EXAMPLE
PROCEDURE
FOR
FIELD­
FILTERING
SAMPLES
FOR
CRYPTOSPORIDIUM
ANALYSIS
USING
PALL
LIFE
SCIENCES
ENVIROCHEK
 
OR
ENVIROCHEK
 
HV
FILTER
JUNE
2003
DRAFT
Draft
June
2003
1
Example
Procedure
for
Field­
Filtering
Samples
for
Cryptosporidium
Analysis
Using
the
Pall
Life
Sciences
Envirochek
 
or
Envirochek
 
HV
Filter
1.0
Required
Materials
Check
to
make
sure
the
following
materials
are
available
before
collecting
sample:

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

Q
Sample
collection
form
Q
Pall
Life
Sciences
Envirochek
 
or
Envirochek
 
HV
sampling
capsule
(
recommend
that
two
filter
capsules
be
kept
on
hand
in
case
the
first
one
clogs
prior
to
filtering
10
L)

Q
Pump
(
if
water
is
not
from
pressurized
source)
­
Electric
centrifugal(
Simer,
cat.
number
M40)
or
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
any
equivalent
pump
that
can
create
a
flow
of
approximately
2
L/
min
Q
Five
lengths
of
12.7­
mm
(
0.5­
in.)
internal­
diameter
clear,
vinyl,
laboratory
tubing
(
Cole
Parmer,
cat.
number,
06405­
19
or
equivalent)

Q
Five
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
(
filter's
maximum
operating
pressure,
original
­
30
psi,
HV
­
60
psi)
(
Cole
Parmer,
cat.
number,
E­
68004­
04,
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
Stopwatch
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
or
gel
ice
pack
(
VWR,
cat.
number,
15715­
105,
or
equivalent)

Q
Three
13
×
18
inch
ziplock
bags
(
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
source
with
the
pressure
gauge.

2.1.4
Assemble
the
sampling
system,
minus
the
Envirochek
 
capsule.
In
place
of
the
Envirochek
 
capsule,
insert
a
0.5­
in.
barbed
connector
between
the
outlet
tubing
from
the
sample
valve
or
the
pressure
regulator
and
the
inlet
tubing
of
the
flow
meter
or
control
valve.
For
high
pressure
(>
20
psig)
sites,
the
sampling
system
should
be
assembled
in
the
following
order:

Q
Reinforced
influent
tubing
Q
Pressure
regulator
Q
Pressure
gauge
Q
Reinforced
inlet
tubing
Q
Envirochek
 
capsule
Q
Reinforced
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
low
pressure
(
1
to
20
psig)
sites,
the
sampling
system
should
be
assembled
at
the
sample
port
valve
in
the
following
order:

Q
Influent
tubing
Q
Envirochek
 
capsule
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
the
connector
in
place
of
the
Envirochek
 
capsule,
to
the
pressurized
water
system,
using
appropriate
fittings
and
clamps.

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
2
L/
min
(
approximately
0.5
gpm).
Using
the
pressure
regulator,
adjust
the
pressure
to
a
maximum
of
30
psi
if
using
the
Envirochek
 
or
60
psi
if
using
the
Envirochek
 
HV.
Observe
the
system
for
leaks
and
take
the
necessary
corrective
action
if
any
are
present.

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
capsule
filter
label
with
a
waterproof
pen:

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

2.1.9
Record
the
following
information
on
the
sample
collection
form:

O
Current
meter
reading
2.1.10
Remove
the
connector
and
in
its
place,
install
the
Envirochek
 
capsule
in
line,
securing
the
inlet
and
outlet
ends
with
the
appropriate
fittings/
clamps.

Note!
Retain
the
vinyl
caps
provided
with
the
Envirochek
 
capsule.
These
caps
will
be
needed
to
seal
the
capsule
for
shipment.
Draft
June
2003
4
2.1.11
Slowly
turn
on
the
pressurized
water
source.
Record
the
following
information
on
the
capsule
filter
label
or
sample
collection
form:

O
Start
time
2.1.12
Vent
the
residual
air
in
the
capsule
using
the
bleed
valve
by
turning
it
counter­
clockwise.
When
the
filter
housing
is
full
of
water,
close
the
bleed
valve.

2.1.13
Monitor
the
water
meter.
When
the
targeted
volume
(
actual
sample
volumes
will
be
dependant
on
the
utility,
but
volumes
are
typically
10
L
[
2.64
gal]
to
50
L
[
13.2
gal])
has
passed
through
the
Envirochek
 
capsule,
shut
off
the
water
source.
Allow
the
pressure
to
decrease
until
the
water
stops.

2.1.14
Record
the
following
information
on
the
capsule
filter
label
and/
or
sample
collection
form:

O
Stop
time
(
when
the
water
was
shut
off)

O
Final
meter
reading
O
Comments
to
laboratory,
if
needed
2.1.15
With
the
capsule
inlet
pointed
up,
loosen
the
outlet
end
of
the
Envirochek
 
capsule
and
allow
water
to
drain
as
much
as
possible.
Water
drainage
from
the
capsule
through
the
outlet
is
acceptable,
as
the
sample
has
passed
through
the
membrane.
Opening
the
vent
valve
during
the
draining
will
speed
the
process.
Be
sure
to
close
the
valve
when
finished.

