EXAMPLE
PROCEDURE
FOR
FIELD­
FILTERING
SAMPLES
FOR
CRYPTOSPORIDIUM
ANALYSIS
USING
IDEXX
FILTA­
MAX
 
FILTERS
JUNE
2003
DRAFT
Draft
June
2003
1
Example
Procedure
for
Field­
Filtering
Samples
for
Cryptosporidium
Analysis
Using
IDEXX
Filta­
Max
 
Filters
1.0
Required
Materials
Check
to
make
sure
the
following
materials
are
available
before
collecting
your
sample:

Q
Several
pairs
of
new,
powder­
free
latex
gloves
(
Lab
Safety
Supply,
cat.
number
16285XL,
or
equivalent)

Q
Sample
collection
form
Q
Filta­
Max
 
foam
filter
module
(
IDEXX,
cat.
number,
FMC
10603)
with
housing
(
IDEXX,
cat.
number,
FMC
10504)

Q
Electric
peristaltic
pump
(
Masterflex
L/
S
variable­
speed
drive,
Cole
Parmer,
cat.
number
LC­
07554­
90,
or
equivalent
with
Masterflex
L/
S
Easy
Load
II
pump
head,
Cole
Parmer,
cat.
number
LC­
77200­
60,
or
equivalent)
(
for
use
only
for
collection
of
samples
from
an
unpressurized
source)

Q
One
length
of
peristaltic
pump
tubing
(
Masterflex
C­
Flex
LS
18
tubing,
Cole
Parmer,
cat.
number
EW­
06424­
18,
or
equivalent)

Q
Four
lengths
of
12.7­
mm
(
0.5­
in.)
internal­
diameter
clear
laboratory
tubing
(
Tygon
formula
R­
3603
[
Cole
Parmer,
cat.
number,
06408­
18],
or
equivalent)

Q
Four
pairs
of
clamps
fit
to
tubing
Q
One
coupling
to
fit
0.5­
in.
internal
diameter
tubing
Q
Pressure
regulator
(
Watts
Regulator,
cat.
number,
263A,
or
equivalent)

Q
Pressure
gauge
(
maximum
operating
pressure
120
psi)
(
Cole
Parmer,
cat.
number,
E­
68004­
05,
or
equivalent)

Q
Water
meter
(
flow
totalizer)
(
Omega
Engineering,
cat.
number,
FTB­
4005,
or
equivalent)

Q
Flow
rate
meter
(
Blue­
White,
cat.
number,
F4437LEA
8,
or
equivalent)

Q
Flow
control
valve
(
Plast­
o­
matic,
model
number
#
FC050B­
1/
2­
PV)

Q
Sample
number
label
Q
Cooler,
approximately
16­
quart
(
Coleman,
model
number,
6216­
703,
or
equivalent)

Q
Two
large
plastic
trash
bags
Q
One
8­
lb
bag
of
ice
Q
Gel
ice
pack
(
VWR,
cat.
number,
15715­
105,
or
equivalent)

Q
Three
13
×
18
inch
ziplock
bag
(
Associated
Bag,
cat.
number,
270­
42,
or
equivalent)

Q
Strapping
tape
Q
Two,
self­
adhesive
plastic
airbill
sleeves
Q
Airbill
for
shipment
Draft
June
2003
2
2.0
Collecting
the
Sample
If
the
sample
will
be
collected
from
a
pressurized
source,
use
the
sample
collection
procedures
in
Section
2.1.
If
the
sample
will
be
collected
from
an
unpressurized
source,
use
the
sample
collection
procedures
in
Section
2.2.

2.1
Sample
Filtration
from
a
Pressurized
Source
2.1.1
Before
connecting
the
sampling
system
to
the
tap
or
source,
turn
on
the
tap
and
allow
the
water
to
flow
for
2
to
3
minutes
or
until
any
debris
that
has
accumulated
in
the
source
water
lines
has
cleared
or
the
turbidity
in
the
water
becomes
visibly
uniform.
Turn
off
the
tap.

2.1.2
Put
on
a
pair
of
powder­
free
latex
gloves
to
prevent
contamination
from
outside
sources.
Any
contamination
of
the
sampling
apparatus
may
bias
the
final
results.

