Document ID: EPA-HQ-OPP-2002-0233-0012
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2002-09-13T04:00Z

[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
1
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
Reviewer:
Esther
Seto
,
Date
Feb.
1,
2001
Peer
Review:
Ibrahim
Barsoum,
PhD
Microbial
Pesticides
Branch
Biopesticides
and
Pollution
Prevention
Division
U.
S.
Environmental
Protection
Agency
_______________

STUDY
TYPE:
Primary
Eye
Irritation
­
Rabbit
PMRA
DATA
CODE
M4.9
/
USEPA
OPPTS
870.2500
TEST
MATERIAL
(PURITY):
Sporodex
WP
­
2000
formulation
5.7
x
10
8
CFU/
g
SYNONYMS:
Pseudozyma
flocculosa,
Sporothrix
flocculosa
CITATION:
Findlay,
John.
(December,
1999).
"Primary
Eye
Irritation
Study
of
Sporodex."
IIT
Research
Institute,
Life
Sciences
Operation,
Chicago,
Illinois.
Laboratory
Project
ID
1178
SN1.
InLife
Study
Dates:
November
1,
1999
­
November
8,
1999.
Unpublished.

SPONSOR:
Université
Laval
Départment
de
phytologie
Cité
universitaire
Québec,
Canada
EXECUTIVE
SUMMARY:
Administration
of
0.
1
g
of
Sporodex
WP
to
the
eyes
of
rabbits
resulted
in
slight
conjunctival
redness
in
5/
6
animals
at
the
1­
hour
scoring
interval
and
in
2/
6
rabbits
at
the
24­
hour
scoring
interval.
By
the
48­
hour
scoring
interval,
all
signs
of
ocular
irritation
had
subsided.
There
were
no
other
adverse
clinical
symptoms
or
mortalities
during
the
7­
day
observation
period.
The
maximum
irritation
score
(MIS)
was
1.
7
at
the
1­
hour
scoring
interval
and
the
maximum
average
score
(MAS),
over
the
24­,
48­
and
72­
hour
scoring
intervals,
was
0.
22.
Based
on
the
MAS,
Sporodex
WP
was
classified
as
minimally
irritating.

The
test
substance
used
in
this
study
was
a
wettable
powder
formulation
containing
a
potential
ocular
irritant
(see
appendix
I).
The
current
formulation,
Sporodex
L,
contains
a
much
lower
level
of
the
potential
irritant
(see
appendix
II).
Therefore,
Sporodex
L
is
expected
to
be
less
irritating
to
the
eye
than
Sporodex
WP.

This
study
fulfills
the
requirements
of
an
acute
eye
irritation
study
(OPPTS
870.2400)
and
is
considered
acceptable.

COMPLIANCE:
Signed
and
dated
GLP
and
Data
Confidentiality
statements
were
provided.
A
Quality
Assurance
statement
was
not
included.

A.
MATERIALS:
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
2
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
1.
Test
Material:
Sporodex
WP
­
2000
formulation
Description:
tan
powder
Lot/
Batch
#:
Lot
No.
CWB1/
23­
08­
99
Purity:
5.7
x
10
8
CFU/
g
CAS
#
TGAI:
n/
a
The
titre
of
the
test
substance
was
determined
by
the
Sponsor
by
using
a
basic
plating
technique.

2.
Vehicle
and/
or
positive
control:
The
test
substance
was
applied
undiluted.

3.
Test
animals:
Species:
Rabbit
Strain:
New
Zealand
White
Age/
weight
at
dosing:
approximately
11.5
weeks
of
age
at
time
of
dosing
1.90
­
2.
30
kg
at
time
of
receipt
body
weight
not
measured
on
day
of
dosing
Source:
Kuiper
Rabbit
Ranch,
Gary,
IN.
Housing:
Animals
were
individually
housed
in
suspended
stainless
steel
cages.
Absorbent
cage
liners
were
placed
in
pans
below
the
floor
of
each
cage
to
absorb
liquids.
Diet:
Each
rabbit
was
provided
with
approximately
150g
of
Certified
Purina
Lab
Rabbit
Chow
HF
#5325
(PMI
Feeds,
St.
Louis,
MO)
daily.
Water:
City
of
Chicago
tap
water
was
provided
ad
libitum.
Environmental
conditions:
temperature:
21.0
­
22.0

C
relative
humidity:
34
­
69%
light
/
dark
cycle:
12
hours
light
/
12
hours
dark
Acclimation
period:
Animals
were
acclimatized
for
11
days.
Each
animal
was
identified
with
an
ear
tag
and
a
cage
card.