2.1.16
Disconnect
the
inlet
end
of
the
Envirochek
 
capsule,
making
sure
not
to
spill
any
of
the
water
remaining
in
the
capsule
through
the
inlet
port.
This
water
is
part
of
your
sample.

2.1.17
Seal
the
inlet
of
the
capsule
with
the
vinyl
end
cap
that
was
previously
saved.

2.1.18
Seal
the
outlet
of
the
capsule
with
the
vinyl
end
cap
that
was
previously
saved.
Place
the
Envirochek
 
capsule
in
a
plastic
ziplock
bag
for
shipment.

2.1.19
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.

2.2
Sample
Filtration
Using
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.
Draft
June
2003
5
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
Envirochek
 
capsule.
In
place
of
the
Envirochek
 
capsule,
insert
a
0.5­
in.
coupling
to
connect
the
influent
tubing
to
the
inlet
tubing
of
the
pressure
regulator.
The
sampling
system
should
be
assembled
in
the
following
order:

G
Influent
tubing
G
Envirochek
 
capsule
G
Outlet
tubing
G
Centrifugal
or
peristaltic
pump
G
Tubing
G
Flow
rate
meter
with
valve
G
Flow
totalizer
(
mechanical
or
graduated
collection
device)

G
Effluent
tubing
to
drain
When
assembling
sample
chain,
make
sure
that
it
is
as
airtight
as
possible
in
order
to
create
a
vacuum.
To
accomplish
this,
make
sure
that
clamps
are
used
at
each
connection
and
that
rubber
washers
are
inserted
into
the
hose
connections
on
the
inlet
and
outlet
ends
of
the
centrifugal
pump.

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
2
L/
min
(
approximately
0.5
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
Draft
June
2003
6
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)

2.2.4.3
After
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.

2.2.6
Record
the
following
information
on
the
capsule
filter
label:

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

2.2.7
Record
the
following
information
on
the
sample
collection
form:

O
Current
meter
reading
2.2.8
Install
the
Envirochek
 
capsule
in
line,
securing
the
inlet
and
outlet
ends
with
the
appropriate
fittings/
clamps.

Note!
Retain
the
vinyl
caps
provided
with
the
Envirochek
 
capsule.
These
caps
will
be
needed
to
seal
the
capsule
for
shipment.

2.2.9
Refill
the
carboy
with
the
sample
to
be
filtered.
As
a
recommendation,
the
carboy
should
not
be
removed
from
the
sampling
chain
and
should
be
refilled
using
a
separate
container.

2.2.10
Turn
on
the
pump.
Record
the
following
information
on
the
capsule
filter
label
and/
or
sample
collection
form:

O
Start
time
2.2.11
Monitor
the
water
meter
continuously
refilling
the
carboy
as
needed.
When
the
targeted
volume
(
actual
sample
volumes
will
be
dependant
on
the
utility,
but
volumes
are
typically
10
L
[
2.64
gal]
to
50
L
[
13.2
gal])
is
reached,
turn
off
spigot
and
remove
tubing
from
the
spigot.
Allow
pump
to
evacuate
water
from
capsule
and
then
shut
off
the
pump.
Draft
June
2003
7
2.2.12
Record
the
following
information
on
the
capsule
filter
label
and/
or
sample
collection
form:

O
Stop
time
(
when
the
pump
was
shut
off)

O
Final
meter
reading
O
Comments
to
laboratory,
if
needed
2.2.13
With
the
capsule
inlet
pointed
up,
loosen
the
outlet
end
of
the
Envirochek
 
capsule
and
allow
water
to
drain
as
much
as
possible.
Water
drainage
from
the
capsule
through
the
outlet
is
acceptable,
as
the
sample
has
passed
through
the
membrane.
Opening
the
vent
valve
during
the
draining
will
speed
the
process.
Be
sure
to
close
the
valve
when
finished.

2.2.14
Disconnect
the
inlet
end
of
the
Envirochek
 
capsule,
making
sure
not
to
spill
any
of
the
water
remaining
in
the
capsule
through
the
inlet
port.
This
water
is
part
of
your
sample.

2.2.15
Seal
the
inlet
of
the
capsule
with
the
vinyl
end
cap
that
was
saved
previously.

2.2.16
Seal
the
outlet
of
the
capsule
with
the
vinyl
end
cap
that
was
saved
previously.
Place
the
Envirochek
 
capsule
in
a
plastic
ziplock
bag
for
shipment.

2.2.17
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
pound
ice
bags
by
putting
half
an
8­
pound
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
bags,
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
capsule
in
a
plastic
ziplock
bag
(
if
this
step
has
not
already
been
performed)
and
place
the
bag
in
the
cooler.
Place
the
ice
packs
around,
but
not
on,
the
bag
containing
the
capsule
to
prevent
freezing.
Draft
June
2003
8
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.

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.
