing
the cotton
the cottonend of end the ofswab,
the swab,
rotating
rotating
the swab
the swab
handlehandle
so that
so all
thatparts
all parts
of theof the
contamination in aviation, marine and ground fuels and is NATO Codified (6640-99-834-3573).
cotton cotton
end contact
end contact the surface.
the surface.
If possible
If possible
swabswab a known a knownarea area
of surface.
of surface.
A.4.4STORAGE AND SHELF LIFE

MicrobMonitor2
STORAGE provides
AND a total
SHELF count of microbial colony forming units (CFU). CFUs are the conventional unit of measure
LIFE
A.4.4
InsertInsert
the cotton
the cotton
end of
endtheofswab
the swab
into the intoneck of theofMicrobMonitor2
the neck the MicrobMonitor2 test bottle
test bottle
and stab
and itstab
intoitthe
intogel
theand
gel agitate
and agitate
for microbial contamination used in a wide range of industries. MicrobMonitor2 detects the important bacteria, yeasts and
for about
for about
15 seconds
15 seconds
and then
and remove
then removeand discard.
and discard.
moulds which can contaminate petroleum products including Hormoconis resinae, Aspergillus, Candida and Pseudomonas
The
The shelf
species. shelf life of MicrobMonitor2
life of MicrobMonitor2 is is 1
1 year
year fromfrom thethe date
date ofof production.
expiry date
date is is printed
printed onon the
the batch
batch label
label ofof each
each
A.4.5MicrobMonitor2
A.4.5
Replace
Replacethe capbox.
the on
cap Store
the
on MicrobMonitor2
the
MicrobMonitor2
bottle between
bottle
and tighten.
and 2 to 22°C,
tighten.
Record in
Record the
the dark.
sample
the The
sample
MicrobMonitor2 box. Store MicrobMonitor2 tests between 2 to 22°C, in the dark. The expiry date applies only to product expiry
details
details
and date
test
and applies
date
test ononly
date theon to product
label
the label
stored
provided
provided
storedandas
as stipulated.
stick
and itstick
on a
stipulated. If
it on
If refrigerated
narrow
a narrow
side of
refrigerated (2
side --of
the
(2 8°C)
thethe
bottle.
8°C) MicrobMonitor2
bottle.
the ProceedProceed
to step
MicrobMonitor2 test
to B.
step
test will
B. usually
will usually still
still be
be useable
useable for
for up
up to
to a
a year
year beyond
beyond the
The MicrobMonitor2
designated test consists
expiry date. of anot
Do screw-capped
store bottle containing
MicrobMonitor2 frozen.a thixotropic
Exposure nutrient
to gel. Atemperatures
freezing sample is added to the
during gel the
transport is
designated
usingacceptable. expiry
a sterile plastic loopdate. Do
or syringe not store
The tests frozen.
bottletoislight
shaken Exposure
and to
the gel freezing
and the during
sample, transport
and anyin the is
acceptable. Avoid
Avoid exposure
exposure of
of MicrobMonitor2
MicrobMonitor2 tests to light during
during storage
storage and
and use.
use. A
A slightly
slightly pink
pink discolouration
discolouration in the
microorganisms in it are dispersed. The geltimeis re-set into a flatbut layer®and ®the bottlethe is performance
incubated in of the dark. The gel
B MicrobMonitor2 this22 22andaffect sample
geldisperse
may develop over during storage ® not ® affect
B Shake Shake
MicrobMonitor2
contains
to disperse
componentsgel
to may sample
which
sample
develop
sustain the
in growth
over the in MICROBMONITOR
time the MICROBMONITOR
during storage
of viable but this
microorganisms
will
willgel.
not gel.
the the performance of the
the test.
itself contributes test.
additional
nutrients. Viable microorganisms in the sample grow into visible spots known as “colonies”, and a reactive compound
B.1 Loosen
B.1 Loosen
and break
and break
up theupgel theinMicrobMonitor2
theingel the MicrobMonitor2
bottlebottle
containing
containing
the dispensed
the dispensed
ALSO
sample,
sample, AVAILABLE
by tapping
by tapping FROM
the bottle
ALSO AVAILABLE FROM ECHAthe bottleECHA
firmly firmly
in theinpalm
the palm
of your
of your or onora on
handhand rubber
a rubber or or
bungbung
MicrobMonitor
MicrobMonitor
“Bumper”
“Bumper”
(available
(available
separately).
separately).
The number of colonies formed is a direct estimate of the number of viable microbial particles (CFU)
present in the sample. The
actualFor use
number with MICROBMONITOR
of colonies 2 ®
is counted or is estimated
For use with MICROBMONITOR2 ® by reference to the Test Results Chart (page 9). The volume tested can
is considered in relation can
to the volume of sample added to the test andprocedure:
expressed as CFU/litre of fuel, or CFU/ml of oil or
The
The following
following items
items can assist
assist in
in performing the MicrobMonitor2
performing the MicrobMonitor2 test
test procedure:
x
x MicrobMonitor
MicrobMonitor Sampling Kit (ECHA16/SB) -- AA sterile, polypropylene bottle for
for sampling fuel
fuel systems and which
MicrobMonitor2 is availableSampling
in packs of Kit5 and
(ECHA16/SB)
44 tests, each sterile, polypropylene
containing everything bottle sampling
that is required to perform systems
the test:and which
enables
enables easy
easy separation
separation of water
of water phase
phase from
from fuel phase;
fueltophase; includes
includes alcohol
alcohol wipes for
for decontaminating sample
decontaminating
B.2 Shake
B.2 Shakethe bottle
the bottle
vigorously
vigorously
for approximately
for approximately
30 seconds
30 seconds liquefy
to liquefy
the gel
theand wipes
gel and sample
points.
points.
xdisperse
disperse theMicrobMonitor2
sample.
the sample. The testThe
testbottle
test can be
bottle
bottles cantapped
be tappedrepeatedly
repeatedlyfor 30forseconds
30 seconds
on a on a
MicrobMonitor
MicrobMonitor “Bumper”“Bumper”to assist
to assist
with shaking.
with shaking.
After After
shaking,shaking,
the geltheshould
gel should
be slightly
be slightly
x
x Sterile syringes*
x
viscous
viscous Incubator
Incubator
but free
but of
free of(ECHA14/IN)
lumps (ECHA14/IN)
lumps
and have
and havea-- uniform
enables
enables optimal,
optimal,
a uniform
consistency consistent
consistency and incubation
consistent
and clarity. incubation
clarity. The temperature
of of of
temperature
The presencepresence of MicrobMonitor2
MicrobMonitor2 tests;
tests; holds
holds 12
12
xbubbles
bubbles Sterile
in the ingel
theloop
MicrobMonitor2
MicrobMonitor2
is geldispensers*
normal test
is normal
and bottles;
test will
andnot
bottles; available
willaffect
not affect battery
the test.or
the test.
available battery or mains
mains (110/230
(110/230 Volt)
Volt) operated.
operated.
x Bottle labels
x
x
x MicrobMonitor
MicrobMonitor
This Bumpers (ECHA16/TB)
instruction leaflet.
Bumpers (ECHA16/TB) -- soft,
soft, rubber-like,
hemispheres whichwhich can
can bebe stuck
stuck to
to a
a table
table top
top and
and enable
enable
easy
easy breaking
breaking and and shaking
shaking of of MicrobMonitor2
MicrobMonitor2 gel. gel.
* The 44 pack is available with or without sterile syringes and loop dispensers.
x
x
B.3 Stop
B.3 Stop
Chlorine
Chlorine
shaking
shaking
Release tablets
theRelease
test testtablets
thebottle bottle
(ECHA21/CP) -- to
(ECHA21/CP)
abruptly
abruptly
so that
so the
to disinfect
thatgel
thecollects
MicrobMonitor2 test
disinfect MicrobMonitor2
gel collects
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MicrobMonitor2 has been developed and patented by ECHA Microbiology Ltd. and it is marketed worldwide by
disposal.
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Proceed
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immediately
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x
x Sterile
Sterile swabs
swabs (ECHA15/SW)
swabs for
for sampling
sampling surface
surface contamination (e.g. tank
contamination (e.g. tank surfaces,
surfaces, filter
filter cartridges
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etc.).