B.
STUDY
DESIGN
and
METHODS:

1.
In
life
dates
­
start
date:
November
1,
1999
termination
date:
November
8,
1999
2.
Animal
Assignment
and
Treatment
­
A
single
group
of
6
female
young
adult
rabbits
were
selected
for
treatment.
Prior
to
treatment,
animals
were
evaluated
for
general
health
and
their
eyes
were
examined
for
corneal
lesions
with
and
without
the
aid
of
2%
fluorescein
and
ultraviolet
light.
On
the
day
of
dosing,
0.
1g
of
Sporodex
WP,
containing
5.
7
x
10
7
CFUof
Pseudozyma
flocculosa,
was
placed
in
the
everted
lower
lid
of
the
right
eye
of
each
animal
and
the
eyelids
were
held
closed
for
approximately
two
seconds.
The
left
eye
of
each
animal
served
as
an
untreated
control.
The
treated
eye
was
rinsed
with
lukewarm
water
24
hours
after
instillation
of
the
test
substance.

3.
Clinical
Observations
­
All
rabbits
were
observed
daily
for
mortality
and
moribundity
for
7
days
following
administration
of
the
test
substance..

4.
Ocular
Examinations
­
At
1,
24,
48,
72
hours,
and
4
and
7
days
after
administration
of
the
test
substance,
treated
and
control
eyes
were
graded
for
ocular
lesions
according
to
the
criteria
of
Draize.
The
cornea
was
examined
for
degree
and
area
of
opacity;
the
iris
for
deepened
folds,
congestion,
swelling,
circumcorneal
hyperemia
and
reaction
to
light;
and
the
conjunctiva
for
redness,
chemosis
and
discharge.
A
slit
penlight
was
used
in
the
scoring
examination.
In
order
to
detect
corneal
lesions,
eyes
were
examined
using
fluorescein
at
the
24­
hour
scoring
interval.
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
3
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
5.
Necropsy
­
After
the
final
observation,
the
rabbits
were
euthanized
by
anaesthetic
overdose
using
sodium
pentobarbital
and
discarded
without
necropsy.

II.
RESULTS
AND
DISCUSSION:

A.
Clinical
Observations
­
No
deaths
occurred
during
the
study.

B.
Ocular
Examinations
­
Individual
eye
irritation
scores
are
presented
in
Table
1.
One
hour
after
test
material
administration,
slight
conjunctival
redness
(grade
1
­
vessels
definitely
injected
above
normal)
was
observed
in
5/
6
animals.
By
the
24­
hour
scoring
interval,
only
2/
6
animals
continued
to
exhibit
slight
conjunctival
redness.
All
signs
of
ocular
irritation
were
absent
at
the
48­
hour
scoring
interval.
Aside
frommild
redness,
no
other
signs
of
ocular
irritation
were
observed
at
any
point
of
the
study.
Based
on
the
Draize
method
of
determining
ocular
irritation,
the
maximumirritation
score,
occurring
at
the
1­
hour
scoring
interval,
was
1.
7
(out
of
a
possible
score
of
110)
and
the
maximum
average
score,
calculated
over
the
24­,
48­
and
72­
hour
scoring
intervals,
was
0.
22.

C.
Reviewer's
Conclusions
­
Administration
of
Sporodex
WP
to
the
eyes
of
rabbits
resulted
in
slight
conjunctival
redness
in
5/
6
animals
at
the
1­
hour
scoring
interval
and
in
2/
6
rabbits
at
the
24­
hour
scoring
interval.
No
other
signs
of
ocular
irritation
were
observed
at
any
point
of
the
study.
There
were
no
other
adverse
clinical
symptoms
or
mortalities
during
the
7­
day
observation
period.
The
maximum
irritation
score
was
1.
7
at
the
1­
hour
scoring
interval
and
the
maximum
average
score
was
0.
22.