2.1.3
Determine
the
pressure
of
the
water
supply
using
a
pressure
gauge.

2.1.4
Assemble
the
sampling
system,
minus
the
Filta­
Max
 
filter.
For
high
pressure
(>
20
psig)
sites,
the
sampling
system
should
be
assembled
at
the
sample
port
valve
in
the
following
order:

Q
Reinforced
influent
tubing
Q
Pressure
regulator
Q
Pressure
gauge
Q
Reinforced
inlet
tubing
Q
Filta­
Max
 
filter
housing
Q
Reinforce
outlet
tubing
Q
Flow
totalizer
(
mechanical
or
graduated
collection
device)

Q
Flow
control
valve
or
flow
rate
meter
with
valve
Q
Effluent
tubing
to
drain
For
a
low­
pressure
(
7.5
to
20
psig)
site,
the
sampling
system
should
be
assembled
at
the
sample
port
valve
in
the
following
order:

Q
Influent
tubing
Q
Filta­
Max
 
filter
housing
Q
Outlet
tubing
Q
Flow
rate
meter
with
valve
Q
Flow
totalizer
(
mechanical
or
graduated
collection
device)

Q
Effluent
tubing
to
drain
2.1.5
Connect
the
sampling
system,
with
an
empty
Filta­
Max
 
filter
housing,
to
the
pressurized
water
system.
Verify
that
the
filter
housing
is
installed
so
that
the
end
closest
to
the
screw
top
cap
is
the
inlet
and
the
opposite
end
is
the
outlet.

2.1.6
Slowly
turn
the
tap
to
fully
open.
Allow
a
minimum
of
20
L
to
flush
the
system.
During
this
period,
perform
the
following
steps:
Draft
June
2003
3
2.1.6.1
Measuring
flow
rate
with
the
flow
rate
meter
or
the
flow
totalizer
and
a
stopwatch,
adjust
the
flow
rate
to
approximately
3
­
4
L/
min
(
approximately
0.8
­
1
gpm).
At
high
pressure
sites,
using
the
pressure
regulator,
adjust
the
pressure
to
a
maximum
of
120
psi.
Observe
system
for
leaks
and
take
the
necessary
corrective
action
if
any
are
present.
A
differential
pressure
of
7.5
psi
is
required
to
create
flow
through
the
filter.
The
recommended
differential
pressure
to
produce
the
flow
rate
of
3
to
4
L/
min
is
75
psi.
Do
not
exceed
the
maximum
operating
pressure
of
120
psig.

2.1.6.2
Record
the
following
information
on
the
sample
collection
form:

O
Public
water
system
(
PWS)
name
O
PWS
address
O
Sampler
name
O
Sample
ID
(
optional)

O
Public
Water
System
Identification
(
PWS
ID)
number
O
Public
Water
System
facility
ID
number
O
Facility
name
O
Sample
collection
point
ID
O
Sample
collection
point
name
O
Sample
collection
date
O
Source
water
type
(
optional
[
but
required
for
E.
coli
sample
forms])

O
Requested
analysis
(
circle
Cryptosporidium
field
sample
for
routine
monitoring
sample;
circle
both
"
Cryptosporidium
field
sample"
and
"
Cryptosporidium
matrix
spike"
sample
if
you
are
sending
an
additional
sample
with
the
monitoring
sample
for
matrix
spike
analysis)

2.1.6.3
After
the
system
has
been
flushed,
any
optional
water
quality
parameters
such
as
temperature,
pH,
and/
or
turbidity
should
be
measured
now.

2.1.7
Turn
off
the
water
at
the
sample
port
valve
when
the
flow
rate
has
been
adjusted
and
the
system
has
been
flushed.

2.1.8
Record
the
following
information
on
the
sample
collection
form:

O
Current
meter
reading
2.1.9
Install
the
Filta­
Max
 
filter
into
the
housing
and
secure
the
housing
cap
by
hand
tightening.
Apply
gentle
pressure
to
create
a
seal
between
the
module
and
the
"
O"
rings
in
the
base
and
lid
of
the
housing.
Excessive
tightening
is
not
necessary,
and
may
shorten
the
life
of
the
"
O"
rings.
A
light
application
of
vacuum
grease
may
be
used
to
lubricate
the
"
O"
rings,
but
too
much
grease
will
produce
a
negative
effect.

Note!
Retain
the
end
caps
provided
with
the
filter
housing.
These
caps
will
be
needed
to
seal
the
housing
for
shipment.
Draft
June
2003
4
2.1.10
Turn
on
the
pressurized
water
source.
Adjust
flow
to
within
3
to
4
L/
min,
if
necessary.

O
Record
start
time
on
the
sample
collection
form.

2.1.11
Monitor
the
water
meter.
When
the
targeted
volume
(
typically
10
L
[
2.64
gal]
or
50
L
[
13.2
gal])
has
passed
through
the
Filta­
Max
 
filter,
shut
off
the
water
source.
Allow
the
pressure
to
decrease
until
the
water
stops.