WHATx IS THE MICROBMONITOR
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x Long, sterile
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The MicrobMonitor2 is used to test for microbial contamination in fuel and oil products, and to monitor microbial growth in
storage tanks and supply facilities. MicrobMonitor2 can be used as a routine monitoring tool, for providing assurances
Other
Other Test Kits & Ancillary Items
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instruction
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x Biocide Rapide
Biocide Rapide (ECHA01/BR)
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test kit for
for assessing presence
on sampling,
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approximate
sample preparation,
and concentration
interpretation of test of
concentration of
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commonly used
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fuelwhen oil
oil biocides.
and contamination
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NotesNotes compliance
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of tests with
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industry standards.
standards.
x
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It is preferable
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see to changes
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www.echamicrobiology.com. content
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likely
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x Technical test support
EP132 Routine theand
Monitoring ofservices
Diesel Fuel Tanks and Distribution Systems with MicrobMonitor2.
Technical
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MicrobMonitor2 support
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facility
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ECHA Microbiology Ltd. provides a full technical service to support our product range. ECHA also offers laboratory testing,ECHA also offers. laboratory testing,
training courses, consultancy and on-site attendance for audit and investigation of microbial contamination and corrosion
training courses, consultancy
Alternatively, MicrobMonitor2
Alternatively, MicrobMonitor2 testand
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flat during
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6

6
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PREPARATION FOR
FOR TESTING
TESTING
Transfer the MicrobMonitor2
Transfer the MicrobMonitor2 test bottle
test bottle
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dark dark
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location
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to maintain
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nominalSampling
nominal
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temperature of 25°C
of 25°C
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In normalIn normal
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for 4 days.
AvoidAvoidexposure
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incubation.
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Sampling should be conducted in a clean and
should be conducted in a clean and consistent
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The
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NotesNotes
on incubation
on incubation
of tests.
of tests.
Sample
Sample type
type and
and recommended
recommended test
test volumes
volumes
Occasional
Occasional
temperature
temperature
fluctuations
fluctuations
(e.g. overnight)
(e.g. overnight)
belowbelow the specified
the specified
temperature
temperature
rangerangeshouldshould
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not affect
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the number
of colonies
of colonies
MicrobMonitor2
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variety petroleum
ofmicrobial
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petroleum products
colonies
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andlonger
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visible and The
visible
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extension
volumean extension
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sample of
which
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specified
which incubation
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tested
incubation
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tested bedepend
should
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depend on
be applied.
on the
If the
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temperature
the
type of sample.
of sample.
temperature falls Refer
falls below below
Refer to the
to the
the specified
the tablerange
specified
range
table below
during
below to determine
during determine
incubation,
to incubation, theincubation
appropriate
the incubation
the
the time time
appropriate
Day _______
shouldshould
be extended
volume beofextended
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sample by test
to a equivalent
time
andequivalent
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which tototal
the time
measuring totaldevice
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timetemperature
thetotemperature
use. is estimated
is estimated
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belowbelow
the specified
the specified
range.range.
If the If the
volume of temperature
samplefalls
to below
test and which measuring device to or
use.
Counted
Colonies
incubation
incubation
temperature falls below
the specified
the specified
rangerangefor a for
period
a period periods
or periods
totaling
totaling
4 days4 ordaysmore,
or more,
microbial
microbial
contamination
contamination
will bewill be
underestimated
underestimated
and the and
result
the should
result should
be considered
be consideredinvalid.invalid.
The temperature
The temperatureshouldshould
not benotallowed
be allowed
to dropto below
drop below
4°C during
4°C during
incubation.
incubation.
Tableof1theof Table 1
Incubation
Incubation culture
the culture
mediummedium
aboveabove
the specified
the specified
temperature
temperature
rangerange
may prejudice
may prejudice
the ability
the ability
to detect
to detect
some some
types types
of microorganism.
of microorganism.
However,
However,
wherewhere
microbial
microbial
contamination
contamination
occursoccurs
in tanks
in tanks
or systems
or systems
wherewhere
the contents
the contents
have a
have
higher
a higher
temperature
Recommended temperature
than
test 25°C,
than 25°C,
the the
Measuring
Sample
contaminating
contaminating temperatures; Recommended
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such cases test
it mayitbe
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Samplemicroorganisms
microorganisms
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for growth
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at higher
volume device
to to
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incubate
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the
attest
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sampled.
sampled. volume device
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW
Results
Results
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count
count
Counted
Colonies
Day 4
Aviation
Aviation
WhenWhen kerosene
kerosene
MicrobMonitor2
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0.5
the mlContamination
ml
IATA
Contamination Limits Limits
shownshown
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in table
2 on 2 on
page page
8), if the
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indicates
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contamination
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is inhighest
the highest
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category
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4 days, it is generally
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acceptable
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Other
and report
and
Other middle
reportresult
themiddle distillate
the without
result without fuels
further
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further
incubation.
fuels diesel, marine
incubation.
(e.g. diesel, marine gas gas oil,
oil, heating
heating oil),
oil), biofuels
biofuels & &
0.25 ml
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gasoline
gasoline
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Examine
Examine
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tank drains
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phase from aircraft fuel 0.1