The
test
substance
used
in
this
study
was
a
wettable
powder
formulation
(see
appendix
I)
and
its
constituent
formulation
ingredients
are
expected
to
be
more
irritating
to
the
eye
than
the
formulation,
Sporodex
L
(see
appendix
II),
that
is
currently
being
submitted
for
registration.
Sporodex
WP
and,
therefore,
Sporodex
L
are
considered
mildly
irritating
to
the
eye
and
no
signal
words
are
required
on
the
label.
Standard
label
statements
instructing
users
to
avoid
contact
with
eyes
are
sufficient.

This
study
fulfills
the
requirements
of
an
acute
eye
irritation
study
(OPPTS
870.2400)
and
is
considered
acceptable.

D.
Deficiencies
­
Data
pertaining
to
the
untreated
control
eyes
were
not
submitted.
This
information,
however,
is
not
critical
since
severe
effects
were
not
noted
in
the
treated
eyes.
No
other
major
or
minor
deficiencies
were
identified.
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
4
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
Table
1
­
Individual
Eye
Irritation
Scores
Cornea:
Density
of
Opacity
(A
=
Degree
of
Denisty,
B
=
Area
of
Cornea
Involved)

Scoring
Interval
1
hour
24
hours
48
hours
72
hours
4
days
7
days
Animal
No.
SexAB
AB
AB
AB
AB
A
B
351F0
0
00
0
0
00
0
0
00
352F0
0
00
0
0
00
0
0
00
353F0
0
00
0
0
00
0
0
00
354F0
0
00
0
0
00
0
0
00
355F0
0
00
0
0
00
0
0
00
356F0
0
00
0
0
00
0
0
00
Iris
Scoring
Interval
Animal
No.
Sex
1
hour
24
hours
48
hours
72
hours
4
days
7
days
351F0
0
00
00
352F0
0
00
00
353F0
0
00
00
354F0
0
00
00
355F0
0
00
00
356F0
0
00
00
Conjunctiva
(A
=
Erythema,
B
=
Chemosis,
C
=
Discharge)

Scoring
Interval
1
hour
24
hours
48
hours
72
hours
4
days
7
days
Animal
No.
SexABCABCAB
CA
BC
A
BCABC
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
5
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
351F1
0
0
00
0
0
00
0
0
00
0
0
00
0
352F0
0
0
00
0
0
00
0
0
00
0
0
00
0
353F1
0
0
00
0
0
00
0
0
00
0
0
00
0
354F1
0
0
10
0
0
00
0
0
00
0
0
00
0
355F1
0
0
10
0
0
00
0
0
00
0
0
00
0
356F1
0
0
00
0
0
00
0
0
00
0
0
00
0
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
6
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
Appendix
I:
Formulation
Ingredients
for
Sporodex
WP
Ingredient
Purpose
%
Weight
Pseudozyma
flocculosa
active
ingredient
2
(1.
5
­
2.
5)

Arabic
Gum
coating
82
(80
­
84)

Lactose
stabilizer
5.5
(3.
7
­
8)

Silicon
Dioxide
dispersing
agent
1.5
(0.
74
­
3)

Water
diluent
9
(5
­
10)

Total
100
[
Sporodex
L
/
2001­
0304
/
PLG
]
~
PROTECTED
~
Acute
Eye
Irritation
Study
/
7
[
Pseudozyma
flocculosa
/
2000­
0680
/
STF]
DACO
M4.
9
/
USEPA
OPPTS
870.2500
Appendix
II:
Formulation
Ingredients
for
Sporodex
L
Ingredient
Purpose
%
Weight
Pseudozyma
flocculosa
active
ingredient
1.
3
(0.
5
­
3.
0)

Arabic
Gum
coating
14.
5
(12
­
17)

Lactose
stabilizer
1.
5
(1
­
2)

Silicon
Dioxide
dispersing
agent
0.
5
(0.
2
­
0.
8)

Water
diluent
82.2
(75
­
88)

Total
100