2.1.12
Record
the
following
information
on
the
sample
collection
form:

O
Stop
time
(
when
the
water
was
shut
off)

O
Final
meter
reading
O
Comments
to
laboratory,
if
needed
2.1.13
Disconnect
the
inlet
end
of
the
filter
housing,
making
sure
not
to
spill
any
of
the
water
remaining
in
the
capsule.
This
water
is
part
of
your
sample.

2.1.14
The
filter
can
either
be
shipped
in
the
filter
housing
or
removed
from
the
filter
housing
and
shipped
alone.

2.1.15
If
the
filter
will
be
shipped
without
the
housing,
open
the
housing
and
dump
the
filter
and
any
water
remaining
in
the
housing
into
a
ziplock
bag.
Place
this
bag
inside
a
second
ziplock
bag
and
seal.

2.1.16
If
the
filter
will
be
shipped
in
the
housing,
seal
the
inlet
and
outlet
of
the
housing
with
the
rubber
stoppers
that
were
previously
saved.
Place
the
filter
housing
containing
the
filter
in
a
plastic
ziplock
bag
for
shipment.

2.1.17
Place
a
label
on
the
outer
ziplock
bag
containing
the
filter
and
using
a
waterproof
pen
record
the
following
information:

O
PWS
ID
O
Facility
name
O
Date
of
sample
collection
Cover
the
label
with
clear
tape
to
keep
the
label
dry.

2.1.18
Immediately
following
sample
collection,
place
the
bag
containing
the
filter
(
with
or
without
filter
housing)
in
a
refrigerator
to
chill
prior
to
packing
the
shipping
cooler
for
shipment.
If
no
refrigerator
is
available,
and
the
sample
will
not
be
shipped
for
several
hours,
place
the
bag
in
the
shipping
cooler
with
ice
to
chill,
and
replace
the
ice
before
shipping.

Note!
Method
1622/
1623
requires
that
the
temperature
of
the
sample
upon
arrival
at
the
laboratory
must
be
<
10
°
C
(
but
not
frozen),
and
the
laboratory
must
have
the
filter
processed
within
96
hours
of
sample
collection.
If
the
sample
temperature
and
holding
time
requirements
are
not
met,
then
the
sample
is
invalid
and
must
be
recollected.

As
a
recommendation,
it
is
best
to
have
the
filter
processed
by
the
laboratory
within
24
hours
of
sampling.
Draft
June
2003
5
2.2
Sample
Filtration
from
an
Unpressurized
Source
2.2.1
Put
on
a
pair
of
powder­
free
latex
gloves
to
prevent
contamination
from
outside
sources.
Any
contamination
of
the
sampling
apparatus
may
bias
the
final
results.

2.2.2
If
sampling
from
a
source
of
unlimited
volume,
it
may
be
desirable
to
pre­
flush
the
sampling
system.
Assemble
the
sampling
system,
minus
the
Filta­
Max
 
filter.
Verify
that
the
filter
housing
is
installed
so
that
the
end
closest
to
the
screw
top
cap
is
the
inlet
and
the
opposite
end
is
the
outlet.
The
sampling
system
should
be
assembled
in
the
following
order:

Q
Influent
tubing
Q
Peristaltic
pump
Q
Filta­
Max
 
filter
housing
Q
Outlet
tubing
Q
Flow
rate
meter
with
valve
Q
Flow
totalizer
(
mechanical
or
graduated
container)

Q
Effluent
tubing
to
drain
2.2.3
Place
the
inlet
end
of
the
inlet
tubing
in
sample
source,
away
from
any
walls,
bottom,
or
other
environmental
surfaces.

2.2.4
Turn
on
the
pump
and
allow
a
minimum
of
20
L
to
flush
the
system.
If
sampling
source
water
from
a
carboy,
continuously
refill
the
carboy
as
necessary
to
flush
the
system.
As
a
recommendation,
the
carboy
should
not
be
removed
from
the
sampling
chain
and
should
be
refilled
using
a
separate
container.
Observe
the
system
for
leaks
and
take
the
necessary
corrective
action
if
any
are
present.
During
this
period,
perform
the
following
steps:

2.2.4.1
Measuring
flow
rate
with
the
flow
rate
meter
or
the
flow
totalizer
and
a
stopwatch,
adjust
the
flow
rate
to
approximately
3
­
4
L/
min
(
approximately
0.8
­
1
gpm)
by
varying
the
pump
speed
or
adjusting
the
valve
(
if
pump
is
not
variable
speed).