Day 3
D.1 IfD.1possible
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Heavy and possible
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and residualexamine
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incubation.
As a As
minimum,
a minimum,
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0.01 on
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least least
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one occasion in theinfirst
the3 first
days 3 days
and again
and again
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the dayfinal ofdayincubation.
of incubation.To examine
To examine
Lubricating,
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hold hold
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background
background
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all visible
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0.01 ml
ml Loop
Loop
colonies.
colonies.A hand A hand
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colonies.
All All
Water phase
Water phase fromfrom fuel
fuel storage
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0.01 ml
purplepurple
colonies colonies
in all in
parts
all parts
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including
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counted
Colonies
gel which
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OnceOnce a colony
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Day 2
evenevenif it volume
The
The grows
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volume larger;
of larger;
of sampleit is the
sample it is number
tested
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can
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of colonies
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in is
that
in order
order important,
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or their
or not their
size. size.
decrease
decrease It is detection
the
the It is
detection level
level of
of the
the test.
test. However,
However,
recommended
recommended
testing
testing volumes that colonies
volumes that colonies
greater
greater are marked
than
than are marked
those
those with awith
recommended
recommendedmarker
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pen on
above
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may
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inbottle
in the
some
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to ensure
to ensure
circumstances
circumstances that that
lead
lead toto underestimation
underestimation ofof microbial
microbial
they are
they not
numbers.
numbers.arecounted
not counted twice.twice.
Ignore Ignore
any airany
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air bubbles
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in theingel.
the gel.
Determining
Determining whether
whether to
to test
test fuel/oil
fuel/oil or
or water
water phase
counted
Colonies
phase
Day 1
D.2 ItD.2
is usually
It is usually
possiblepossible
to count
to count
up toupabout
to about
250 colonies.
250 colonies.
If theIfnumber
the number
of colonies is toois too
of colonies
Depending
Depending
numerous
numerous toon
to count, on the
the
count, system
system
visually sampled
sampled
visually
compare
comparetheand
and test
thethe
theto sampling
sampling
test
the the location,
to Test location,
Results samples
Test Results
Chart (seemay
samples
Chart may
(see
page contain
contain
page
9) fuel/oil
fuel/oil and/or
9) and/or free
free water
water phase.
phase.
MicrobMonitor2
MicrobMonitor2 can be used to test either fuel/oil or associated water. Water phase will usually contain far
holding
holding
the test
the against
test can
against
a be used
white
a to
white test
background. either
background. fuel/oil
The chart
The or
chartassociated
provides
provides
an water.
estimate
an Water
estimate
of the
of phase
the will usually contain far more
more
colonymicroorganisms
count. than
microorganisms
colony
count. than areare present
present in
in fuel
fuel or or oil
oil phase
phase andand therefore
smaller volume
volume is is usually
usually tested.
tested. Because
Because free
free water
water
phase
phase may may not not always
always be be recovered
ensure consistency
consistency whenwhen conducting
conducting regular
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
microbiological
monitoring,
monitoring, it it is
is usually
usually recommended
recommended that that fuel
fuel or
or oil
oil phase
phase from
from above
above any
any free
free water
water is is tested.
tested. However,
However, inin some
some cases
cases
it
it isis recommended
recommended to to additionally
additionally test
test water
water phase
phase (e.g. (e.g. samples
samples from
from aircraft
aircraft fuel
fuel tanks,
tanks, and
and when
when investigating
investigating
suspected
suspected microbial
microbial growth
growth inin facilities).
facilities). Further
Further information
information is is provided
provided inin the
the relevant
relevant technical
technical guidance
guidance documents
documents
NotesNotes
on examining
listed on examining tests tests
listed on on page
page 2. 2.
Colonies
Colonies
are usually
are usually
circularcircular
but maybut have
may have
irregular
irregular
edges. edges.
Different
Different
types types
of microorganisms
of microorganisms
can grow
can grow
at different
at different
rates inrates
the in
gel
the
culture
gel culture
To determine
To and
determine if
if the
the sample contains free water hold it
it up to the light
light and visually examine it.
it. It
It may help to gently swirl
swirl the
mediummedium therefore
and therefore the sample
colonies contains
the colonies
may be may offree
be water
different
of holdsizes.
different
sizes. At up to
theAt therecommended
recommended
the andincubation
visually examine
incubation
temperature, may
temperature,
colonieshelp ofto
colonies gently
bacteria and
of bacteria the
and
sample
sample
yeastsyeasts to
usuallyusually
develop create
to create
develop
withina vortex.
awithin
1vortex.
to 2 1days In order
to 2Inand
order
days remainto enable
andtoremain
enable
quite access
access
small.
quite small.
Mouldsto
to the
Moulds
developsample
the sample
develop with
more more
slowly the
with slowly
the measuring
measuring
but eventually
but eventuallydevices
devices
produce produce provided,
provided,
large large
colonies it may
it may
colonies
which be
be
which
necessary
necessary
may have
may have
a powdery to transfer
atopowdery
transfer some
some
or "fuzzy"
or sample,
sample,
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after
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shaking,
Generally,
Generally,
theto a
tomore
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smaller
more clean
clean
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container.
there there
are inare
theintest
thebottle, the smaller
test bottle, the smaller
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the colonies
will be.
will be.
IgnoreIgnore
any microbial
any microbial coloniescolonies
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after the
afterspecified
the specified
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incubation
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is complete
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Preparing
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Colonies
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quickly
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If the MicrobMonitor2
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Occasionally, MicrobMonitor2
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some some
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test
antioxidant bottles
bottles
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ambient
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temperature
and produce
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light light
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)

Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Location
Location
Location
Location
Location

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
they
peachpeach are
or orange
they used.
or orange
are Remove
colourcolour
used. in the in
Remove gelthethe
gel seal
(usually
the (usually
seal from
within
from 12 the
within
the 12cap
hours).
cap of
hours).
This the
ofcolour
the bottle
This colour
change
bottle before
change
will not
before using
will
interfere
using with MicrobMonitor2
each
not interfere
each the
withgrowth
the growth
MicrobMonitor2 of anyof test. Avoid
Avoid prolonged
microorganisms
any microorganisms
test. and and
prolonged
in mostexposure
in cases
most cases
exposure of the
microbial
ofmicrobialMicrobMonitor2
thecoloniescolonies
can be
MicrobMonitor2 can betest
counted bottles
counted
test or estimated
bottles to
to direct
or estimated
ignoring
direct sunlight
ignoring
sunlight or
the background
the other
orbackground bright
other colour.
bright light
colour.
Inlight Induring
exceptional
during preparation,
exceptional
casescases
preparation, theincubation
the interference
incubationmay and
interference may
and
examination colonies.
colonies.
In such In cases
examination of of tests.
be so be
strong
so strong
that users
that may
users find
mayit find
difficult
it difficult
to distinguish
to distinguish
colourcolour
interference
interference
from anfromestimated
an estimated
countcountof 10,000of 10,000 such cases
the the
tests.
fuel should
fuel should
be retested
be retested
using using
a smaller test volume
a smaller test volume
(e.g. 0.01
(e.g. ml)
0.01soml)
that
so the
thatinterference
the interference
effecteffect
is diluted
is diluted
out; ifout;
the iforiginal
the original
result result
was was
genuinely
genuinely
due todue
heavy
to heavy
microbial
microbial
contamination
contamination it wouldit would
normallynormally
be expected
be expected
that the
thatretest
the retest
will showwill a
show
discernible
a discernible
number number
of red/purple
of red/purple
Cleanliness
Cleanliness
colonies
colonies
and the
and the during
during
background
background testing
testing
colour colour
will bewill
lessbeintense.
less intense.
Some Some
bacteria
bacteria
can, oncan,
prolonged
on prolonged
MicrobMonitor2
MicrobMonitor2 tests can incubation,
be incubation,
conductedspreadspread
through
through
on-site, the gel
in
in an
theproducing
office gel producing
or in a large
a a large
irregularly
laboratory.irregularly
shaped
However, shaped
colony,
colony,
streakstreak
precautions or patchor patch
should of redof
be orred or
taken to
purplepurple
colourcolour
in the in thetests
gel. gel. can
These Thesebe
bacteria conducted
bacteria grow on-site,
usuallyusually quickly
grow quickly
andan office
thus
and testsor
ifthus in examined
ifare aare
tests laboratory.
examined However,
while while
the colonies precautions
the colonies
are stillare still should
small be1after
small after
(e.g. (e.g. taken
or 2 1 orto2
ensure
ensure
days incubation)
days testing
testing
incubation)
they areis
they conducted
is conducted
more
are more in
easily easilya
counted. reasonably
in acounted.
reasonably
The centre clean
clean
The centre
of each area
ofarea
each
streakto avoid
tostreak
oravoid
patch contamination
contamination
or patch
shouldshould
be counted of
be countedthe
of as
the sample
sample
a single
as a single
colony.and
and the test
the Continue
colony. by
test by
Continue
microorganisms
microorganisms
incubation
incubation
and count
and count
any other
anythan
other
new new those
than those
colonies
coloniesin the
in which
which the test
test
develop. sample.
sample. Wash
develop. Wash hands
hands before
before and
and after
after testing.
testing. During
During testing
testing wear wear clean,
clean, nitrile,
nitrile,
vinyl
vinyl or
or polythene
polythene gloves.
gloves. Avoid
Avoid touching
touching areas
areas of
of the
the syringe
syringe and
and loop
loop dispenser
dispenser which
which come
come into
into direct
direct contact
contact with
with the
the
the MicrobMonitor2
sample of MicrobMonitor2
of MicrobMonitor2
sample and
leafletavoid
and avoid touching
touching the
the inside
inside of
of the MicrobMonitor2 test test bottle
bottle cap
cap and
and neck
neck when
when dispensing
dispensing sample.
See technical
See technical leaflet
EP157 EP157
Technical
Technical
Assistance
Assistance
Reading
Reading
ResultsResults for further
for further
information
information
on reading
on reading
and interpreting
and interpreting
tests tests
sample.
with unusual
with unusual
or atypical
or atypical
appearance.
appearance.
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW
Results
Results
that that
count
count
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW
7

7

MICROBMONITOR2
® ®
MICROBMONITOR
MICROBMONITOR2
MICROBMONITOR ® ® ® ®
2 2
E Calculate
DISPOSAL
TEST the number of microbial CFU in the sample.
PROCEDURE
x
Wear gloves For avoid
and tests oftouching
fuel, numbersinside
of microbes are conventionally handsasafter
expressed number of CFU litre.
perMicrobMonitor2
A. Add sample to thethe of the bottle
MICROBMONITOR or
® cap
2 2® testand wash
bottle. handling any test
bottles x
showingFormicrobial growth.
tests of water phaseBefore disposal,ofdisinfect
or oil, numbers microbesall areMicrobMonitor2 test bottles
conventionally expressed as which
number show growth
of CFU by
per ml.
immersion of the opened bottles in a strong disinfectant solution (e.g. household bleach) overnight or by incineration or by
Follow the appropriate steps below to add distillate fuel or biofuel (A.1), heavy/residual fuel or oil (A.2), water (A.3)
using
a) chlorine release
Results tablets
of test of (available
fuel phase separately).
sample:
or surfaces deposits (A.4) to the MicrobMonitor2 test bottle.
Decontaminated
If 0.5 A.1 Aviation
ml of fuel hastests,
beenunused
kerosene, tests
other
tested, or those
middle
multiply showing
distillate
the number fuels no
ofand microbial
gasoline.
colonies growth
counted or can be disposed
estimated of
by 2000 to as normal
give waste of
the number in
accordance
microbial CFU with
perlocal
litrewaste regulations.
of fuel. If there are no colonies after the specified incubation time there are less than 2000
microbial
A.1.1CFU per litre ofprior
fuel.to testing, shake the sample for 30 seconds. If fuel samples contain
Immediately
visible free water, the sample should be allowed to stand for 12 minutes to enable water to
STORAGE
If 0.25settle AND
ml offrom
fuel theSHELF
has beenLIFE
sample tested,
to be multiply
tested. theFor number of colonies
consistency, it may becounted or estimated
advisable to apply bythis4000 to give the number of
microbial CFU
settling timeper
forlitre of samples
all fuel fuel. If even
thereifare
freeno colonies
water after the specified incubation time there are less than 4000
is not observed.
microbial
The CFU
shelf life of per litre of fuel. is 1 year from the date of production. The expiry date is printed on
MicrobMonitor2 the batch label of each
MicrobMonitor2 box. Store MicrobMonitor2 tests between 2 to 22°C, in the dark. The expiry date applies only to product
If a different
stored volume of
as stipulated. fuel has been
If refrigerated (2 tested, then
- 8°C) the a calculation can
MicrobMonitor2 test be
will made
usuallyas
stillfollows:
be useable for up to a year beyond the
designated expiry date. Do not store MicrobMonitor2 frozen. Exposure to freezing temperatures during transport is
acceptable.Number of microbial
Avoid exposure CFU per litre = Number
of MicrobMonitor2 testsoftocolonies counted
light during or estimated
storage and use. xA1000slightly pink discolouration in the
MicrobMonitor2 gel may develop over time during storage Volumebut of fuel tested
this will not (ml)
affect the performance of the test.
A.1.2 Remove the cap of the MicrobMonitor2 test bottle taking care not to touch the inside
b) and Results
placeof testaof
it on water
clean phase or oil:
surface.
ALSO AVAILABLE FROM ECHA
If 0.1 ml of water has been tested, multiply the number of colonies counted or estimated by 10 to give the number of
microbial CFU per ml of water. If there are no colonies after the specified incubation time there are less than 10 microbial
For
CFU use with
per ml MICROBMONITOR2
of water. ® ®
2
If 0.01
The ml of water
following itemsor oil assist
can has been tested, multiply
in performing the number oftest
the MicrobMonitor2 colonies counted or estimated by 100 to give the number
procedure:
of microbial CFU per ml of water or oil. If there are no colonies after the specified incubation time there are less than 100
microbial CFU per ml of water or oil.
x MicrobMonitor Sampling Kit (ECHA16/SB) - A sterile, polypropylene bottle for sampling fuel systems and which
A.1.3 Open
enables theseparation
easy peel packofcontaining
water phase a syringe at the
from fuel handle
phase; end and
includes remove
alcohol for decontaminating sample
wipesthe
If a different
syringe volumecare
taking of water
not toortouch
oil hasthe
been tested,
lower then
barrel andanozzle.
calculation can be made as follows:
points.
Number of microbial CFU per ml = Number of colonies counted or estimated
x Incubator (ECHA14/IN) - enables optimal, consistent
Volume of waterincubation temperature
or oil tested (ml) of MicrobMonitor2 tests; holds 12
MicrobMonitor2 test bottles; available battery or mains (110/230 Volt) operated.
Note
x MicrobMonitor Bumpers (ECHA16/TB) - soft, rubber-like, hemispheres which can be stuck to a table top and enable
easy breaking
If the number of coloniesandisshaking of MicrobMonitor2
too numerous to count and gel.
a sample volume of 0.5 ml or 0.25 ml of fuel or 0.1 ml or 0.01 ml
of water or oil was tested, the Test Results Chart can be used directly to determine the approximate number of
A.1.4 Using the syringe, draw fuel from approximately 3 cm below the surface of the
microbial
x CFU per litre of tablets
fuel or per ml of water- or oil.
Chlorine
sample. Release
If there is less than(ECHA21/CP)
6 cm depth oftofuel phase,MicrobMonitor2
disinfect test bottles
draw sample from showing microbial growth for easy
approximately
disposal.
halfway down the fuel phase. Avoid the transfer of visible interfacial particulate, water
INTERPRETATION OF TEST
droplets or emulsion in theRESULTS
aliquot to be tested.
x Sterile swabs (ECHA15/SW) - swabs for sampling surface contamination (e.g. tank surfaces, filter cartridges etc.).
There are no universally accepted limits or specifications for microbiological contamination in fuels or oils. The number of
x
microbialLong,
CFUsterile
which bulb pipettes
define moderate(ECHA07/PP/EL23) - to enable water
and heavy contamination to will
levels be removed
depend on from the bottom
a number of samples.
of factors including the
fuel type and intended use, the sampling location, whether the sample is taken from supply and distribution facilities or
from point of end use, the intended fuel storage time and specific operational circumstances. Some guidance limits are
Other
shown Test
in
A.1.5 Kits2fuel
Table
Draw & Ancillary
below. These
into the Items
limitsuntil
syringe areit intended
is more thanto half
provide early
full and indication
then, with thethat microbial
syringe nozzle growth
is occurring in the
facility sampled. press
uppermost, Higherthelevels of contamination
plunger to expel air. Expelwill often befuel
surplus required before
until the end the onset
of the of operational problems or any
plunger
detrimental
x aligns effect
with
SigTestsΠ:
SigTests®:theon
Sig fuelfor
mark fitness
Sulphide the for use. volume
required
(ECHA02/SS) For
— further information
of sample
simple test kit(see see the
table
for Sulphate1). technical
Reducing guidance leaflets
Bacteria (SRB) listedcan
which on cause
page 2.serious
corrosion and sulphide spoilage in fuel, oil and water systems. Can be used on-site or in the laboratory.
Table 2
x Sample
Biocide Rapide (ECHA01/BR) — simple Moderate kit for assessing presence
rapid test Heavy and approximateComment
concentration of
commonly used fuel and oil biocides.
Fuel from aircraft fuel tank drain 4000 to 20,000 CFU / litre >20,000 CFU / litre IATA
Contamination
x Any level and bottom sampling devices (ECHA23) — for safe sampling from fuel and oil storage tanks in
Limits
Water from
A.1.6 aircraft fuel tank drain 1000 to 10,000 CFUMicrobMonitor2
/ ml >10,000 CFU /and
ml
Insert the tip of the syringe
compliance with industry standards. into the neck of the test bottle
Bulkdispense the sample
representative by sample
fuel fully depressing
from the syringe plunger.
x supplyFor&our 4000 see
full range of test kits and ancillaries
distribution to 20,000 CFU / litre >20,000 CFU / ml
e.g. Aviation fuel,
Fuel from bottom / drain of supply &
10,000 to 100,000 CFU / litre >100,000 CFU / litre diesel, marine gas
Technical
distributionsupport and services
tank / filter
oil
Water phase from bottom / drain of
ECHA Microbiology Ltd.fuel
provides 100,000 to 1,000,000 CFUour/ product
ml >1,000,000 CFU /also
ml offers laboratory testing,
supply & distribution tank /afilter
full technical service to support range. ECHA
training courses, consultancy and on-site attendance for audit and investigation of microbial contamination and corrosion
associated with
Lubricating petroleum
/ hydraulic oilproducts and petroleum,
in circulation marine,
100 to 10,000 CFUaviation
/ ml and other industrial
>10,000 CFUfacilities.
/ ml
A.1.7 Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample
details and test date on the label provided and stick it on a narrow side of the bottle.
Lubricating / hydraulic oil sump 1000 to 10,000 CFU / ml >10,000 CFU / ml
Proceed to step B.