2.2.4.2
Record
the
following
information
on
the
sample
collection
form:

O
Public
water
system
(
PWS)
name
O
PWS
address
O
Sampler
name
O
Sample
ID
(
optional)

O
Public
Water
System
Identification
(
PWS
ID)
number
O
Public
Water
System
facility
ID
number
O
Facility
name
O
Sample
collection
point
ID
O
Sample
collection
point
name
O
Sample
collection
date
O
Source
water
type
(
optional
[
but
required
for
E.
coli
sample
forms])
Draft
June
2003
6
O
Requested
analysis
(
circle
Cryptosporidium
field
sample
for
routine
monitoring
sample;
circle
both
"
Cryptosporidium
field
sample"
and
"
Cryptosporidium
matrix
spike"
sample
if
you
are
sending
an
additional
sample
with
the
monitoring
sample
for
matrix
spike
analysis)

2.2.4.3
Once
the
system
has
been
flushed,
any
optional
water
quality
parameters
such
as
temperature,
pH,
and/
or
turbidity
should
be
measured
now.

2.2.5
Turn
off
the
pump
when
the
flow
rate
has
been
adjusted
and
the
system
has
been
flushed.
Following
flushing,
or
if
flushing
is
not
performed,
proceed
with
the
following
steps:

2.2.6
Record
the
following
information
on
the
sample
collection
form:

O
Current
meter
reading
2.2.7
Install
the
Filta­
Max
 
filter
into
the
housing
and
secure
the
housing
cap
by
hand
tightening.
Apply
gentle
pressure
to
create
a
seal
between
the
module
and
the
"
O"
rings
in
the
base
and
lid
of
the
housing.
Excessive
tightening
is
not
necessary,
and
may
shorten
the
life
of
the
"
O"
rings.
A
light
application
of
vacuum
grease
may
be
used
to
lubricate
the
"
O"
rings,
but
too
much
grease
will
produce
a
negative
effect.

Note!
Retain
the
end
caps
provided
with
the
filter
housing.
These
caps
will
be
needed
to
seal
the
housing
for
shipment.

2.2.8
Turn
on
the
peristaltic
pump.
Record
the
following
on
the
sample
collection
form:

O
Start
time
2.2.9
Monitor
the
water
meter.
When
the
targeted
volume
(
typically
10
L
[
2.64
gal]
or
50
L
[
13.2
gal])
has
passed
through
the
Filta­
Max
 
filter
or
the
container
is
depleted,
remove
the
inlet
tubing
from
the
source.
Shut
off
the
pump
when
all
the
water
has
been
forced
through
the
filter.
Allow
the
pressure
to
decrease
until
the
water
stops.
If
sampling
from
a
container,
add
1
L
of
reagent
water
per
every
10­
L
container
volume
and
rinse
the
sides
of
the
container.
Resume
pumping
until
the
rinse
volume
is
filtered.

2.2.10
Record
the
following
information
on
the
sample
collection
form:

O
Stop
time
(
when
the
pump
was
shut
off)

O
Final
meter
reading
or
measured
total
volume
sample,
not
including
reagent
rinse
volume,
if
any
O
Comments
to
laboratory,
if
needed
2.2.11
Disconnect
the
inlet
end
of
the
filter
housing,
making
sure
not
to
spill
any
of
the
water
remaining
in
the
capsule.
This
water
is
part
of
your
sample.

2.2.12
The
filter
can
either
be
shipped
in
the
filter
housing
or
removed
from
the
filter
housing
and
shipped
alone.

2.2.13
If
the
filter
will
be
shipped
without
the
housing,
open
the
housing
and
dump
the
filter
and
any
water
remaining
in
the
housing
into
a
ziplock
bag.
Place
this
bag
inside
a
second
ziplock
bag
and
seal.
Draft
June
2003
7
2.2.14
If
the
filter
will
be
shipped
in
the
housing,
seal
the
inlet
and
outlet
of
the
housing
with
the
rubber
stoppers
that
were
saved
previously.
Place
the
filter
housing
containing
the
filter
in
a
plastic
ziplock
bag
for
shipment.

2.2.15
Place
a
label
on
the
outer
ziplock
bag
containing
the
filter
and
using
a
waterproof
pen
record
the
following
information:

O
PWS
ID
O
Facility
name
O
Date
of
sample
collection
Cover
the
label
with
clear
tape
to
keep
the
label
dry.