10

8 4 TTRRUUTSSRTTU +

+
DDT E D I INNNNO
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+

+
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+
+
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E I V E AACC
VET

CURATE DEPENDABLE
CAUCRCAUTREAT E D E PDEENPDEANBDLAEB L E
MICROBMONITOR22 Test Results Chart
® ®
A.2 Heavy andFOR

PREPARATION residual fuels and oils.
TESTING
Number of colonies in
Microbial Content of
Sampling Appearance
A.2.1 Immediately after incubation
prior to testing,
considerations test bottle
shake the sample for 30 seconds. Volume tested
Sample
(counted or estimated)
A.2.2 Remove the cap of the MicrobMonitor2 test bottle and place it on a clean surface.
Sampling should be conducted in a clean and consistent manner which prevents introduction of background contamination
into the sample. Once fuel samples have been taken, any microbes present will tend 0.5 mltoofslowly die and it is important
fuel (syringe) <2000 CFU toper litre
A.2.3
test Open as
samples the peel
soon aspack containing
possible. a loop
If the test dispenser
is carried and than
out more remove
24 hitafter
taking care not
sampling, it istopossible
touch the
thatloop end and
the results the
will not
lower part of the handle.
reflect the viable microbial content present at the time of sampling. To avoid this, part or all of the MicrobMonitor2 test
procedure can be conducted at the sampling location. 0.25 ml of fuel (syringe) <4000 CFU per litre
A.2.4 Immerse the loop end into the fuel/oil to approximately 3 cm below the surface of the sample or, if there is less than 6
None
cm depth of fuel/oil phase, to halfway down the fuel/oil phase. Remove the loop from the sample and allow surplus fuel/oil
Sample type
to drain off; and
ensure recommended
a film testwithin
of fuel/oil is trapped volumes
the loop. 0.1 ml of water (syringe) <10 CFU per ml
MicrobMonitor2 can be
A.2.5 Insert the loop intoused to test
the neck of athe
wide variety of petroleum
MicrobMonitor2 products
test bottle and
and stab associated
it into the ml
0.01 water.
gelofand Theoilvolume
agitate
water or it briefly of
to sample
transfer
which should be tested will depend on the type of sample. Refer to the table below to determine the <100 CFU per ml
appropriate
the sample. (loop dispenser)
volume of sample to test and which measuring device to use.
A.2.6 Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample details and test date on the label
provided
Table 1 and stick it on a narrow side of the bottle. Proceed to step B.
Recommended test Measuring
Sample
Sample
A.3 Water phase associated with fuel/oil. 10 Colonies
volume
volume device
device
A.3.1 If akerosene
Aviation test of water phase is required, allow the sample
kerosene to stand
If possible, untilthe
count freeexact
water0.5hasml
mlsettled to the bottom of the sample. It
is recommended the water phase test is conducted after any fuel/oil
number phase and
of colonies test. The test procedure is essentially the same as
described
Other above
middle but an
distillate appropriate
fuels technique
(e.g. diesel, marine for
gasseparating/removing
calculate
oil, heatingtheoil),
actual water&from0.1
number
biofuels themlsample
of water
needs to be used. 100 CFU per ml
(syringe)
of CFU present in the sample0.25 ml ml Syringe
Syringe
gasoline
gasoline
A.3.2 As appropriate for the volume of sample being tested (see table 1) use the loop 0.01 dispenser or or
ml of water syringe
oil to measure the
required volume ofaircraft
water. fuel tank drains 1000 CFU per ml
Water phase from drains ml(loop dispenser)
0.1 ml
a) To test
Heavy and0.1residual
ml of water
fuelsusing the syringe;
fuels ml
0.01 ml
hydraulic 100 Colonies

Lubricating,
The water phase can and
be drawnother oils
oils
directly from the bottom of the sample and added ml the MicrobMonitor2
0.01 ml
0.5toml of fuel (syringe)
Looptest bottle.
Loop
c. 105 CFUToper litre
enable easy access to the
Water phase from fuel storage tanks water it
tanksmay be necessary to first decant off some fuel from
0.01 the
ml
ml sample and/or transfer the water
If possible, count the exact
phase to a separate small, sterile container using a syringe or a long sterile pipette (supplied separately). Mix gently by
number of colonies and
swirling but avoid mixing the fuel and water layers together. 0.25 ml of fuel (syringe) 105 CFU per litre
The volume of sample tested can be changed in order calculate the actual
to increase number the detection level of the test.c.However,
or decrease
testing volumes of CFUmay present in the sample
Open the peel greater than thosea recommended
pack containing above
syringe at the handle end and in some
removecircumstances
the syringelead to underestimation
taking care not to touch of the
microbial
lower
numbers.
barrel and nozzle. Insert the barrel of the syringe into the sample so that the nozzle 0.1isml
in of
thewater
water(syringe)
phase. Drawc.water 103 CFU
intoper ml
or
the syringe, remove from the sample and then, with the syringe nozzle uppermost, expel air. Expel surplus water until the end
Determining
of the plunger aligns whether with tothetest
0.1 mlfuel/oil
mark. or water
Insert the tipphaseof the syringe into the neck0.01
Estimate 100 colonies if
of the MicrobMonitor2
ml of water or oil test bottle and
dispense the sample by fully depressing the syringe plunger. Replace the cap on the(loop MicrobMonitor2 c. 104 CFU per ml
result is similar to picture dispenser) bottle and tighten.
Record the sample details and test date on the label provided and stick it
Depending on the system sampled and the sampling location, samples may contain fuel/oil and/or free water phase.on a narrow side of the bottle. Proceed to step
B.
microorganisms than are present in fuel or oil phase and therefore a smaller volume is usually tested. Because free water
b) To test
phase may 0.01not
ml of water using
always the loop dispenser;
be recovered in samples, to ensure consistency when 0.5conducting regular microbiological
ml of fuel (syringe) c. 106 CFU per litre
monitoring, it is usually recommended that fuel or oil phase from above any free water is tested. However, in some cases
itA issyringe can be used
recommended to to draw watertest
additionally fromwater
the bottom
phase of the samples
(e.g. sample and fromthen a drop
aircraft fuelof tanks,
water placed
and when into the loop and
investigating
added to the
suspected MicrobMonitor2
microbial growth intest bottle. Alternatively,
facilities). transfer is
Further information the water phase
provided in thetorelevant
a0.25
separate small,
ml technical
of fuel sterile
guidance
(syringe) container
c. 10using
6 CFUaper litre
documents
syringe
listed onorpage
a long2. sterile pipette (supplied separately). Estimate Avoid transfer of fuel. ifInvert the container with the separated water
1000 colonies
three times to mix. result is similar to picture
To determine if the sample contains free water hold it up to the light and visually examine 0.1 ml ofit.water
It may(syringe)
help to gently c.10 4 CFU
swirl theper ml
Removeto
sample the cap ofathe
create MicrobMonitor2
vortex. In order totest bottleaccess
enable and place to theit on a clean
sample withsurface. Open the devices
the measuring peel pack containing
provided, it may a loop
be
dispenser to
necessary and remove
transfer some it taking
sample, care not
after to touch
shaking, to athe loop clean
smaller end and the lower part of the handle. Immerse the loop end
container.
into the separated water or fill the loop with a drop of water using a syringe. Allow surplus 0.01 ml of water
water or oil
to drain away but c.10 ensure theper ml
5 CFU
circle of the loop remains filled with water; ®note, if a residue of fuel is present this can impair (loop thedispenser)
ability of the loop to fill with
Preparing the MICROBMONITOR2 test bottles
water. Insert the loop into the neck of the MicrobMonitor2 test bottle and stab it into the gel and agitate it briefly to transfer
the sample. Replace the cap on the MicrobMonitor2 bottle and tighten. Record the sample details and test date on the
If the MicrobMonitor2 test bottles have been stored refrigerated, allow them to equilibrate to ambient temperature before
label provided and stick it on a narrow side of the bottle. Proceed to step B.
they are used. Remove the seal from the cap of the bottle before using each MicrobMonitor2 test. Avoidc.10 prolonged
7 CFU per litre or
exposure of the MicrobMonitor2 test bottles to direct sunlight or other bright light 0.5during
ml of fuel (syringe) incubation and
preparation,
Note The MicrobMonitor Sampling Kit (supplied separately) is a sample bottle which has a side siphon and water collection above
chamber to enable easy separation of water from fuel samples.
c.107 CFU per litre or
Estimate 10,000 or more 0.25 ml of fuel (syringe)
Cleanliness during testing above
colonies if result similar to
picture
MicrobMonitor2 tests can be conducted on-site, in an office or in a laboratory. However, 0.1 ml of precautions
water (syringe)should c.10be taken
5 CFU pertoml or above
ensure testing is conducted in a reasonably clean area to avoid contamination of the sample and the test by
microorganisms other than those in the test sample. Wash hands before and after testing. During testing wear clean, nitrile,
0.01 ml of water or oil
vinyl or polythene gloves. Avoid touching areas of the syringe and loop dispenser which come into direct contact c.106 CFU with the
per ml or above
(loop dispenser)
sample and avoid touching the inside of the MicrobMonitor2 test bottle cap and neck when dispensing sample.
5HVXOWVWKDWFRXQW
Results that that
count
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW
Results
Results
that count
count
3

5
9
®
®2® ®2®
MICROBMONITOR
MICROBMONITOR
MICROBMONITOR2
MICROBMONITOR2
MICROBMONITOR 2 ®
MICROBMONITOR2
DISPOSAL
DISPOSAL
DISPOSAL
WHAT IS MICROBMONITOR
THE MICROBMONITOR ®
2 TEST?
WHATDISPOSAL
IS THE 2 ® TEST?
Wear gloves
WearWear and
andis avoid
MicrobMonitor2
gloves
MicrobMonitor2
gloves and
avoid atouching
is avoid simple
touching the
the inside
and
touching easy
the
inside toof
inside the
use
theof
oftest bottle
test
the
bottle kit or
bottlecap
orwhich
cap or and
enables
cap wash
and hands
the
wash after
quantitative
hands handling
after any
assessment
handling MicrobMonitor2
of the
any viable
MicrobMonitor2
MicrobMonitor2 test
microbial
test test
Wear gloves a simple
and avoid and easy
touching to
theuse
inside ofkitthewhichbottle or and
enables capthe wash hands
andquantitative
wash afterassessment
hands handling
after any
of the
handling anyviable microbial
MicrobMonitor2 test
bottles showing
content microbial
in petroleum growth.
products Before
including disposal,
liquid disinfect
fuels, oils all
and MicrobMonitor2
associated test
water. bottles which
MicrobMonitor2 show growth
enables by
on-site or
bottles
bottles
content showing
showing
in petroleum
bottles showing microbial
microbial products
microbial growth.growth.Before
including
growth. Before
disposal,
liquid
Before disposal,
fuels,
disposal, oils disinfect
disinfect
and all
disinfect all MicrobMonitor2
all
MicrobMonitor2
associated water.
MicrobMonitor2 test
test bottles
whichwhich
MicrobMonitor2
test bottles
bottles show
enables
which show
growthgrowth
on-site
show or by
by
growth by
immersion
immersion
laboratory of
immersion
of the
laboratory
the
testingopened
testing
of the
opened
in opened
accordance bottles
in accordance
bottles in
bottles
in a
a
withstrong
inwith
a
strong
ASTM disinfectant
ASTM
strong
and and
disinfectant
disinfectant
IP Standard solution
IP solution
Standard (e.g.
solution
(e.g.
Methods household
Methods
(e.g.
ASTM ASTM bleach)
household
household
D7978 D7978 overnight
and
bleach)
bleach)
and IP613 or
-or
overnight
overnight
IP613 - by
by or
Determinationincineration
Determination
by or
incineration
incineration
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CodifiedCodified (6640-99-834-3573).
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the MicrobMonitor2
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test
test
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procedure:
test
test procedure:
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as CFU/litre of fuel, CFU/ml
waterwater associated
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with fuel.
x
x x MicrobMonitor
MicrobMonitor Sampling Sampling Kit (ECHA16/SB)
(ECHA16/SB)
Kit (ECHA16/SB) -- 44
A sterile,
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x x MicrobMonitor2
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test bottles
x SterileSterile syringes* - enables optimal, consistent incubation temperature of MicrobMonitor2 tests; holds 12
x x
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x x BottleBottlelabelslabels
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leaflet. (ECHA16/TB)
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x
x x Chlorine Release
Chlorine tablets
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tablets -- to
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patented by ECHA ECHA Microbiology
Microbiology Ltd. and and
it marketed
is marketed worldwide
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disposal.
ECHAECHA and
and its its distributors.
distributors.
x
x x Sterile swabs (ECHA15/SW) -- swabs for
for sampling surface contamination (e.g. tank
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x SterileSterile
swabs
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swabs swabs for
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The MicrobMonitor2 is usedis used
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Other
Other Test
Other Kits
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about Other
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x
x x SigTests®: Sig
Sig Sulphide
SigTestsΠ: Sig (ECHA02/SS) —
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test kit for Sulphate Reducing Bacteria (SRB) which can
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(ECHA02/SS) —
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MicrobMonitor2
MicrobMonitor2 in specific
in specific applications
applications is provided
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the technical application
application leaflets
leaflets listedlisted
below below
(free (free to download
to download
x
x x
from fromBiocide
Biocide Rapide
Rapide (ECHA01/BR)
Biocide Rapide (ECHA01/BR)
(ECHA01/BR) —
— simple
These These
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leaflets
simple
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include
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x Biocide Rapide (ECHA01/BR) — simple rapid test for assessing presence and approximate concentration of
resultsresults
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oil contamination
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x
x x EP096
x Any EP096
level
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(ECHA23) in Accordance
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tanks
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with industry
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compliance with industry
industry standards.
standards.
industry standards.
x x EP119EP119 Routine
Routine Monitoring
Monitoring of Aviation
of Aviation Fuels Fuels in Supply
in Supply and and Distribution
Distribution Facilities,
Facilities, Airport
Airport Depots
Depots and and Into-plane
Into-plane
x
x For our full
x Operations
Forfull
For our our range
Operations
with
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withtest kits and
MicrobMonitor2.
full range
range ofkits
of test testand ancillaries
MicrobMonitor2.
kits ancillaries see
and ancillaries
ancillaries www.echamicrobiology.com.
x For our full of test kits and see
x x EP132EP132
Technical Routine
support
Technical and
support Monitoring
services of Diesel Fuel Tanks and Distribution Systems with MicrobMonitor2.
with MicrobMonitor2.
Technical Routine
support
Technical andand
Monitoring
support services
of
services
and
Diesel
services
Fuel Tanks and Distribution Systems
x
ECHAxMicrobiology
EP166 EP166 Routine
Ltd.
Routine Monitoring
provides
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serviceonto
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our productInstallations
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also
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laboratory
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ECHAECHA
ECHA Microbiology
Microbiology Ltd.
Microbiology Ltd.
Ltd. provides
provides full
provides a fullDiesel
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for audit and
and investigation of
of microbial contamination and
and corrosion
training courses,
training consultancy
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on-site attendance for
for audit
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and investigation
investigation of
of microbial
microbial
investigation contamination
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microbial contamination and
and corrosion
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x x EP157
associated
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with petroleum
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PREPARATION
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Sampling
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Sampling should be
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litre or CFU/ in
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Sampling should conducted clean consistent manner which prevents introduction background contamination
into the
the sample. OnceOnce
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Contamination
into into the sample. fuel samples taken, any microbes present will tend
into the sample. Once fuel samples have been taken, any microbes present will tend to slowly die andimportant
sample. Once fuel have been taken, any microbes present will tend to to
slowly slowly
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Microbial
test samples as soon as possible. If the test is carried out more than 24 h after sampling, it is possible that the results will
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Microbial
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h after
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not
sample
test samples
test as soon
as as possible.
as If theIf the carried
is out more
out than than
24 h after
24 sampling, it is possible
sampling, it that the
that the not
will
sample
reflect the viable