2.2.16
Immediately
following
sample
collection,
place
the
bag
containing
the
filter
housing
in
a
refrigerator
to
chill
prior
to
packing
the
shipping
cooler
for
shipment.
If
no
refrigerator
is
available,
and
the
sample
will
not
be
shipped
for
several
hours,
place
the
bag
in
the
shipping
cooler
with
ice
to
chill.
Replace
the
ice
before
shipping.

Note!
Method
1622/
1623
requires
that
the
temperature
of
the
sample
upon
arrival
at
the
laboratory
must
be
<
10
°
C
(
but
not
frozen),
and
the
laboratory
must
have
the
filter
processed
within
96
hours
of
sample
collection.
If
the
sample
temperature
and
holding
time
requirements
are
not
met,
then
the
sample
is
invalid
and
must
be
recollected.

As
a
recommendation,
it
is
best
to
have
the
filter
processed
by
the
laboratory
within
24
hours
of
sampling.

3.0
Packing
the
Sample
3.1
Insert
two
large
plastic
trash
bags
into
the
shipping
cooler
to
create
a
double
liner.
Immediately
before
packing
the
cooler,
create
two
4­
lb
ice
packs
by
putting
half
of
an
8­
lb
bag
of
ice
into
two
separate
ziplock
bags.
To
prevent
leaks
place
each
ice
pack
into
an
additional
ziplock
bag.
Gel
packs
or
blue
ice
may
be
used
instead
of
wet
ice,
as
long
as
the
sample
is
maintained
in
the
appropriate
temperature
range.
Seal
the
ziplock
bag,
expelling
as
much
air
as
possible,
and
secure
top
with
tape.

Note!
Shipping
companies
may
delay
sample
shipments
if
leakage
occurs.
Double
liners
and
ziplock
bags
around
ice
will
prevent
leakage
and
delays.

3.2
Place
the
bag
containing
the
filter
housing
into
the
shipping
container.
Place
the
ice
pack
around,
but
not
on,
the
sample
bag
to
prevent
freezing.
Inflated,
empty
sample
bags
can
be
placed
between
the
filter
housing
and
the
ice
packs
to
prevent
the
filter
from
freezing.

3.3
If
you
will
be
monitoring
sample
temperature
during
shipment,
place
in
the
cooler
the
temperature
monitoring
device
(
e.
g.,
extra
sample
bottle
for
measuring
sample
temperature
upon
receipt
at
the
laboratory,
thermometer
vial,
or
Thermochron
 
iButton).
Seal
each
liner
bag
by
twisting
top
of
bag
and
tying
in
a
knot.
Draft
June
2003
8
3.4
Peel
the
backing
off
one
of
the
plastic
airbill
sleeves
and
attach
the
sleeve
to
the
inside
of
the
cooler
lid.

O
Sign
and
date
the
sample
collection
form
Fold
the
completed
sample
collection
form,
and
place
it
inside
the
plastic
sleeve.

3.5
Close
the
cooler
lid,
seal
the
horizontal
joints
with
duct
tape,
and
secure
the
lid
with
tape
by
taping
the
cooler
at
each
end,
perpendicular
to
the
seal.

Note!
Shipping
companies
may
delay
sample
shipments
if
leakage
occurs.
Be
sure
to
seal
the
cooler
joints.

3.6
Peel
the
backing
off
of
the
second
airbill
sleeve
and
attach
the
sleeve
to
the
outside
of
the
cooler
lid.
Complete
the
shipping
airbill
with
the
laboratory
address,
billing
information,
sample
weight,
and
shipping
service.
Remove
the
shipper's
copy
of
the
airbill,
and
place
the
remainder
of
the
airbill
inside
the
plastic
sleeve.

4.0
Shipping
and
Tracking
4.1
Ship
samples
on
the
day
of
collection
and
use
a
reliable
shipping
service
for
next­
day
delivery.
If
samples
are
not
shipped
the
day
of
collection,
the
sample
must
be
maintained
at
<
10
°
C
(
but
not
frozen)
by
chilling
in
a
refrigerator
or
cooler
filled
with
ice.

4.2
Contact
the
laboratory
to
notify
them
of
the
sample
shipment.
Request
that
the
laboratory
contact
you
the
next
day
if
the
sample
is
not
received.

4.3
Using
the
airbill
number
on
the
shipper's
copy
of
the
airbill,
track
the
sample
shipment
using
the
shipping
company's
web
page
or
by
contacting
the
shipping
company
over
the
phone.

4.4
If
problems
are
encountered
with
the
shipment,
communicate
with
the
shipping
company
to
resolve,
and
update
the
laboratory
regarding
the
status
of
the
shipment.