viable microbial content present at the
the time of sampling.
sampling. To avoid
avoid this, part
part or allorofall MicrobMonitor2
theofMicrobMonitor2 test test
reflect
reflect the
reflect the
the viable
viable microbial
microbial content
microbial content
present
content present
at
present at the
the time
attime of
time of
of sampling.
To
sampling. To
To avoid
this,
avoid this,
this, orpart
partall of
or the
all the MicrobMonitor2
of the MicrobMonitor2 test test
procedure
procedure can
can be
procedure can
be conducted at
at the
be conducted
conducted
conducted atsampling
theat the sampling
sampling location.
sampling location.
location.
procedure can be the location.
Sample type
Sample and
typetype recommended
and and test
test volumes
recommended test
Sample
Sample type recommended
and recommended test volumes
volumes
volumes
CFU/
MicrobMonitor2 can
can be
MicrobMonitor2 canused
be to
to test a wide variety of
of petroleum products and
and associated water. The
The volume of
of sample
MicrobMonitor2
MicrobMonitor2 be be used
canused used to
toatest
test a
a wide
wide
test variety
variety
wide of
of petroleum
petroleum
variety products
products
petroleum products and
and associated
associated water.
water.
associated water. The
The volume
volume
volume of
of sample
sample
sample
which should
which
whichwhich
should be
shouldtested
be tested will
be tested
tested depend
will depend on
will depend
depend the
on the type
on type of
the type
typesample. Refer
of sample.
sample.
of sample. to the
Refer
ReferRefer table
to table
to the below
the table
table
below to
belowdetermine the
to determine
determine
to determine appropriate
the appropriate
appropriate
the appropriate
should be will on the of to the below to the
Day _______
_______
volume of
volumesample
of to test and which measuring device to use.
of sample to test and
and which measuring device to
to use.
Counted
volume of sample to test

toand
test which measuring device to use.
Colonies
volume sample which measuring device use.

Counted
Colonies
Table 1
TableTable
1 1
Table 1
Day
Recommended
Recommended test
Recommended Measuring
test test Measuring
Measuring
Sample
Sample
Sample Recommended test Measuring
Sample volume
volume
volume device
device
device
volume device
Counted
Colonies
2 test results
Counted
Colonies
Day 4
Aviation kerosene 0.5

0.5 ml
Day 4
Aviation
Aviation kerosene
kerosene
Aviation kerosene 0.5 ml
ml0.5 ml
Other middle
Other
OtherOther
middle distillate
middle
distillate fuels
distillate (e.g.
fuels
fuels fuels diesel,
(e.g.
(e.g. (e.g. marine
diesel,
diesel, marine gas
gas oil,
marine gasheating
oil, oil),
oil), biofuels
oil, heating
heating
heating &
oil), biofuels
biofuels
biofuels & 0.25& ml
middle distillate diesel, marine gas oil, oil), &
0.25 0.25 ml
ml ml Syringe
Syringe
Syringe
gasoline
gasoline
gasoline 0.25 Syringe
gasoline
Counted
Colonies
Water phase from aircraft fuel

fuel tank
fuel drains 0.1
0.1 ml
Counted
Water phase from aircraft tank drains

drains 0.1 ml
ml
Colonies
WaterWater
phase from from
phase aircraft
aircraft tank
fuel drains
tank ml0.1
Day 3
Day 3
Heavy
Heavy and
and residual
Heavy fuels
and residual
residual
residual fuels
fuels fuels 0.01
0.01 ml
0.01 ml
ml ml
Heavy and 0.01
®
MICROBMONITOR
Lubricating,
Lubricating,
hydraulic and
and other
hydraulic other oils
and other
other oils
oils oils 0.01
0.01 ml
0.01 ml
ml ml LoopLoop
LoopLoop
Lubricating, hydraulic and 0.01
Water phase
Water
WaterWater
phase from
phase fuel
fuel storage
from
from from tanks
fuel storage
storage
storage tanks
tankstanks 0.01
0.01 ml
0.01 ml
ml ml
phase fuel 0.01
counted
Colonies
counted
Colonies
Day 2
Day 2
The
The volume
The of
of sample tested can
can be changed in
in order to
to increase or
or decrease the
the detection level of
of the test. However,
The volume
volumevolume of
of sample
sample tested
tested
sample tested can
be be
be changed
canchanged
changed in
in order
order to
to increase
increase
order increase or
or decrease
decrease
decrease the
the detection
detection level level
detection theof
level the
the test.
oftest. However,
However,
test. However,
testing volumes
testing greater
volumes than
greater those
than recommended
those above
recommended may
above in
may some
in circumstances
some lead
circumstances to underestimation
lead to underestimationof
of microbial
of
testing volumes
testing greater
volumes than than
greater thosethose
recommended above
recommended may may
above in some circumstances
in some lead lead
circumstances to underestimation
to underestimation of microbial
microbial
microbial
numbers.
numbers.
numbers.
numbers.
counted
Determining
Colonies
whether
Determining to
to test
whether to fuel/oil
test or
or water phase
test fuel/oil or
or water phase
counted
Colonies
Determining whether
Determining whethertest
to fuel/oil water
fuel/oil phase
water phase
Day 1
Day 1
Depending
Depending on
on the on system
the sampled and
and the sampling location, samples may contain fuel/oil and/or free
free water phase.
Depending
Depending the the system
onsystem sampled
system sampled
sampled and
the the
the sampling
andsampling location,
location,
sampling samples
samples
location, may may
samples contain
contain
may fuel/oil
fuel/oil
contain and/or
fuel/oil and/or
and/or free
free water
water phase.
water phase.
phase.
MicrobMonitor2
MicrobMonitor2 can can
be be
used used
to to
test test
either either
fuel/oilfuel/oil
or or associated
associated water.
water. Water Water
phase phase
will will usually
usually contain
contain far far
more more
microorganisms
microorganisms than than are present
present
are in fuel
fuelinor
or oil
oil phase
phase and therefore
therefore a smaller
smaller volume is usually
usually tested. Because free water
water
microorganisms
microorganisms than are
than are present
in
present in fuel
fuel oror oil
oil phase
and
phase and
and therefore
a
therefore a
a smaller
volume
smaller volume
is
volume is
is usually tested.
tested.
usually Because
Because
tested. free
Because free
free water
water
phase may
phase not
may always
not alwaysbe
always recovered
be recovered
recovered in samples,
in samples,
samples,to ensure
to ensureconsistency
ensure when
consistency conducting
when conducting regular microbiological
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
phase may may not always be recovered in samples, to ensure consistency whenwhen conducting regular microbiological
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
0.5/0.25/0.1/0.01/other__
phase not be in to consistency conducting regular microbiological
monitoring, it is
monitoring, is usually
usually
it recommended that fuel
fuel or oil or
phase from from
above any free
free water is tested.
tested. However, in some
some casescases
monitoring, it
monitoring, it is
is usually
usually recommended
recommended
recommended that that
thator fuel
oil
fuel oil
oil phase
phase
or from
phase above
from above
any
above any free
free water
any water is
water is
is tested. However,
However,
tested. in
However, in
in some
cases
some cases
it
it is
is recommended
it is
is recommended
recommended
recommended to
to additionally
to additionally
additionally test
test water
additionally test water
water phase
water
phase (e.g.
phase samples
(e.g.
(e.g. (e.g.
samples from
samples aircraft
from
from from fuel
fuel tanks,
aircraft
aircraft and
fuel tanks,
tanks, tanks, when
and
and and investigating
when
whenwhen investigating
investigating
it to test phase samples aircraft fuel investigating
suspected
suspectedmicrobial
microbialgrowth in
growth facilities).
in Further information is provided in the relevant technical guidance documents
suspected
suspectedmicrobial
microbialgrowthgrowth in facilities).
in facilities). Further
facilities). Further information
information
Further is
is provided
is provided
information in thein
provided the
the relevant
inrelevant technical
technical
relevant guidance
guidance
technical documents
documents
guidance documents
listed on
on page
listed
listedlisted on page
page 2.
page
2. 2.
on 2.
To
To determine
To if
if the sample contains free
free water hold it
it up
up to the light and visually examine it.
it. It
It may help to
to gently swirl
swirl the
To determine
determine
determine the if the
the sample
ifsample contains
contains
sample contains free
free water
water hold hold
water hold it
it up
to theto
up the
tolight light
the and and
and visually
light visually examine
examine
visually examine it.
it. It
may It may
help help
may to
to gently
gently
help gently swirl
swirl the
the the
sample
sample to
sample
to create
to
create a
create
a vortex.
a
vortex. In
vortex.
In order
In
order to
order
to enable
to
enable access
enable
access to
access
to the
theto sample
the
sample with
sample
with the
with
the measuring
the devices
measuring
measuring devices provided,
devices it
provided,
provided, it may
mayit be
may
be be
sample to create a vortex. In order to enable access to the sample with the measuring devices provided, it may be
necessarytransfer
to somesome
sample, after after
shaking, to a
a smaller
smaller cleanclean
container.
necessary to
some
transfer some sample,
sample, after
sample, shaking,
shaking,
after to
shaking, to
to a
a smaller
clean
smaller container.
container.
clean container.
®
Preparing
Preparing the
the MICROBMONITOR
Preparing
Preparing the
MICROBMONITOR 2
2 ® test
the MICROBMONITOR
MICROBMONITOR 2 ® bottles
2 ®
test test
test bottles
bottles
bottles
If
If the MicrobMonitor2 test
test bottles have been stored refrigerated, allow them to
to equilibrate to
to ambient temperature before
theIf the MicrobMonitor2
the
IfMicrobMonitor2
MicrobMonitor2 test
test bottles
bottles havehave
bottles beenbeen
have stored
been stored refrigerated,
refrigerated,
stored allowallow
refrigerated, themthem
allow them to
to equilibrate
equilibrate to
to ambient
ambient
equilibrate temperature
temperature
ambient before
before
temperature before
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)

Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Sample Ref.
Location
Location
Location
Location
Location

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
MicrobMonitor2
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
Volume tested (ml)
they are used.

Sample Ref.
Remove the seal from

Sample Ref.
the cap of the bottle

Sample Ref.
before using each

Sample Ref.
test. Avoid
Sample Ref.
prolonged
Location
Location
Location
Location
Location

Type
Type
Type
Type
Type
Date tested
Date tested
Date tested
Date tested
Date tested
they
they they are used.
are used.
are used.
Remove Remove the from
the seal
Remove the seal from
seal from the cap
the cap
the cap
of the of bottle
of the bottle
the bottle
beforebefore using
usingusing
before each
eacheach MicrobMonitor2
MicrobMonitor2 test.
test. test.
MicrobMonitor2 Avoid
AvoidAvoid prolonged
prolonged
prolonged
exposure of the MicrobMonitor2 test bottles to direct sunlight or other bright light during preparation, incubation and
exposure
exposure of the
exposure of the MicrobMonitor2
the
of MicrobMonitor2
MicrobMonitor2 test
test bottles
test bottles to
to direct
to direct
bottles sunlight
sunlight
direct or
or other
or other
sunlight brightbright
other light light
bright during
light during preparation,
preparation,
during incubation
incubation
preparation, and and
incubation and
examination tests.
of
examination of
tests.
Cleanliness
Cleanliness during
Cleanliness testing
during
during testing
testing
Cleanliness during testing
tests can
MicrobMonitor2 tests be
can conducted
be on-site, in
in an office or
or in a
a laboratory. However, precautions should be
be taken to
MicrobMonitor2
MicrobMonitor2 can
tests be be conducted
canconducted
conducted on-site,
on-site, anin
on-site, an
an office
inoffice in or
office or in
in a
a laboratory.
laboratory. However,
However,
laboratory. precautions
precautions
However, should
precautions should
should be
be taken
taken to to
taken to
ensure testing
ensure is conducted in a reasonably clean area to avoid contamination of the sample and the test by
ensureensure testing is conducted in a reasonably clean area to avoid contamination of the sample and the test by
testing
testing is is conducted
conducted in a in a reasonably
reasonably clean clean
area area
to to
avoid avoid contamination
contamination of of
the the
sample sample
and and
the the
test test
by by
microorganisms otherother
microorganisms than than
thosethose
in the
theintest
test sample.
sample. WashWashhandshands
before and after
after testing. During testing wear clean, nitrile,
microorganisms other
microorganisms than
other those
than in
those in the
the test
test sample.
Wash
sample. hands
Wash before
hands before
and
before and
and after testing.
testing.
after During
testing. During testing
testing
During wearwear
testing clean,
clean,
wear nitrile,nitrile,
clean, nitrile,
vinyl
vinyl or
or polythene
vinyl
polythene gloves.
or polythene
polythene
gloves. Avoid
gloves. touching
Avoid
AvoidAvoid
touching areas
touching of
of the
areas
areasareas ofsyringe
theof and
and loop
the syringe
syringe
syringe dispenser
and loop
loop loop which
dispenser
dispenser come
which
whichwhich into
into direct
come
comecome contact
into direct
directdirect with
with the
contact
contact with the
the the
vinyl or gloves. touching the and dispenser into contact with
sample and avoid
avoid touching the inside
inside of the MicrobMonitor2
theofMicrobMonitor2 test bottle
bottle cap and
and neck whenwhendispensing sample.
samplesample
and
sample and
and avoid
avoid touching
touching the
touching the
the inside
of
inside the MicrobMonitor2
of the test
MicrobMonitor2 test
test bottle
cap
bottle cap and
and neck
capneck when
neck dispensing
dispensing
when sample.
sample.
dispensing sample.
Results that count

5HVXOWVWKDWFRXQW 3
3
Results
Results
that that
count
count
5HVXOWVWKDWFRXQW
5HVXOWVWKDWFRXQW 11

MICROBMONITOR® 2
12 TRUSTED + INNOVATIVE + ACCURATE + DEPENDABLE

EP066 MM2 Instruction Leaflet 130318

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For Detection

Instructions for Use

Water phase from aircraft fuel tank drains

Water from aircraft fuel drains

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

x Results that count

x EP157 Technical Assistance Reading Results of MicrobMonitor2.

The MicrobMonitor2 test consists of anot

x MicrobMonitor2 test bottles

Number of microbial CFU per ml = Number of colonies counted or estimated

Volume tested (ml)

Volume tested (ml)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

lower part of the handle.

0.25 ml of fuel (syringe) 4 x 104 CFU per litre

Water phase from drains ml

Water phase from aircraft fuel tank drains 0.1 ml

number of colonies and

Estimate 100 colonies if 104 CFU

consistency when conducting regular microbiological 0.5/0.25/0.1/0.01/other__

three times to mix. result is similar to picture

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

they are used. Remove the seal from

A.4.4STORAGE AND SHELF LIFE

Water aircraft fuel tank

phase from aircraft fuel 0.1

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Volume tested (ml)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

Incubation Temp. (°C)

A.2 Heavy andFOR

hydraulic 100 Colonies

reflect the viable

volume of sample to test

volume sample which measuring device use.

Aviation kerosene 0.5

Water phase from aircraft fuel

Water phase from aircraft tank drains